indian journal of research in pharmacy and biotechnology (ijrpb) vol-31 issue -3

Volume 1 Issue 3 www.ijrpb.com May-June 2013

Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2320-3471 (Online) ISSN: 2321-5674 (Print)

Editor B.Pragati Kumar, M.Pharm, Assistant Professor,

Nimra College of Pharmacy

Consulting editor

Dr. S Duraivel, M.Pharm, Ph.D., Principal, Nimra College of Pharmacy

Associate Editors

Mr. Debjit Bowmick, M.Pharm., (Ph.D) Assistant Professor, Nimra College of Pharmacy

Mr. Harish Gopinath, M.Pharm., (Ph.D) Assistant Professor, Nimra College of Pharmacy

Dr. M. Janardhan, M.Pharm., Ph.D. Professor, Nimra College of Pharmacy

Dr. A. Ravi Kumar, M.Pharm., Ph D. Professor, Bapatla College of Pharmacy

Editorial Advisory Board

Dr.Y.Narasimaha Reddy, M. Pharm., Ph D.

Principal, University college of Pharmaceutical Sciences, Kakatiya University, Warangal.

Dr. Biresh Kumar Sarkar, Asstt.Director (Pharmacy),

Kerala

Dr.V.Gopal, M. Pharm., Ph D.

Principal, Mother Theresa Post Graduate & Research Institute of Health Sciences,Pondicherry-6

Dr. M.Umadevi, M.Sc. (Agri), Phd Research Associate, Tamil Nadu Agricultural

University, Coimbatore

Dr. J.Balasubramanium, M. Pharm., Ph D. General Manager, FR&D

R A Chem Pharma Ltd., Hyderabad

Dr. V.Prabhakar Reddy, M. Pharm., Ph D.

Principal, Chaitanya College of Pharmacy Education & Research, Warangal

Dr.P.Ram Reddy, M. Pharm., Ph D.

General Manager, Formulation, Dr.Reddy’s Laboratory, Hyderabad

Dr. S.D.Rajendran, M. Pharm., Ph D.

Director, Pharmacovigilance, Medical Affairs, Sristek Consultancy Pvt. Ltd, Hyderabad


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S.No. Contents Page No.

1. Development and evaluation of drotoverine taste masked tablets with improved

dissolution efficiency using solid dispersion technique

Anusha P, Nirajana V.A, Syed Mohammed, Shaik Jilani, Ch. Murali Krishna, Harish.G

275-280

2. Effect of different disintegrants on Ciprofloxacin conventional tablets

Harish.G, Ch.Bhargavi, A.Riyajune, Md.Yasmeen, Syed Mohammed, Ch.Murali Krishna,

Sk.Jilani, Nirajana.V.A

281-287

3. Chronotherapy for nocturnal asthama

Papola Vibhooti , Rajan G, Bisht Seema , Dr. Kothiyal Preeti 288-298

4. Recent trends in scope and opportunity of clinical research in India

Hemant Singh, Abhinav Srivastva

299-304

5. Sustained release drug delivery system

Navin Dixit, Sheo Dutt Maurya, Bhanu P.S.Sagar

305-310

6. Antibacterial activity of ethanolic extracts of Nyctanthes arbortristis and Nerium

oleander

A.Ravi Kumar, Ch.S.D.Phani Deepthi Yadav

311-313

7. Food poisoning and its safety precaution

M. Umadevi, K.P.Sampath Kumar, Sai Pavan, Sd.Gosia Sultana, D. Bhowmik

314-323

8. Microencapsulation technology

K.P.Sampath Kumar,Tejbe.Sk, Shameem Banu, P.Naga Lakshmi , D.Bhowmik 324-328

9. An overview about pharmacy education in India Praneta Desale

329-332

10. Phytochemical evaluation of Nyctanthes arbortristis, Nerium oleander and Catharathnus roseus Ch.S.D. Phani Deepthi Yadav, N.S.P. Bharadwaj, M. Yedukondalu, Ch. Methushala, A. Ravi Kumar

333-338

11. Angina pectoris epidemic in India: a comprehensive review of clinical features, differential diagnosis, and remedies Shravan Paswan, Ranjan Kumar Sharma, Alok Ranjan Gaur, Avinash Sachan, Mahendra Singh Yadav, Preeti Sharma, Mrinmoy Gautam

339-345

12. Stability Indicating HPLC method for the estimation of Cinacalcet hydrochloride API Manikandan Krishnan, Santhana Lakshmi Karunanidhi, Gayathri Sola, Akshitha .Y

346-350

13. Bioavailability: criteria for approving a drug product for marketing Sandhya Singh, Faheem Ajmal Ansari, Shravan Paswan, Rnjan Kumar Sharma, Alok Ranjan Gaur

351-359

14. The effect of superdisintegrants on the dissolution of calcium carbonate fast dissolving tablets Mohammed Farhana, J.Preeti, Md Faizulla, Budda Chellibabu, Harish.G, Rajnesh Kumar Singh

360-364

15. Phytochemical and pharmacological studies on whole plant of Asystasia gangetica T.K.Gopal, Megha.G, D.Chamundeeswari, C.Umamaheswara Reddy

365-370

16. Investigation of in-vitro anti-oxidant, anti-inflammatory and anti-arthritic activity of aerial parts of Securinega leucopyrus (willd.) Muell T.K.Gopal, T.Sheela, D.Chamundeeswari, C.Umamaheswara Reddy

371-378



17. Transdermal sonophoresis technique- an approach for controlled drug delivery

K.P.Sampath Kumar, Debjit Bhowmik, M.Komala

379-381

18. A comprehensive review of Eladi Vati Navin Dixit, Sheo Dutt Maurya , Bhanu P.S.Sagar

382-384

19. Preparation and characterization of some herbal ointment formulations with evaluation of antimicrobial property

Pulak Majumder and Susmita Majumder

385-390

20. The effects of air pollution on the environment and human health

Shyam Bihari Sharma, Suman Jain, Praveen Khirwadkar, Sunisha Kulkarni 391-396

21. Formulation and evaluation of orodispersible tablets of Cinnarizine by super disintegrants addition method Praveen Khirwadkar, Kamlesh Dashora, Shyam Bihari Sharma

397-400

22. Effective hypoglycemic action of metformin combinations against Dexamethasone induced diabetes mellitus in rats Mohanraghupathy.S, Jayabharath N, Bhuvana Tejay, Hameera Khanam B, Lavanya Lahari B

401-403

23. A review on medicinal plants having antioxidant potential Prof.S.K Sharma, Mr. Lalit Singh, Suruchi Singh

404-409

24. Invitro anti-inflammatory activity of Strychnos potatorum linn seed by HRBC membrane stabilization V.Vijayakumar, Dr C.K.Hindumathy

410-412

25. Synthesis and characterization of 1, 3, 4-oxadiazole and 1,3,4- thiadiazole Ramanji Naik

413-419

26. Preparation, characterization and evaluation of Olmesartan medoxomil β- cyclodextrin complexes V. Prudhvi Raj, Subhashis Debnath, Maleswari, M. Niranjan Babu

420-427

27. Wafers technology – a newer approacah to smart drug dilevery system Papola Vibhooti*, Kothiyal Preeti

428-439

28. Evaluation of anti-ulcer effects of ethanolic extract of Delonix regia flower

Samaresh Pal Roy, Kamlesh Prajapati, Ramji Gupta, Dipanwita Bhadra, Nikunj Patel,

Archana Batiwala, Gautam Sonara, Neerav Gheewala, T. Kannadasan

440-445

29. A study on medication non-adherence in ambulatory diabetic patients and need for

pharmacist intervention for improving patient adherence

Dr. Praveen Kumar G

446-447

30. Recent trends in positive and negative aspects of food on bioavalabilty of drugs

Gowthami B, Sk Nahida Fazilath, Sanaulla Md, K Prudhvi Raj, Dastagiriah G, Tabassum Sk

448-460

31. A review on collagen based drug delivery systems Sahithi B, Ansari Sk, Hameeda Sk, Sahithya G, Durga Prasad M, Yogitha Lakshmi

461-468

ISSN: 2321-5674(Print)

ISSN: 2320 – 3471(Online)

Anusha P et.al Indian Journal of Research in Pharmacy and Biotechnology

Volume 1(3) May-June 2013 Page 275

DEVELOPMENT AND EVALUATION OF DROTOVERINE TASTE

MASKED TABLETS WITH IMPROVED DISSOLUTION EFFICIENCY USING

SOLID DISPERSION TECHNIQUE Anusha P*

1, Nirajana V.A

2, Syed Mohammed

1, Shaik Jilani

1, Ch. Murali Krishna, Harish.G

1

1. Nimra college of Pharmacy, Jupudi, Vijayawada, Andhra Pradesh, India

2. Faculty of Pharmacy, Sri Ramachandra University, Porur, Chennai.

*Corresponding author: E.Mail:[email protected]

ABSTRACT

The purpose of this research is to mask the bitter taste of the drug, Drotoverine using solid dispersion

technique. The taste-masked drug is formulated in to a conventional tablet by direct compression method for ease

of administration. Taste masking was done by solid dispersion using polymer such as urea and mannitol by

melting/fusion method Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM)

were performed to identify the physicochemical interaction between drug and carrier, hence its effect on

dissolution. Conventional taste masked tablets were evaluated for weight variation, disintegration time, hardness

and friability. In vitro drug release studies were performed for conventional tablets of drotoverine. Bitterness

score was evaluated on volunteers. FTIR spectroscopy and SEM showed no interaction between drug and carriers.

Conventional tablets prepared using solid dispersion, showed faster disintegration and complete bitter taste

masking of drotoverine. In addition the prepared tablets exhibited better dissolution profile. Taste evaluation of

taste masked tablets in human volunteers rated tasteless with a score of 0. Thus, results conclusively demonstrated

successful masking of taste and oral disintegration of the formulated tablets in the oral cavity with improved

dissolution.

Key words: Drotoverine, conventional tablet, solid dispersion, taste masking

INTRODUCTION

The oral route is the most convenient, appropriate and acceptable way to administer medications. Several

oral active pharmaceuticals ingredients and bulking agents have Unpleasant bitter taste; hence this often times

results to non compliance to medications by Patients. Taste masking is a means of masking the bitter taste of drug

in order to improve the Palatability of the drug, which in turn improves patience compliance. Drotoverine is a

novel non-Anticholinergic smooth muscle antispasmodic drug. Its Chemical name is 1- [(3, 4-diethoxy phenyl)

methylene]-6, 7-diethoxy-1, 2, 3, 4-tetrahydroisoquinoline with a molecular formula of C24 H31NO4. HCl. It

decreases the influx of active calcium into smooth muscles due to inhibiting of phosphodiesterase and

intracellular increase of cAMP level. Its oral bioavailability is about 100% with a biologic half- life of about 7 to

12 h. It adult dose is about 40 to 80mg one to three times a day.

Drotoverine has poor aqueous solubility, and thus resulting in incomplete absorption after oral

administration. This is due to a large fraction of the dose remaining undissolved for absorption. Under such

conditions, the bioavailability can be increased by using, a more water soluble formulation. Solid dispersion is an

efficient means of improving the dissolution rate and hence the bioavailability of a range of poorly soluble drug.

Further drotoverine has an extremely unpleasant bitter taste. The exact mechanism of bitterness is unknown.

Masking of bitter taste of the drotoverine is an extremely important factor in the formulation of tablets to ensure

patient compliance. Taster masked tablets were useful in patients, such as pediatric, geriatric, who may face

difficulty in swallowing conventional tablets or capsules and liquid orals or syrup.

Many reported techniques such as polymer coating, microencapsulation, use of lecithins and related

substances, liposomes and various polymeric materials mask the bitterness by controlling drug release at salivary

pH. However it is a major challenge to develop taste masked conventional tablets with improved drug release.

Thus in the present study an attempt has been made to formulate taste masked conventional tablets of drotoverine

with improved dissolution so as to prepare a “patient-friendly dosage form”. Furthermore, the study undertakes to

investigate solid-state characterization, and attempts to see the possible mechanism of taste masking and

improved dissolution rate.

2. MATERIALS AND METHODS

2.1. Materials: Drotoverine was obtained as a gift sample from Apex laboratory ltd, Chennai; sodium starch

glycolate, aspartame, magnesium stearate and micro crystalline cellulose was obtained as a gift sample from Intex

chemicals, Chennai.





2.2. Methods

2.2.1. Preparation of Drotoverine Solid Dispersion Using Mannitol and Urea as Carriers: The solid

dispersion of drotoverine with urea and mannitol in 1:1, 1:2 and 1:3 ratios were carried out using melting or

fusion method. The preparation of physical mixture of a drug and a water-soluble carrier and heating it directly

until it melted. The melted mixture is then solidified orally in an ice-bath under vigorous stirring. The final solid

mass is crushed, pulverized and sieved.

2.2.2. Characterization of solid dispersion: The prepared drotoverine and mannitol, drotoverine and urea solid

dispersions were characterized for solubility studies, FTIR studies and SEM studies.

2.2.3. Tablet Formulation: Oral conventional tablets containing equivalent of 80 mg of drotoverine were

compressed on an eight-station single rotary tabletting press (GMC, using a flat punch with break line by direct

compression technique.

3. Characterization of Prepared Tablets: The prepared tablets were evaluated for its physical characteristics

like hardness thickness, weight variation, friability, disintegration test.

3.1. In vitro dissolution study: One tablet of F1 or F2 or MKT formulation were placed in a cylindrical basket

(aperture size 425μm: diameter 20mm; height 25mm), and immersed in 900ml of leaching fluid (Stimulated

gastric fluid maintained at 37 ± 2oC). The fluid was stirred at 100rpm (Model Disso 2000, Lab India). Samples of

the leaching fluid (5ml) were withdrawn at selected time intervals with a syringe fitted with a cotton wool plug

and replaced with an equal volume of drug-free dissolution fluid. The samples were suitably diluted with blank

dissolution fluid and were analyzed for content of drotaverine hydrochloride spectrophotometrically at λmax,

302.8 nm by using an ElicoSL 210 UV-Visible double beam spectrophotometer (Elico, India). The amounts

released were expressed as a percentage of the drug content in each dissolution medium. The dissolution test was

carried out in quadruplicate and the mean results reported.

3.2. Taste evaluation: Taste evaluation was done on 6 volunteers by using time intensity method. One tablet was

held in mouth for 10 seconds bitterness levels were recorded instantly and after 10 seconds,

30seconds, 1 minute and 2 minutes, bitterness levels were recorded.

4. RESULTS

Table.1. Formulation table for solid dispersion Table.2. Formulation Table for Tablets

Formulation code Polymer Ratio

F1 Mannitol 1:1

F2 Mannitol 1:2

F3 Mannitol 1:3

F4 Urea 1:1

F5 Urea 1:2

F6 Urea 1:3

*for all the formulations

Name of the ingredient Quantity per single tablet*

Drug polymer SD 80mg

Sodium starch glycolate 6.6mg

Aspartame 1.1mg

Microcrystalline cellulose 16.8mg

Magnesium stearate 5.5mg

Total 110mg

Table.3. Pre-compression parameters for drug polymer solid dispersion

Formulation code F1 F2 F3 F4 F5 F6

Angle of repose 25°71’ 26°42’ 28°93’ 24°32’ 25°43’ 29°47’

Bulk Density(gm/ml) 0.74 0.72 0.69 0.64 0.75 0.78

Tapped Density(gm/ml) 0.86 0.82 0.87 0.85 0.89 0.89

Compressibility Index (%) 13.95 12.19 20.68 24.70 15.73 12.35

Hausners ratio 1.18 1.14 1.25 1.30 1.16 1.14

Table.4. Post compression studies

Formulation

code

Weight

variation (mg)

Thickness

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

F1 Complies 1.9 4 0.05 99.56

F2 Complies 1.9 4 0.06 98.45

F3 Complies 2.0 5 0.03 99.12

F4 Complies 1.9 4 0.05 98.22

F5 Complies 2.1 5 0.06 97.44

F6 Complies 2.0 4 0.01 99.61





Table.5. In-vitro dissolution study for Drotoverine solid dispersion Time in

min

Percent drug release formulation code

F1 F2 F3 F4 F5 F6

0 0 0 0 0 0 0

5 14.11 14.75 15.39 16.56 22.25 26.03

10 25.03 26.14 30.01 32.78 39.56 45.35

15 34.22 36.78 40.56 43.06 54.09 69.46

20 41.86 45.67 52.88 55.12 67.03 78.75

25 52.45 55.76 61.75 67.95 73.25 86.95

30 58.56 65.20 67.25 72.15 84.02 92.10

Table.6. In-vitro dissolution study for

marketed drotoverine tablets

Table.7.Comparison of dissolution profile for formulated

and marketed products Time in min Percent drug release

0 0

5 24.36

10 35.12

15 54.32

20 67.75

25 75.55

30 82.36

Time in

min

%Drug Release

Marketed drotoverine

tablets

Formulated

drotoverine tablets

0 0 0

5 24.36 26.03

10 35.12 45.35

15 54.32 69.46

20 67.75 78.75

25 75.55 86.95

30 82.36 92.10

Table.8. Bitterness evaluations of prepared drotoverine tablets: volunteers Formulation code and bitterness

F1 F2 F3 F4 F5 F6

1 0 0 0 0 0 0

2 0 0 X 0 0 0

3 0 0 0 X 0 0

4 x 0 0 0 x 0

5 0 0 0 0 0 0

0=No bitterness; x=Threshold bitterness

Figure.1. IR spectro of the pure drug, Drotaverine Hcl Figure.2.IR spectro of the formulation F1

Figure.3.IR spectro of the formulation F2 Figure.4. IR spectro of the formulation F3





Figure.5.IR spectro of the formulation F4 Figure.6.IR spectro of the formulation F5

Figure.6. IR spectro of the formulation F6

Figure.7.SEM image of the pure drug,

Drotaverine Hcl

Figure.8.SEM image of the formulation F1

Figure.9.SEM image of the formulation F2 Figure.10.SEM image of the formulation F3

Figure.11. SEM image of the formulation F4 Figure.12.SEM image of the formulation F5

Figure.13. SEM image of the formulation F6





Figure.14. In-Vitro Dissolution studies of the

formulated batches

Figure.15. In-Vitro Dissolution studies Optimized

and marketed

5. DISCUSSION

Fourier Transform Infrared Spectroscopy: Infrared spectra matching approach was used for thedetection of

any possible chemical reaction between the drug and the polymer. The IR spectrum of the physical mixture was

done to detect any appearance or disappearance of peaks. The compatibility between the drug and the polymer

were evaluated using FTIR matching method. The IR spectra of pure drug and polymer are shown in (figure 1).

Pre-Formulation Studies: The angle of repose of prepared drotaverine tablet mixture was in the range 20°-30°.

Normally if the value falls between 20°-30°, it shows good flow property. The bulk density and tapped density

were found to be in the range of 0.7 to 0.8 g/cm3. A Hausner’s ratio was within the range of 1.13 to 1.32, lesser

than 1.25 is considered to be an indication of

good flow property. The compressibility index was within the range of 10-25 hence falls within the good range.

Post-Compressional studies: The post compressional characteristic for all the formulated batches was found to

be within the limits as per Indian pharmacopeia. The hardness was found to be within 4-5 Kg/cm2 in all the

formulations. In all the formulations, the friability value is less than 1% giving an indication that tablets

formulated are mechanically stable. All the tablet formulations compile the weight variation test. The weight of

all the formulations was found to be within the limits. The assay of all the formulations was found to be within the

pharmacopoeial (Table 4).

In-vitro dissolution study: All the formulation was subjected to dissolution studies and it was absorbed that the

batch F6 showed about 92.10% of release and was found to be maximum when compared to other batches.

Formulated batch F1 and F2 showed a slow release pattern with about only 58.56% and 65.20% of drug release at

the end of 30mins. (table 5), and the batches F3, F4, F5 showed a release of about 67.25% to 84.02% of drug

release. For the formulation f7 the percentage amount of drug release was found to be with the pharmacopeial

limits.the comparission of dissolution profiles for formulated drotaverine and marketed drotaverine tablets were

shown in the table no 8 and it was concluded that percent drug release for formulated drotaverine tablets was

more when comparared to marketed drotaverine tablets.

Taste evaluation of all formulations: The time intensity study for taste in human volunteers of the formulated

drotaverine hydrochloride with the polymer SD revealed considerable masking of the bitter taste of drotaverine

HCl with degree of bitterness below the threshold value within 120seconds (See Table 8). Sensory evaluation of

the tablets with both polymers proved good palatability.

6. CONCLUSION

This study has established effective taste masking of drotaverine HCl with the use of the solid dispersion

techniques using urea and mannitol as carriers. Taste masking and rapid dissolution of drotaverine HCl tablets

formulated in this investigation may possibly help in the administration of drotaverine HCl in a more palatable

form in the absence of water and more importantly since drotaverine HCl solid dispersion tablet formulations are

not presently in the market. Hence, “patient-friendly dosage form” of bitter drugs, especially for pediatric and

geriatric patients, can be developed using this technique.

7. REFERENCES

A. Rajpoot, Formulation and In-vitro Evaluation of Immediate release tablets of Drotaverine HCl, J. Chem.

Pharm. Res, 3(4), 2011, 333-341





Srikanth MV, Uhumwangho MU, Sunil SA, Design and evaluation of taste masked Drotaverine HCl

orodispersible tablets using polymethacrylate polymers, Der Pharmacia Lettre, 2(6), 2010, 223-231.

Vijayanand P, Formulation, Development and Evaluation of Novel Dosage Form Containing Silk Fibroin for

Elderly Patients, RJPBCS, 3(1), 2010, 524,



Harish G et.al Indian Journal of Research in Pharmacy and Biotechnology


EFFECT OF DIFFERENT DISINTEGRANTS ON CIPROFLOXACIN

CONVENTIONAL TABLETS Harish.G

1*, Ch.Bhargavi

1, A.Riyajune

1, Md.Yasmeen

1, Syed Mohammed

1, Ch.Murali Krishna

1, Sk.Jilani

1,

Nirajana.V.A2

1.Nimra College of Pharmacy, Jupudi, Vijayawada.

2. Faculty of pharmacy, Sri Ramachandra University, Chennai.

*Corresponding author: E.Mail: [email protected]

ABSTRACT

The objective of the present study is to design and evaluate the effect of disintegrating agents such as

Starch, Cross Caramellose Sodium and Sodium starch glycolate on ciprofloxacin tablet. The nature of the

Ciprofloxacin which is slightly soluble in water which affects the drug disintegration process there by inhibits the

drug release from the Conventional Tablet. Hence in the present study the effect of disintegrating agents at

different concentrations is carried out on the ciprofloxacin hcl to find out the optimized concentration followed by

stability studies for a period of 3 months.

INTRODUCTION

Despite increasing interest in controlled-release drug delivery systems, the most common tablets are those

intended to be swallowed whole and to disintegrate and release their medicaments rapidly in the gastrointestinal

tract (GIT). The proper choice of disintegrant and super-disintegrant to improve its consistency of performance is

of critical importance to the formulation development of such tablets. Drug release from a solid dosage form can

be enhanced by addition of suitable disintegrants. In more recent years, increasing attention has been paid to

formulating fast dissolving and/or disintegrating tablets that are swallowed, but also orally disintegrating tablets

that are intended to dissolve and/or disintegrate rapidly in the mouth. The present study is an attempt to select best

possible combination of drug and disintegrating agent to formulate rapidly disintegrating tablet of ciprofloxacin

conventional tablets which disintegrates faster thereby reducing the time of onset of action. Lactose is selected as

diluents, Starch, Sodium starch glycolate, CCS and Crospovidone were selected as disintegrants. PVP K 30M

paste was used as a binder in all formulations, Magnesium stearate and Talc as a Lubricant, Aerosil as a Glidant.

The percentage Drug content of all tablets was found to be between 95% - 105%, which was within the limit.

From the data obtained, it is observed that the formulation containing crosprovidone disintegrant disintegrate

rapidly when compared to other disintegrating agents such as Starch, SSG, and CCS with ciprofloxacin.

MATERIALS AND METHODS

Ciprofloxacin obtained as a gift sample from Intex chemicals Pvt ltd, Chennai. Starch, Cross Caramellose

Sodium and Sodium Starch Glycolate was obtained from Fischer Ltd, Chennai. All other excipients which are

used are of high standard analytical grade.

Pre-Formulation Studies:

Drug-excipients compatibility studies: Compatibility of drug with excipients was determined by FTIR using

kBr pellet technique, in the wavelength region of 4000-400cm-1

.

Table 1: Formulation Table of Ciprofloxacin Hcl conventional tablet

Formulation FA1 FA2 FA3 FB1 FB2 FB3 FC1 FC2 FC3 FD1 FD2 FE1 FE2

Ciprofloxacin

(mg)

500 500 500 500 500 500 500 500 500 500 500 500 500

Starch (%) 50 10 15 - - - - - - - - - -

SSG (%) - - - 4 5 6 - - - - - - -

CCS (%) - - - - - - 10 20 30 - - - -

BCD (%) - - - - - - - - - 40 80 40 80

Aerosil (%) 200 200 200 200 200 200 200 200 200 200 200 200 200

Lactose 220 260 255 246 265 264 260 250 240 230 90 230 90

PVP (%) 50 50 50 50 50 50 50 50 50 50 50 50 50

Talc (%) 50 50 50 50 50 50 50 50 50 50 50 50 50

Magnesium

Sterate (%)

30 30 30 30 30 30 30 30 30 30 30 30 30

Total Weight

(mg)

1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000






Quantity sufficient of Ciprofloxacin for a batch of 50 tablets was separately mixed to ensure complete

mixing. A tablet containing 500 mg equivalents of ciprofloxacin was compressed. All ingredients were weighed

and passed through 40# sieve, blended in a Poly Bag except Magnesium Stearate for 10 minutes. Mix half the part

of the disintegrated with the above mixture after passing through the sieve. The resultant mixture was wet massed

using suitable binder (qs) for granulation. This wet mass was passed through 20# sieve in order to form granules.

These granules were dried and the dried granules were passed through 30# sieve. These dried granules were

lubricated with Magnesium stearate, which was previous, passed through 60# Sieve. The lubricated granules were

punched to tablets using single punching machine.

Drug content: The estimation of drug content for ciprofloxacin tablets was performed by crushing three tablets

and quantity equivalent to 45mg was taken and determined using 0.1M Hcl using UV spectrophotometer at about

276nm

Weight Variation: The USP weight variation test was run by weighing 20 tablets individually, calculating the

average weight, and comparing the individual tablet weights to the average. The tablets met the USP tests that

were not more than 2 tablets were outside the percentage limit and no tablets differed by more than 2 times the

percentage limit.

Hardness: Hardness of the tablets was determined by breaking it between the second and third fingers with

thumb being as a fulcrum. There was a sharp snap the tablet was deemed to have acceptable strength. Hardness of

the tablets was determined by Stokes Monsanto Hardness Tester and the hardness should be found within the

range of 3.5-5.5 kg/cm².

Friability: The friability of tablets was determined by Roche Friablator. 20 tablets were taken and weighed. After

weighing the tablets were placed in the Roche Friablator and subjected to the combined effects of abrasion and

shock by utilizing a plastic chamber that revolves at 25RPM for minutes dropping the from a distance of six

inches with each revolution. After operation the tablets were de-dusted and reweighed.

Content Uniformity: In this test, 30 tablets were randomly selected contained for sample, Ciprofloxacin

Hydrochloride should contain not less than 98.0 per cent and not more than 102.0 percent.

Thickness: The thickness of a tablet was the only dimensional variable related to the process. 10 tablets were

measured for their thickness and diameter with a Caliper, Thickness Gauge. Average thickness and diameter were

calculated.

Disintegration Test: Disintegration time is considered to be one of the important criteria in selecting the best

formulation. For most tablets the first important step toward solution is break down of tablet into smaller particles

or granules, a process known as disintegration. Place one tablet into each tube and suspend the assembly in to the

1000ml beaker containing water maintained at 37 ± 2oC and operate the apparatus for 30 seconds. Remove the

assembly form the liquid. Observe the tablets, if one or two tablets fail to disintegrate completely; repeat the test

on 12 additional tablets. The requirement is met if not less than 16 of the total of 18 tablets tested are

disintegrated.

In-Vitro Drug Release Studies: In our case to study the release kinetics of drug we used USP II apparatus

(Paddle type, 2) with 900 ml, pH 6.8 phosphate buffer as the dissolution medium. The paddle was rotated 50 rpm

and 5ml of aliquots were withdrawn at pre-determined time intervals and an equal amount of thee medium was

replaced to maintain sink conditions. The aliquots were diluted suitably and the amount of drug(s) released was

determined spectrophotometrically using U.V. at wavelength 271 nm.

Evaluation of Formulated Ciprofloxacin Tablet:

Evaluation of blend characteristics: Ciprofloxacin Tablet was prepared by using wet granulation method.

Various formulations were made as shown in table: 6. The Formulated Ciprofloxacin tablet were evaluated for

Pre-formulation parameters like angle of repose, bulk density, tapped density, Compressibility index and

Hausner’s Ratio.

The results of disintegration time of all the tablets prepared by wet granulation found to be varied with

change in concentration of disintegrating agents from few seconds to several minutes. Formulations FD 1 and FE1

disintegrated within 3min and found to be more effective. The disintegration time of the tablets using different

disintegrants decreases in the following order BC > croscarmellose sodium > SSG > Starch. It is observed that,

when BC is used as disintegrant, tablets disintegrate rapidly with in less time compared to other tablets prepared

using croscarmellose sodium, starch and sodium starch glycolate disintegrants. Though tablets prepared by intra

and extra granulation method found to be more effective in comparison with formulation prepared by only extra





granulation. When concentration of Starch, SSG, CCS and BC is increased, the disintegration time was reduced

The angle of repose of prepared Ciprofloxacin tablet was in the range of 20°-30°. Normally if the value falls

between 20°-30°, it shows good flow property. The bulk density and tapped density were found to be in the range

of 0.37 to 0.38 g/cm3 and 0.44 to 0.45g/cm

3 respectively. A Hausner ratio was within the range of 1.16 to 1.17,

lesser than 1.25 is considered to be an indication of good flow property. The compressibility index was within the

range of 5-15 hence falls within the excellent range.

The results were tabulated in table 15. The pre-formulation characteristics results for all the

formulation of ciprofloxacin tablets using FB as disintegrating agent found to be within the range, compressibility

index for FB1 and FB3 was found to be with in good range of 12-16 were as FB2 was in excellent range. The

angle of repose of prepared tablet was in the range of 20°-30°. Normally if the value falls between 20°-30°, it

shows good flow property. The bulk density and tapped density were found to be in the range of 0.34 to

0.36g/cm3 and 0.39 to 0.40 g/cm

3 respectively. A Hausner ratio was within the range of 1.07 to 1.18, lesser than

1.25 is considered to be an indication of good flow property. The compressibility index was within the range of 5-

15 hence falls within the excellent range. The results were tabulated in table 16.

The pre-formulation characteristics results for all the formulation of ciprofloxacin tablets using FC as

disintegrating agent found to be within the range, angle of repose and compressibility index was found to be

within good range. The angle of repose of prepared ciprofloxacin tablet was in the range of 20°-30°. Normally if

the value falls between 20°-30°, it shows good flow property. The bulk density and tapped density were found to

be in the range of 0.35 to 0.36 g/cm3 and 0.39 to 0.41 g/cm

3 respectively. A Hausner ratio was within the range of

1.08 to 1.18, lesser than 1.25 is considered to be an indication of good flow property. The compressibility index

was within the range of 5-15 hence falls within the excellent range. The results were tabulated in table 17. The

formulation of ciprofloxacin using 4% B.C disintegrant found to be within the limits for both FD1 and FD2 and

falls in good range. The angle of repose of prepared ciprofloxacin tablet was in the range of 20°-30°. Normally if

the value falls between 20°-30°, it shows good flow property. The bulk density and tapped density were found to

be in the range of 0.36 to 0.38g/cm3 and 0.40 to 0.41 g/cm

3 respectively. A Hausner ratio was within the range of

1.07 to 1.16, lesser than 1.25 is considered to be an indication of good flow property. The compressibility index

was within the range of 5-15 hence falls within the excellent range.

The angle of repose of prepared ciprofloxacin using FE as disintegrant was in the range of 20°-30°.

Normally if the value falls between 20°-30°, it shows good flow property. The bulk density and tapped density

were found to be in the range of 0.36 to 7 g/cm3 and 0.340 to 0.41g/cm

3 respectively. A Hausner ratio was within

the range of 1.10 to 1.11, lesser than 1.25 is considered to be an indication of good flow property. The

compressibility index was within the range of 5-15 hence falls within the excellent range.

The post compressional characteristic for all the formulated batches was found to be within the limits as

per Indian pharmacopeia 2007. The hardness was found to be within the range of 3.5 to 5.5 Kg/cm2 in all the

formulations indicating good mechanical strength with an ability indicating physical and mechanical strength with

an ability to withstand physical and mechanical stress conditions while handling. In all the formulations, the

friability value is less than 1% giving an indication that tablets formulated are mechanically stable. All the tablet

formulations passed the weight variation test. The weight of all the formulations was found to be within the limits.

The assay of all the formulations was found to be with in the 90% to 110% acceptable limits.

The disintegration time of the entire Formulated batch varies with change in concentration of

disintegrating agents from few seconds to several minutes. Formulations FD2 and FE2 disintegrated within 3min

and found to be more effective. The disintegration time of the tablets using different disintegrants decreases in the

following order Starch > CCS > SSG >CP. It is observed that, when BC is used as disintegrant, tablets

disintegrate rapidly with in less time compared to other tablets prepared using croscarmellose sodium, starch and

sodium starch glycolate disintegrants. Though tablets prepared by intra and extra granulation method found to be

more effective in comparison with formulation prepared by only extra granulation. When concentration of Starch,

SSG, CCS and BC is increased, the disintegration time was reduced significantly show in table 2..

In vitro drug release studies were conducted for the formulation using USP dissolution apparatus type-II

(paddle), at 50 rpm. The percentage drug release at the end of 30 min was found in the range 48 – 73% using FA

as disintegrant and 67-77% using FB as disintegrant. Invitro drug release studies were conducted for the

formulation using USP dissolution apparatus type-II (paddle), at 50 rpm. The percentage of drug release was

determined at a time interval of 0, 5, 10, 15, 20, 25, 30 min and at the end of 30 min it was found in the range 80-





95% using FC as disintegrant. Invitro drug release studies were conducted for the formulation using USP

dissolution apparatus type-II (paddle), at 50 rpm. The percentage of drug release was determined at an time

interval of 0, 5, 10, 15, 20, 25, 30 min and at the end of 30 min it was found in the range 81-95% using 4% BC as

disintegrant and 78-98% using 8% BC as disintegrant shown in figure 9, 10 and 11.

Stability Studies: Drug molecules are of reactive naturally, the additives are considered to be inert substance but

this may not be true for all additives in a formulations. Hence, in developing any formulation, when additive are

selected the same must be super imposed on to drugs and with other additives present in the formulation, to see

how compatible they are with the other formulation ingredients. A lot of literature has got piled up on drug-

excipients incompatibilities, which is a handy tool in the hand of a formulation man, to avoid possible drug-

excipient interactions. But even with this literature at the back, formulation may be highly individualistic and each

formulation may have problems of its own. There are methods called FTIR, differential scanning calorimetric,

thin layer chromatography for investigating these interactions in short period of time.

On many occasions, even with the sum total of knowledge available, it is not possible to envisage, all the

interactions and a formulation while remaining good for a certain period of time, may go bad in the long run.

There is not ready made answer for such situation and all that is possible is to “wait and watch”, the method

called “Real time study”. As per ICH guide line for stability study, which advice the formulation to store their

products at 30 ° c and 65 % RH to find out actual shelf life period or to assure the product quality free from

unwanted interactions. Real time study of ICH guidelines involves storage of products at 40° C& 65 % RH for the

complete proposed shelf life period, and analyzing the product sample every month in the first 3 months, every 3

months from 4th month onwards up to one year, every 6 month in the second year of storage, afterwards once in a

year till shelf life is completed. ICH guidelines also demands for storing samples at 40 ° c and 75 % RH (stress

condition or accelerated stability study) for relatively short period of time (3-6 months) which depends on claimed

shelf life period as well as the zone (zone 1/2/3/4 of the world) to which the product is meant to be exported. This

later study (with stress conditions) is to mine the alternating climates condition during the shelf life of the product.

The stability parameters for all the formulation were evaluated after 15, 30, 45, 60, and 90 days for 40 °C at 75%

RH.

Tab 2: Disintegration Time of the Ciprofloxacin Fast disintegrating tablet

Formulation With Disk Without Disk

I II III I II III

FA1 11min 43 sec 10 min 30 sec 10min 52sec 14min 32 sec 15min 11sec 15min 48 sec

FA2 8min 2sec 9min 33 sec 8min 18 sec 11min 14sec 12min 31 sec 11min 56 sec

FA3 4min 41 sec 5min 8 sec 4min 55sec 9min 23sec 9min 51 sec 8min 50sec

FB1 11min 41 sec 10min 21 sec 10min 54 sec 14min 11sec 14min 56sec 13min 34sec

FB2 8min 43sec 9min 21sec 9min 5sec 12min 37sec 14min 12sec 12min 44sec

FB3 4min 21sec 5min 32 sec 4min 13sec 7min 23sec 7min 47 sec 6min 43sec

FC1 9min 21sec 8min 55 sec 10min 12 sec 11min 15 sec 11min 24 sec 10min 55min

FC2 7min 43sec 8min 11 sec 8 min 5sec 9min 22 sec 9min 17 sec 10 min 31 sec

FC3 5min 22 sec 5min 42sec 6min 31sec 6 min 4 sec 7min 41sec 7min 18sec

FD1 4min 45 sec 4min 52 sec 3min 21sec 7min 19 sec 7min 47 sec 6min 14 sec

FD2 2 min 51 sec 2min 11 sec 1min 33sec 3min 46sec 4min 23 sec 4min 11sec

FE1 4min 31sec 5min 55sec 4min 14sec 9min 50sec 8min 14sec 8min 19sec

FE2 3min 11 sec 2 min 47 sec 3min 17sec 5min 11sec 6min 42sec 5min 4 sec





Fig 1: IR spectra of the pure drug, Ciprofloxacin

Hcl.

Fig 2: IR spectra of FA

Fig 3: IR spectra of FB Fig 4: IR spectra of FC

Fig 5: IR spectra of the FD&FE Disintegrant

Fig 6: IR spectra of the mixture of the

Ciprofloxacin and FA

Fig 7: IR spectra of the mixture of the Ciprofloxacin

and FB

Fig 7: IR spectra of the mixture of the

Ciprofloxacin and FC





0

20

40

60

80

100

0 10 20 30 40

% C

um

ula

tive

am

ou

nt

of

dru

g

rele

ase

Time in Minutes

FA1

FA2

FA3

FB1

FB2

FB3

0

20

40

60

80

100

0 20 40 % C

um

ula

tive

am

ou

nt

of

dru

g

rele

ase

Time in minutes

FC1

FC2

FC3

0

20

40

60

80

100

120

0 20 40

% C

um

ula

tive

am

ou

nt

of

dru

g

rele

ase

Time in minutes

FD1

FD2

FE1

FE2

Fig 8: IR spectra of the mixture of the Ciprofloxacin and FD &FE

Fig 9: Dissolution profile of Ciprofloxacin tablets

Using FA and FB as disintegrant

Fig 10: Dissolution profile of Ciprofloxacin tablets

Using FC disintegrants

Fig 11: Dissolution profile of Ciprofloxacin tablets Using FD disintegrants

CONCLUSION

Selecting appropriate formulation excipients and manufacturing technology can obtain the design

feature of fast disintegrating tablet. The disintegrants have the major function to oppose the efficiency of the tablet

binder and the physical forces that act under compression to form the tablet. The stronger the binder, the more

effective must be the disintegrating agents in order for the tablet to release its medication. Ideally, it should cause

the tablet to disrupt, not only into the granules from which it was compressed, but also into powder particles from

which the granulation was prepared. From this study, it is concluded that the disintegrants such as Starch, SSG,

CCS was compared with combination of disintegrants and in this study optimized combination of disintegrant

prepared by intra and extra granulation method was found to be the most effective as they disintegrate rapidly

when compared to other disintegrants, and the percentage drug release also shows a higher dissolution profile.

REFERENCES Budavani S O, Neil N J, Smith A, The Merck Index, An Encyclopedia of Chemicals, Drugs and Biologicals,

29th Ed, Published by Merck Research Laboratories, Division of Merck & Co. Inc, 1996, 181

http://www.pharmainfo.net/excipients





Caramella C, Colombo P, Conte U, La Manna A, Tablet disintegration update: the dynamic approach, Drug.

dev.Ind.Pharm, 13 (12), 1987, 2111-2145.

Chaudhary K P R, Sujata Rao, Formulation and Evaluation of Dispersible tablets of poorly soluble drugs, Indian

J. Pharm. Sci, (2), 1992, 31-32.

Cohen Y, Lach JL, Factors affecting the Effect of Disintegrating Agent, J. Pharm. Sci, 52, 1963, 122.

Cousin, Rapidly Disintegrable multiparticular Tablets, U S Patent, 5, 464, 632 (1995).

E Sallam, Ibrahim H, R Abu Dahab, M Shubair, Enam Khalil, Drug.Dev and Industrial Pharmacy, Vol. 24(6),

1998, 501–507.

Garnpimol C, Ritthidej, Parichat Chomto, Sunibhond Pummangura, Piamsak Menasveta, Chitin and Chitosan as

Disintegrants in Paracetamol Tablets, Drug Dev. Ind. Pharm, 20(13), 1994, 2109-2134.

Grasono Alesandro, US Patent 6,197,336 2001

Gupta G D, Gaud R S, Formulation and Evaluation of Nimesulide Dispersible Tablets Using Natural

Disintegrants, Indian.J. Pharm Sci., 62(5), 2000, 339-342.

H N Bhargava, D Shah, A Anaebonam, B Oza, An Evaluation of Smecta as a Tablet Disintegrant and Dissolution

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Papola Vibhooti Indian Journal of Research in Pharmacy and Biotechnology


CHRONOTHERAPY FOR NOCTURNAL ASTHAMA Papola Vibhooti

* Rajan G, Bisht Seema, Dr.Kothiyal Preeti

Shri Guru Ram Rai Institute of Technology & Sciences Dehradun, Uttarakhand, India

*Corresponding author:[email protected]

ABSTRACT Chronotherapeutic drug delivery system is useful in the treatment of disease, in which drug

availability is timed to match rhythm of disease, in order to optimize therapeutic effect and minimize side

effect. Nocturnal asthma is defined as any sleep related worsening of reversible airway disease.

Approximately 80 percent of severe asthmatic attacks occur between midnight and 8 a.m. Nocturnal

asthma is associated with critical symptoms and urgent need for proper medications. The onset of

nocturnal asthmatic attacks is rare in the first part of night, 80% of asthmatic attacks occur between

midnight and 8 a.m., and deaths from asthma are more common during these hours. In a study of asthma

mortality, 79% of the patients who died had a disturbed sleep before the death. In a survey of almost 8000

patients with varying degrees of asthma found that approximately - 75% of asthmatics attacks happened

once a week with symptoms, 64 % three times a week, and 39 % every night. Nocturnal asthma is

currently controlled by taking either a long-acting β2 agonists like salmetrol inhalers, sustained release

theophylline. All the current sustained release formulation has a shortcoming of inability to maintain high

blood levels for that long period. This may lead to leaving the patient unprotected against the worse

events of nocturnal asthma. Thus, a smart drug delivery that is administrated before sleep and maintains

high blood levels for longer period (from midnight to 8 am in the morning) could be very much beneficial

for proper management of nocturnal asthma.

Key words: Chronotherapy, cardian rhythm, nocturnal, morbidity, diurnal

INTRODUCTION

In order to increase the effectiveness of drug there are many approaches which have been applied. The

pharmaceutical formulations which are for direct ingestion for oral administration and orally administrated drugs

are generally absorbed from the gastrointestinal tract. Many functions of the human body show considerable

change in a day. These variations cause changes both in disease state and in plasma drug concentrations (Lin,

2011). Coordination of biological rhythms with medical treatment is called chronotherapy. Chronotherapy

considers a person’s biological rhythms in determining the timing and amount of medication to optimize a drug’s

desired effects and minimize the undesired ones. Study of influence of biological rhythm on the effects of

medication is known as chronopharmacology while the science of study of biological rhythms is known as

chronobiology. With the understanding of biological time keeping the idea came that these rhythms must affect

how the body responds to drugs administered over the course of the day (Hizli, 2009) (Ohdo, 2006). Appropriate timing of administration can improve efficacy and diminish toxicity. Chronotherapy is

relevant when the risk or intensity of the symptoms of disease vary with time as in the case of allergic rhinitis,

arthritis, asthma, myocardial infarction, congestive heart failure, stroke and peptic ulcer disease (Haus, 2007).

The human circadian rhythm is based on sleep activity cycle, is influenced by our genetic makeup and hence,

affects the body’s functions day and night (24-hour period) (Devdhawala, 2010).

Coordination of biological rhythms with medical treatment is called Chronotherapy. Chronotherapy

considers a person’s biological rhythms in determining the timing and amount of medication to optimize a drug’s

desired effects and minimize the undesired ones. Study of influence of biological rhythm on the effects of

medication is known as chronopharmacology while the science of study of biological rhythms is known as

chronobiology. To understanding the biological time keeping the idea came that these rhythms must affect how

the body responds to drugs administered over the course of the day (Awasthi, 2010) (Hizli, 2009).

raditionally, drug delivery systems have focused on constant/sustained drug output with the objective of

minimizing peaks and valleys of drug concentrations in the body to optimize drug efficacy and to reduce adverse

effects. A reduced dosing frequency and improved patient compliance can also be expected for the

controlled/sustained release drug delivery systems, compared to immediate release preparations (Saigal N, 2009).

Some of the rhythms that affect our body are ultradian (cycles shorter than a day like firing of neurons take

milliseconds), circadian (cycles lasting 24 h such as sleeping and waking pattern), infradian (cycles longer than a

day like menstrual cycles) and seasonal rhythms (such as seasonal affective disorders causing more depression in





susceptible individuals in winter). Circadian rhythm governs every process of our body. The term circadian

rhythm was first given by Halberg and Stephens in 1959.

Chronotherapy: The term "chrono" basically refers to the observation that every metabolic event undergoes

rhythmic changes in time. Perhaps the best known and studied chronobiology frequency is the circadian rhythm

which approximates the earth's 24-hour rotation around the sun. Researchers have recently concluded that both

disease states and drug therapy are affected by a multitude of rhythmic changes that occur within the human body.

Chronotherapeutic refers to a treatment method in which in vivo drug availability is timed to match rhythms of

disease in order to optimize therapeutic outcomes and minimize side effects. It is based on the observation that

there is an interdependent relationship between the peak-thorough rhythmic activity in disease symptoms and risk

factors, pharmacologic sensitivity and pharmacokinetics of many drugs. Biological rhythms concern to the control

of biological functions including those of the autonomic nerve system, endocrine system, and immune system, are

fundamental in homeostasis and in protection against various diseases.

Chronotherapeutics: The first chronotherapy to be widely applied in clinical practice was introduced in the

1960s the alternate-day morning schedule of conventional tablet corticosteroid medication. Other chronotherapies

have since been widely used in clinical medicine in the U S, Europe, and Asia; These include special evening

theophylline systems for chronic obstructive pulmonary disease, conventional evening H2 -receptor ant agonists

for peptic ulcer disease, and conventional evening cholesterol medications for hyperlipidemia.

Chronotherapeutics refers to a treatment method in which in vivo drug availability is timed tomatch

rhythms of disease in order to optimize therapeutic outcomes and minimize side effects. It is based on the

observation that there is an interdependent relationship between the peak-to trough rhythmic activity in disease

symptoms and risk factors, pharmacologic sensitivity, and pharmacokinetics of many drugs .

Figure.1.Human biological lock

Circadian Time Structure: Circadian rhythms are the rhythm in the chronotherapeutic and the dysfunction of

circadian rhythms can affect the brain functioning and it can be improved by the chronotherapeutic approach.

Circadian rhythms are self-sustaining, endogenous oscillations that occur with a periodicity of about 24

hours.Circadian rhythm regulates several body functions such as metabolism, physiology, behavior, sleep

patterns, hormone production, and so on. The circadian rhythm not only affects most physiological functions but

also influences the absorption, distribution, metabolism, and elimination (ADME) of drugs, leading to changes in

drug availability and target cell responsiveness.





Figure.2. Human circadian time structure

Circadian rhythms can change the sleep-wake cycles, hormone release, body temperature, and other important

bodily functions driving the alteration of various physiological, biochemical, and behavioral processes.

Circadian Rhythms of Diseases: The biological rhythm studies help in defining the temporal organization of

human beings. One means of illustrating the human circadian time structure is to depict the peak time of 24-h

rhythms on a clock--like diagram. The 24 h clock pattern of diseases showing prominent day-night patterns when

symptoms of disease are most frequent .Variation in the severity of many diseases over a 24-hour period is well

known diseases such as bronchial asthma, myocardial infarction, angina pectoris, rheumatic disease, ulcers 43,

diabetes, and attention deficit syndrome, hypercholesterolemia and hypertension show symptomatic changes due

to circadian rhythm city. Aggravation of asthmatic attacks occur after midnight or in the early morning due to

limited lung function promoted by circadian changes at that time. Many common diseases also display a marked

circadian variation during onset or exacerbation of symptoms,

Figure.3.The circadian pattern of diseases

Asthma: Asthma may be the most common disease with the largest circadian variation. It is considered as a

chronic condition where airways are hyperreactive to certain irritants which can constrict them, and so making

difficulty in breathing. Such a constriction is often called as bronchospasm and is followed by excess production

of mucus and inflammation in the membranes lining the walls of the airways. “Breathing through a straw” is a

commonly description that can explain the situation. Allergens, fumes, smoke, and/or dry and cold air are

common irritants. Laughing or exercise can also cause the constriction of airways. Asthmatics vary considerably

in the impact of illness on their life, response to treatment and severity of symptoms. Because asthma has such a

striking circadian variation, several types of chronotherapy have been tried. In one study, use of a timed-release

formulation of theophylline (Theo- 24) achieved therapeutic drug concentrations during the night and avoided

toxic levels during the day. Asthma is well suited for chronotherapy, with beta 2- agonists and oral

corticosteroids.





Nocturnal asthma: Some biological rhythms come about monthly or even annually, asthma changes fairly

predictably on a circadian cycle or 24 hour. Even in normal, lung function differs between day and night. The

activity of the lung exhibits a circadian rhythm with a maximum around 4 p.m. and a minimum around 4 a.m. In

asthmatic patients, the intensity of variation in lung function is as much as 50% in a day. Bronchial reactivity

generally follows the same circadian cycle in Asthmatic patients. It can be defined as any sleep-related Worsening

of reversible airway disease. Shortness of breath or wheezing at night is symptoms generally shown. Nocturnal

asthma is associated with critical symptoms and urgent need for proper medications. The onset of nocturnal

asthmatic attacks is rare in the first part of night 80% of asthmatic attacks occur between midnight and 8 a.m., and

deaths from asthma are more common during these 9 hours. In a study of asthma mortality, 79% of the patients

who died had a disturbed sleep before the death. In a survey of almost 8000 patients with varying degrees of

asthma found that approximately 75% of asthmatics attacks happened once a week with symptoms, 64 % three

times a week, and 39 % every night.

Causes of Nocturnal Asthma: Nocturnal asthma is probably because of multiple factors than to a single cause.

Asthma attacks are aggravated mainly by irritants. Exposure to allergens daytime can be as important as exposure

to allergens in the bedroom during sleep. A series of physiological events from three to eight hours are poorest

about 4 a.m after the initial exposure called late-asthma response (LAR); it may match to the night time for some

people and it can persist over nights. An increase in a susceptible patient's risk of LAR from 40 to 90 % by

allergen exposure in the evening. In some people, the inflammation worsens correspondingly with circadian

changes in peak expiratory flow rates in night. Another contributing factor to nocturnal asthma may be airway

secretions. About 70 % of asthmatics experience postnasal drip and/or chronic sinusitis. Asthma often improves

when the sinuses are cleared in daytime. Influence of airway temperature on onset of symptoms was studied.

Bronchospasm is produced after a brief exposure to cold and dry air. Breathing warm humidified air can reverse

this.

Diagnosis of Nocturnal Asthma: If asthma symptoms worsen at night it is important to inform clinician.

Monitoring lung function using a peak flow meter is necessary. Peak flow meter is a portable device that

measures the lung volume and how time by which air can be expelled from the lungs. Low peak flow meter

values indicates that there is a tightening of the airways, and can be an early warning of impending respiratory

symptoms, such as shortness of breath and wheezing. Recording peak flow rates at bedtime serves as a document

for nocturnal asthma. During any awakening at night and in the morning also serves as a record .

Chronotharapy of asthma:Asthma and Sleep: In asthma, the relative role of circadian and sleep systems has

been a subject of controversy, and this issue remains unresolved.Initially, it was suspected that sleep systems

played the major role. In a study of shift-workers, there appeared to be an immediate phase shift in the circadian

rhythm of peak expiratory flow (PEF) when subjects rotated shifts, such that the decline in airway function

remained coupled to the sleep period. In asthma, the resistance increases progressively across the night, whether

subjects sleep or not, although the increase is much greater during sleep. These results are supported by the

observations that the onset of asthmatic attacks is less common in the first part of the night. These data allow us to

reach certain conclusions. First, it seems likely that both circadian and sleep factors play a role in asthma. Also,

the progressive decline in airway function across the night does not suggest a typical change in a neuronal control

process coupled to sleep. Notably, airway function is maximal at the time of increased sleepiness during the

afternoon, and declines as sleep pressure dissipates during sleep.

Effect of nocturnal asthma on daytime performance, morbidity, and mortality: With the decreased sleep

efficiency in asthma, and reports of daytime tiredness/sleepiness, the possibility exists that performance at work

or school will be affected. A study of nocturnal asthmatic and control subjects demonstrated that the asthmatic

subjects had increased scores for subjective sleepiness. This reflected decreased objective sleep quality.

Interestingly, daytime cognitive performance was worse in the nocturnal-asthma group. This area of research

needs further investigation. The morbidity of ventilatory failure, and also the mortality in asthma, are linked to the

nocturnal worsening of lung function, which may be related to a blunted arousal mechanism caused by

fragmented sleep. Interestingly, in one study of asthma mortality, 79% of the 168 patients who died had a sleep

disturbance reported prior to the terminal event This contrasts with the usually accepted mortality risk factors of a

previous ICU admission (5% in the study cited ), more than two hospitalizations/emergency- room (ER) visits in

the preceding year (28%), or psychologic disturbance (13%).





Circadian/sleep physiologic and challenge response:

Lung function and thoracic blood volume: It has been shown that sleep enhances the nocturnal increases in

airway resistance and also leads to marked reduction in the volume of hyperinflated lungs in patients with

nocturnal asthma. Such volume changes do not account directly for all of the nocturnal change in airway

resistance. However, artificially reducing lung volumes in awake asthmatic individuals to their typical levels

during sleep did trigger worsening of airway obstruction, suggesting that the effects of sleep on lung volume

could contribute to the nocturnal worsening of asthma. The effects of sleep on lung volume could be mediated by

several different mechanisms including:a sleep associated reduction in inspiratory muscle tone, a decrease in

pulmonary compliance; and an increase in intrapulmonary blood pooling. In particular, the effects of sleep on

intrapulmonary blood volume (IPBV) are intriguing, since there is evidence that such pooling of blood can

promote airway narrowing. Using capillary volume (Vc) as a surrogate marker of IPBV, it has been shown that

Vc increased overnight in asthmatic individuals with nocturnal worsening of lung function .

Gastrointestinal Function and the Lung: There is significant variation in gastrointestinal (GI) function during

sleep. The circadian rhythm of human basal gastric-acid secretion is characterized by a peak in the early evening

and a nadir between 5:00 and 11:00 in the morning. There are conflicting data as to whether esophageal acid

causes a decrease in airway function. In one study of sleeping individuals with nocturnal asthma, no significant

acute or sustained change was observed in airflow resistance relative to periods of increased esophageal acid

content, suggesting that gastroesophageal reflux (GER) contributed little to the nocturnal worsening of asthma.

Although it appears that asthma is more responsive to the ets of GER during the diurnal cycle than during the

nocturnal cycle, the exact role of circadian/sleep effects in esophageal acid-induced bronchoconstriction remains

unclear.

Nasal-Sinus–Lung Interaction: There is evidence that upper-airway disease (i.e., allergic rhinitis, sinusitis, and

nasal polyps) influences and may contribute to the intensity of lower-airway disease. Allergic rhinitis, for

example, can intensify airway responsiveness and even provoke asthma symptoms. Data indicate that treatment of

allergic rhinitis diminishes bronchial responsiveness and asthma. Active sinusitis can also cause an increase in the

asthma process as shown in animal models, which appears to involve drainage of nasal mediators into the lower

airway. Other processes that link the nasal sinus to the lung have been identified in studies of viral infections of

the nose that produce an increase in lower-airway reactivity. Also, there is a day– night cycle in nasal patency and

perhaps in inflammation. All of these data suggest an important interaction between the nasal sinus and lower-

airway function.

Chronotherapy

General Principles: Bodily functions have been incorrectly assumed to be relatively constant throughout the 24 h

of the day and other periods of time. Numerous studies have shown that the kinetics and dynamics of

pharmacotherapies vary significantly according to the biologic time of administration during the 24 h-cycle,

menstrual cycle, or annually, owing to the cumulative effect of endogenous rhythms in crucial physiologic and

biochemical functions. Chronotherapeutics is the synchronization of medication levels in time with reference to

need, taking into account biologic rhythms in the pathophysiology of medical conditions, and/or rhythm-

dependencies in patient tolerance for given chemical interventions. Chronotherapeutics can sometimes be

achieved by the judicious timing of conventional sustained- release (SR) formulations, although reliance on

special drug-delivery systems seems to constitute a more dependable means of matching drug level to biologic

need and tolerance.

β2-Agonists, Theophylline, and Anticholinergic Therapy: Certain SR formulations of theophylline can be

administered so that a rising blood level of the drug occurs when airway obstruction is increasing, while adverse

effects are reduced. For this purpose, SR theophylline is administered once daily, in the evening, for the

management of nocturnal asthma. Various tablet formulations for the sustained-release of b-agonists have been

used in a chronotherapeutic fashion for the management of asthma. As with theophylline, very little information

exists about comparing the effects of or adding a long-acting b2-agonist oral preparation to an inhaled

corticosteroid using chronotherapeutic techniques. Salmeterol has been shown to control symptoms of nocturnal

asthma to a substantial degree, and to improve sleep quality and daytime cognitive performance in patients with

chronic asthma. Drugs that antagonize the vagal nervous system should be useful in the management of nocturnal

asthma as a means of counteracting the enhanced nocturnal parasympathetic tone that occurs in the disease.





Corticosteroids and Leukotriene-active Drugs: Corticosteroids have been used in a chronotherapeutic manner,

with the finding that their long-term oral administration at 8:00 A.M. and 3:00 P.M. was more effective in

controlling nocturnal asthma than the same doses given at 3:00 P.M. and 8:00 P.M. Other studies have shown that

a single 3:00 P.M. dose of prednisone improved lung function and reduced airway inflammation more effectively

than the same single dose given at 8:00 A.M. and 8:00 P.M (Beam, 1992). Not only can oral steroids be dosed

chronotherapeutically, but inhaled corticosteroids can also be efficacious when used in this manner (Pincus,

1995). Although the leukotriene-active drugs, including zileuton, zafirlukast and montelukast, are new in the

treatment of asthma, they have been shown to alleviate the symptoms and the decrement in lung function seen in

nocturnal asthma. It has been shown that zileuton in particular decreased nighttime increases in leukotriene B4

(LTB4) and (LTE4) while improving lung function (Wenzel, 1995). Zafirlukast has also been shown to decrease

nighttime awakenings and improve morning PEF rates . Although these agents have only been studied at set doses

and times regardless of the presence or absence of nocturnalasthma, the improvements observed were significant,

and it is likely that these agents will prove very useful in the treatment of nocturnal asthma when used

chronotherapeutically.

Ideal Characteristics of Chronotherapeutic Drug Delivery System should:

be non-toxic within approved limits of use,

have a real-time and specific triggering biomarker for a given disease state,

have a feed- back control system (e.g. self-regulated and adaptative capability to

circadian rhythm and individual patient to differentiate between awake – sleep status),

be biocompatible and biodegradable, especially for parenteral administration,

be easy to manufacture at economic cost,

be easy to administer in to patients in order to enhance compliance to dosage regimen. Chronotherapeutic drug delivery systems: Controlled release formulations can be divided into subgroups of

rate-controlled release,delayed-release and pulsed-release formulations. Delayed-release formulations include

time controlled release and site specific dosage forms. When constant drug plasma levels need to be avoided, as in

chronotherapy, time-controlled or pulsed-release formulations are preferable, especially in the treatment of early

morning symptoms. By timing drug administration, plasma peak is obtained at an optimal time and the number of

doses per day can be reduced. Saturable first-pass metabolism and tolerance development can also be avoided.

Various technologies to develop timecontrolled peroral drug delivery systems have been extensively studied in

recent decades. Some of these systems are discussed in the following subsections.

Enteric-coated systems: Enteric coatings have traditionally been used to prevent the release of a drug in the

stomach Enteric coatings are pHsensitive and drug is released when pH is raised above 5 in the intestinal fluid.

These formulations can be utilised in time-controlled drug administration when a lag time is needed. Because of

the unpredictability of gastric residence, such systems cannot be the first choice when a time-controlled release is

required. In the treatment of nocturnal asthma, a salbutamol formulation containing a barrier coating which is

dissolved in intestinal pH level above about 6, has been successfully used. The system contains a core which is

film coated with two polymers, first with HPMC and then with a gastro-resistant polymer (Eudragit® L30D). In

this system the duration of the lag phase in absorption can be controlled by the thickness of the HPMC layer.

Figure.4.Schematic representation of Enteric coated system

Layered systems: These are one or two impermeable or semipermeable polymeric coatings (films or compressed)

applied on both sides of the core. To allow biphasic drug release, a three-layer tablet system was developed . The

two layers both contain a drug dose. The outer drug layer contains the immediately available dose of drug. An

intermediate layer, made of swellable polymers,separates the drug layers. A film of an impermeable polymer





coats the layer containing the otherdose of drug. The first layer may also incorporate a drug-free hydrophilic

polymer barrier providing delayed (5 h) drug absorption. Conte et al has also studied a multi-layer tablet system

(Geomatrix®).It consists of a hydrophilic matrix core containing the drug dose. This kind of three layer device

has been used in the treatment of Parkinsonian patients using L-- dopa/benserazide. Nighttime problems and

early-morning symptoms of Parkinsonism can be avoided by using a dualrelease Geomatrix@ formulation, which

allows daily doses of drug to be reduced and leads to extent of bioavailability 40 % greater than when a traditional

controlled release formulation is employed.

Figure.5. Geminex TIMERx technology based bilayered dual release tablet

Time-controlled explosion systems (TES): These have been developed for both single and multiple unit dosage

forms [80],[81]

. In Both cases, the core contains the drug, an inert osmotic agent and suitable disintegrants.

Individual units can be coated with a protective layer and then with a semipermeable layer, which is the rate

controlling membrane for the influx of water into the osmotic core. As water reaches the core, osmotic pressure is

built up. The core ultimately explodes, with immediate release of the drug. The explosion of the formulation can

also be achieved through the use of swelling agents. Lag time is controllable by varying the thickness of the outer

polymer coating.

Sigmoidal release systems (SRS): For the pellet-type multiple unit preparations, SRS containing an osmotically

active organic acid have been coated with insoluble polymer to achieve different lag-times. By applying different

coating thicknesses, lag times in vivo of up to 5 hours can be achieved. Release rates from SRS, beyond the lag

time, has been found to be independent of coating thickness.

Press-coated systems: Delayed-release and intermittent-release formulations can be achieved by press-- coating.

Presscoating, also known as compression coating, is relatively simple and cheap, and may involve direct

compression of both the core and the coat, obviating the need for a separate coating process and the use of coating

solutions. Materials such as hydrophilic cellulose derivatives can be used and compression is easy on a laboratory

scale. On the other hand, for large-scale manufacture, special equipment is needed. The major drawbacks of the

technique are that relatively large amounts of coating materials are needed and it is difficult to position the cores

correctly for the coating process. Conte et al have developed a press coated device in which the inner core

contains the drug and the outer coat is made of different types of polymers. The outer barrier, which controls drug

release, can be either swellable or erodible. Lag times can be varied by changing the barrier formulation or the

coating thickness (Halsas M, 1998). Hydrophilic polymers such as hydroxypropyl methylcellulose and sodium

alginate have been used in the coat to control drug release.

In recent years, various controlled release, especially time-controlled release; drug delivery systems based

on compression coating technology have been studied. Most such Formulations release drug after a lag phase,

followed by a rapid dissolution of a core. Tablets formulated with Penwest's TIMERx ®

oral controlled release

technology comprise an inner core containing drug and an outer layer compression-coated with TIMERx, a

hydrophilic matrix of the heteropolysaccharides xanthan and locust bean gum (Baichwal A, 2002).

Figure.6. TIMERx based drug delivery system Examples of chronopharmaceutical technologies: Currently key technologies in chronopharmaceutics includes:

CONTINR, physico-chemical modification of the active pharmaceutical ingredient (API), OROSR, CODASR,





CEFORMR, DIFFUCAPSR, chronomodulating infusion pumps, TIMERxR, threedimensional printing,

controlled-release (CR) erodible polymer and CR microchip strategies. Readers may find advantages and

disadvantages of each technology depending on their specific needs on the website of each developer/marketer

website before selection. Informations on FDA approval status and dosage formed were compiled from the FDA

electronic orange book.

For asthma (CONTINR technology): In this technology, molecular coordination complexes are formed between

a cellulose polymer and a non-polar solid aliphatic alcohol optionally substituted with an aliphatic group by

solvating the polymer with a volatile polar solvent and reacting the solvated cellulose polymer directly with the

aliphatic alcohol, preferably as a melt. This constitutes the complex having utility as a matrix in controlled release

formulations since it has a uniform porosity (semipermeable matrixes) which may be varied. This technology has

concretely enabled the development of tablet forms of sustained-release aminophylline, theophylline, morphine,

and other drugs. Research suggested that evening administration of UniphylR (anhydrous theophylline) tablets

represented a rational dosing schedule for patients with asthma who often exhibit increased bronchoconstriction in

the morning. Patients demonstrated improved pulmonary function in the morning compared with use of twice-

daily theophylline when once-daily UniphylR was administered in the evening. Thus, evening administration of

once-daily theophylline may block the morning dip in lung function commonly seen. CONTINR technology

provides for closer control over the amount of drug released to the bloodstream, and benefits patients in terms of

reducing the number of doses they need to take every day, providing more effective control of their disease

(particularly at night), and reducing unwanted side effects.

Marketed preparation for asthma till now

FDA approval date – Sept.01.1982

API- Theopylline

Proprietary name dosage form - Uniphyl

Chronopharmaceutical technology- CONTIN

CONCLUSION

Chronopharmaceutics will certainly improve patient outcome and optimize disease management in the

future. Research in chronopharmacology has demonstrated the importance of biological rhythms in drug therapy

and this has led to a new approach to the development of drug delivery systems. Optimal clinical outcome cannot

be achieved if drug plasma concentrations are constant. If symptoms of a disease display circadian variation, drug

release should also vary over time. Different technologies have been applied to develop time-controlled, pulsed,

triggered and programmed drug delivery devices in recent years. Since it is seems that timing of drug

administration in disease therapy has significant impact upon treatment success, chronotherapeutics remains an

important area for continuing research. It is concluded that the treatment of asthma with the Chrono-optimized

preparation over night is more effective than treatment with a conventional preparation in twice-daily dosage. In

addition, lung function showed greater stability, throughout the day, with once-daily evening therapy than with

traditional 12 hr dosing. It is well known that human body temperature, blood pressure, and pulse rate reach high

values during the day and fall at night. Similarly, all other physiological functions and activities are subject to a

daily cyclical variation known as their circadian rhythm. The respiratory function is no exception and is known to

experience a trough in activity from late night until early morning. In other words, in application once daily at

bedtime could be expected to prevent asthma attacks for practically the entire 24-hour period and, as maximum

blood concentration is reached in the early morning, would be particularly effective against attacks caused by

morning dip.

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Hemant Singh Indian Journal of Research in Pharmacy and Biotechnology

Volume 1(3) May-June 2013 9

RECENT TRENDS IN SCOPE AND OPPORTUNITY OF CLINICAL

RESEARCH IN INDIA Hemant Singh

*, Abhinav Srivastva

I.E.C. College of Eng & Technology, Greater Noida.


ABSTRACT

Clinical Research is one of the most knowledge-intensive industry. It is complete biography of drug

from its inception in the lab to its introduction to the consumer market and beyond. Any molecule is

identified is subjected to pre-clinical and clinical trials before entering in to market. Though post

marketing surveillance is also part of the same. Pre-clinical studies are associated with effect of drug on

animals. All the toxicological studies, tests for teratogenicity, carcinogenicity are carried out.After this,

the data obtained from the studies are submitted as an IND (Investigational Drug Application) to take

permission for human studies. Then, it is enter in to clinical trials. There are 4 phases in it. It is very clear

that India has become a very preferred destination for clinical research. The industry is growing

exponentially and is expected to reach Rs. 7000 Cr by 2010. Statics shows that if India's clinical trial

business grows to 10% of that in US by 2015, the industry will need approximately 50,000 clinical

research professionals.

Key Words: Clinical Research, healthy volunteers, Clinical Trial Phases.

INTRODUCTION

Clinical Research is a systematic study to evaluate the effectiveness and safety of medications or medical

devices by monitoring their effects on large groups of people. Clinical research has become a multi-billion and

multidisciplinary industry. A number of factors favor India as a clinical research hub. There are numerous

government-funded medical and pharmaceutical institutions with state-of-the-art facilities, which can serve as

ideal centers for multi-centered clinical trials.India boasts of well-trained and qualified manpower, well versed in

English. The research and the development process in India can be done at a more affordable price. More

importantly, there is vast clinical material, which can be utilised, due to the prevalence of a large variety of

diseases, including widespread cases of cancer and diabetes, India is viewed the world over as the ideal location

for clinical research trials for the pharmaceutical industry.India is becoming a hub for clinical research; the

demand for professionals in this field is growing rapidly. Clinical research business in India will be worth $1

billion by 2010. Thus, there will soon be a massive demand for clinical research professionals, making it an

interesting career option with massive growth potential. According to a survey, there are 2,50,000 vacancies

available worldwide 50,000 job openings in India by 2010. Highest remuneration packages owing to the shortage

of skilled professionals.There are high demand for trained professionals in this field; the pay package is

impressive at the entry level. Freshers can expect a pay packet of around three lakhs or more per anum. If you

have a master’s degree backing your qualifications, then the amount is almost doubles. Clinical research is an

industry where experience counts, thus the longer you are in this field; higher the salary you can expect.So there is

really a very good scope for clinical research in India.

HUMAN CLINICAL TRIAL PHASES

Phase I: Studies assess the safety of a drug or device. This initial phase of testing, which can take several months

to complete, usually includes a small number of healthy volunteers (20 to 100), who are generally paid for

participating in the study. The study is designed to determine the effects of the drug or device on humans

including how it is absorbed, metabolized, and excreted. This phase also investigates the side effects that occur as

dosage levels are increased. About 70% of experimental drugs pass this phase of testing.

Phase II: Studies test the efficacy of a drug or device. This second phase of testing can last from several months

to two years, and involves up to several hundred patients. Most phase II studies are randomized trials where one

group of patients receives the experimental drug, while a second "control" group receives a standard treatment or

placebo. Often these studies are "blinded" which means that neither the patients nor the researchers know who has

received the experimental drug. This allows investigators to provide the pharmaceutical company and the FDA

with comparative information about the relative safety and effectiveness of the new drug. About one-third of

experimental drugs successfully complete both Phase I and Phase II studies.




Volume 1(3) May-June 2013 0

Phase III: Studies involve randomized and blind testing in several hundred to several thousand patients. This

large-scale testing, which can last several years, provides the pharmaceutical company and the FDA with a more

thorough understanding of the effectiveness of the drug or device, the benefits and the range of possible adverse

reactions. 70% to 90% of drugs that enter Phase III studies successfully complete this phase of testing. Once

Phase III is complete, a pharmaceutical company can request FDA approval for marketing the drug.

Phase IV: Studies, often called Post Marketing Surveillance Trials, are conducted after a drug or device has been

approved for consumer sale. Pharmaceutical companies have several objectives at this stage to compare a drug

with other drugs already in the market; to monitor a drug's long-term effectiveness and impact on a patient's

quality of life; and to determine the cost-effectiveness of a drug therapy relative to other traditional and new

therapies. Phase IV studies can result in a drug or device being taken off the market or restrictions of use could be

placed on the product depending on the findings in the study.

SCOPE FOR CLINICAL RESEARCH IN SOUTH ASIAN NATIONS

With a very large number of drugs, worth more than $50 billion in annual revenues, coming off patent in

the next few years, many western pharma companies are now increasingly relinquishing business activities that

are not considered core, such as clinical research, and are moving towards contract research. Since in the western

countries, clinical research is characterized by extremely high costs and long gestation periods and the same work

can be performed in developing countries at a fraction of the cost and much faster, many global pharma

companies are increasingly turning their attention to Asia to benefit from low R&D costs in the region and also to

gain access to Asia's drug markets. Clinical trials have thus gone ‘global,’ because CROs find it easier to conduct

them in underdeveloped countries, as this is cheaper or has less ethical encumbrances or legal risks. Naïve patient

populations, English speaking doctors, low costs and other advantages offered by developing countries have

opened up new avenues for the clinical trials market. Hence there is lot of scope for clinical research activities for

the South Asian nations if they make use of the upcoming opportunities in this field.

India: The future of clinical research in India continues to be hazy. While on the one hand there are predictions

that India will be the next hotspot for clinical trials, on the other , industry forums speak of ongoing challenges

and stagnant growth for a variety of reasons, such as regulatory delays in approval, escalating costs, inconsistent

quality, ethical irregularities etc. While several CROs have started operations in India basing their future on the

rosy predictions, it is equally true that existing players are struggling to grow or even sustain their current

business.

Since registration of clinical trials in Clinical Trial Registry of India (CTRI) was made mandatory from

mid-2009 onwards, close to 1800 trials have been registered in this database. Of these, about 69 per cent are

sponsored by the pharmaceutical industry (both multinational and Indian companies), the rest being funded by

grants from various government and not-for-profit institutions (e.g. DBT, CSIR, AIIMS, ICMR, Ministry of

Defence,).

Indian CROs can now look forward for more opportunities to conduct clinical studies within the generic

space. For drugs that are not absorbed in the GI tract, plasma concentrations are not useful to determine delivery

of the drug to the site of activity, and hence bioequivalence of the generic product to the innovator cannot be

established by BA/BE studies. In such cases, one needs to demonstrate therapeutic equivalence with clinical or

pharmaco-dynamic studies.

Assocham had predicted that India would garner about 15 per cent of the global clinical trials opportunity.

However, out of over 1,00,000 human trials being conducted in 178 countries, less than 2,000 (two per cent) are

being done in India compared to over 9,000 (nine per cent) in China.

Some of the advantages that India offers as a clinical trial destination are : availability of diverse patient

population across major therapeutic segments such as oncology, metabolics, neurology etc; high degree of

compliance to international guidelines such as the ICH GCP and the regulations laid down by the US FDA;

availability of well qualified, English speaking research professionals including physicians; lower costs compared

to the west; increasing prevalence of illnesses common to both developed and developing countries; availability

of good infrastructure and changes in Patent Laws since January 2005.

Bangladesh: According to an article in The Pharma World, leading health Journal in Bangladesh,

notwithstanding its potential, clinical research is still underdeveloped in Bangladesh. There is a lack of capacity

for bioequivalence studies, no analytical capacity (samples have to be shipped to foreign countries such as

Singapore for any analytical treatment) and limited bio-banking and documentation of clinical specimens. There





is no CRO activity in the country. This cripples the ability of the health and pharmaceutical industry to move

forward, severely stunts the professional growth of health care personnel and limits their ability to become

competitive for funding in the global arena.

Fortunately, with appropriate input from global scientists and consultants, Bangladesh appears very well

placed to fill this void and develop world-class clinical research capacity. There is a significant foundation for

such clinical research capacity in the health care sector in Bangladesh. Hospitals in the country have the

advantage of access to a large population base presenting a range of clinical conditions. These hospitals have

talented physicians who are fluent in English and eager to take advantage of opportunities to expand their

expertise and scope of activities into state-of-the-art health care research.

Moreover Bangladesh has a significant generic pharma industry, currently marketing its products mainly

in the domestic market and in non-regulated international markets. Building Contract Clinical Research capacity

in Bangladesh will serve the local pharmaceutical companies and allow Bangladesh to take its deserved position

in the rapidly-growing global clinical research products and services market.

Bangladesh can be the next frontier in the global CRO industry. Based on GDP and the volume of the

pharma industry compared to other Asian economies such as India and China, Bangladesh could potentially

capture five per cent of the Asian CRO market by 2020. The country is well poised to launch a CRO for multiple

reasons that involve both its own fledgling pharma industry as well as the increasing demands in the global

market, the article says.

The development of a globally-competitive CRO will have an impact beyond the domestic pharma

industry. It will attract foreign companies, seeking high quality research at more affordable prices. It will also

have a multiplier effect on other areas of the economy that transcend the pharma industry, and contribute to the

transition from low-wage labour-intensive activities into higher-wage knowledge-based industries. Building

capacity for upscale knowledge industries, including biotechnology and health science, historically has had a

remarkable impact on wealth building and human development in the West and is expected to have a similar

positive impact in emerging economies.

Global Health Expansion (GHE) LLC is a Washington DC-based company that is currently partnering

with Bangladeshi entities to develop a world class CRO in Bangladesh. The goal of GHE is to team international

know-how in regulatory issues, clinical pharmacology and contract clinical research with the current capacity in

Bangladesh in clinical research and drug manufacturing. This could be done by building synergies between

academia and industry in order to develop the capacity (human and physical) for a true knowledge-based industry

in Bangladesh. Driving such efforts through an international partnership will create unique opportunities for all

partners, the article points out.

Pakistan: The clinical research industry is still in its infancy in Pakistan despite having a large pool of patients, a

large number of English-speaking physicians, low value of rupee, a network of high volume medical centres and a

good know how of this business as many physicians have conducted clinical trials in other countries

The country is the sixth most populous country in the world and being a developing country, the patients’

population in Pakistan is also very large.

Industry experts are of the view that unless the government encourages research culture and streamlines

public-sector universities, the country would continue to lose opportunities in this important arena.

Some experts believe that the local private sector is not motivated to conduct research. This is in contrast to China

and India, which have emerged as the top destinations for researchers in the global pharmaceutical industry.

If Pakistan is interested in attracting more clinical trials to the country, it must take bold steps in these areas,

publicize them and inspire the confidence of international sponsors in the measures taken. The speed of current

healthcare and regulatory reforms will be crucial in this respect, they point out.

Sri Lanka: Sri Lanka is an emerging market for clinical trial business with some contract research organizations

already operational in the country. Although Sri Lanka is getting involved more and more in clinical research, it

significantly lacks professionals formally trained in conducting trials. The need for clinical research training in Sri

Lanka for medical and life science graduates has been stressed upon by various research papers published by Sri

Lankan authors.

Clinical trials are sets of tests in medical research and drug development that generate safety and efficacy data

(or more specifically, information about adverse drug reactions and adverse effects of other treatments) for health

interventions (e.g., drugs, diagnostics, devices, therapy protocols). They're conducted only after satisfactory

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information has been gathered on the quality of the nonclinical safety, and health authority/ethics

committee approval is granted in the country where approval of the drug or device is sought.

The U.S. National Institutes of Health (NIH) organizes trials into five different types:

Prevention trials look for better ways to prevent disease in people who have never had the disease or to prevent a

disease from returning. These approaches may include medicines, vitamins, vaccines, minerals, or lifestyle

changes.

Screening trials test the best way to detect certain diseases or health conditions.

Diagnostic trials are conducted to find better tests or procedures for diagnosing a particular disease or condition.

Treatment trials test experimental treatments, new combinations of drugs, or new approaches to surgery or

radiation therapy.

Quality of life trials (supportive care trials) explore ways to improve comfort and the quality of life for

individuals with a chronic illness.

Compassionate use trials or expanded access trials provide partially tested, unapproved therapeutics to a small

number of patients who have no other realistic options. Usually, this involves a disease for which no effective

therapy exists, or a patient who has already attempted and failed all other standard treatments and whose health is

so poor, he does not qualify for participation in randomized clinical trials. Usually, case-by-case approval must be

granted by both the FDA and the pharmaceutical company for such exceptions. The clinical trial, in its simplest

form, involves the application of the experimental variable – treatment to a person or group of persons – and

observation during or following application of the treat ment to measure its effect. That measure (outcome

measure) may be death, occurrence or recurrence of some morbid condition, or a difference indicativeof change

(e.g. difference in blood pressure measured for each person just prior to the start of treatment and again at some

point during or after treatment). There is no way to “test” a treatment or to “prove” its effectiveness in the absence

of some absolute or relative measure of success. Trials are said to be controlled if the effect of a treatment is

measured against a comparison treatment administered over the same time period and under similar conditions.

That comparison treatment may be another test treatment or, depending on circumstances, a control treatment

consisting of an accepted standard form of therapy, a placebo or sham treatment, or observation only (no

treatment). A trial is said to be uncontrolled if it does not have a comparison treatment or if the enrolment to and

administration of the test and comparison treatments is not concurrent.

History: Prove thy servants, I beseech thee, ten days; and let them give us pulse to eat, and water to drink. Then

let our countenances be looked upon before thee, and the countenance of the children that eat of the portion of the

King’s meat: and as thou seest, deal with thy servants. So he consented to them in this matter, and proved them

ten days. And at the end of ten days their countenances appeared fairer and fatter in flesh than all the children

which did eat the portion of the King’s meat. Fortuitous events can produce conditions reminiscent of the features

of a trial. One such account is that given by Ambroise Paré (surgeon, 1510–1590) during the battle in 1537 for

the castle of Villaine. The treatment for gunshot wounds in Paré’s time was boiling oil poured over the wound.

Because of the intensity of the battle, Paré ran out of oil and resorted to using an ointment made of egg yolks, oil

of roses, and turpentine. The result of his “trial” is summarized by his observation the morning after the battle:

Treatment protocol: The treatment protocol (the general term, study protocolor trial protocol has broader

meaning and refers to the constellation of activities involved in conducting a trial) of the trial specifies the

treatments being studied, the manner and methodof usage and administration, and conditions under which other

treatments are called for when needed for the well-being of those enrolled.Protocols for all research involving

human beings are subject to review and approval by institutional review boards (IRBs) or ethics review boards

(ERBs) before implementation and at periodic intervals thereafter until the research is finished Only patients

judged eligible (as determined by specified may be enrolled, and among those, only those who consent to

participate in the trial. Persons are under no obligation to enroll or to continue once enrolled, and must be so

informed prior to being enrolled. All trials involve data collection at various time points over the course of

enrollment and follow-up of persons. The amount collected per person depends on the nature of the disease or

condition being treated and on the nature of the treatment process implied by the study treatments being used.

Classes of trials: Most clinical trials involve parallel treatment designs, i.e. designs where an assignment unit

(usually a person) is assigned to receive only one of the treatments under study. The word parallel indicates that

two or more groups of assignment units are proceeding through the trial side by side, with the only ostensible

difference (other than baseline differences in the composition of the groups) being the treatment administered.

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The goal in trials with parallel treatment designs is for each person enrolled to receive the assigned treatment and

to have no exposure to any of the other treatments under study in the trial (except where the requirements for

proper care are overriding and make such exposure necessary).

Drug trials: Compounds, no matter how promising or impassioned the pleas for use, have to go through a series

of tests in animals before they can be tested in humans. Those considered to lack promise after animal testing do

not come to testing in humans. Most drug trials are done under an INDA held by the sponsor of the drug. The

“sponsor” in the vernacular of the Food and Drug Administration (FDA) is typically a drug company, but can be a

person or agency without “sponsorship” interests in the drug. Regulations require investigators to report adverse

events to the FDA. The general guidelines regarding consent are similar, but not identical, to those promulgated

by the Office for Protection from Research Risks (OPRR) for IRBs.

Randomized trials: A randomized trial is a trial having a parallel treatment design in which treatment

assignment for persons (treatment units) enrolled is determined by a randomization process similar to coin flips or

tossings of a die. The trialist’s purpose in randomization is to avoid selection bias in the formation of the

treatment groups. Randomization does not guarantee comparability of the treatment groups with regard to the

various entry characteristics of interest. Indeed, one can, by chance, have differences among the treatment groups.

A large difference (one yielding a small P value) can arise by chance and, hence, cannot be taken as prima facie

evidence of a “breakdown” (e.g. “peeking” or other purposeful acts aimed at determining assignment before

issue) of the randomization process, unless supported by other evidence of a “breakdown”. The randomization

may be simple (complete) or restricted. The purpose of restriction is to force the assignments to satisfy the

specified assignment ratio at intervals during enrolment.

Masking: Masking is the purposeful concealment of some fact or condition and is done to keep knowledge of

that fact or condition from influencing the behavior, observation, or reporting of persons so masked. Masking, in

the context of trials, is imposed to reduce the likelihood of a treatment-related bias due to knowledge of treatment

assignment. Masked treatment administration has been used as a mark of “quality” for trials. There is, therefore, a

tendency to view results from masked trials as more reliable than those from unmasked trials. In truth, however,

masked treatment administration is rarely 100% effective. All forms of treatment, and especially those involving

drugs, produce side-effects and telltale signs that may serve to unmask treatment. Hence, the protection provided

by masking can be illusory. As a result, it is better to make assessments of “quality” in terms of the risk of

treatment-related bias and the likely effect of such bias, if present, on the results reported. The risk of treatment-

related bias is low for “hard” outcome measures and with explicitly defined treatment protocols, even in the

absence of masked treatment administration.

Analysis: The protection provided against treatment-related bias by the assignment process is futile if the analysis

is biased. Treatment comparisons, to be valid, must be based on analyses that are consistent with the design used

to generate them. Analyses involving arrangements of data related to treatment administered may be performed,

but only as supplements to the primary analyses. They should not and cannot serve as replacements for those

analyses.

TYPES OF CLINICAL TRIALS

We often think of clinical trials as a method of studying new drugs, but many different types of trials are

in process to evaluate cancer.

Prevention trials:Prevention trials look at substances and lifestyle factors that may raise or lower the risk of

developing cancer

Screening trials: Screening trials study methods to diagnose cancer in its early stages when it is often more

curable

Diagnostic trials: Diagnostic trials are done to look for the best methods of finding out if a person has cancer, or

to accurately determine the stage of cancer that is present.

Treatment trials: Treatment trials evaluate the ability of drugs, radiation, surgery, or other measures to treat

cancer.

Supportive care trials: Supportive care trials are also called quality-of-life trials. They study the ability of a drug

or procedure to lessen the symptoms of cancer or symptoms related to the treatment of cancer.

CONCLUSION

Clinical trials are conducted to collect data regarding the safety and efficacy of new drug and device

development. There are several steps and stages of approval in the clinical trials process before a drug or device





can be sold in the consumer market, if ever.Drug and device testing begins with extensive laboratory research

which can involve years of experiments in animals and human cells. If the initial laboratory research is successful,

researches send the data to the Food and Drug Administration (FDA) for approval to continue research and testing

in humans.Once approved, human testing of experimental drugs and devices can begin and is typically conducted

in four phases. Each phase is considered a separate trial and, after completion of a phase, investigators are

required to submit their data for approval from the FDA before continuing to the next phase.

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Medicine, 12, 1991, 1373–1534.

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Berlin, J.A, Laird, N.M., Sacks, H.S. & Chalmers, T.T, A comparison of statistical methods for combining event

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Blackwelder, W.C, Proving the null hypothesis in clinical trials, Controlled Clinical Trials, 3, 1982, 345–353.

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Trials, 5, 1984, 97–105.

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clinical trials: summary of a panel discussion, Controlled Clinical Trials 10, 1989, 254–281.

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Brosgart, C.L, Mitchell, T, Charlebois, E., Coleman, R., Mehalko, S., Young, J. & Abrams, D.I, Off-label drug

use in human immunodeficiency virus disease, Journal of Acquired Immune Deficiency Syndrome and Human

Retrovirology, 12, 1996, 56–62.

Brown, B.W Jr, Designing for cancer clinical trials: Selection of prognostic factors, Cancer Treatment Reports,

64, 1980, 499–502.

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Navin Dixit et.al Indian Journal of Research in Pharmacy and Biotechnology


SUSTAINED RELEASE DRUG DELIVERY SYSTEM Navin Dixit*, Sheo Dutt Maurya, Bhanu P.S.Sagar

I.E.C.College of Eng & Technology, Greater Noida (U.P)

*Corresponding author: [email protected]

ABSTRACT

The oral route is the most popular route used for administration of drugs, which is due in part

to the ease of administration and to the fact that gastrointestinal physiology offers more flexibility

in dosage form design than most other routes. The terms Sustained release, prolonged release,

modified release, extended release or depot formulations are used to identify drug delivery systems

that are designed to achieve or extend therapeutic effect by continuously releasing medication over

an extended period of time after administration of a single dose. There are several reasons for

attractiveness of these dosage forms: provides increased bioavailability of drug product, reduction

in the frequency of administration to prolong duration of effective blood levels, reduces the

fluctuation of peak trough concentration and side effects and possibly improves the specific

distribution of the drug.

Key words: Sustained Release, Drug delivery system, Matrix, Drug Release

INTRODUCTION

Probably the earliest work in the area of sustained drug delivery dosage forms can be traced to the 1938

patent of Israel Lipowski. This work involved coated pallets for prolonged release of drug and was presumably

forerunner to the development of the coated particle approach to sustained drug delivery that introduced in the

early 1950s. Ideally, a drug should arrive rapidly at the site of action (receptor) in the optimum concentration,

remain for the desired time, be excluded from other sites, and be rapidly removed from the site when indicated i.e.

the basic goal of the therapy is to achieve a steady state blood level that is therapeutically effective and non-toxic

for an extended period of time. Generally, the time course of a dosage form (pharmacokinetics) in man is

considered to be controlled by the chemical structure of the drug. Decreasing the rate of absorption and/ or

changing the dosage form provide a useful adjunct. When it is feasible or desirable to modify the drug compound

on a molecular level, often sought is a product that will requireless frequent administration to obtain the required

biologic activity time profile; for example, a tablet that has the same clinical effect when administered every

twelve hours. In another instance, it may be desirable to decrease the absorption rate in order to obtain a more

acceptable clinical response (Girish K Jani, 2009).

Tablets are one of the most stable and commonly administered oral dosage forms. Since the later part of

nineteen-century, tablets have been widespread and their popularity continues. Tablets remain popular as dosage

form because of the advantages afforded both to the pharmaceutical manufacturers and patients. These includes:

simplicity and economy of preparation, stable and convenient in packing, ease of transporting and dispensing,

accuracy of single dosage regimen, compactness and portability, and blandness of taste and ease of

administration. The goal in designing sustained or controlled delivery systems is to reduce frequency of dosing or

to increase the effectiveness of the drug by localization at the site of action, reducing the dose required, providing

uniform drug delivery. If one were to imagine the ideal drug delivery system, two prerequisites would be

required. First, it would be a single dose for duration of treatment, whether it is for days of weeks, as with

infection, or for lifetime of the patient, as in hypertension or diabetes. Second, it should deliver the drug directly

to the site of action, thereby minimizing or eliminating side effects. This may necessitate delivery to specific

receptors or to localization to cells or to specific areas of the body.

Oral ingestion has long been the most convenient and commonly employed route of drug delivery.

Indeed, for sustained release systems, oral route of administration has received most of the attention with respect

to research on physiological and drug constraints as well as design and testing of products. This is because of the

fact that there is more feasibility in dosage form design for oral route than for parenteral or any other route. The

design of oral sustained release delivery systems is subject to several intercalated variables of considerable

importance. Among these are the types of delivery systems, the disease being treated, the patient and the length

of therapy and the properties of the drug. In conventional drug therapy, it can be seen from the Figure 1.1 that the

administration of drug by either intravenous injection or an extravascular route e.g. Orally, intramuscularly, or





rectally does not maintain drug blood level within the therapeutic range for an extended period of time. The short

action is due to the inability of conventional dosage forms to control temporal delivery (Banks Michael, 1991).

DRUG SELECTION FOR ORAL SUSTAINED RELEASE DRUG DELIVERY SYSTEMS

The biopharmaceutical evaluation of a drug for potential use in controlled release drug delivery system

requires knowledge on the absorption mechanism of the drug form the G. I. tract, the general absorbability, the

drug’s molecular weight, pKa, solubility at different pH and apparent partition coefficient.

Table.1. Parameter for drug selection

Parameter Preferredvalue

Molecular weight/ size < 1000

Solubility > 0.1 µg/ml for pH 1 to pH 7.8

Pka Non ionized moiety > 0.1% at pH 1 to pH 7.8

Apparent partition coefficient High

Absorption mechanism Diffusion

General absorbability From all GI segments

Release Should not be influenced by pH and enzymes

The pharmacokinetic evaluation requires knowledge on a drug’s elimination half- life, total clearance, absolute

bioavailability, possible first- pass effect, and the desired steady concentrations for peak and trough.

Table.2. Pharmacokinetic parameter for drug selection

Parameter Comment

Elimination half life Preferably between 0.5 and 8 h

Total clearance Should not be dose dependent

Elimination rate constant Required for design

Apparent volume of distribution Vd The larger Vd and MEC, the larger will

be the required dose size.

Absolute bioavailability Should be 75% or more

Intrinsic absorption rate Must be greater than release rate

Therapeutic concentration Css av The lower Css av and smaller Vd, the

loss among of drug required

Toxic concentration Apart the values of MTC and MEC,

safer the dosage form. Also suitable for

drugs with very short half-life.

Matrix diffusion controlled drug delivery system: In this type of controlled drug delivery system, the drug

reservoir results from the homogeneous dispersion of the drug particles in either a lipophilic or a hydrophilic

polymer matrix.

Mode of action of hydrophilic matrix dosage form: Hydrophilic matrix dosage forms essentially consist of a

compressed blend of hydrophilic polymer and drug. According to the generally accepted mechanism, the drug

release from hydrophilic matrix dosage forms starts when the tablet comes in contact with gastrointestinal fluid.

The surface of the tablet hydrates to release exposed drug and at the same time form a viscous polymer mucilage

or gel. This gel fills the interstices within the tablet, retarding further ingress of liquid. The concentration of

polymer within the hydrated layer ranges from dilution at the outer surface to around 90% at the boundary with

the drug core. Within this layer, drug in various states of dissolution (undissolved in dilute solution; in saturated

solution) is distributed amongst the other ingredients of the tablets.





Fig.1.Matrix diffusion controlled drug delivery system

Drug release occurs immediately from the surface (burst effect) followed by diffusion through, and / or

erosion of, the hydrated layer. The relative proportions of drug released by diffusion and erosion are determined

by the drug’s solubility properties and by the physical and chemical nature of the hydrated polymer. This in turn

is influenced by other factors, including drug characteristics, dissolution medium and other, which continue to be

investigated.

Modified-release drug delivery systems: In order to overcome the drawbacks of conventional drug delivery

systems, several technical advancements have led to the development of modified release drug delivery systems.

The modified-release delivery systems may be divided conveniently into four categories4:

1. Delayed release

2. Sustained release

3. Site-specific targeting

4. Receptor targeting

Delayed release system: Delayed-release systems are those that use repetitive, intermittent dosing of a drug from

one or more immediate-release units incorporated into a single dosage form. Examples of delayed-release

systems include repeat-action tablets and capsules, and enteric-coated tablets where timed release is achieved by

a barrier coating.

Sustained release system: Sustained-release systems include any drug-delivery system that achieves slow release

of drug over an extended period of time. If the systems can provide some control, whether this is of a temporal or

spatial nature, or both, of drug release in the body, or in other words, the system is successful at maintaining

constant drug levels in the target tissue or cells, it is considered a controlled-release system.

Site-specific targeting: Site-specific and receptor targeting refer to targeting of a drug directly to a certain

biological location. In the case of site-specific release, the target is adjacent to or in the diseased organ or tissue.

Receptor targeting: For receptor release, the target is the particular receptor for a drug within an organ or tissue.

Both of these systems satisfy the spatial aspect of drug delivery and are also considered to be controlled drug-

delivery systems.

Sustained release drug delivery systems: During the past few years, conventional dosage forms of drugs are

rapidly being replaced by the new and the novel drug delivery systems. Amongst, these the controlled

release/sustained release dosage forms have become extremely popular in modern therapeutics. The basic

rationale for sustained release drug delivery is to alter the pharmacokinetics and pharmacodynamics of drugs by

using novel drug delivery systems or by modifying the molecular structure or physiological parameters inherent in

a selected route of administration. It is desirable that the duration of drug action becomes more a design property

of a rate controlled dosage form and less or not at all a property of the drug molecule’s inherent kinetic properties.

Thus, optimal design of a sustained/ controlled release system necessitates a thorough understanding of the

pharmacokinetics and pharmacodynamics of the drug. When the drug is administered in a conventional dosage

form, it results in a fluctuation of drug concentration at the site of action (peak and valley pattern) and therefore in

systemic circulation and tissue compartment. Figure 1.2 shows the difference between the conventional and

sustained release dosage forms.

Advantages of sustained release drug delivery: Following are the potential advantages of sustained release

products

1. Decreased local and systemic side effects reduced gastrointestinal irritation.

2. Better drug utilization reduction in total amount of drug used.





3. Improved efficiency in treatment, optimized therapy, more uniform blood concentration.

4. Reduction in fluctuation in drug level and hence more uniform pharmacological response, cure of control

of condition more promptly, less reduction in drug activity with chronic use.

5. Method by which sustained release is achieved can improve the bioavailability of some drugs e.g. drugs

susceptible to enzymatic inactivation can be protected by encapsulation in polymer systems suitable for

sustained release.

6. Improved patient compliance, less frequent dosing, reduced night-time dosing, reduced patient care time.

The importance of patient compliance in successful drug therapy is well recognized. It has been found

that there is an inverse relationship between the number of dosages per day and the compliance rate.

7. Although the initial unit cost of sustained release products is usually greater than that of conventional

dosage forms because of the special nature of these products, the average cost of treatment over an

extended time period may be less. Economy may also result from a decrease in nursing time and

hospitalization time.

Disadvantages of sustained release drug delivery: The disadvantages of sustained release drug delivery system

are

1. Decreased systemic availability in comparison to immediate release conventional dosage forms, which

may be due to incomplete release, increased first-pass metabolism, increased instability, insufficient

residence time complete release, site specific absorption, pH dependent stability, etc.

2. Poor in vitro – in vivo correlation.

3. Retrieval of drug is difficult in case of toxicity, poisoning or hypersensitivity reactions.

4. Reduced potential for dose adjustment of drugs normally administered in varying strengths.

Classification of oral sustained/controlled release systems

Diffusion controlled Systems

Reservoir devices: A core of drug (reservoir) surrounded by a polymeric membrane characterizes them. The

nature of the membrane determines the rate of drug release. The characteristics of reservoir diffusion systems are

1. Zero order drug release is possible.

2. The release rate is dependent on the type of polymer.

3. High molecular weight compounds are difficult to deliver through the device.

Matrix devices: It consists of drug dispersed homogenously in a matrix. The characteristics of matrix diffusion

systems are

1. Zero order release cannot be obtained.

2. Easy to produce than reservoir devices.

3. High molecular weight compounds are delivered through the device.

Dissolution controlled systems

Matrix dissolution controlled systems: Aqueous dispersions, congealing, spherical agglomeration, etc. can be

used.

Encapsulation dissolution controlled systems: Particles, seeds, granules can be coated by techniques such as

microencapsulation.

Diffusion and dissolution controlled systems: In a bioerodible matrix, the drug is homogenously dispersed in a

matrix and it is released either by swelling controlled mechanism or by hydrolysis or by enzymatic attack.

Sustained release matrix tablets: One of the least complicated approaches to the manufacture of sustained

release dosage forms is the direct compression of drug, release retardant, and additives to form a tablet in which

drug is embedded in a matrix core of retardant. Alternatively drug retardant blend may be granulated prior to

compression. Such tablets are called as matrix tablets. Three classes of release retarding materials are used for the

formulation of matrix tablets.

They include

1. Insoluble or ‘skeleton’ matrices

2. Water insoluble, erodable matrices

3. Hydrophilicmatrices.





Table1.1. Some important materials used for preparing sustained release tablets.

Matrix characteristics Release retarding material

Insoluble, inert

Polyethylene, Polyvinyl Chloride

Methyl acrylate-methacrylate copolymer

Ethyl Cellulose

Insoluble, erodible

Carnauba wax, Steryl alcohol

Stearic acid, Polyethylene glycol

Polyethylene glycol monostearate

Triglycerides

Hydrophillic

Methyl cellulose, Hydroxyethyl cellulose

Hydroxypropyl methycellulose

Sodium carboxymethylcellulose

Carboxypolymethylene

Sodium alginate, Galactomannose

Tablets prepared from these materials are egested intact and not break apart in GI tract. The rate-liming

step in controlling the release of drug from these formulations is liquid penetration into the matrix unless

channeling agents are included in the formulation to promote permeation of water into the matrix. This allows

drugs dissolution and diffusion from the channel created in the matrix. In these tablets, drug bioavailability is

dependent on polymer-ratio. The bioavailability may be modified by addition of diluents such as lactose. These

forms of matrix tablets are not useful if dose of drug is high or if the drug is insoluble in water. Waxes, lipids and

related materials form matrices that control the release through both pore diffusion and erosion. Release

characteristics are more sensitive to digestive fluid composition than the tablets preparation by insoluble material.

Total release of drug from the wax-lipid matrices is not possible, since a certain fraction of the dose is coated with

impermeable wax films. For dispersion of drug with the base, three methods are used, which include the fusion

technique. In absence of additives, the drug release is non-linear from these systems. Additives like polyvinyl

pyrrolidone or polyoxyethylene lauryl esters can lead to apparent zero-order release.

The third group of matrix formers represents non digestible materials, which form gels in situ. The release

of drug from these systems is controlled by penetration of water through a gel layer produced by hydration of

polymer and diffusion of drug through the swollen, hydrated matrix, in addition to the erosion of gelled layer. The

extent to which the erosion or diffusion controls the release depends on polymer selected as well as on the drug-

polymer ratio used in the formulation. High drug polymer ratios results in formulations from which drug release is

controlled attrition.

Mechanism of drug release: On exposure to aqueous fluid, hydrophilic matrices take up water, and polymer

starts hydrating to form a gel layer. An initial burst of soluble drug may occur due to surface leaching when a

matrix containing a swellable glassy polymer comes in contact with an aqueous medium, there is an abrupt

change from a glassy to a rubbery state which is associated with swelling process with time, water infiltrator deep

into the case increasing the thickness by the gel layer. Concomitantly the outer layer becomes fully hydrated and

states dissolving or eroding. When water reaches the center of the system and the concentration of drug fells

below the solubility value, the release rate of drug begins to reduce. At the same time, an increase in thickness of

the barrier layer with time increases the diffusion path length, reducing the rate of drug release. Drug release

kinetic associated with these gel – layer dynamic, range initially from Fickian to annomalous (Non – Fickian) and

subsequently from quasi – Constant ( near Zero order ) to constant. In general, two major factors control the

drug release from swelling controlled matrix system. They include many processes given below.

1. The rate of aqueous medium infiltration into the matrix, followed by a relaxation process (hydration,

gelatin or swelling).

2. The rate of matrix erosion

As a result of these simultaneous processes, two front are evident, a swelling front, where the polymer get

hydrated, and an eroding front. The distance between these two fronts are called diffusion layer thickness.

Diffusion layer thickness depends on the selective rate at which the swelling and eroding fronts move in relation

to each other. If the polymer gets slowly, solvent can penetrate deep into the glassy matrix the dissolving the

drug; there for gel layer thickness and it stability are council in controlling drug release. Swelling of HPMC





matrix tablet was higher for higher a molecular weight. They attributed this to the large hydrodynamic volume

occupied by higher molecular weight chain when hydrated. As the polymer chain becomes more hydrated and the

gel becomes more dilute, the disentanglement concentration may be reached that is the critical polymer

concentration below which the polymer chain disentangle and detached from gelled matrix.

CONCLUSION

There are several reasons for attractiveness of these dosage forms: provides increased

bioavailability of drug product, reduction in the frequency of administration to prolong duration of

effective blood levels, reduces the fluctuation of peak trough concentration and side effects and possibly

improves the specific distribution of the drug. If one were to develop an ideal drug delivery system, two pre-

requisites would be required: Firstly single dose for the duration of treatment whether for days or weeks as

with infection, diabetes or hypertension. Second it should deliver the active entity directly to the site of

action minimizing the side effects.

REFERENCES

Anoop Kumar singh et al. Isolation, Characterization and formulation properties of a new plant gum obtained

from mangifera indica, Int J Pharm Biomed Res, 1(2), 2010, 35-41.

Banks Michael and E.alulton Michael, Fluidized-bed Granulation: A Chronology, DIP, 17(11), 1991, 1437-

1463.

Davies WK, Gloor WT, Batch production of pharmaceutical granulations in a fluidized bed: Effects of process

variables on physical properties of final granulation.Journal of Pharmaceutical Sciences, 60(12), 1971, 1869-

1874.

Davies WL, Gloor WT. Batch production of pharmaceutical granulations in a fluidized bed 2: effect of various

binders and their concentrations on granulations and compressed tablets. Journal of Pharmaceutical sciences

1972, 61(4), 1972, 618-622.

Georgakopoulos PP, Malamataris S, Dolamidis G, The Effect of using different grades of PVP and gelatin

as binder in the fluidized bed granulation and tableting of lactose, Pharmazie, 38(4), 1983, 240-243.

Girish K Jani, Dhiren P Shah, Vipul D Prajapati, Vineet C Jain. Gums and Mucilages, AJPS, 4(5), 2009, 309-323.

Heng PWS, Roller compaction of crude plant material: Influence of process variables, PVP and Co-

milling. Pharmaceutical Development and Technology, 9(2), 2004, 135-144.

Leon L, Herbert AL, Joseph LK, The theory and practice of industrial pharmacy, 3rd

ed. Bombay, Varghese

Publishing House, 1991.

Lowenthal W, Disintegration of tablets, Journal of Pharmaceutical Sciences, 61(11), 2006, 1695-1711.

Odeku OA, Itiola OA, Evaluation of the effects of khaya gum on the mechanical and release properties

of paracetamol tablets, DIP, 29(3); 2003, 311-320.

Pereira de Souza T et al. Eudragit E as excipient for production of granules and tablets from phyllanthus niruri l

spray-dried extract, AAPS Pharm Scitech, 8(2), 2007, A-34.



Ravi Kumar and Deepthi Yadav et.al Indian Journal of Research in Pharmacy and Biotechnology


ANTIBACTERIAL ACTIVITY OF ETHANOLIC EXTRACTS OF

NYCTANTHES ARBORTRISTIS AND NERIUM OLEANDER A.Ravi Kumar, CH.S.D.Phani Deepthi Yadav*

Department of Pharmacognosy, Bapatla College of Pharmacy, Bapatla, Andhra Pradesh, India

*Corresponding author: E-Mail:[email protected]

ABSTRACT

In the present study, an attempt was made to investigate the anti-bacterial activity of Nyctanthes

arbortristis and Nerium oleander. The crude drug powder extracts of the leaves of the above plants were

taken for the study. The antibacterial activity was performed by using both gram positive and gram

negative organisms viz., B.subtilis and E.coli respectively. The Phytochemical Screening was done for the

selected plants. Phenolic compounds, tannins, flavonoids, cardiac glycosides, saponins and alkaloids

were present in Nyctanthes arbortristis. Steroids, alkaloids, flavanoids, carbohydrates, cardiac glycosides

and tannins were present in Nerium oleander.

Key words: Anti-bacterial activity, Nyctanthes Nerium species plants

INTRODUCTION

Herbal medicine – It is also called botanical medicine or phytomedicine-refers to using plants seeds,

flowers, roots for medicinal purpose. Herbal has a long tradition of use of outside of conventional medicine. It is

becoming more main stream as improvements in analysis and quality control along with advances in clinical

research show the value of herbal medicine in the treating and preventing disease. The medicinal action of plants

is unique to a particular plant species, consistent with the concept that the combination of secondary metabolites

in a particular plant is taxonomically distinct. This article discusses the description and uses of two medicinal

plants Nyctanthes arbor and Nerium oleander. Nyctanthes arbor-tristis is commonly known as Night-flowering

Jasmine, Coral Jasmine and Parijat. It is used for its antibacterial, anthelmintic, anti-inflammatory,

hepatoprotective, immunopotential, anti pyretic, antioxidant and anti fungal activity Nerium oleander is an

evergreen shrub or small tree in the dogbane family Apocynaceae and it is toxic in all its parts. It is used

traditionally in treating dermatitis, abscesses, eczema, psoriasis, sores, warts, corns, ringworm, scabies, herpes,

skin cancer, asthma, dysmenorrheal, epilepsy, malaria, abortifacients, emetics, heart tonics, and tumor.


Plant Materials: The leaves of plants Nerium and Nyctanthes species were authentified by Prof.

V.Satyanarayana, department of plant breeding, Bapatla Agricultural College, Bapatla Andhra Pradesh, India.

They were collected from different places of Andhra Pradesh, India.

Solvent Extraction: The leaves of Nerium oleander, and Nyctanthes arbortristis were collected, washed, dried

and powdered separately. 50g of dried powder of the leaves was weighed and transferred into a conical flask and

it was macerated with sufficient amount of ethanol for about a week days. The whole mixture was filtered and

filtrate was collected, concentrated in a china dish on a hot plate till the residue was obtained. The extract was

collected, labelled and stored for further experimental use.

Fig 1: Nyctanthes arbortristis Plant Fig 2: Nerium oleander Plant

Microorganisms: The test organisms used were E.coli (ATCC 25922) a Gram –ve strain and B.subtilis (ATCC

21332) a Gram +ve strain which were obtained from PG and Research Department of Biotechnology Bapatla






College of Pharmacy Bapatla Andhra Pradesh India The strains were sub-cultured on nutrient agar slants and

were incubated for 24 hrs.

ANTIBACTERIAL ACTIVITY

Agar well diffusion method: Required glass ware was washed and dried in a hot air oven. The sterilized agar

medium was transferred into the Petri dishes, was allowed to solidify at room temperature. The selected test

organism was spread over the solidified agar with the help of a swab stick. Sterile borer was used to make wells

of 8mm diameter .The dilutions of ethanolic extracts of Nyctanthes arbortristis, Nerium oleander solutions of

combined ethanolic extracts of Nyctanthes arbortristis with Nerium oleander respectively were poured in the

wells with the help of a sterile syringe needle. In each Petri plate a well was prepared for standard i.e.,

ciprofloxacin 10µg/ml solution. The Petri plates were placed in a refrigerator for 5min to allow diffusion. Later

the Petri plates were incubated in inverted position at 370 C for 24 hours in the incubator. After 24hours the zone

of inhibition was observed and diameter in mm was measured and recorded.

Qualitative analysis: The extracts and crude dried powders of Nyctanthes arbortristis, Nerium oleander were

subjected to qualitative analysis for presence of chemical constitutens of Nytanthes arbortristis Nerium oleander

Table.1.Anti-bacterial activity of Nyctanthes Arbortristis and Nerium Oleander

COMPONENT

DOSE

Zone of Inhibition (mm)

E.COLI B.SUBTILIS

CIPROFLOXACIN (STD) 10 µg/ml 20mm 22mm

Ethanolic extract of

Nyctanthes arbortristis

500µg/ml - -

750µg/ml

-

-

1000µg/ml 2mm

3mm

Ethanolic extract of Nerium

oleander

500 µg/ml - -

750µg/ml

- -

1000µg/ml 4mm 5mm

Combined ethanolic extracts

of Nerium oleander and

Nyctanthes arbortristis

1000µg/ml - -

1500µg/ml

8mm

10mm

2000µg/ml 12mm 15mm

RESULTS AND DISCUSSION

The study of the chemical constituents and the active principles of the medicinal plants have acquired a

lot of importance all over the world. The present study includes the antibacterial activity of extracts of leaves of

Nyctanthes arbortristis in combination with the leaf extracts of Nerium oleander separately were performed.

Earlier studies on Nyctanthes arbortristis indicated that the ethylacetate and chloroform extracts showed

significant activity on both Gram +ve and Gram –ve strains. But in present study ethanolic leaf extract showed

that the activity on bacterial strains was significant. But comparably the activity on B.subtilis was more than that

of E.coli. As the activity obtained for the leaf extract was significant the combined leaf extract of Nyctanthes

arbortristis and Nerium oleander were used which showed a synergistic effect i.e., Nerium oleander increased the

antibacterial activity of the Nyctanthes arbortristis. The results were given in Table 1

CONCLUSION

The Screening of Phytochemical constituents of the plants Nyctanthes arbortristis Nerium oleander

analysis indicated the presence of carbohydrates, glycosides and alkaloids. The combined ethanolic extract of

Nyctanthes arbortristis and Nerium oleander exhibited significant antibacterial activity of 2000µg/ml

concentration.





AKNOWLEDGEMENTS The authors are thankful to Management Principal, Bapatla College of Pharmacy, Bapatla, Andhra

Pradesh, India for permitting and providing necessary facilities for carrying out to do the project work.

REFERENCES

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49-55.

Amarite O, Bhuskat P, Patel N and Gadgoli C,Evaluation of antioxidant activity of carotenoid from

Nyctanthes arbortristis, Int J Pharmacol Biol Sci, 2, 2007, 57-59.

Andrews JM, BSAC Standardised disc susceptibility testing method, J Antimicrob Chemother, 48, 2001, 5–

16.

Chattopadhyay RR, Sarkar SK, Ganguli S, Banerjee RN, Basu TK, Antiinflammatory and acute toxicity

studieswith leaves of Vinca rosea linn. in experimental animals. Indian Journal of Physiology and

Pharmacology, 36, 1992, 291–292.

Chattopadhyay RR, Sarkar SK, Ganguli S, Banerjee RN, Basu TK, Hypoglycemic and antihyperglycemic

effect ofleaves of Vinca rosea Linn, Indian Journal of Physiology andPharmacology, 35, 1991, 145–151.

El-Sayed, A. and G.A. Cordell, Catharanthamine: A new antitumor bisindole alkaloid from Catharanthus

roseus, J. Nat. Prod, 44, 1981, 289-293.

Evans WC and Trease, Pharmacognosy, Elsevier publication, 19th Edn, 1983, 96-98.

Hassan KA, Brenda AT, Patrick V, Patrick OE, In vivo antidiarrheal activity of the ethanolic leaf extract of

Catharanthus roseus Linn. (Apocyanaceae) in Wistar rats, Afr J Pharm Pharmacol, 5(15), 2011, 1797-1800.

Jaleel CA, R Gopi and R Paneerselvam, Alterationsin non-enzymatic antioxidant componentsof

Catharanthus roseus exposed to paclobutrazol gibberellic acid and Pseudomonas fluorescens, Plant Omics J,

2, 2009, 30-40.

Kirtikar KR and Basu BD, Indian Medicinal Plants, Vol.VII, (Sri Satguru Publications, New Delhi,) 2000,

2110-2113.

Kokate C K, purohit A P, Gokhale S B, A Textbook of Pharmacognosy, Nirali Prakashan, 2009, 27-28

Kokate CK, Purohit AP and Gokhale SB., Pharmacognosy. Nirali prakashan, 2006, 35th Edn: 99-100

Kusum S and Akki, Phytochemical investigation and in vitro evaluation of Nyctanthes arbortristis leaf extract

for antioxidant property, J Pharm Res, 2(4), 2009, 752-755

Lokesh R,Leonard Barnalas, Madhuri P,Saurav K,Sundar K, Larvicidal activity of Trigonella, foenumand

Nerium oleander Linn, Current research journal of biological sciences, 2(3), 2010, 154-160.

Mathuram V and Kundu AB, Occurrence of two new ester of 6-Hydroxyloganin in Nyctanthes arbortristis.

J Indian Chem Soc, 68, 1991, 581-584

Muhammad LR, N Muhammad A, Tanveer andS.N. Baqir, 2009. Antimicrobial activity of different extracts

of Catharanthas roseus, Clin. Exp. Med J, 3, 2009, 81-85

Ohadoma SC, Micheal HU, Effects of co-administration of methanol leaf extract of Catharanthus roseus on

the hypoglycemic activity of metformin and glibenclamide in rats, Asian Pac J Trop Biomed, 4(6), 2011, 475-

477

Omkar A, Jeeja T and Chhaya G, Evaluation of anti-inflammatory activity of Nyctanthes arbortristis and

Onosma echiodes, Phrmacog mag, 8, 2006, 258-260.



Sampath Kumar et.al Indian Journal of Research in Pharmacy and Biotechnology


FOOD POISONING AND ITS SAFETY PRECAUTION M. Umadevi

1, K.P.Sampath Kumar

*2, Sai Pavan

3, Sd.Gosia Sultana

3, D. Bhowmik

3

1. Tamil Nadu Agricultural University, Coimbatore

2. Coimbatore Medical College, Coimbatore

3. Nimra College of Pharmacy,Vijayawada


ABSTRACT

Food poisoning occurs when you swallow food or water that contains bacteria, parasites, viruses, or

toxins made by these germs. Most cases of food poisoning are from common bacteria such as

Staphylococcus or E. coli. Food poisoning, also known as acute gastroenteritis, is an acute inflammation

of the lining of the stomach and small bowel. Food poisoning is a common, usually mild, but sometimes

deadly illness that occur suddenly (within 48 hours) after consuming a contaminated food or drink. Most

of the common contaminants cause nausea, vomiting, diarrhea, and abdominal cramping. Depending on

the contaminant, fever and chills, bloody stools, dehydration, and nervous system damage may follow.

Food poisoning comes from eating foods that contain germs like bad bacteria or toxins, which are

poisonous substances. Bacteria are all around us, so mild cases of food poisoning are common.

Key words: Food poisoning, Infectious agents, Food handling, Infectious organisms.

INTRODUCTION

Food poisoning, also called food-borne illness, is illness caused by eating contaminated food. Infectious

organisms including various bacteria, viruses and parasites or their toxins are the most common causes of food

poisoning. Infectious organisms or their toxins can contaminate food at any point during its processing or

production. Contamination can also occur at home if food is incorrectly handled or cooked. Food poisoning

symptoms often include nausea, vomiting or diarrhea, which can start just hours after eating contaminated food.

Most often, food poisoning is mild and resolves without treatment. But some cases are severe, requiring

hospitalization. Food poisoning is a common, usually mild, but sometimes deadly illness. Typical symptoms

include nausea, vomiting, abdominal cramping, and diarrhea that occur.

FOOD POISONING CAUSES

More than 250 known diseases can be transmitted through food. The estimates unknown or undiscovered

agents cause 68% of all food-borne illnesses and related hospitalizations. Many cases of food poisoning are not

reported because people suffer mild symptoms and recover quickly. Also, doctors do not test for a cause in every

suspected case because it does not change the treatment or the outcome.

The known causes of food poisoning can be divided into two categories: Infectious agents and toxic

agents. Infectious agents include viruses, bacteria, and parasites. Toxic agents include poisonous mushrooms,

improperly prepared exotic foods (such as barracuda - ciguatera toxin), or pesticides on fruits and vegetables.Food

usually becomes contaminated from poor sanitation or preparation. Food handlers who do not wash their hands

after using the bathroom or have infections themselves often cause contamination. Improperly packaged food

stored at the wrong temperature also promotes contamination.

Food poisoning can affect one person or a group of people who all ate the same contaminated food. It

more commonly occurs after eating at picnics, school cafeterias, large social functions, or restaurants.

The germs may get into the food you eat (called contamination) in different ways:

Meat or poultry can come into contact with bacteria from the intestines of an animal that is being

processed

Water that is used during growing or shipping can contain animal or human waste

Food handling or preparation in grocery stores, restaurants, or homes

Food poisoning often occurs from eating or drinking:

Any food prepared by someone who does not wash their hands properly

Any food prepared using cooking utensils, cutting boards, and other tools that are not fully cleaned

Dairy products or food containing mayonnaise (such as coleslaw or potato salad) that have been out of the

refrigerator too long

Frozen or refrigerated foods that are not stored at the proper temperature or are not reheated properly

Raw fish or oysters


http://www.emedicinehealth.com/script/main/art.asp?articlekey=4510








Raw fruits or vegetables that have not been washed well

Raw vegetables or fruit juices and dairy products (look for the word "pasteurized," which means the food

has been treated to prevent contamination)

Undercooked meats or eggs

Water from a well or stream, or city or town water that has not been treated

Many types of germs may cause food poisoning, including:

Campylobacter enteritis

Cholera

E. coli enteritis

Fish poisoning

Staphylococcus aureus

Salmonella

Shigella

Infants and elderly people are at the greatest risk for food poisoning. You are also at higher risk if:

You have a serious medical condition, such as kidney disease or diabetes

You have a weakened immune system

You travel outside of the United States to areas where you are exposed to germs that cause food poisoning

Pregnant and breastfeeding women have to be especially careful to avoid food poisoning.More than 250

known diseases can be transmitted through food. Many cases of food poisoning are not reported because

people suffer mild symptoms and recover quickly. Also, doctors do not test for a cause in every suspected

case because it does not change the treatment or the outcome.

The known causes of food poisoning can be divided into two categories:

Infectious agents include viruses, bacteria, and parasites. Toxic agents include poisonous mushrooms, improperly

prepared exotic foods or pesticides on fruits and vegetables.Food usually becomes contaminated from poor

sanitation or preparation. Food handlers who do not wash their hands after using the bathroom or have infections

themselves often cause contamination. Improperly packaged food stored at the wrong temperature also promotes

contamination.

FOOD POISONING SYMPTOMS

Symptoms of food poisoning depend on the type of contaminant and the amount eaten. The symptoms

can develop rapidly, within 30 minutes, or slowly, worsening over days to weeks. Most of the common

contaminants cause:

nausea

vomiting

diarrhea

abdominal cramping

fever

Usually food poisoning is not serious, and the illness runs its course in 24-48 hours.

Viruses: Viruses account for most food poisoning cases where a specific contaminant is found. Noroviruses are a

group of viruses that cause a mild illness (often termed "stomach flu") with nausea, vomiting, diarrhea, abdominal

pain, headache, and low-grade fever. These symptoms usually resolve in two to three days. It is the most common

viral cause of adult food poisoning and is transmitted from water, shellfish, and vegetables contaminated by feces,

as well as from person to person. Outbreaks are more common in densely populated areas such as nursing homes,

schools, and cruise ships (hence the viral infection is also known as the "Cruise Ship Illness"). The term

Norovirus has been approved as the official name for this group of viruses. Several other names have been used

for noroviruses, including Norwalk-like viruses, caliciviruses (because they belong to the virus

family Caliciviridae), and small round structured viruses.

Rotavirus: Causes moderate to severe illness with vomiting followed by watery diarrhea and fever. It is the most

common cause of food poisoning in infants and children and is transmitted from person to person by fecal

contamination of food and shared play areas.

Hepatitis A: Causes moderate illness with sudden onset of fever, loss of appetite, abdominal pain, and feeling of

tiredness followed by jaundice, which is a yellowing of the eyes and skin. Symptoms usually last less than two

http://www.ncbi.nlm.nih.gov/pubmedhealth/n/pmh_adam/A000224/

















months, but can be prolonged or relapse for up to six months. It is transmitted from person to person by fecal

contamination of food.

Bacteria: Bacteria can cause food poisoning in two different ways. Some bacteria infect the intestines, causing

inflammation and difficulty absorbing nutrients and water, leading to diarrhea. Other bacteria produce chemicals

in foods (known as toxins) that are poisonous to the human digestive system. When eaten, these chemicals can

lead to nausea and vomiting, kidney failure, and even death.

Salmonellae: Salmonellae are bacteria that may cause food poisoning; the illness itself is often referred to

as Salmonella or Salmonella infection. TheCDC estimates that each year 1 million people are infected with

Salmonella, amounting to $365 million in direct medical costs annually. Salmonellaecause a moderate illness with

nausea, vomiting, crampy diarrhea, andheadache, which may come back a few weeks later as arthritis (joint

pains). In people with impaired immune systems (such as people with kidney disease,HIV/AIDS, or those

receiving chemotherapy for cancer), Salmonellae can cause a life-threatening illness. The illness is transmitted by

undercooked foods such as eggs, poultry, dairy products, and seafood.

Campylobacter: Causes mild illness with fever, watery diarrhea, headache, and muscle aches. Campylobacter is

the most commonly identified food-borne bacterial infection encountered in the world. It is transmitted by raw

poultry, raw milk, and water contaminated by animal feces.

Staphylococcus aureus: Causes moderate to severe illness with rapid onset of nausea, severe vomiting, dizziness,

and abdominal cramping. These bacteria produce a toxin in foods such as cream-filled cakes and pies, salads

(most at risk are potato, macaroni, egg, and tuna salads, for example) and dairy products. Contaminated salads at

picnics are common if the food is not chilled properly.

Bacillus cereus: Causes mild illness with rapid onset of vomiting, with or without diarrhea and abdominal

cramping. It is associated with rice (mainly fried rice) and other starchy foods such as pasta or potatoes. It has

been speculated that this bacteria may also be used as a potential terrorist weapon.

Escherichia coli (E coli): Causes moderate to severe illness that begins as large amounts of watery diarrhea,

which then turns into bloody diarrhea. There are many different types of this bacteria. The worst strain can cause

kidney failure and death (about 3% to 5% of all cases). It is transmitted by eating raw or undercooked hamburger,

unpasteurized milk or juices, or contaminated well water. Outbreaks of food poisoning due to E. coli have also

occurred following ingestion of contaminated produce.

Shigella (traveler's diarrhea): Causes moderate to severe illness with fever,diarrhea containing blood or mucus

or both, and the constant urge to have bowel movements. It is transmitted in water polluted with human wastes.

Listeria monocytogenes: Listeriosis is a moderate to severe illness with nausea and vomiting. Some affected

individuals can progress to developmeningitis from Listeria. It is transmitted on many tips of uncooked foods

such as meats, fruits, vegetables, soft cheeses, unpasteurized milk, and cold cut meats. Pregnant woman and

newborns are at increased risk for serious infections. In 2011, in an outbreak caused by tainted cantaloupe, 25

people died and 123 people were infected in 26 states.

Clostridium botulinum (botulism): Causes severe illness affecting the nervous system. Symptoms start

as blurred vision. The person then develops problems talking and overall weakness. Symptoms then progress to

breathing difficulty and the inability to move arms or legs. Infants and young children are particularly at risk. It is

transmitted in foods such as home-packed canned goods, honey, sausages, and seafood.

Because botulism can be released in the air, it is considered a potential biological weapon for terrorists.

Vibrio cholerae: Causes mild to moderate illness with crampy diarrhea, headache, nausea, vomiting, and fever

with chills. It strikes mostly in the warmer months of the year and is transmitted by infected, undercooked, or raw

seafood.

Vibrio parahaemolyticus: Causes moderate to severe abdominal cramping, nausea, vomiting, and fever. In

immunocompromised individuals, it can cause severe or deadly disease. It is transmitted by eating raw or

undercooked fish, particularly oysters.

Parasites rarely cause food poisoning. When they do, they are usually swallowed in contaminated or untreated

water and cause long-lasting but mild symptoms.

Giardia (beaver fever): Causes mild illness with watery diarrhea often lasting one to two weeks. It is transmitted

by drinking contaminated water, often from lakes or streams in cooler mountainous climates.

The infection can also be spread from person to person by food or other items contaminated with feces from an

infected person.

















Cryptosporidium: Causes moderate illness with large amounts of watery diarrhea lasting two to four days. May

become a long-lasting problem in people with poor immune systems (such as people with kidney disease or

HIV/AIDS or those on chemotherapy for cancer). It is transmitted by contaminated drinking water.

Toxoplasma: The CDC estimates that more than 60 million people in the U.S. carry the Toxoplasma parasite, but

few have symptoms because the immune system keeps the parasite from causing illness. When it does cause

disease, symptoms include headache, blurred vision, and eye pain. It is transmitted by eating undercooked or raw

meat, contaminated water, or contact with contaminated cat feces. Pregnant women and those with compromised

immune systems infected with Toxoplasma can have severe health complications.

Toxic agents are the least common cause of food poisoning. Illness is often an isolated episode caused by poor

food preparation or selection (such as picking wild mushrooms).

Mushroom toxins: Illness can range from mild to deadly depending on the type of mushroom eaten. Often there

is nausea, vomiting, and diarrhea. Some types of mushrooms produce a nerve toxin, which causes sweating,

shaking, hallucinations, and coma.

Ciguatera poisoning: Caused by eating fish that contains toxins produced by a marine algae

called Gambierdiscus toxicus. It can cause moderate to severe illness with numbness of the area around the mouth

and lips that can spread to the arms and legs, nausea, vomiting, muscle pain and weakness, headache, dizziness,

and rapid heartbeat. The toxin may cause sensory problems in which hot things feel cold and cold things feel hot.

It is transmitted by eating certain large game fish from tropical waters-most specifically barracuda, grouper,

snapper, and jacks. According to the CDC, ciguatera has no cure. Symptoms may disappear in days or weeks, but

may persist for years.

Scombroid: Causes mild to moderate illness with facial flushing, burning around the mouth and lips, peppery-

taste sensations, a red rash on the upper body, dizziness, headache, and itchy skin. Severe symptoms may include

blurry vision, respiratory distress, and swelling of the tongue and mouth. Symptoms typically last from four to six

hours, and rarely more than one or two days. It is transmitted in seafood, mostly mahi-mahi and tuna, but can also

be in Swiss cheese.

Pesticides: Cause mild to severe illness with weakness, blurred vision, headache, cramps, diarrhea, increased

production of saliva, and shaking of the arms and legs. Toxins are transmitted by eating unwashed fruits or

vegetables contaminated with pesticides.

MEDICAL CARE

Contact your doctor if any of the following situations occur:

Nausea, vomiting, or diarrhea lasts for more than two days.

The ill person is a child younger than three years of age.

The abdominal symptoms are associated with a low-grade fever.

Symptoms begin after recent foreign travel.

Other family members or friends who ate the same thing are also sick.

The ill person cannot keep any liquids down.

The ill person does not improve within two days even though they are drinking large amounts of fluids.

The ill person has a disease or illness that weakens their immune system (for example, HIV/AIDS, cancer

and undergoing chemotherapy, kidney disease).

The ill person cannot take their normal prescribed medications because of vomiting.

The ill person has any nervous system symptoms such as slurred speech, muscle weakness, double vision,

or difficulty swallowing.

The ill person is pregnant.

Go to the nearest hospital's emergency department if any of the following situations occur:

The ill person passes out or collapse, become dizzy, lightheaded, or has problems with vision.

A fever higher than 101 F (38.3 C) occurs with the abdominal symptoms.

Sharp or cramping pains do not go away after 10-15 minutes.

The ill person's stomach or abdomen swells.

The skin and/or eyes turn yellow.

The ill person is vomiting blood or having bloody bowel movements.

The ill person stops urinating, have decreased urination, or have urine that is dark in color.














The ill person develops problems with breathing, speaking, or swallowing.

One or more joints swell or a rash breaks out on the ill person's skin.

The ill person or caretaker considers the situation to be an emergency.

FOOD POISONING DIAGNOSIS

If the person visits a doctor or a hospital emergency department because they think they may have food

poisoning, a thorough examination will be performed, including measurements of blood pressure, pulse, breathing

rate, and temperature. The doctor will perform a physical exam, which screens for outward signs and symptoms

of the illness. They will assess how dehydrated the patient is and examine the abdominal area to make sure the

illness is not serious.The doctor may need to do a rectal examination. The doctor performs this test by inserting a

lubricated and gloved finger gently into the rectum. The purpose is to make sure there are no breaks in the rectal

wall. A sample of stool is taken and tested for blood and mucus. In some cases, a sample of stool or vomit can be

sent to the laboratory for further testing to find out which toxin caused the illness. In a majority of cases, a

specific cause is not found.A urine sample helps assess how dehydrated the patient is and may indicate possible

kidney damage.Blood tests may be performed to determine the seriousness of the illness. An X-ray of the

abdomen or a CT scan may be taken if the doctor suspects the patient's symptoms may be caused by another

illness.

FOOD POISONING SELF-CARE AT HOME

Short episodes of vomiting and small amounts of diarrhea lasting less than 24 hours can usually be cared

for at home. Do not eat solid food while nauseous or vomiting but drink plenty of fluids. Small, frequent sips of

clear liquids (those you can see through) are the best way to stay hydrated. Avoid alcoholic, caffeinated, or sugary

drinks. Over-the-counter rehydration products made for children such as Pedialyte and Rehydralyte are expensive

but good to use if available. Sports drinks such as Gatorade and Powerade are fine for adults if they are diluted

with water because at full strength they contain too much sugar, which can worsen diarrhea. Home remedies to

treat nausea or diarrhea such as tea with lemon and ginger can be used for relief from symptoms. There are no

proven herbal food poisoning cures. Consult a health care practitioner before taking any natural food poisoning

remedies. After successfully tolerating fluids, eating should begin slowly, when nausea and vomiting have

stopped. Plain foods that are easy on the stomach should be started in small amounts. Initially consider eating rice,

wheat, breads, potatoes, low-sugar cereals, lean meats, and chicken (not fried). Milk can be given safely, although

some people may experience additional stomach upset due to lactose intolerance. Most food poisonings do not

require the use of over-the-counter medicines to stop diarrhea, but they are generally safe if used as directed. It is

not recommended that these medications be used to treat children. If there is a question or concern, always check

with a doctor.

SIGN AND TEST

Your health care provider will examine you for signs of food poisoning, such as pain in the stomach and

signs your body does not have as much water and fluids as it should. This is called dehydration. Tests may be

done on your stools or the food you have eaten to find out what type of germ is causing your symptoms. However,

tests may not always find the cause of the diarrhea. In more serious cases, your health care provider may order a

sigmoidoscopy. A thin, hollow tube with a light on the end is placed in the anus to look for the source of bleeding

or infection.

MEDICAL TREATMENT SITUATIONS

Medical treatment is necessary if following situations occur:

Nausea, vomiting, or diarrhea lasts for more than two days.

The abdominal symptoms associated with a low-grade fever.

Other family members or friends who ate the same thing are also sick.

The ill person cannot keep any liquids down.

The ill person does not improve within two days even drinking large amounts of fluids.

The ill person has a disease or illness that weakens immune system (for example, HIV/AIDS, cancer and

undergoing chemotherapy, kidney disease).

The ill person cannot take normal prescribed medications because of vomiting.

The ill person has any nervous system symptoms such as slurred speech, muscle weakness, double vision,

or difficulty swallowing.









Tests: If the patient visits a doctor or a hospital emergency department because they think they may have food

poisoning, a thorough examination will be performed, including

Measurement of blood pressure

Pulse

Breathing rate

Temperature

Examination of the abdominal area

Rectal examination

Sampling of stool for blood and mucus testing

Sample of stool or vomit for testing to find out which toxin caused the illness

A urine sample helps assess how dehydrated the patient is and may indicate possible kidney damage

Blood tests may be performed to determine the seriousness of the illness

An x-ray of the abdomen or a CT scan may be taken

RISK

Higher risk foods include:

1. Meat, especially undercooked mince and rolled, formed or tenderised meats

2. Raw or undercooked poultry such as chicken, duck and turkey

3. Raw or lightly cooked eggs including foods made from raw egg such as unpasteurised mayonnaise

4. Small goods such as salami and hams

5. Seafood

6. Cooked rice not kept at correct temperatures

7. Cooked pasta not kept at correct temperatures

8. Prepared salads such as coleslaw, pasta salads and rice salads

9. Prepared fruit salads

10. Unpasteurised dairy products.

FOOD POISONING TREATMENT

Self-Care at Home: Short episodes of vomiting and small amounts of diarrhea lasting less than 24 hours can

usually be cared for at home.

Do not eat solid food while nauseous or vomiting but drink plenty of fluids. Small, frequent sips of clear

liquids are the best way to stay hydrated. Avoid alcoholic, caffeinated, or sugary drinks. Over-the-counter

rehydration products made for children such as Pedialyte and Rehydralyte are expensive but good to use

if available. Sports drinks such as Gatorade and Powerade are fine for adults if they are diluted with water

because at full strength they contain too much sugar, which can worsen diarrhea.

After successfully tolerating fluids, eating should begin slowly, when nausea and vomiting have stopped.

Plain foods that are easy on the stomach should be started in small amounts. Consider eating rice, wheat,

breads, potatoes, low-sugar cereals, lean meats, and chicken (not fried) to start. Milk can be given safely,

although some people may experience additional stomach upset due to lactose intolerance.

Most food poisonings do not require the use of over-the-counter medicines to stop diarrhea, but they are

generally safe if used as directed.

Medical Treatment: The main treatment for food poisoning is putting fluids back in the body (rehydration)

through an IV and by drinking. The patient may need to be admitted to the hospital. This depends on the severity

of the dehydration, response to therapy, and ability to drink fluids without vomiting. Children, in particular, may

need close observation.

Anti-vomiting and diarrhea medications may be given.

The doctor may also treat any fever to make the patient more comfortable.

Antibiotics are rarely needed for food poisoning. In some cases, antibiotics worsen the condition. Only a

few specific causes of food poisoning are improved by using these medications. The length of illness with

traveler's diarrhea can be decreased with antibiotics, but this specific illness usually runs its course and

improves without treatment.





With mushroom poisoning or eating foods contaminated with pesticides, aggressive treatment may

include pumping the stomach (lavage) or giving medications as antidotes. These poisonings are very

serious and may require intensive care in the hospital.

Prevention: Safe steps in food handling, cooking, and storage are essential to avoiding food-borne illness.

Bacteria cannot be seen, smelled, or tasted, which may be on any food.

Illness can be prevented by:

1) Controlling the initial number of bacteria present.

2) Preventing the small number from growing.

3) Destroying the bacteria by proper cooking.

4) Avoiding re-contamination.

Follow the food safety guidelines to keep contaminants away:

Safe shopping

Buy cold foods last during shopping trip. Get them home fast.

Never choose torn or leaking packages.

Do not buy foods past their expiration dates.

Keep raw meat and poultry separate from other foods.

Safe storage

Keep it safe, refrigerate.

Place raw meat, poultry, or fish in the coldest section of refrigerator.

Check the temperature of appliances. To slow bacterial growth, the refrigerator should be at 40°F, the

freezer at 0°F.

Cook or freeze fresh poultry, fish, and meats within 2 days.

Safe food preparation

Keep everything clean.

Wash hands before and after handling raw meat and poultry.

Sanitize cutting boards often in a solution of one teaspoon chlorine bleach in one quart of

water.

Keep raw meat, poultry, fish, and their juices away from other food. After cutting raw meats, wash hands,

cutting board, knife, and counter tops with hot, soapy water.

Safe cooking

Keep hot foods hot and cold foods cold.

Use cooked leftovers within four days.

Complication: Dehydration is the most common complication. This can occur from any causes of food

poisoning. Less common, but much more serious complications depend on the bacteria that are causing the food

poisoning. These may include:

Arthritis

Bleeding problems

Damage to the nervous system

Kidney problems

Swelling or irritation in the tissue around the heart

Prevention: Here are steps you can take to prevent food poisoning at home:

Wash your hands, utensils and food surfaces often. Wash your hands well with warm, soapy water before and

after handling or preparing food. Use hot, soapy water to wash the utensils, cutting board and other surfaces you

use.

Keep raw foods separate from ready-to-eat foods. When shopping, preparing food or storing food, keep raw

meat, poultry, fish and shellfish away from other foods. This prevents cross-contamination.

Cook foods to a safe temperature. The best way to tell if foods are cooked to a safe temperature is to use a food

thermometer. You can kill harmful organisms in most foods by cooking them to the right temperature. Ground

beef should be cooked to 160 F (71.1 C), while steaks and roasts should be cooked to at least 145 F (62.8 C). Pork






needs to be cooked to at least 160 F (71.1C), and chicken and turkey need to be cooked to 165 F (73.9 C). Fish is

generally well-cooked at 145 F (62.8 C).

Refrigerate or freeze perishable foods promptly. Refrigerate or freeze perishable foods within two hours of

purchasing or preparing them. If the room temperature is above 90 F (32.2 C), refrigerate perishable foods within

one hour.

Defrost food safely. Do not thaw foods at room temperature. The safest way to thaw foods is to defrost foods in

the refrigerator or to microwave the food using the "defrost" or "50 percent power" setting. Running cold water

over the food also safely thaws the food.

Throw it out when in doubt. If you aren't sure if a food has been prepared, served or stored safely, discard it.

Food left at room temperature too long may contain bacteria or toxins that can't be destroyed by cooking. Don't

taste food that you're unsure about — just throw it out. Even if it looks and smells fine, it may not be safe to eat.

Food poisoning is especially serious and potentially life-threatening for young children, pregnant women and their

fetuses, older adults, and people with weakened immune systems. These individuals should take extra precautions

by avoiding the following foods:

Raw or rare meat and poultry

Raw or undercooked fish or shellfish, including oysters, clams, mussels and scallops

Raw or undercooked eggs or foods that may contain them, such as cookie dough and homemade ice

cream

Raw sprouts, such as alfalfa, bean, clover or radish sprouts

Unpasteurized juices and ciders

Unpasteurized milk and milk products

Soft cheeses (such as feta, Brie and Camembert), blue-veined cheese and unpasteurized cheese

Refrigerated pates and meat spreads

Uncooked hot dogs, luncheon meats and deli meats

REMEDIES OF FOOD POISONING

Here are the top five easy and quick home remedies to get instant relief from food poising:

1. Ginger: Ginger is a well known remedy for various gastrointestinal distresses. Loaded with potent anti-

inflammatory compounds ginger helps quell nausea and gastric distress. Chewing a piece of ginger tossed in

honey helps relieve the severity of nausea. Ginger tea relieves stomach cramps and upset caused by food

poisoning.

2. Cumin: Add a tablespoon of crushed cumin seeds to the soup to soothe the inflammation in your stomach.

Crushed cumin with fenugreek powder mixed with a glass of water or half a cup crud helps relive abdominal pain

and vomiting. Cumin seeds are of great benefit to the digestive system as they help stimulate the secretion of

pancreatic enzymes, compounds necessary for proper digestion and nutrient assimilation.

3. Basil: Basil is another excellent home remedy to cure stomach infection due to its anti-bacterial properties.

Taking juice of basil leaves with water empty stomach in the morning enhances digestive power. Basil juice helps

stop vomiting immediately. Strain the juice of a few basil leaves and add it to a tablespoon of honey to get instant

relief. Mix chopped basil leaves, sea salt and one shake of black pepper to three tablespoon of crud. Take the

mixture three times in a day till you are totally cured of food poisoning. It will also cure any cramps or gas

problem associated with food poisoning.

4. Lemon: The acidity of the lemon juice kills the micro-organism and toxins in the gastrointestinal tract.

Squeeze juice of a lemon and add a pinch of sugar to it and drink, or you can even add lemon to your tea.

Since fluid intake is very important as one tends to lose more water through diarrhea taking lemon juice in short

intervals helps you keep hydrated.

5. Peppermint tea: Peppermint oil helps relieve symptoms of irritable bowel syndrome, including indigestion,

dyspepsia, and colonic muscle spasms. It is extremely beneficial for people suffering from stomach spasms due to

food poisoning. Add a few drops to your tea; your cramps will vanish in a couple of hours.

TREATMENTS

You will usually get better in a couple of days. The goal is to make you feel better and make sure your

body has the proper amount of fluids.

Getting enough fluids and learning what to eat will help keep you or your child comfortable. You may need to:





Manage the diarrhea

Control nausea and vomiting

Get plenty of rest

If you have diarrhea and are unable to drink or keep down fluids, you may need fluids given through a

vein (by IV). This is especially true for young children. If you take diuretics, ask your health care provider if you

need to stop taking the diuretic while you have diarrhea. Never stop or change medications without first talking to

your health care provider. For the most common causes of food poisoning, your doctor will NOT prescribe

antibiotics. You can buy medicines at the drugstore that help slow diarrhea. Do not use these medicines without

talking to your health care provider if you have bloody diarrhea, a fever, or the diarrhea is severe. Do not give

these medicines to children

Treatments and drugs: Treatment for food poisoning typically depends on the source of the illness, if known,

and the severity of your symptoms. For most people, the illness resolves without treatment within a few days,

though some types of food poisoning may last a week or more.Treatment of food poisoning may include:

Replacement of lost fluids: Fluids and electrolytes — minerals such as sodium, potassium and calcium that

maintain the balance of fluids in your body — lost to persistent diarrhea need to be replaced. Children and adults

who are severely dehydrated need treatment in a hospital, where they can receive salts and fluids through a vein

(intravenously), rather than by mouth. Intravenous hydration provides the body with water and essential nutrients

much more quickly than oral solutions do.

Antibiotics: antibiotics if you have certain kinds of bacterial food poisoning and your symptoms are severe. Food

poisoning caused by listeria needs to be treated with intravenous antibiotics in the hospital. And the sooner

treatment begins, the better. During pregnancy, prompt antibiotic treatment may help keep the infection from

affecting the baby.

CONCLUSION

Food poisoning is a common infection that affects millions of people in the India each year. Most

commonly, patients complain of vomiting, diarrhea, and crampy abdominal pain. People should seek medical care

if they have an associated fever, blood in their stool, signs and symptoms of dehydration, or if their symptoms do

not resolve after a couple of days. Treatment focuses on keeping the patient well hydrated. Most cases of food

poisoning resolve on their own. Prevention is key and depends upon keeping food preparation areas clean, good

hand washing, and cooking foods thoroughly. Food poisoning is the name for the range of illnesses caused by

eating or drinking contaminated food or drink. It is also sometimes called food borne illness.

REFERENCES

Food and Agriculture Organization of the United Nations, GASGA Technical Leaflet - 3 Mycotoxins in Grain,

Retrieved 12 August 2007.

World Health Organization, Chapter 2 Foodborne Hazards in Basic Food Safety for Health Workers Retrieved 12

August 2007.

Food and Drug Administration, Sec. 683, 100 Action Levels for Aflatoxins in Animal Feeds (CPG 7126.33),

Retrieved 13 August 2007.

Henry, Michael H, Mycotoxins in Feeds: CVM’s Perspective , Retrieved 1 January 2012.

Webley DJ, Jackson KL, Mullins JD, Hocking AD, Pitt JI, Alternaria toxins in weather-damaged wheat and

sorghum in the 1995–1996 Australian harvest , Australian Journal of Agricultural Research, 48 (8), 1997, 1249-

56.

Centers for Disease Control and Prevention, Diagnosis and management of foodborne illness, A primer for

physicians and other health care professionals, MMWR, 53(RR-4), 2004, 1-32.

Centers for Disease Control and Prevention (2008). Toxoplasmosis. www.cdc.gov/toxoplasmosis/factsheet.html.

Centers for Disease Control and Prevention (2009). Foodborne infections.

www.cdc.gov/nczved/divisions/dfbmd/diseases/foodborne_infections.

http://www.who.int/entity/foodsafety/publications/capacity/en/2.pdf

http://www.fda.gov/ora/compliance_ref/cpg/cpgvet/cpg683-100.html

http://www.fda.gov/AnimalVeterinary/Products/AnimalFoodFeeds/Contaminants/ucm050974.htm





Centers for Disease Control and Prevention (2009). Listeriosis. Available online:

www.cdc.gov/nczved/divisions/dfbmd/diseases/listeriosis.

Centers for Disease Control and Prevention (2009). Noroviruses and drinking water from private wells.

www.cdc.gov/healthywater/drinking/private/wells/disease/norovirus.html.

Centers for Disease Control and Prevention (2009).Salmonellosis.:

www.cdc.gov/nczved/divisions/dfbmd/diseases/salmonellosis.

Centers for Disease Control and Prevention (2009). Shigellosis.

www.cdc.gov/nczved/divisions/dfbmd/diseases/shigellosi

http://www.webmd.com/a-to-z-guides/listeriosis-topic-overview

http://children.webmd.com/norovirus-symptoms-and-treatment

http://www.webmd.com/food-recipes/food-poisoning/tc/salmonellosis-topic-overview





MICROENCAPSULATION TECHNOLOGY K.P.Sampath Kumar

1*,Tejbe.Sk

2, Shameem Banu

2, P.Naga Lakshmi

2, D.Bhowmik

3

1. Coimbatore Medical College, Coimbatore

2. Nimra Pharmacy College,Vijayawada

3. Karpagam University,Coimbatore


ABSTRACT

Microparticulate drug delivery systems provide tremendous opportunities for designing new

controlled and delayed release oral formulations, thus extending the frontier of future pharmaceutical

development. The Microparticulate offers a variety of opportunities such as protection and masking,

reduced dissolution rate, facilitation of handling, and spatial targeting of the active ingredient. It is the

process by which individual particles or droplets of solid or liquid material (the core) are surrounded or

coated with a continuous film of polymeric material (the shell) to produce capsules in the micrometer to

millimeter range, known as microcapsules. Microencapsulation technology can protect active materials

against environment, stabilize them, prevent or suppress volatilization. Microencapsulation technology

can provide new forms and features and many polymeric drug delivery systems, biodegradable polymers

have been used widely as drug delivery systems because of their biocompatibility and biodegradability.

Microencapsulation is a powerful technique to achieve targeted delivery and on-demand release of

different active ingredients. Many synthetic and natural biodegradable polymers present exciting

opportunities in tailor-making the micro particle formulations for long-term drug release with specific

release rates. Hence finally concluded that continuous knowledge up gradation is required in order to

make desired drug delivery system and minimization of problems associated with physicomechanical

techniques and complete knowledge about selection of raw materials and method for their

microencapsulation to get desire goal of study.

Key words: Microencapsulation technology, targeted delivery, protection, masking.

INTRODUCTION

Microencapsulation is a rapidly expanding technology. It is the process of applying relatively thin

coatings to small particles of solids or droplets of liquids and dispersions. Microencapsulation provides the

means of converting liquids to solids, of altering colloidal and surface properties, of providing environmental

protection and of controlling the release characteristics or availability of coated materials. Microencapsulation

is receiving considerable attention fundamentally, developmentally and commercially. The term microcapsule is

defined as a spherical particle with size varying from 50nm to 2mm, containing a core substance. Microspheres

are in strict sense, spherical empty particles. However the terms microcapsule and microsphere are often

used synonymously. The microspheres are characteristically free flowing powders consisting of proteins

or synthetic polymers, which are biodegradable in nature, and ideally having a particle size less than 200µm.

Solid biodegradable microcapsules incorporating a drug dispersed or dissolved throughout the particle matrix

have the potential for the controlled release of drug. These carries received much attention not only for prolonged

release but also for the targeting of the anticancer drug to the tumour. The concept of miroencapsulation was

initially utilized in carbonless copy papers. More recently it has received increasing attention in pharmaceutical

and biomedical applications. The first research leading to the development of micro encapsulation procedures for

pharmaceuticals was published by Bungenburg de Jong and Kass in 1931 and dealt with the preparation of gelatin

spheres and the use of gelatin coacervation process for coating. In the late 1930s, Green and co-workers of

National cash register co. Dayton, Ohio, developed the gelatin coacervation process. Since then may other

coating materials and processes of application have been developed by the pharmaceutical industry for the

microencapsulation of medicines. Over the last 25 years pharmaceutical companies for microencapsulated drugs

have taken out numerous patents.

Reasons for microencapsulation: The primary reason for microencapsulation is found to be either for sustained

or prolonged drug release. This technique has been widely used for masking taste and odor of many drugs to

improve patient compliance. This technique can be used for converting liquid drugs in a free flowing powder.The

drugs, which are sensitive to oxygen, moisture or light, can be stabilized by microencapsulation. Incompatibility

among the drugs can be prevented by microencapsulation. Vaporization of many volatile drugs e.g. methyl

salicylate and peppermint oil can be prevented by microencapsulation. Many drugs have been microencapsulated






to reduce toxicity and GI irritation including ferrous sulphate and KCl. Alteration in site of absorption can also be

achieved by microencapsulation. Toxic chemicals such as insecticides may be microencapsulated to reduce the

possibility of sensitization of factorial person. Bakan and Anderson reported that microencapsulated vitamin A

palmitate had enhanced stability.

Microparticles offer various significant advantages as drug delivery systems, including:

i. An effective protection of the encapsulated active agent against (e.g. enzymatic) degradation

ii. The possibility to accurately control the release rate of the incorporated drug over periods of hours to

months.

iii. An easy administration (compared to alternative parenteral controlled release dosage forms, such as

macro-sized implants).

iv. Desired, pre-programmed drug release profiles can be provided which match the therapeutic needs of the

patient.

Microparticulate drug delivery systems are an interesting and promising option when developing an oral

controlled release system.Microcapsules are finally dispersed in various dosage forms, such as hard gelatin

capsules, which may be enteric coated, soft gelatin capsules, or suspensions in liquids, all of which allow

dispersion of individual microcapsules on release.Microcapsules continue to be of much interest in controlled

release because of relative ease in design and formulation and partly on the advantages of microparticulate

delivery systems. The latter include sustained release from each individual microcapsule and offer greater

uniformity and reproducibility. Additional advantage over monolithic systems containing multiple doses is the

greater safety factor in case of a burst or defective individual in subdivided dosage forms. Finally, it has been

argued that multiple particle systems are distributed over a great length of gastro-intestinal tract, which should

result in, (a) lowered local concentrations and hence reduced toxicity or irritancy, and (b) reduced variability in

transit time and absorption rate

Basic consideration of microencapsulation technique: Microencapsulation often involves a basic

understanding of the general properties of microcapsules, Such as the nature of the core and coating materials,

the stability and release characteristics of the coated materials and the microencapsulation methods. The

intended physical characters of the encapsulated product and the intended use of the final product must also be

considered.

a. Core material: The core material, defined as the specific material to be coated, can be liquid or solid in nature.

The composition of the core material can be varied as the liquid core can include dispersed and/or dissolved

material. The solid core can be a mixture of active constituents, stabilizers, diluents, excipients and release rate

retardants or accelerators.

b. Coating materials: The coating material should be capable of forming a film that is cohesive with the core

materials, be chemically compatible and non reactive with the core material and provide the desired coating

properties such as strength, flexibility impermeability, optical properties and stability. The total thickness of

the coatings achieved with microencapsulation techniques is microscopic in size.

c. Stability, release and other properties: Three important areas of current microencapsulation application are

the stabilization of core materials, the control of the release or availability of core materials and separation of

chemically reactive ingredients within a tablet or powder mixture. A wide variety of mechanisms is available to

release encapsulated core materials; such as disruption of the coating can occur by pressure, shear or abrasion

forces, permeability changes brought about enzymatically etc., improved gastro tolerability of drugs can be

obtained by microencapsulation.

d. Physical character of the final product: Microcapsules should have desirable physical properties like

ability to flow, to be compacted or to be suspended and the capsule wall must be capable of resisting the

pressure during compression etc.

e. coating materials: A number of different substances both biodegradable as well as non-biodegredable have

been investigated for the preparation or microcapsules. These materials include the polymers of natural and

synthetic origin and also modified natural substances. Some of the polymers used in the preparation of the

microcapsules are classified and listed below.





SYNTHETIC POLYMERS

Non-biodegradable

1. PMMA

2. Acrolein

3. Glycidyl methacrylate

4. Epoxy polymers

Biodegradable

1. Lactides and glycolides and their copolymers

2. Polyalkyl cyano acrylates

3. Polyanhydrides

4. Corbopol

NATURAL MATERIALS

A. Proteins

B. Albumins

C. Gelatin

D. Collagen

E. Carbohydrates

F. Starch , Agarose

G. Carrageenan

H. Chitosan

I. Chemically modified carbohydrates

J. DEAE cellulose

K. Poly (acryl) dextran

L. Poly (acryl) starch

METHODS OF MICROENCAPSULATION Preparation of microecapsules as prolonged action dosage form can be achieved by various techniques under

following headings.

1. Coacervation phase separation

a. By temperature change

b. By incompatible polymer addition

c. By non-solvent addition

d. By salt addition

e. By polymer-polymer interaction

f. By solvent evaporation

2. Multi orifice centrifugal process.

3. Pan coating

4. Air suspension coating

5. Spray drying and spray congealing

6. Polymerization

7. Melt dispersion technique

1. Coacervation phase separation: Microencapsulation by coacervation phase separation is generally

attributed to the national cash register (NCR) corporation and the patents of green et.al. The general outline of

the processes consists of three steps carried out under continuous agitation.

1. Formation of three immiscible chemical phases

2. Disposition of the coating, and

3. Rigidization of the coating

a. By thermal change: phase separation of the dissolved polymer occurs in the form of immiscible liquid droplets,

and if a core material is present in the system, under proper polymer concentration, temperature and agitation

conditions, the liquid polymer droplets coalesce around the dispersed core material particles, thus forming the

embryonic microcapsules. As the temperature decreases, one phase becomes polymer-poor (the

microencapsulation vehicle phase) and the second phase. (The coating material phase) becomes polymer-rich.

b. By incompatible polymer addition: it involves liquid phase separation of a polymers coating material and





microencapsulation can be accomplished by utilizing the incompatibility of dissimilar polymers existing in a

common solvent.

c. By non-solvent addition: a liquid that is a non-solvent for a given polymer can be added to a solution of the

polymer to induce phase separation. The resulting immiscible liquid polymer can be utilized to effect

microencapsulation of an immiscible core material.

d. By salt addition: there are two types of coacervation: simple and complex. Simple coacervation involves the

use of only on colloid, e.g. gelatin in water, and involves removal of the associated water from around the

dispersed colloid by agents with a greater affinity for water, such as various alcohols and salts. The

dehydrated molecules of polymer tend to aggregate with surrounding molecules to form the coacervate.

Complex coacervation involves the use of more than one colloid. Gelatin and acacia in water are most frequently

used, and the coacervation is accomplished mainly by charge neutralization of the colloids carrying opposite

charges rather than by dehydration.

e. By polymer-polymer interaction: the interaction of oppositely charged poly electrolytes can result in the

formation of a complex having such reduce solubility that phase separation occurs.

f. By solvent evaporation: the processes are carried out in a liquid manufacturing vehicle. The

microcapsule coating is dispersed in a volatile solvent, which is dispersed in volatile solvents, which is

immiscible with the liquid manufacturing vehicle phase. A core material to be microencapsulated is dissolved or

dispersed in the coating polymer solution. With agitation, the core material mixture is dispersed in the liquid

manufacturing vehicle phase to obtain the appropriate size microcapsule. The mixture is then heated if necessary

to evaporate the solvent for the polymer. In the case in which the core material is dissolved in the coating polymer

solution, matrix type microcapsules are formed. The solvent evaporation technique to product microcapsules is

applicable to a wide variety of core materials. The core materials may be either water soluble or water insoluble

materials.

2. Multiorifice centrifugal process: The South-West research institute (SWRI) has developed a mechanical

process for producing microcapsules that utilizes centrifugal forces to hurl, a core material particle through an

enveloping microencapsulation membrane therapy effecting mechanical microencapsulation. Processing

variables include the rotational speed of the cylinder, the flow rate of the core and coating materials, the

concentration and viscosity of the coating material an the viscosity and surface tension of the core material. This

method is capable of microencapsulating liquids and solids of varied size ranges, with diverse coating

materials.

3. Pan coatings: The microcapsulation of relatively large particles by pan coating method are generally

considered essential for effective coating. The coating is applied as a solution or as an automized spray to

the desired solid core passed over the coated materials during coatings is being applied in the coating pans.

4. Air suspension coating: The process consists of the dispersing of solid particulate core materials in a

supporting air stream and the spray coating of the air suspended particles. Within coating chambers,

particles are suspended on an upward moving air stream. The design of the chamber and its operating parameters

effect a re-circulating flow of the particles through the coating zone portion of the chamber, where is a coating

material, usually a polymer solution is spry-applied to the moving particles.

5. Spray drying and spray congealing: Spray drying and spray congealing processes are similar in that both

involve dispersing the core material in liquefied coating substance and spraying or introducing the core coating

mixture into some environmental condition, whereby relatively rapid solidification of the coating is

effected. The principle difference between the two methods is the means by which coating solidification

is accomplished. Coating solidification in the case of spray during is effected by rapid evaporation of solvent in

which the coating material is dissolved. Coating solidification in spray congealing method, however, is

accomplished by thermally congealing a molten coating material or by solidifying the dissolved coating by

introducing the coating core material mixture into a nonsolvent. Removal of the nonsolvent or solvent from the

coated product is then accomplished by sorption extraction or evaporation techniques.

6. Polymerization: The method involves the reaction of monomeric unit located at the interface existing between

a core material and a continuous phase in which the core material is dispersed. The continuous or core material

supporting phase is usually a liquid or gas and therefore the polymerization reaction occurs at a liquid-liquid,

liquid-gas, solid-liquid or solid-gas interface e.g., microcapsules containing protein solutions by incorporating the

protein in the aqueous diamine phase.





7. Melt-dispersion technique: In this technique the coating material is melted by heating upto 80oC. The

drug is suspended in it and then emulsified in water containing emulsifying agent at 80oC under stirring.

Microcapsules are formed as the temperature of the system reaches to room temperature.

CONCLUSION

Microencapsulation is one of the quality preservation techniques of sensitive substances and a method for

production of materials with new valuable properties. Microencapsulation is process of enclosing micron sized

particles in a polymeric shell.Significances of microencapsulation For Sustained or prolonged drug release For

Masking test and odour of many drugs Converting liquid into free flowing properties Drugs which are sensitive to

Light,oxygen, moisture they are easily stabilized. Microencapsulation technologies are applied in any area of the

industry. It can be found in: Cell immobilization, Beverage production, Protection of molecules from other

compounds, Drug delivery, Quality and safety in food, agricultural & environmental sectors, pharmaceuticals etc.

REFERENCES

Dziezak JD, Microencapsulation and encapsulated ingredients, Food Technology, 42(4), 1988, 136-51.

Fergason JL, Polymer encapsulated nematic liquidcrystals for display and light control applications, SID Int.

Symp Digest, 16, 1985, 68-70.

Green BK & Schleicher L, The National Cash Register Company, Dayton, Ohio, Oil containing microscopic

capsules and method of making them, US Patent 2,800,457.23 July 1957, 11.

Green BK, The National Cash Register Company, Dayton, Ohio, Oil containing microscopic capsules and

methodof making them, US Patent 2,800,458, 23 July 1957,

Jackson LS & Lee K, Microencapsulation and encapsulated ingredients, Lebensmittel WissenschaftTechnol, 24,

1991, 289-97.

Mars GJ & Scher HB, Controlled delivery of cropprotecting agents, Wilkens, R.M. (Ed.) Taylor and Francis,

London, 1990, 65-90.

Scher, H. B. In Proceedings of the 5th InternationalCongress of Pesticides Chemistry, edited by Miyamoto, J.&

Kearney, P C Pergamon Press, Oxford. 1982, 295-300.

Schnoring H, Dahm M & Pampus G, Fed. Rep. of Germany, Process for the Production of Microcapsules, US

Patent 4,379,071, 5 April 1983. 9pp.

Shahidi F & Han XQ, Encapsulation of food ingredients, Crit Rev. Food Sci. Nutr, 33, 1993, 501-47.

Zhang MQ, Yin T, Rong MZ & Yang GC, Selfhealing epoxy composites–preparation and effect of the healant

consisting of microencapsulated epoxy and latent curing agent. Composites Sci. Technol, 67(2), 2007, 201-12.



Praneta Desale Indian Journal of Research in Pharmacy and Biotechnology


AN OVERVIEW ABOUT PHARMACY EDUCATION IN INDIA Praneta Desale*

ISPOR Asia Consortium Education Committee, Maharashtra-424304, India.

* Corresponding author: Email- [email protected]

ABSTRACT

If the current trend toward automation continues, there will be severe cuts in the employment

of pharmacists in pharmaceutical industries in India. To survive in society as a professional, the

pharmacist will have to find suitable alternative avenues. The World Health Organization (WHO)

has organized three meetings on the potential role of pharmacists in the health care system: in New

Delhi in 1988, in Tokyo in 1993, and in Vancouver in 1997. The first meeting outlined the various

activities of pharmacists, namely regulatory control and drug management, legislation, procurement,

storage and distribution of drugs, drug information, quality control, community and hospital

pharmacy, industrial pharmacy, and academic activities, the second meeting introduced the concept

of ph armaceutical care, and the third meeting was on developing curriculums for the education of

future pharmacists. The International Pharmaceutical Federation has developed good pharmacy

practice standards for the community and hospital settings many pharmacy institutions in India have

started offering programs in pharmacy practice and clinical pharmacy. The Pharmacy Council of

India (PCI) has initiated steps to raise the minimum qualification of registered pharmacists. This

ambitions plan materializes there will be ample opportunity for pharmacists to serve society.

Key words: Pharmacy education, Pharmacy council of India, Education Regulations

INTRODUCTION

The beginning of pharmaceutical education in India was initiated at the Banaras Hindu University

(BHU) in 1932 by Professor M. L. Schroff. From there it has been a long journey of almost 80 years for this

profession in this country. The enactment of the Pharmacy Act 1948 established the statutory regulation of

pharmacy institutions in India. The Pharmacy Council of India (PCI) was established in 1949 under “Ministry

of Health” and the first education regulations (ER) framed in 1953, which were subsequently amended in

1972, 1981 and 1991. On the other hand, the pharmacy education has never been part of paramedical team and

hence, its development has been quite unique and quite different from rest of the world. Pharmacy Council of

India and Pharmacy Act were created to establish minimum qualification required to be a pharmacist. The role

of pharmacist in the society was never been given its due place and did not grow due to less paying job

compared to job in industry. This would have been the reason for transfer of pharmacy education from PCI to

All India Council of Technical Education (AICTE) under the “Ministry of Human Resource Development”.

Currently, PCI and AICTE regulate pharmacy profession and education respectively in India. Both of

these regulatory bodies have been doing a regulatory function without bothering to create a permanent

mechanism of updating curriculum along with development in the field. In short, it can be said that evolution

of pharmacy education has been quite confusing and developed like a vagabond. Hence, evolution of

pharmacy education has been primarily due to evolution of pharmaceutical industries and has lot of impact

under curriculum of “Bachelor and Master in Pharmacy” programmes. Similarly, medical education in India

grew with less focus on research and development and hence, India produced medical graduates more with

clinical sense acquired through experience and less of a doctors with analytical bent of mind. Due to tight

junctions at the entry point, integration of the thoughts of medical sciences, pharmaceutical sciences, nursing,

engineering sciences and basic sciences have never taken place. Primarily, this resulted in isolated

development of medical education without integration with other sciences including pharmaceutical. It is also

true in case of pharmacy education. It may also one of the reason of pharmacy education not to be a part of

healthcare system.

Today, the global institutes are moving towards excellence in research and capability building in order

to better meet the requirements of 21st century. This forces us to evaluate status of pharmacy education in

India. There is a rapid transition in pharmacy profession worldwide and in the era of globalization, we cannot

be silent spectators. If we have to compete with the rest of the world and become guiding torch for rest of the

world, we will have to become proactive. It means, we have to define the goals of pharmacy education for

present and future and re-frame our curriculum according to defined goals to meet the global challenges. In

the past decade, the technical education in India has spread its roots at an amazing rate. On the other hand,

there is sharp decrease in interest and overall admissions to undergraduate programme (B. Pharmacy) in

pharmacy during the last three academic years. This decline may be attributed due to changed trends in

pharmaceutical industry which has become primarily research and marketing oriented from production

oriented. Role of knowledge in giving in increasing employability of the students has become need of the





hour. Simultaneously, other facets of pharmacy profession should be given adequate attention in curriculum

development and in creation of knowledge based manpower for service of the society. The products of this

form of education lack the much needed professionalism and rational thinking required for problem solving.

So, the situation demands a nudge for the system to ensure its revival in order to better meet the needs of 21st

century.

Pharmacy is related to health sciences. It is the profession responsible for the preparation, dispensing

and appropriate use of medication and which provides services to achieve optimal therapeutic outcomes. A

Pharmacists job is to prepare, mix, compound or dispense drugs and medicines, ointments, powder, pills,

tablets and injections on the prescription of a medical practitioner, dentist or veterinarian. In detail, they are

concerned with production of pharmaceutical products, development of the methods or processes of

production and quality control. Those in research concern themselves with synthesis of new drugs (what is

commonly referred to as molecules), new processes, clinical testing of the effects of such drugs on animals

and humans, and obtaining the required License from the drug control authorities. A pharmacist is required to

explain the mode and precautions regarding the use of medicines dispensed in a hospital pharmacy, prepare

special formulations normally not available in the market, assist the physician in rendering necessary

information about various drugs, their contra-indications, incompatibility etc.

PHARMACY PROFESSION IN INDIA

Currently there are over a million pharmacists in India with around 55% of them in community, 20%

in hospital, 10 % in industry & regulatory. And 2 % in academia in India, formal pharmacy education leading

to a degree began in 1937, with the introduction of a 3 year industry – oriented Bachelor of Pharmacy course.

To meet the varying needs of the profession at different levels the following pharmacy programs are offered in

India today: Diploma in Pharmacy (D.Pharm.), Bachelor of Pharmacy (B.Pharm.), Master of Pharmacy

(M.Pharm.), practice- based Doctor of Pharmacy (Pharm.D.), and Doctor of Philosophy in Pharmacy (Ph.D.).

To practice as a pharmacist in India, one needs at least a diploma in pharmacy, which is awarded after 2 years

and 3 months of pharmacy studies & practical training. These diploma-trained pharmacists are currently the

mainstay of pharmacy practice in India. Every year nearly 20000 D. Pharm, 30,000 B. Pharm, 6000 M.Pharm

and 700 Pharm.D. students graduate in the Country. Pharmacy Council of India (PCI) is the statutory body

established in 1949, for regulating pharmacy education and practice of pharmacy profession in India. PCI is

actively working towards strengthening and upgrading the curriculum to produce competent workforce that is

able to meet the growing demands of the industry & community. In 2003, the Pharma Vision 2020 Charter

was released by the then President of India, Dr. A.P.J. Abdul Kalam, at the 55th Indian Pharmaceutical

Congress at Chennai. The Vision 2020 is focused on promoting the highest professional ethical standards of

pharmacy, focusing the image of pharmacists and competent healthcare professionals, sensitizing the

community, government and others on vital professional issues and supporting pharmaceutical education and

sciences in all aspects. Indian Pharmaceutical Association once again, with the support of the leaders of the

pharmacy profession presented the road map to Pharma Vision 2020 at the 58th Indian Pharmaceutical

Congress held in December 2006 at Mumbai. The themes of the subsequent Congresses in the country have

been centered on Pharma Vision 2020.

PHARMACEUTICAL INDUSTRY IN INDIA

It is exciting to be part of the Pharmaceutical industry in India today, In 2007, the turnover of the

Indian Pharmaceutical Industry was USD 8.4 bn, with additional USD 5.8 bn generated from exports. If we

take a look at the top 10 pharmaceutical companies in India, more than 50% of their annual turnover comes

from exports. India exports to over 100 countries & also boasts of having the largest USFDA approved

facilities outside USA. The number is rising every year as Indian companies keep on adding facilities to cater

to the increasing demand. India is the 3rd

largest manufacturer of pharmaceuticals in the global market in

terms of volume of sales. In terms of value, India is 14th globally which clearly shows that the prices of the

medicines in India are lowest in the world. India has slowly but surely emerged as the global outsourcing hub-

be it for manufacturing or R&D, Clinical research or basic drug discovery. The reasons are not hard to fathom.

India has a huge pool of qualified & well trained professionals, While maintaining stringent quality standards,

it is possible to establish GMP- compliant manufacturing units at around 30% cheaper rates as compared to

any of the regulated markets. All these factors make outsourcing various pharmaceutical operations to India a

very lucrative option for the big pharma that are trying hard to manage their profit margins.Indian

pharmaceutical companies have also been making increasing forays into the global generics market, which is

continuously increasing because of many of the blockbuster drugs going off-patent, Identification of

molecules going off-patent, well thought out time lines, speed of doing development work and good





regulatory understanding are some of the key factors that define success in the generic industry; attributes that

are the forte of Indian companies.

TOTAL QUALITY MANAGEMENT IN PHARMACY EDUCATION

Applying principles of TQM to pharmacy education in India leads to the development of pharmacy

education in India. The concept of Total Quality Management (TQM) although developed by an American

was successfully implemented by Japan in their recovery from World War II. The concept of TQM is

applicable to academics. Many educators believe that the concept of TQM provides guiding principles for

needed educational reform. Education is a fast moving commodity in the market and is mainly business

oriented which means it should give some profit to the undertaker. TQM is a philosophy for perfection and

continuous improvement in services offered to someone or one's own performance. The TQM principles

which are most salient to educational reform are as follows: Synergistic relationship: According to this

principle, an organization must focus, first and foremost, on its “suppliers” and “customers”. In other words,

teamwork and collaboration are essential. The concept of synergy suggests that performance and production is

enhanced by pooling the talent and experience of individuals. In a classroom, teacher-student teams are the

equivalent of industry’s front-line workers. The product of their successful work together is the development

of the student’s capabilities, interests, and character.

Continuous improvement and self evaluation: TQM emphasizes self-evaluation as part of a continuous

improvement process. In addition, this principle also laminates to the focusing on students’ strengths,

individual learning styles, and different types of intelligences. A system of ongoing process: The recognition

of the organization as a system and the work done within the organization must be seen as an ongoing process.

Quality speaks to working on the system, which must be examined to identify and eliminate the flawed

processes that allow its participants to fail.

Leadership: The upper level provides proposes basic way of functioning, provides quality staff, while the

lower level are directly linked to the students as lecturers who perform the most important functions of the

whole system. The school teachers must establish the context in which students can best achieve their

potential.

REGULATION BODIES OF PHARMACY EDUCATION

There is no doubt that currently there is enormous gap existing between education and practice of

pharmacy. Most of the academic institutions providing education in pharmacy are away from practice

environment. The overall basis of pharmacy education is still extra biological synthesis, physicochemical

studies, analysis, and manufacturing aspects of drug. It is a common feeling that the medical practitioner is

better placed for pharmacists' job than the pharmacists themselves. The dispensing services are poor. The

syllabus and duration of the two-year diploma course in pharmacy education in India is completely outdated

and irrelevant in the present industry context. It is a heterogeneous mixture of clinical and industrial subjects.

Since clinical subjects are there PCI comes into the picture and AICTE came in because of industrial

orientation of pharmacy syllabus. Pharmacy as a nascent science developed like this in the last century.

During 1940s and 50s, hospitals and industries were established in large numbers in India.

Consequently, pharmacists and pharmaceutical chemists were required in huge numbers. Hence

pharmacy education was developed in such away to satisfy the requirement of industry and hospital. Short-

term compounders and or D. Pharm. course to satisfy the needs of hospital and medical shops and B. Pharm.

course for the industry were started. This is proved by the fact that in the last few decades D. Pharm. holders

are not employed by the industry and B. Pharm. holders are not in many numbers in hospitals or medical

shops. In the West, pharmacy education is patient-oriented and is responsible for Healthcare Management,

while in India pharmacy education is industry-oriented.Nearly 55 per cent of the jobs are available in the

industry sector while 30per cent in education.

There are only three per cent jobs in healthcare. There must be revolutionary changes in the healthcare

system e.g. making laws for appointing pharmacists at each Primary Health Centre and government hospitals.

There should be adequate staff in the state drugs control departments for better control of drug distribution

system. It is crystal clear that separation and improvement of clinical and industrial subjects in the pharmacy

syllabus is a compulsion of the time. But it is yet to be completed, that is why there is such a situation and a

lot of infighting among government authorities. Present. Pharm. syllabus can be divided into 2 major courses

like B. Pharm.(Clinical) and B. Pharm. (Industrial) as it has been already decided to abolish D. Pharmacy

course. Such an arrangement will increase the confidence and competitive skills of pharmacy graduates

among health care team and technocrats and some sort of specialization during under graduation itself.

If two B. Pharm.courses are created as above, needless to say clinical course can be controlled by PCI

and industrial course by AICTE. Private college managements can opt for





any one of the courses. If any college wants to run both the courses theyshould accept both masters, there is

no other go. Existing D. Pharm. colleges who are in the verge of closure can adopt B. Pharm. (Clinical) and

continue to serve the profession. This stunted growth of professional pharmacy in our country is the result of

misplaced belief that profession is same as vocation. This belief has kept Indian pharmacy academics

completely focused on industrial pharmacy at the cost of real - communitypharmacy.

While the justification for focusing pharmacy education on Industrial

Pharmacy after attaining national freedom was valid, its review to make it relevant in contemporary scenario

is already too late. Our present system has produced half a million''qualified'' pharmacists but not many

''trained'' professionals. This has effectively led to a situation where neither there is a need felt by the society

nor is there anyone available to fulfill that "professed" need. This situation feeds on itself to such an extent

that any attempt to keep one's knowledge updated and work professionally has strong economic disincentives

in Indian retail pharmacy practice. Gravity of the situation dawns upon us when we think about petitions filed

in High courts that propose scrapping of the Pharmacy Act because the pharmacists - according to petitioners -

do not play any role other than selling the drugs like all other commodities. There is virtually a complete lack

of any training or incentive to professionalize – as a result of which even the most enthusiastic pharmacists

gradually convert into mere traders. The uninspiring implementation of statutory provisions has led to

a cancerous proliferation of retail drug shops and the situation now threatens the profession itself. The retail

pharmacist shall be relevant to the society ‘only'' if he can make a difference to the patient - by providing him

information about drug usage to achieve better outcome than the patient obtains by uninformed usage of

drugs.

CONCLUSION

Each pharmacy institute should operate a model pharmacy; this would not only improve the image of

pharmacists in Indian society but provide an opportunity for pharmacy students to train in community

practice. The minimum wages established by state governments for pharmacists working in drugstores should

be properly implemented and periodically revised. Even though medicines are now dispensed in the

manufacturer’s original pack wherever possible, additional labeling should include generic name and strength,

dose and frequency, date of dispensing, name of patient, name and address of dispenser and pharmacy, and

date after which the product in not to be used. Finally, to improve patient compliance, oral or written

instructions should be provided by the pharmacist. Although raising the minimum qualification of registered

pharmacists to the B.Pharm, degree is desirable, the economics of employing pharmacists in drug stores,

particularly in remote rural areas, need to be considered. Even if standards for good pharmacy practice are set

in India, it will take years to meet them fully, until then, pharmacists in hospital and community setting need

to take steps on their own to improve their image and protect the health of patients and the public.

REFERENCE

Good pharmacy practice in community and hospital pharmacy settings, WHO/FARM/ DAPS, 1996, 5-10.

Mohanta GP, Manna PK, Valliappan K, Manavalan R, Pharmacy education in India ,Am J Health-Syst

Pharm, 2001, 809-810.

Seth PD, A responsible pharmacist of the 21st century in India through education and service, Pharma

Times, 1999, 31 (Jan), 30.

The role of pharmacists in the healthcare system, WHO/FARM, 1994, 30-60.



Phani Deepthi Yadav et.al Indian Journal of Research in Pharmacy and Biotechnology


PHYTOCHEMICAL EVALUATION OF NYCTANTHES ARBORTRISTIS,

NERIUM OLEANDER AND CATHARATHNUS ROSEUS CH.S.D. Phani Deepthi Yadav*, N.S.P. Bharadwaj, M. Yedukondalu, CH. Methushala, A. Ravi Kumar

*PG & Research Department of Pharmacognosy, Bapatla College of Pharmacy,

Bapatla-522 101, Andhra Pradesh, India

*Corresponding Author: [email protected]

ABSTRACT

In the present study, an attempt was made to investigate phytochemical evaluation of Nyctanthes

arbortristis, Nerium oleander and Catharathus roseus. The crude drug powder extracts of the leaves of

the above plants were taken for the study. The Phytochemical Screening was done for the selected plants.

Phenolic compounds, tannins, flavonoids, cardiac glycosides, and alkaloids were present in Nyctanthes

arbortristis. Alkaloids, flavanoids, carbohydrates, glycosides and tannins were present in Nerium

oleander. Alkaloids, saponins, flavanoids, carbohydrates and anthraquinone glycosides were present in

Catharanthus roseus.

Key words: Phytochemical screening, Nyctanthes, Nerium and Catharanthus, Plant species.

INTRODUCTION

Herbal medicine also known as botanical medicine or phytomedicine-refers to using plants seeds, flowers,

roots for medicinal purpose. Herbalism has a long tradition of use of outside of conventional medicine. It is

becoming more main stream as improvements in analysis and quality control along with advances in clinical

research show the value of herbal medicine in the treating and preventing disease. The medicinal action of plants

is unique to a particular plant species, consistent with the concept that the combination of secondary metabolites

in a particular plant is taxonomically distinct for three medicinal plants and their description and uses

respectively. Nyctanthes arbor-tristis is commonly known as Night-flowering Jasmine, Coral Jasmine and Parijat.

It is used for its antibacterial, anthelmintic, anti-inflammatory, hepatoprotective, immunopotential, anti pyretic,

antioxidant and anti fungal activity Nerium oleander is an evergreen shrub or small tree in

the dogbane family Apocynaceae, toxic in all its parts. Used traditionally in treating dermatitis, abscesses,

eczema, psoriasis, sores, warts, corns, ringworm, scabies, herpes, skin cancer, asthma, dysmenorrheal, epilepsy,

malaria, abortifacients, emetics, heart tonics, and tumor. Catharanthus roseus commonly called Madagascar

periwinkle is an evergreen shrub or herbaceous plant which exhibits the anti cancer activity due to the presence of

vincristine and vinblastine. Here in the present study the above three plants were taken for Phytochemical

screening of plants extracts crude dried powdered drug were taken and evaluated. The phytochemical constituents

were studied by qualitative analysis for performing various chemical tests.


Plant Materials: The leaves of plants Nerium, Nyctanthes and Catharanthus species were authentified by Prof.

V.Satyanarayana Department of Plant Breeding, Bapatla Agricultural College, Bapatla, Andhra Pradesh, India

.They were collected from different areas of Guntur, Prakasham and Krishna districts of Andhra Pradesh, India .

Solvent Extraction: The leaves of Nerium oleander, Catharanthus roseus and Nyctanthes arbortristis were

collected, washed, dried and powdered separately. 50g of dried powder of the leaves was weighed and transferred

into a conical flask and it was macerated with sufficient amount of ethanol for about a week days. Process is

repeated with water. The whole mixture was filtered and filtrate was collected, concentrated in a china dish on a

hot plate till the residue was obtained. The extracts was collected, labeled and stored for further experimental use.

Qualitative analysis for detection of Carbohydrates Alkaloids Cardiac Anthraquinone Saponin Glycosides

Flavonoids Tannins: The extracts and crude dried powders of Nyctanthes arbortristis, Nerium oleander and

Catharanthus roseus were subjected to qualitative analysis for presence of chemical constituents of Nyctanthes

arbortristis, Nerium oleander and Catharanthus roseus by performing various chemical tests.





TEST FOR CARBOHYDRATES

TEST FOR ALKALOIDS To 250 mg of each extracts, 10 ml of dilute HCl was added, mixed and filtered. To the filtrate the following

reagents were added and tested.

TEST PROCEDURE

WAGNER’S TEST

2 ml of Wagner’s reagent was added to the above filtrate solution and

observed.

HAGER’S TEST To the 2 ml of above filtrate solution, 2 ml of picric acid was added and

observed.

TEST FOR GLYCOSIDES

The extract was tested for the presence of saponin glycosides, cardiac glycosides and anthraquinone glycosides

TEST FOR SAPONIN GLYCOSIDES

TEST PROCEDURE

FOAM TEST To 200 mg of each extracts, 15 ml of distilled

water was added, shake it well and observed.

TEST FOR CARDIAC GLYCOSIDES

TEST PROCEDURE

LEGAL’S TEST

To 50 mg of each extracts, 1 ml of pyridine, 1 ml of

Sodium nitro prusside solution were added and observed.

KELLER-KILIANI TEST To 50 mg of each extracts, 2 ml of glacial acetic acid, 1

ml FeCl3 solution were added, heated and then cooled.

This was transferred to a test tube containing 2ml conc.

H2SO4and observed.

TEST FOR ANTHRAQUINONE GLYCOSIDES

TEST PROCEDURE

BORNTRAGER’S TEST To 200 mg of each extracts, dil. H2SO4was added and

boiled. Then it was filtered and cooled. To the cold

filtrate, 3 ml of benzene was added and mixed. The

benzene layer was separated and to it, ammonia (2 ml)

was added and ammonical layer was observed.

TEST PROCEDURE

MOLISCH’S TEST

200 mg of extracts were dissolved separately in 5ml of water and filtered. 2

ml of the above sample solution is placed in a test tube. Two drops of the

Molisch reagent is added. The solution is then poured slowly into a tube

containing 2 ml of concentrated sulphuric acid and observed.

FEHLING’S TEST

1ml of Fehling’s solution A and 1ml of Fehling’s solution B were added to

100mg of extracts separately. They were heated on a boiling water bath for 5

min and observed.

BENEDICT’S TEST To the 150 mg of each extracts, 2ml of Barfoed’s reagent was added. Then

the mixture was heated on a boiling water bath for 5 min, cooled and

observed.





TEST FOR FLAVANOIDS

TEST PROCEDURE

LEAD ACETATE TEST To the 100 mg of each extracts, lead acetate (5 ml) was

added and observed.

TEST FOR TANNINS

To 100 mg of each extracts, the following reagents were added and observed.

1. 5 ml of 5% w/v FeCl3 solution.

2. 5 ml acetic acid solution.

3. 5 ml dil. KMnO4 solution.

TEST FOR STEROIDS

TEST PROCEDURE

SALKOWSKI TEST To 100 mg of each extracts, 2 ml of CHCl3, 2 ml of

conc. H2SO4were added, mixed thoroughly and both the

layers were observed for color.

LIBERMAN AND BURCHARD TEST To 200 mg of each extracts, 5ml CHCl3, 5 ml acetic

anhydride were added. Two drops of H2SO4 was added

from the sides of test tube and observed.

PHYTOCHEMICAL EVALUATION OF NYCTANTHES ARBORTRISTIS

Table 1. Phytochemical evaluation of Nyctanthes arbortristis

Chemical tests Result Chemical tests Result

Test for carbohydrates A. Molisch's test

B. Fehling's test

C. Benedict's test

D. Barfoed's test

Positive

Positive

Positive

Positive

Test for alkaloids A. Hager's test

B. Wagner's test

Positive

Positive Test for flavanoids Lead acetate test

Positive Test for saponins A. Foam test

Negative Test for cardiac glycosides A. Legal test

B. Keller-killiani test

positive

positive

Test for steroids A. Lieberman burchard test

B. Salkowski test

Negative

Negative

Test for anthraquinone

glycosides A. Borntrager's test

Negative

PHYTOCHEMICAL EVALUATION OF NERIUM OLEANDER

Table 2. Phytochemical evaluation of Nerium oleander



B. Fehling's test

C. Benedict's test

Positive

Positive

Positive


B. Salkowski test

Positive

positive


B. Wagner's test

Positive Test for cardiac glycosides A. Legal test


Positive

Positive

Test for flavanoids Lead acetate test

Positive Test for anthraquinone glycosides

Borntragers test

Negative

Test for saponins Foam test

Negative

Test for tannins A. Fecl3test

B. Acetic acid test

C. Kmn04 test

C. Kmn04 test

Positive

Positive

Positive





PHYTOCHEMCIAL EVALUATION OF CATHARANTHUS ROSEUS

Table. 3. Phytochemcial evaluation of Catharanthus roseus



B. Fehling's test

C. Benedict's test

D. Barfoed's test

Positive

Positive

Positive

Positive

Test for saponins A. Foam test

Positive


B. Salkowski test

Negative

Negative


B. Wagner's test

Positive Test for cardiac glycosides A. Legal test


Negative

Negative

Test for flavanoids Lead acetate test

Positive Test for anthraquinone

glycosides A. Borntrager's test

Positive


The study of the chemical constituents and the active principles of the medicinal plants have acquired a

lot of importance all over the world. The present study includes the phytochemical screening of the plants

Nyctanthes arbortristis, Nerium oleander and Catharanthus roseus. The plants were collected and were

authentified. Then they were shade dried and powdered and were subjected to phytochemical screening. The

dried powdered leaves of Nyctanthes arbortistis, Nerium oleander and Catharanthus roseus were subjected to

extraction with ethanol separately. The qualitative chemical tests for the ethanolic extracts were performed.

The investigation showed that Nyctanthes arbortristis contains carbohydrates, alkaloids, flavanoids and cardiac

glycosides. The screening showed that Nerium oleander possesses carbohydrates, flavanoids, alkaloids, steroids,

cardiac glycosides and tannins. The screening showed that Catharanthus roseus possesses carbohydrates,

flavanoids, saponins, and alkaloids. The results were given in Table 1 and Table 2 and Table 3 respectively.

CONCLUSION

The screening of phytochemical constituents of plants Nyctanthes arbortristis, Nerium oleander and

Catharanthus roseus indicated the presence of carbohydrates, flavonoids and alkaloids in common. Nerium

oleander does not contain saponins and Catharanthus roseus lacks tannins and cardiac glycosides and Nyctanthes

arbortristis do not have steroids, saponins and tannins. These plants contain more metabolites and there is a need

for further investigations using fractionated extracts and purified chemical components.

AKNOWLEDGEMENTS

The authors are thankful to Management and Principal of Post Graduate Research Centre Division, Bapatla

College of Pharmacy, Bapatla, Andhra Pradesh, India in permitting and providing necessary facilities for carrying

out to do the project work.

Nyctanthes arbortristis (Aerial Parts of thePlant) Nerium oleander(Aerial Parts of the Plant)





Catharanthus roseus (Aerial Parts of the Plant)

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Shravan Paswan et.al Indian Journal of Research in Pharmacy and Biotechnology


ANGINA PECTORIS EPIDEMIC IN INDIA: A COMPREHENSIVE REVIEW OF

CLINICAL FEATURES, DIFFERENTIAL DIAGNOSIS, AND REMEDIES Shravan Paswan

1*, Ranjan Kumar Sharma

1, Alok Ranjan Gaur

2, Avinash Sachan

1,

Mahendra Singh Yadav1, Preeti Sharma

3, Mrinmoy Gautam

4

1. Advance Institute of Pharmaceutical Education & Research, Kanpur, India

2. Department of Pharmacy, Pranveer Singh Institute of Technology Kanpur, India

3. Department of Pharmaceutical Science, Banasthali University, Rajasthan, India

4. Royal College of Pharmacy and Health Sciences, Berhampur, Odisha, India

*Corresponding author: E-mail: [email protected]

ABSTRACT

The community pharmacist is the globally accepted professional to cater the pharmaceutical care to

the patients at the time of dispensing the medicine it self. This will correct the wrong practices in

medicine usage and make the patient a partner in his improvement of health. Angina pectoris is the

medical term for chest pain or discomfort due to coronary heart disease. Angina is a symptom of a

condition called myocardial ischemia. It occurs when the heart muscle (myocardium) doesn't get as much

blood (hence as much oxygen) as it needs. This usually happens because one or more of the heart's

arteries (coronary blood vessels that supply blood to the heart muscle) is narrowed or

blocked. Insufficient blood supply is called ischemia. Major risk factors for angina include cigarette

smoking, diabetes, high cholesterol, high blood pressure, sedentary lifestyle and family history of

premature heart disease. The distal part of the heart does not receive any more blood and individuals will

develop chest pain. Initially when the coronary disease is mild the angina will occur during exercise,

eating heavy meals, extreme heat or stress. As the coronary disease worsens, the angina will come on

with minimal work and may even occur at rest. The first approach in the treatment of angina pectoris is

to prevent the progression of heart disease. By addressing the known causes of heart disease, such as

reducing high cholesterol levels, controlling high blood pressure, stopping smoking, losing weight,

exercising and eating a “heart-healthy” diet, the symptoms can be reduced. Most people can live a

productive life if they make the necessary lifestyle changes. By following medical advice, taking doctor-

prescribed medication, maintaining a good physical condition and eating well, angina can be

controlled. There are also natural alternatives to conventional medicine such as herbal and homeopathic

remedies useful in controlling angina without the harsh side effects associated with prescription drugs.

Herbal and homeopathic remedies are safe and gentle to use, while at the same time addressing the

underlying causes of the condition.

Key Words: Community pharmacist, Angina pectoris, Coronary Heart Disease, Ischemia, Myocardium.

INTRODUCTION Angina pectoris: Angina pectoris is a medical condition that occurs when the heart receives a decreased amount

of oxygenated blood. Often, this occurs due to deposits of cholesterol, clogging the blood vessels that carry blood

to the heart. Patients who have angina pectoris are at an risk for having a heart attack, chest pain behind the

breastbone is the most common sign of angina pectoris. The discomfort may feel like pressure, squeezing, burning

or tightness, reports the National Heart Lung and Blood Institute. The chest pain is most common during exercise,

physical work or sexual activity. Emotional stress, cold weather and nightmares may also trigger an attack of

chest pain, explains Cedars-Sinai Medical Center.

Patients with severe angina pectoris may also develop the pain during rest without the presence of any

stress. An episode of chest pain caused by angina pectoris may last from 5 to 30 minutes. Angina is a common

presenting symptom (typically, chest pain) among patients with coronary artery disease. Statistics reveal that

close to 7 million American suffer from angina and countless more do not even know if they have it. Each year

close to half a million new cases are diagnosed. The common condition affects nearly ¼ individuals over the age

of 55. Each year, there are more than 1 million cases of recurrent acute angina, with a morality rate of close to

40%. In addition, there are a significant number of individual who die suddenly and have no symptoms. No race

or ethnic groups are immune from coronary artery disease.

Angina pectoris is far more common in women than men. In addition, angina in women can present in an

atypical fashion. The pain may not be in the chest area and the pain may have a different quality. To improve the


http://en.wikipedia.org/wiki/Tobacco_smoking

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http://en.wikipedia.org/wiki/Hypercholesterolemia

http://en.wikipedia.org/wiki/Hypertension

http://en.wikipedia.org/wiki/Sedentary_lifestyle

http://en.wikipedia.org/wiki/Family_history_(medicine)

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prognosis, it is essential that one change the lifestyle. One should adhere to these changes, otherwise angina will

recur. The best preventive measures one can take include - start an exercise program, stop smoking, decrease

alcohol consumption, avoid stressful situation, avoid heavy and fatty meals. The best treatment of angina is drug

therapy. The most common group of drugs used to treat angina is nitroglycerines. Nitroglycerin is available in

various formulations. It can be taken by mouth, placed under the tongue and can even be given intravenously.

Nitroglycerin has the ability to open up (dilate) blood vessel and allows more blood flow to the heart.

Nitroglycerin is usually taken when one feels the chest pain occuring. A tablet is placed underneath the tongue

and within a few minutes the pain will disappear. In some cases, two tablets may be required. Nitroglycerin

formulations are also available as an oral pill which must be taken 2-3 times a day. Beta blockers have been used

to treat coronary disease for decades. They act by decreasing the work of the heart and thus decrease oxygen

utilization. Unfortunately they work in the long term and do not work immediately, like nitroglycerin. Beta

blockers have to be taken every day and have a few side effects like a decrease in libido. Calcium channel

blockers are very effective in the treatment of angina, but like the beta blockers they have to be taken daily.

Causes of angina pectoris: In most cases, the cause of angina is coronary atherosclerosis: the thickening of

arteries that supply blood, oxygen and nutrients to the heart. This happens when fatty deposits, called plaques or

atheroma, narrow the arteries over time and reduce blood flow to the heart. Symptoms may only appear at times

when the heart needs more blood supply, such as in stressed condition, exercising or climbing stairs. As the heart

tries to pump blood faster to meet the body's increased demands, the narrowed arteries struggle to keep up. The

heart then receives less oxygen, which causes pain in the heart that is felt as chest pain.In severe cases this can

also happen when the heart is at rest.

Types of angina pectoris: Angina is classified as one of the following two types:

1. Stable angina

2. Unstable angina

1. Stable Angina: Stable angina is the most common angina, and the type most people mean when they refer to

angina. People with stable angina usually have angina symptoms on a regular basis.The episodes occur in a

pattern and are predictable. For most people, angina symptoms occur after short bursts of exertion. Stable angina

symptoms usually last less than five minutes. They are usually relieved by rest or medication, such as

nitroglycerin under the tongue.

2. Unstable Angina: Unstable angina is less common. Angina symptoms are unpredictable and often occur at

rest.This may indicate a worsening of stable angina, but sometimes the first time a person has angina it is already

unstable. The symptoms are worse in unstable angina - the pains are more frequent, more severe, last longer,

occur at rest, and are not relieved by nitroglycerin under the tongue. Unstable angina is not the same as a heart

attack, but it warrants an immediate visit to the healthcare provider or a hospital emergency department. The

patient may need to be hospitalized to prevent a heart attack. If the patient has stable angina, any of the following

may indicate worsening of the condition. An angina episode that is different from the regular pattern being

awakened at night by angina symptoms.

Coronary Heart Disease: The most common cause for the heart not getting enough blood is coronary heart

disease, also called coronary artery disease. In this disease, the coronary arteries become blocked, narrowed, or

otherwise damaged. They can no longer supply the heart with all of the blood it needs. Most cases of coronary

heart disease are caused by atherosclerosis (hardening of the arteries). Atherosclerosis is a condition in which a

fatty substance/cholesterol builds up inside the blood vessels. These buildups are called plaques, and they can

block blood flow through the vessels partially or completely. Multiple risk factors, particularly: diabetes, high

blood pressure, smoking, high cholesterol, and genetic predisposition may accelerate this build up.

Coronary Artery Spasm: Another cause of unstable angina is coronary artery spasm. Spasm of the muscles

surrounding the coronary arteries causes them to narrow or close off temporarily. This blocks the flow of blood to

the heart muscle for a brief time, causing angina symptoms. This is called variant angina or Prinzmetal angina.

This is not the same as atherosclerosis, although some people have both conditions. The symptoms often come on

at rest (or during sleep) and without apparent cause. Cocaine use/abuse can cause significant spasm of the

coronary arteries and lead to a heart attack.

Risk factors: All of us have fatty deposits in our arteries to some degree. Atherosclerosis can start as early as our

20s and increases with age. But there are risk factors that are known to increase the development of fatty deposits

that can cause the arteries to narrow.

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A family history of atherosclerosis

High levels of LDL cholesterol in the blood

High blood pressure

Smoking

Being male

Diabetes

Obesity

Stress

Lack of regular exercise

Symptoms of angina pectoris: Symptoms typically start during physical exertion or emotional stress. They are

often worse in cold or windy weather and sometimes after big meals.

A squeezing or heavy pressing sensation on the chest.

Increased shortness of breath on exercise.

A sense of heaviness or numbness in the arm, shoulder, elbow or hand, usually on the left side.

A constricting sensation in the throat.

The discomfort can radiate into arms, the jaw, teeth, ears, stomach and in rare cases between the shoulder

blades.

Unstable angina is associated with the same symptoms at rest. In some cases the fatty deposits that restrict

blood flow can rupture. Blood then clots around the rupture, and the clot may be large enough to block the artery

and seal off the blood supply. This may cause unstable angina or a heart attack.

Pathophysiology: Myocardial ischemia develops when coronary blood flow becomes inadequate to meet

myocardial oxygen demand. This causes myocardial cells to switch from aerobic to anaerobic metabolism, with a

progressive impairment of metabolic, mechanical, and electrical functions. Angina pectoris is the most common

clinical manifestation of myocardial ischemia. It is caused by chemical and mechanical stimulation of sensory

afferent nerve endings in the coronary vessels and myocardium. These nerve fibers extend from the first to fourth

thoracic spinal nerves, ascending via the spinal cord to the thalamus, and from there to the cerebral cortex.

Studies have shown that adenosine may be the main chemical mediator of anginal pain. During ischemia,

ATP is degraded to adenosine, which after diffusion to the extracellular space, causes arteriolar dilation and

anginal pain. Adenosine induces angina mainly by stimulating the A1 receptors in cardiac afferent nerve endings.

Heart rate, myocardial inotropic state, and myocardial wall tension are the major determinants of myocardial

metabolic activity and myocardial oxygen demand increases in the heart and myocardial contractile state result in

increased myocardial oxygen demand. Increases in both after load (ie, aortic pressure) and preload (ie, ventricular

end-diastolic volume) result in a proportional elevation of myocardial wall tension and, therefore, increased

myocardial oxygen demand. Oxygen supply to any organ system is determined by blood flow and oxygen

extraction. Because the resting coronary venous oxygen saturation is already at a relatively low level

(approximately 30%), the myocardium has a limited ability to increase its oxygen extraction during episodes of

increased demand. Thus, an increase in myocardial oxygen demand (eg, during exercise) must be met by a

proportional increase in coronary blood flow. The ability of the coronary arteries to increase blood flow in

response to increased cardiac metabolic demand is referred to as coronary flow reserve (CFR). In healthy people,

the maximal coronary blood flow after full dilation of the coronary arteries is roughly 4-6 times the resting

coronary blood flow.

CFR depends on at least 3 factors: large and small coronary artery resistance, extravascular (ie,

myocardial and interstitial) resistance, and blood composition.

Myocardial ischemia can result from (1) a reduction of coronary blood flow caused by fixed and/or

dynamic epicardial coronary artery (ie, conductive vessel) stenosis, (2) abnormal constriction or deficient

relaxation of coronary microcirculation (ie, resistance vessels), or (3) reduced oxygen-carrying capacity of the

blood.

Atherosclerosis is the most common cause of epicardial coronary artery stenosis and, hence, angina

pectoris. Patients with a fixed coronary atheroscerotic lesion of at least 50% show myocardial ischemia during

increased myocardial metabolic demand as the result of a significant reduction in CFR. These patients are not able

to increase their coronary blood flow during stress to match the increased myocardial metabolic demand, thus

they experience angina. Fixed atherosclerotic lesions of at least 90% almost completely abolish the flow reserve;

http://www.netdoctor.co.uk/diseases/facts/hypercholesterolaemia.htm

http://www.netdoctor.co.uk/diseases/facts/hypertension.htm

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http://www.netdoctor.co.uk/hearthealth/diabetes.htm

http://www.netdoctor.co.uk/hearthealth/obesity.htm

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http://www.netdoctor.co.uk/diseases/facts/coronarythrombosis.htm

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patients with these lesions may experience angina at rest.Coronary spasm can also reduce CFR significantly by

causing dynamic stenosis of coronary arteries.

Prinzmetal angina is defined as resting angina associated with ST-segment elevation caused by focal

coronary artery spasm. Although most patients with Prinzmetal angina have underlying fixed coronary lesions,

some have angiographically normal coronary arteries. Several mechanisms have been proposed for Prinzmetal

angina: focal deficiency of nitric oxide production, hyperinsulinemia, low intracellular magnesium levels,

smoking cigarettes, and using cocaine. Approximately 30% of patients with chest pain referred for cardiac

catheterization have normal or minimal atherosclerosis of coronary arteries. A subset of these patients

demonstrates reduced CFR that is believed to be caused by functional and structural alterations of small coronary

arteries and arterioles (ie, resistance vessels). Under normal conditions, resistance vessels are responsible for as

much as 95% of coronary artery resistance, with the remaining 5% being from epicardial coronary arteries (ie,

conductive vessels).

The former is not visualized during regular coronary catheterization. Angina due to dysfunction of small

coronary arteries and arterioles is called microvascular angina. Several diseases, such as diabetes mellitus,

hypertension, and systemic collagen vascular diseases (eg, systemic lupus erythematosus, polyarteritis nodosa),

are believed to cause microvascular abnormalities with subsequent reduction in CFR. The syndrome that includes

angina pectoris, ischemia like ST-segment changes and/or myocardial perfusion defects during stress testing, and

angiographically normal coronary arteries is referred to as syndrome X. Most patients with this syndrome are

postmenopausal women, and they usually have an excellent prognosis Syndrome X is believed to be caused by

microvascular angina. Multiple mechanisms may be responsible for this syndrome, including; (1) impaired

endothelial dysfunction (2) increased release of local vasoconstrictors, (3) fibrosis and medial hypertrophy of the

microcirculation, (4) abnormal cardiac adrenergic nerve function, and/or (5) estrogen deficiency.

A number of extravascular forces produced by contraction of adjacent myocardium and intraventricular

pressure can influence coronary microcirculation resistance and thus reduce CFR. Extravascular compressive

forces are highest in the subendocardium and decrease toward the subepicardium. Left ventricular (LV)

hypertrophy together with a higher myocardial oxygen demand (eg, during tachycardia) cause greater

susceptibility to ischemia in subendocardial layers. Myocardial ischemia can also be the result of factors affecting

blood composition, such as reduced oxygen-carrying capacity of blood, as is observed with severe anemia

(hemoglobin, <8 g/dL), or elevated levels of carboxyhemoglobin. The latter may be the result of inhalation of

carbon monoxide in a closed area or of long-term smoking. Ambulatory ECG monitoring has shown that silent

ischemia is a common phenomenon among patients with established coronary artery disease. In one study, as

many as 75% of episodes of ischemia (defined as transient ST depression of >1 mm persisting for at least 1 min)

occurring in patients with stable angina were clinically silent. Silent ischemia occurs most frequently in early

morning hours and may result in transient myocardial contractile dysfunction (ie, stunning). The exact

mechanism(s) for silent ischemia is not known. However, autonomic dysfunction (especially in patients with

diabetes), a higher pain threshold in some individuals, and the production of excessive quantities of endorphins

are among the more popular hypotheses.

Exams and tests: Upon hearing about the patient's symptoms, the primary healthcare provider or the provider in

the emergency department will immediately think of angina and other heart problems. Time is of the essence-

treatment will probably begin as the evaluation continues. An electrocardiogram (ECG) will be done. This

painless test checks for abnormalities in the beating of the heart. Electrodes are attached to the chest and other

points on the body. The electrodes read the electrical impulses linked to the beating of the heart. The ECG looks

for signs of a heart attack or of impaired blood flow to the heart. For many patients with angina, the ECG result is

normal. The patient may have a chest x-ray. This will show any fluid buildup in the lungs. It can also rule out

some other causes of chest pain. There is no blood lab test that can tell with certainty that someone is having

angina. There are certain blood tests that suggest that a person may be having a heart attack. These tests may be

done if a heart attack is suspected.While these tests are going on, the healthcare provider will be asking questions

to help with the diagnosis.The questions will be about the symptoms and about the patient's medical history,

previous operations, medications, allergies, and habits and lifestyle. The physical exam will include listening to

the heart and lungs and feeling the heart through the chest. If, after these tests, the healthcare provider suspects the

patient may have coronary heart disease, additional tests will be performed to confirm the possibility.







Exercise stress test: An ECG is taken before, during, and after exercise (usually walking on a treadmill) to detect

inadequate blood flow to the heart muscle indirectly by changes on the ECG. This usually is done only for stable

angina.

Thallium stress test: This is a more complex and expensive test that injects a radioisotope into the circulation

and indirectly detects parts of the heart that may not be getting enough blood during "stress" (usually walking on a

treadmill, or after administration of a drug that mimics exercise in those unable to walk on the treadmill). This

information indicates more accurately whether any of the coronary arteries may be narrowed, causing inadequate

blood flow to the heart muscle or ventricle. Again, this is usually done only for stable angina.

Dobutamine echocardiogram stress test: This is done for people who cannot walk on a treadmill. A drug called

dobutamine stimulates and speeds up the heart, creating an increased demand or need for blood flow to the left

ventricle or muscle. If the muscle shows a slowing of function on the ultrasound image of the heart muscle, then it

indirectly indicates inadequate blood flow to the muscle.

Coronary angiogram (or arteriogram): This test of the coronary arteries is the most accurate but also the most

invasive. It is a type of x-ray. A thin plastic tube called a catheter is threaded through an artery in the arm or groin

to one of the main coronary arteries. A contrast or harmless dye is injected into the arteries. The dye depicts the

arteries directly and shows any blockage more accurately than the above or more noninvasive procedures.

The healthcare provider will make the decision about whether these tests or any treatment need to be done on an

urgent basis. If so, the patient will be admitted to the hospital. If not, the tests will be scheduled for the next few

days, and the patient may be allowed to go home

TREATMENT OF ANGINA PECTORIES

The patient may need to take several medicines to control symptoms and improve angina.

Aspirin in low doses reduces the tendency of small blood cells called platelets to stick together, which

helps prevent the formation of a blood clots.

Glyceryl trinitrate relaxes the arteries of the heart and relieves angina attacks. GTN comes in sublingual

tablet or spray form.

Long-acting nitrates reduce the frequency of angina attacks. These can be in the form of tablets or patches

and are very effective. Their main side-effect is headache, but this often disappears once the nitrate has

been taken for some weeks.

Beta-blockers block the effect of the hormone adrenaline so that the pulse is slowed and blood pressure

lowered. This reduces the heart's need for oxygen and improves the supply of blood to the heart muscle.

They are also important in protecting the heart after heart attacks.

Calcium-channel blockers reduce the muscle tension in the coronary arteries, expanding them and

creating more room. They also slightly relax the heart muscle, reducing the heart's need for oxygen and

reducing blood pressure.

The potassium-channel activator nicorandil (Ikorel) reduces muscle tension in the blood vessel walls,

expanding them and improving the flow of blood and the supply of oxygen.

Surgery: If the patient have severe angina that is not responding to medication, a cardiologist may decide to need

surgery to restore heart function to an adequate level and reduce the likelihood of a heart attack. This can be done

by one of the following operations.

Angioplasty: the narrowed coronary artery is dilated (opened up) with a balloon. A small tube called a

stent may also be inserted into the artery at this time to help prevent it narrowing down again in the future.

Bypass operation: a superficial blood vessel is taken from another part of the body, usually the leg, and

joined to the coronary artery to bypass the obstruction to blood flow.

Transmyocardial revascularization: Transmyocardial revascularization is a procedure for patients who cannot

undergo angioplasty or surgery. A simple incision is made in the chest, and a laser is used to "drill" small holes

through the outside wall of the heart into the left ventricle. About 20-40 holes are made. Bleeding from these

holes is minimal and usually stops after a few minutes of pressure. It is not clear why this helps relieve angina.

One theory is that it stimulates growth of new blood vessels that improve blood flow to the heart. Other

investigators believe it is a placebo effect. Current research is focusing on trying to find growth factors that could

be injected into coronary arteries or directly into the left ventricle to encourage growth of new blood vessels.

Diet plan for preventing angina pectoris: Obviously, a healthy diet goes a long way in preventing angina

pectoris. The following basic guidelines need to be followed with meticulous care.


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Avoid fatty foods to the maximum possible extent. This includes fried food, milk products such as butter

and cheese, full cream milk, oils, etc. Fermented milk products are good for people with angina. This includes

curds. Use only vegetable oils for cooking. This includes sunflower, olive, groundnut and rapeseed oils. Avoid

salt in the diet. Do not consume foods that are too much salty. In meats, red meats such as mutton, beef and pork

must be avoided. White meats such as poultry and fish are beneficial. Fishes with high body oil content must be

preferred. This includes sardines, tunas, mackerels, salmons, herrings, etc. Canned fish must be strictly avoided.

There should be at least two to three fish consumptions per week. Carbohydrates should form the major part of

the food. This includes cereals, wheat, rice, bread, potatoes and pasta. It is found that a little bit of alcohol is

actually beneficial for angina, but excess is harmful. The safe limit of alcohol is as follows:- For men: 21 units per

week, and not more than 4 units on any one day .For women: 14 units per week, and not more than 3 units on any

one day.

Prevention of angina pectoris: Many of the risk factors for angina can be tackled by lifestyle changes.

Eat a varied and healthy diet with plenty of leafy vegetables. Avoid sugary foods and saturated fats found

in meat and full-fat dairy products.

Stop smoking.

Lose weight if you are overweight.

Exercise more: aim for a half-hour walk each day.

If you have diabetes or high blood pressure, maintain treatment for these conditions.

Ayurvedic herbs: Ayurveda is a treasure-house of remedies for angina pectoris. There is a long list of herbs that

have been used since ancient times for the treatment of the condition. The following is a list of these herbs with

their actions on the human body.

Table.1. Ayurvedic herbs useful in the treatment of heart diseases Ayurvedic

Name of the

Herb

Biological

Name of the

Herb

Common

English Name

of the Herb

Action on the Human Body

Alfalfa Medicago

satina

Alfalfa Juice of the alfalfa grass is used for people with arterial and heart

problems. The benefits of this juice are improved by taking it in a

mixture with carrot juice.

Amla Emblica

officinalis

Indian

Gooseberry

Amla tones up all the organs of the body, and that includes the

heart. Thus it betters the pumping action of the heart.

Chachinga Trichosanthes

anguina

Snake Gourd The leaves of the snake gourd have been traditionally used as

medicine for treating pain in the heart due to physical exertion.

Haldi Curcuma longa Turmeric Curcumin is a chemical compound present in turmeric. This

compound is known to lower the amount of serum cholesterol and

even the blood sugar level.

Kahu Terminalia

arjuna

Arjuna Arjuna is perhaps the most beneficial herb used by Ayurvedic

practitioners in the treatment of angina-related problems. The bark

of the arjuna tree is known to have stimulant action on the heart.

Lahsoona Allium sativa Garlic Garlic is beneficial for people with angina pectoris as it is a known

blood thinner. For this reason, garlic must be included in the diet.

Peepal Ficus religiosa Peepal The leaves of the peepal tree are effective in treatment of heart

ailments. They are known to strengthen the heart and thus keep

angina pectoris at bay.

Pyaaza Allium cepa Onion Trials have shown that regular usage of onion for five months is

beneficial in decreasing serum cholesterol. It is also beneficial in

decreasing thrombosis.

Rojmari Achillea

millefolium

Blood Wort The herb of blood wort is beneficial in the treatment of circulatory

problems due to its stimulant action. It can bring down high blood

pressure, which is a leading cause of angina pectoris.

Tilpushpi Digitalis

purpurea

Digitalis Treatment of angina pectoris is one of the most elemental purposes

digitalis is put to. Digitalis stimulates the muscle activity of the

heart and makes it pump better. Thus it forces more blood into the

heart and improves nourishment.

http://www.netdoctor.co.uk/health_advice/facts/smokehealth.htm

http://www.netdoctor.co.uk/health_advice/facts/bodymassindex.htm

http://www.netdoctor.co.uk/hearthealth/diabetes.htm

http://www.netdoctor.co.uk/diseases/facts/hypertension.htm





Guggul is an age-old remedy used by Ayurvedic exponents for treating angina pectoris and its

complications. Guggul is in fact a mixture of several substances that have been extracted from the Commiphora

mukul plant. This medicine is effective in treating atherosclerosis, which is a leading cause of angina. This is

because of guggulsterone, which is a compound found in the guggul plant.

CONCLUSION

Angina Pectoris is a term for chest pain caused by an inadequate supply of blood and oxygen to the heart.

More than 9 million people in the United States have angina. With angina, the affected patient’s heart may get

sufficient blood for daily activities, but the arteries may not be able to respond appropriately to increased demands

for oxygen during exercise, times of emotional or physical stress, and with extremes of temperature. Treatments

for angina depend on the type and severity of the angina. For most people with stable angina, symptoms disappear

after a period of time. Rest and nitroglycerin may be sufficient to treat the attack. For people with unstable angina,

other medications such as antiplatelet medications, beta blockers, calcium channel blockers and blood thinners

may be given. In severe cases of angina, or where the risk of a heart attack is high, surgical procedures such as a

coronary artery bypass grafting and percutaneous transluminal angioplasty may be recommended.

REFERENCES

Droste C, Roskamm H, Experimental pain measurement in patients with asymptomatic myocardial ischemia, J

Am Coll Cardiol, 1, 1983, 340–345.

Falcone C, Sconocchia R, Guasti L, et al. Dental pain threshold and angina pectoris in patients with coronary

artery disease. J Am Coll Cardiol,12, 1998, 348–352.

Gettes LS, Winternitz SR, Monitoring to detect "silent" ischemia. In: Stern S, ed. Ambulatory ECG Monitoring.

Chicago, Ill: Year Book Medical Publishers, Inc; 1978: 93–106.

Kannel WB. Unrecognized myocardial infarction.In: Stern S, ed. Silent Myocardial Ischemia. London, UK:

Martin Dunitz Ltd; 1998; 47–53.

Laukkanen JA, Kurl S, Lakka TA, et al. Exercise-induced silent myocardial ischemia and coronary morbidity and

mortality in middle-aged men, J Am Coll Cardiol, 38, 2001, 72–79.

Mark DB, Hlatky MA, Califf RM, et al. Painless exercise ST deviation on the treadmill: long term prognosis. J

Am Coll Cardiol, 14, 1989, 885–888.

Mazzone A, Cusa C, Mazzucchelli I, et al. Increased production of inflammatory cytokines in patients with silent

myocardial ischemia, J Am Coll Cardiol, 38, 2001, 1895–1901.

Miranda CP, Lehmann KG, Lachterman B, et al. Comparison of silent and symptomatic ischemia during exercise

testing in men. Ann Intern Med, 114, 1991, 645–656.

Pepine CJ, Sharaf B, Andrews TC, et al. Relation between clinical, angiographic and ischemic findings at baseline

and ischemia-related adverse outcomes at 1 year in the Asymptomatic Cardiac Ischemia Pilot study. J Am Coll

Cardiol, 29, 1997, 1483–1489.

Pierdomenico SD, Bucci A, Costantini F, Circadian blood pressure changes and myocardial ischemia in

hypertensive patients with coronary artery disease, J Am Coll Cardiol, 31, 1998, 1627–1634.

Schang SJ, Pepine CJ, Transient asymptomatic ST segment depression during daily activity, Am J Cardiol, 39,

1977, 396–402.

Stern S, Tzivoni D, Early detection of silent ischemic heart disease by 24-hour electrocardiographic monitoring of

active subjects, Br Heart J, 36, 1974, 481–486.



Manikandan K et.al. Indian Journal of Research in Pharmacy and Biotechnology


STABILITY INDICATING HPLC METHOD FOR THE ESTIMATION OF

CINACALCET HYDROCHLORIDE API Manikandan Krishnan*, Santhana Lakshmi Karunanidhi, Gayathri Sola, Akshitha .Y

Department of Pharmaceutical Analysis, SRM College of Pharmacy,

SRM University, Kattankulathur, Tamilnadu 603203, India

*Corresponding author: E.mail: [email protected]

ABSTRACT

A stability-indicating liquid chromatography method has been developed and validated for

the determination of Cinacalcet hydrochloride in a laboratory mixture as well as in a tablet

formulation developed in-house. Efficient chromatographic separation was achieved on phenomenex

C18 column (150 mm×4.6 mm, 5.0 μm) with mobile phase containing Methanol: Water (70:30v/v) pH

adjusted to 3.6 with dilute Orthophosphoric acid at a flow rate of 1.3 mL/min and the eluent was

monitored at 271 nm using Shimadzu LC-10AT-VP & LC-20 AD with Spinotech (Winchrome)

software. Linearity range was found to be between 50-300μg/ml and the linear regression coefficient

was not more than 0.999. The values of % RSD are less than 2% indicating accuracy and precision of

the method. The percentage recovery varies from 97-103%w/w. LOD and LOQ were found to be

within limit. The system suitability parameters like tailing factor, number of theoretical plates,

Asymmetry were found to be within limit. Forced degradation studies were performed by using this

method. There was a significant degradation in the presence of 0.1M HCl, 0.1M NaOH, 30%H2O2,

heat and light. The proposed method is simple, accurate and rapid and hence can be employed for

routine quality control analysis.

Keywords: Cinacalcet Hydrochloride, Stability-indicating, ICH guidelines, HPLC

INTRODUCTION

Cinacalcet Hydrochloride is N-[(1R)-1(Naphthyl) ethyl] - 3 - [3(trifluoromethyl) phenyl] Propan - 1-

amine hydrochloride an oral calcimimetic agent that increases the sensitivity of the calcium-sensing receptor

to activation by extracellular calcium. The calcium sensing receptors on the surface of parathyroid gland

regulate parathyroid hormone (PTH) secretion. Increased PTH stimulates osteoclastic activity resulting in

cortical bone resorption and marrow fibrosis. Increasing the sensitivity of these receptors results in a lowering

of PTH which subsequently lowers serum calcium levels.

Fig 1: Chemical structure of Cinacalcet hydrochloride

EXPERIMENTAL

Instrumentation: Quantitative High Performance Liquid Chromatographic analysis was performed on a

Shimadzu LC-10AT-VP, LC-20 AD pumps and UV Detector with Spinotech (Winchrome) software.

Reagents and Chemicals: Cinacalcet hydrochloride reference standard was obtained as a gift sample from

Actavis Pharma Manufacturing Pvt. Ltd. Chennai, India and has been used as reference drug without prior

treatment. Synthetic mixture of Cinacalcet Hydrochloride was prepared by mixing Reference standard drug,

Magnesium Stearate, Talc, Starch which were procured from local supplier. Analytical / HPLC grade

chemicals and solvents used were obtained from Ranbaxy Fine Chemicals Limited (Delhi, India).

Chromatography: Chromatography was performed on phenomenex C18 column (150 mm×4.6 mm, 5.0 μm)

with mobile phase containing Methanol: Water (70:30v/v) pH adjusted to 3.6 with dilute Orthophosphoric

acid at a flow rate of 1.3 mL/min and the eluent was monitored at 271 nm

Calibration plot for Cinacalcet hydrochloride: A Stock solution of Cinacalcet hydrochloride (1000µg/ml)

was prepared in methanol. The gradient dilutions were prepared by taking 0.5, 1, 1.5, 2, 2.5 and 3ml of the

solution and made up to 10 ml with mobile phase. 20μl of each concentration was injected twice. Calibration

cure was constructed by plotting mean peak areas against the corresponding drug concentration. From the

graphs it was found that the Cinacalcet Hydrochloride obeys Beer’s law and the Range lies between 50-

300μgml.





VALIDATION

Precision: Precision of an analytical method is the degree of agreement among individual test results when

the procedure is applied repeatedly to multiple samplings of a homogeneous sample. Precision of analytical

method is usually expressed as the standard deviation (or) relative standard deviation.

Cinacalcet Hydrochloride of concentration 200μg/ml was prepared and injected for six times and analyzed

under optimized chromatographic conditions.

Limit of detection and limit if quantification: Cinacalcet hydrochloride at concentration in the lower part of

the linear range of calibration plot was used to determine limit of detection (LOD) and Limit of

quanfication(LOQ). They were determinated from the slope of calibration plot and standard deviation of the

blank sample by use of the equations

LOD = 3.3×Standard deviation / Slope and

LOQ = 10×Standard deviation / Slope

Recovery: Recovery studies were carried out by mixing standard quantity of drug with pre analyzed sample

formulation and the contents were analyzed by proposed method. From the sample stock solution a dilution of

concentration of 50μg/ml was prepared and to this standard solution of series of concentrations from 0, 50,

100, 150, 200, 250 μg/ml was added and scanned.

Analysis of Synthetic mixture: Due to non- availability of formulation in the local pharmacies, a synthetic

mixture was prepared using drug with suitable excipients. Synthetic mixture of Cinacalcet hydrochloride was

used for the assay. Accurately weighed quantity of Cinacalcet Hydrochloride equivalent to 100mg and

transferred into a 100ml volumetric flask. To this 30ml of methanol was added and ultrasonicated for half an

hour until the drug was extracted. The solution was filtered by using Whatmann filter paper No:45μ and

finally made up to the mark with mobile phase to give a concentration of 1mg/ml. Different aliquots of 1.0,

1.5and 2.0ml was taken from the above prepared stock and made up to 10ml with mobile phase to get

concentrations of 100, 150, 200μg/ml respectively. 20μl of the aliquots were injected and the chromatograms

were recorded.

Forced degradation of Cinacalcet hydrochloride

Acid and base induced degradation: Cinacalcet hydrochloride (25 mg) of standard was separately taken in a

two 50 ml volumetric flask, dissolved in 25 ml methanol and diluted to volume with 0.1M HCl and 0.1M

NaOH. The solution was refluxed for 24 hours at 40C to 80C, and sampling was done at different intervals

of time and diluted with the mobile phase to the final concentration of 100 µg/ml and are analyzed by

proposed HPLC method

Hydrogen peroxide induced degradation: Cinacalcet hydrochloride (25 mg) of was taken in a 50 ml

volumetric flask, dissolved in methanol and diluted to volume with 1% H2O2, 3%H2O2 and 30 % H2O2.The

solution was collected at different time intervals. Then these solutions were diluted with the mobile phase to

the final concentration of 100 µg/ml and are analyzed by proposed HPLC method.

Thermal Degradation: Cinacalcet hydrochloride (25 mg) was taken in petri dish (10cm in diameter) and

spread to a thickness of 1mm and the sample is exposed to a temperature of 105ºC for24 hrs and for wet heat

degradation drug was kept in reflux at 80°C, for 24 hrs. Then the samples were taken at different time

intervals and diluted with the mobile phase to the final concentration of 100 µg/ml for further analysis by

HPLC.

Photochemical Degradation: Cinacalcet hydrochloride (25 mg) was taken on Petri dish (10cm in diameter)

and spread to a thickness of 1mm and exposed to Sunlight, UV-light and dark place for 24 hrs.

Simultaneously the drug solutions (1mg/ml) were taken in different volumetric flasks and exposed to Sunlight,

UV-light and dark place for 24 hrs. Then these samples are taken at different time intervals and diluted with

the mobile phase to the final concentration of 100 µg/ml for further analysis using HPLC

RESULT AND DISCUSSION

Chromatographic conditions: The present study of stability indicating high performance liquid

chromatographic method of Cinacalcet hydrochloride in bulk and synthetic mixture, different

chromatographic condition were applied. Among the different mobile phase employed the mobile phase of

Methanol: Water (70:30 v/v) pH adjusted to 3.6 with dilute Orthophosphoric acid at a flow rate of 1.3 mL/min

and the eluent was determined in UV detection at 271 nm.

Linearity: The calibration curve showed the concentrations ranging from 50-300μg/ml of Cinacalcet

Hydrochloride were prepared and 20μl of each concentration was injected two times. The Correlation

Coefficient values were found to be within the acceptance criteria for the drug. The Limit of Detection and

Quantitation values were calculated.

The results were shown in Figure 1, 2 and tabulated in Table-1, 2





Precision: Cinacalcet Hydrochloride of concentration 200μg/ml was prepared and injected for six times and

analyzed under optimized chromatographic conditions. The standard deviation, Relative Standard Deviation

for Cinacalcet Hydrochloride was found to be within the acceptance criteria. (< 2%) and hence the method

was found to be precise. The results are tabulated in Table 2.

Limit of detection and limit if quantification: The signal and noise ratios of 3:1 and 10:1 were considered

as LOD and LOQ respectively. From the results of linearity the LOD and LOQ were found to be 5.063 μg/ml

and 15.20μg/ml respectively. . The results are tabulated in Table 2.

Recovery studies:

A known amount of the drug was added to formulation and subjected to check the reproducibility of the

method. The estimated percentage recovery was found to be within the normal range (97% – 103%), this

proves the reproducibility of the method

Forced degradation studies: HPLC studies of samples obtained on stress testing of Cinacalcet hydrochloride

under different conditions using Methanol: Water: (70:30 v/v) and pH adjusted to 3.6 with O-Phosphoric acid

as the mobile solvent system suggested the following degradation behavior.

During the study it was observed than upon treatment of Cinacalcet hydrochloride with acid(0.1M HCl), base

(0.1M NaOH), hydrogen peroxide (1%, 3%, and 30%), heat(dry and wet heat) and photo

degradation(Sunlight, UV light(254nm) and Dark light. Table 3 indicated the extent of Cinacalcet

hydrochloride degradation was observed in various Stress condition. Figure 4 to 8 shows the chromatogram of

forced degraded sample.

Table 1: Linearity data of the HPLC method Cinacalcet hydrochloride Concentration (μg/ml) Peak Area (m V .s)

50

100

150

200

250

300

12171129

21982505

32616363

42370019

53819060

63894844

Table 2: Summary of HPLC Results Parameter Result found Limit

Retention time (min) 5,52 -

Correlation coefficient (R2) 0.999 >0.996

Slope (m) 21097 -

Intercept (c) 0.999 -

LOD 5.063 -

LOQ 15.20 -

SD 0.9799 -

RSD (%) 0.478 <2%

SE 0.4004 -

% Purity(w/w) 97-103 97-103

Table 3: Result of force degradation studies of Cinacalcet hydrochloride API Stress Condition / Duration % Degradation

Acid Degradation(0.1M HCl, 40˚C, 24h) 88.6

Base Degradation(0.1M NaOH, 80˚C, 12h) 92.2

Oxidative degradation(1%H2O2, 25˚C, 8h) 1.3



Thermal degradation(Soild sample, 105˚C, 24h) 28.8

Thermal degradation(Solution, 80˚C, 24h) 92.2

Photo degradation(Soild sample, Sunlight, 24h) 29.5

Photo degradation(Solution, Sunlight, 24h) 65.2

Photo degradation(Soild sample, UV light, 24h) 13.7

Photo degradation(Solution, UV light, 24h) 18.2

Photo degradation(Soild sample, Dark light, 24h) 9.3

Photo degradation(Solution, Dark light, 24h) 11.1





Figure 1: Chromatogram of Cinacalcet

Hydrochloride at 271nm

Figure 2: Calibration curve of Cinacalcet

Hydrochloride using HPLC


Hydrochloride standard (100 µg/ml) at 271nm. Figure 4: Chromatogram of Cinacalcet

Hydrochloride under acid stress condition at 24 hr


Hydrochloride under alkali stress condition at

12hrs.

Figure 7: Chromatogram of Cinacalcet Hydrochloride

under wet heat stress condition at 24hrs

Figure 8: Chromatogram of Cinacalcet Hydrochloride Solution under UV light at 24 hrs

CONCULTION





The proposed HPLC method is precise, specific, accurate and stability indicating. The sample

recoveries in Synthetic mixture were in good agreement with their respective amount taken which suggested

noninterference of synthetic mixture excipients in the estimation. The method can be used to determine the

purity of the drug obtained from different sources by detecting related impurities. Hence, the proposed method

stands validated and may be used for routine and stability sample analysis

ACKNOWLEDGEMENT

The authors are grate full to Actavis Pharma Manufacturing Pvt. Ltd. Chennai, India for providing the pure

sample of Cinacalcet hydrochloride

REFERENCES

Fen Hang, Hangyun wang, Qianzhoo, Hongzong liu, Pei Hu and JiJIang, Determination of Cinacalcet

Hydrochloride in Human plasma by Liquid Chromatgraphy - tandem mass spectrometry, Journal of

Pharmaceutical & Biomedical Analysis, 10, 2011, 2-18.

Ibrahim A Darwish, Mona M Al- Shehri and Manal A El – Gendy, Novel spectrophotometric method for

determination of Cinacalcet Hydrochloride in its tablets via derivitization with 1,2 – naphthoquinone 4 –

Sulphonate, Chemistry Central Journal, 6,11, 2012, 655-658.

ICH Harmonized Tripartite guideline, validation of analytical procedures, Text and Methodology Q2 (R1), 4th

version, 27oct 1994, 01-17.

ICH Stability testing of new drug substances and products (Q1 A R2), 4thversion, 2003, 5.

Julia A, Barmon Balfour, Scott and Lesley J, Cinacalcet Hydrochloride, Drugs, 65(2), 2005, 271-281.

Martindale, the Complete Drug Reference, 28, 770-772.

Narogi R ., Vishvottam K, Prashanth Komarneni, Raghupati Aleti, Naga Surya Prakash Padala and

Ilayaraja.K, Quantification of Cinacalcet by LC-MS/MS using Liquid-Liquid Extraction from 50μl of plasma,

Journal of Pharmaceutical and Biomedical Analysis, 56(5), 5, 2011, 373-81.

Nuin E, Andreu I, Torres MJ, Miranda MA, Miguel A, Jime Nez and Consueloo M, Enhanced Photo safety of

Cinacalcet Hydrochloride upon complexation with serum albumin, The Journal of Physical Chemistry, B, 115,

5, 2011, 1158-1164.

Ravi Bhushan and Ritual Dubey, Indirect Reversed Phase HPLC and Direct Thin layer Chromatographic

enantioresolution of (R,S)- Cinacalcet, Biomedical Chromatography, 25(6), 2011, 674-679.

Satinder Ahuja, Michael W Dong, Hand book of pharmaceutical analysis by HPLC, 25, 336-345.

Sethi P D, High Performance Liquid Chromatography Quantitative Analysis of Pharmaceutical Formulations,

1, 2001, 02-07.

Sigala Ashok, Raghunath Babu CH V, Satish Varma M and BalaSwamy G, A new validated liquid

chromatographic method for the determination of impurities in Cinacalcet Hydrochloride, Analytical

Chemistry-An Indian Journal, 8(4), 2009, 278-297.

Vadde ravinder, Sigala Ashok, Satish varma M, Raghunaath babu C V and Gubba Balaswamy, A validated

chiral LC method for the enantiomeric separation of (S)-Cinacalcet from (R) – Cinacalce, Chromatographia,

70, 2009, 229-232.



Shravan Paswan et.al. Indian Journal of Research in Pharmacy and Biotechnology


BIOAVAILABILITY: CRITERIA FOR APPROVING A DRUG PRODUCT FOR

MARKETING Sandhya Singh

1, Faheem Ajmal Ansari

1, Shravan Paswan

2*,

Rnjan Kumar Sharma2, Alok Ranjan Gaur

3

1Azad Institute of Pharmacy and Research Lucknow, India

2Advance Institute of Pharmaceutical Education & Research, Kanpur, India

3Department of Pharmacy, Pranveer Singh Institute of Technology, Kanpur, India


ABSTRACT

FDA ensure that the drug product for marketing should be safe, effective and meet all applicable

standards, for this FDA requires bioavailability/pharmacokinetic studies and, where necessary,

bioequivalence studies for all drug products (FDA Guidance for Industry, 2003). The U.S. Food and

Drug Administration (FDA) define bioavailability as "the rate and extent to which the active drug

ingredient or therapeutic moiety is absorbed from a drug product and becomes available at the site of

drug action". Generally direct and indirect methods use to assess drug bioavailability. For assessing

bioavailability or clinical availability of a drug, its rate and extent of absorption and its first-pass

metabolism must be evaluated. The clinical response of the patient or the amount of active drug at the

target site of action at different time periods should also be assessed. In order to achieve targeted

minimum level for therapeutic or clinical effect, the medical practitioner must understand various

contributing factors that could affect the bioavailability. For the scientists, they must also be aware of

some essential intrinsic factors that influence the formulation. There are basically three factors, which

affect bioavailability physiological factors, physicochemical factors and pharmacological factors.

Key Words: FDA, Bioavailability, Bioequivalence, Pharmacokinetic, Physicochemical

INTRODUCTION

In approving a drug product for marketing, the FDA ensures that the drug product is safe and effective for

its labeled indications for use. Moreover, the drug product must meet all applicable standards of identity, strength,

quality, and purity. To ensure that these standards are met, the FDA requires bioavailability/pharmacokinetic

studies and, where necessary, bioequivalence studies for all drug products (FDA Guidance for Industry, 2003).

Bioavailability may be considered as one aspect of drug product quality that links in-vivo performance of the drug

product used in clinical trials to studies demonstrating evidence of safety and efficacy. For unmarketed drugs that

do not have full NDA approval by the FDA, in-vitro and/or in-vivo bioequivalence studies must be performed on

the drug formulation proposed for marketing as a generic drug product.

Bioavailability: The U.S. Food and Drug Administration (FDA) define bioavailability as "the rate and extent to

which the active drug ingredient or therapeutic moiety is absorbed from a drug product and becomes available at

the site of drug action". Because in practice it is rare that drug concentrations can be determined at the site of

action (e.g., at a receptor site), bioavailability is more commonly defined as "the rate and extent that the active

drug is absorbed from a dosage form and becomes available in the systemic circulation."

Objectives of bioavailability studies: Bioavailability studies are important as

1. Primary stages of development of a suitable dosage form for a new drug entity.

2. Determination of influence of excipients, patient related factors & possible interaction with other drugs on

the efficiency of absorption.

3. Development of new formulations of the existing drugs.

4. Control of quality of a drug product during the early stages of marketing in order to determine the

influence of processing factors, storage & stability on drug absorption.

Relative and absolute bioavailability: The area under the drug concentration–time curve (AUC) is used as a

measure of the total amount of unaltered drug that reaches the systemic circulation. The AUC is dependent on the

total quantity of available drug, FD0, divided by the elimination rate constant, k, and the apparent volume of

distribution, VD. F is the fraction of the dose absorbed. After IV administration, F is equal to unity, because the

entire dose enters the systemic circulation. Therefore, the drug is considered to be completely available after IV

administration. After oral administration of a drug, F may vary from a value of 0 (no drug absorption) to 1

(complete drug absorption).






Relative availability: Relative (apparent) availability is the availability of the drug from a drug product as

compared to a recognized standard. The fraction of dose systemically available from an oral drug product is

difficult to ascertain. The availability of drug in the formulation is compared to the availability of drug in a

standard dosage formulation, usually a solution of the pure drug evaluated in a crossover study. The relative

availability of two drug products given at the same dosage level and by the same route of administration can be

obtained using the following equation.

(1.1)

Where drug product B is the recognized reference standard. This fraction may be multiplied by 100 to give

percent relative availability. When different doses are administered, a correction for the size of the dose is made,

as in the following equation:

(1.2)

Urinary drug excretion data may also be used to measure relative availability, as long as the total amount of intact

drug excreted in the urine is collected. The percent relative availability using urinary excretion data can be

determined as follows:

(1.3)

Where [D u]∞ is the total amount of drug excreted in the urine.

Absolute Availability

The absolute availability of drug is the systemic availability of a drug after extravascular administration (eg, oral,

rectal, transdermal, subcutaneous) compared to IV dosing. The absolute availability of a drug is generally

measured by comparing the respective AUCs after extravascular and IV administration. This measurement may

be performed as long as VD and k are independent of the route of administration. Absolute availability after oral

drug administration using plasma data can be determined as follows:

(1.4)

Absolute availability, F, may be expressed as a fraction or as a percent by multiplying F x 100. Absolute

availability using urinary drug excretion data can be determined by the following:

(1.5)

The absolute bioavailability is also equal to F, the fraction of the dose that is bioavailable. Absolute availability is

sometimes expressed as a percent, ie, F = 1, or 100%. For drugs given intravascularly, such as by IV bolus

injection, F = 1 because the entire drug is completely absorbed. For all extravascular routes of administration,

such as the oral route (PO), the absolute bioavailability F may not exceed 100% (F > 1). F is usually determined

by Equation 1.4 or 1.5, where PO is the oral route or any other extravascular route of drug administration.

Example: The difference between absolute and relative bioavailability is illustrated by the following hypothetical

example.

Assume that an intravenous injection (Product A) and two oral dosage forms (Product B and Product C),

all containing the same dose of the same drug, are given to a group of subjects in a crossover study. Furthermore,

suppose each product gave the values for AUC indicated in Table 1.

Table.1. Data for Absolute and Relative Bioavailability

Drug Product Are Under the Curve (mcg/ml) x hr

A. Intravenous injection 100

B. Oral dosage form, brand or reference standard 50

C. Oral dosage form, generic product 40





The F for Product B and Product C is 50% (F = 0.5) and 40% (F = 0.4), respectively. However, when the two oral

products are compared, the relative bioavailability of Product C as compared to Product B is 80% [3].

FACTOR AFFECTING BIOAVAILABILITY Drug is absorbed from the gastrointestinal (GI) tract after being dissolved according to its intrinsic

absorbability. If the test product shows the same pattern of drug dissolution in the GI tract in vivo as the reference

one, that product must be equivalent unless other ingredients do not modulate the absorption of active drug. In

other words, physicochemical properties of drugs such as water solubility and membrane permeability do not

affect the bioequivalency of oral product. A wide range of factors can influence the bioavailability of a drug.

Basically, the availability of the drug or its metabolite to the target organ or receptor is controlled by three

principal factors:

1. The rate and extent of drug release from its formulation, and its subsequent absorption.

2. The first-pass effect while passing through the liver after absorption.

3. The conjoint effect of plasma protein binding, drug distribution to various body fluids, metabolism

and excretion.

For assessing bioavailability or clinical availability of a drug, its rate and extent of absorption and its first-pass

metabolism must be evaluated. The clinical response of the patient or the amount of active drug at the target site

of action at different time periods should also be assessed. In order to achieve targeted minimum level for

therapeutic or clinical effect, the medical practitioner must understand various contributing factors that could

affect the bioavailability. For the scientists, they must also be aware of some essential intrinsic factors that

influence the formulation.

In general, the following factors were discussed.

Physiological Factors: High variability in oral drug absorption is caused by several factors. Deviations in

physiological conditions in the GI tract of volunteers would affect dissolution and permeation of drugs even in the

same individual. For example, bile acid secretion into the small intestine promotes the dissolution of poorly

soluble drugs to enhance the bioavailability. On the other hand, it was reported that food intake often reduced the

oral absorption of BCS class 3 drugs. These facts indicate that low solubility and low permeability of drugs may

cause not only the incomplete oral absorption but also the high variability in it.

Metabolism in the intestine and liver affects the oral bioavailability as the first-pass effects after

absorption. Also deviations of the metabolic activity in pharmacokinetic parameters due to the change in total

body clearance of drugs. High clearance of drugs therefore, might be one of the risk factors for high variability in

human bioequivalence study.

In short, the following physiological factors are known to affect bioavailability:

1. These include the effect of gastrointestinal fluids such as pH, mucus, bile salts, complexing components.

2. Gastric motility such as gastric emptying, presence of food, rest and exercise.

3. Gastrointestinal transit time which can be affected by a large number of drugs.

4. Metabolism of drugs by the gut wall, liver, skin and bronchial mucosa.

5. The pharmacogenetic factors determining the rate of hepatic metabolism.

6. Various disease states such as malabsorption, achlorhydria, thryrotoxicosis and celiac disease.

7. Other factors include the gut flora, age, sex, weight and physical status of the patients.

Physicochemical Factors: There are various physicochemical factors that may influence absorption of drugs into

the bloodstream. A dissolution testing is essential to establish a profile of each generic product with specific

physicochemical characteristic of the solid dosage form. This testing will ensure the permeability and solubility of

drugs. At the drug development stage, these factors are essential to be evaluated and should not be the direct

causes of failure in bioequivalence study. The physicochemical factors which point to the need for dissolution

testing include:

1. Low drug solubility – to establish the evidence that the drug has a low aqueous solubility.

2. Poor product dissolution – to establish from the literature that the dissolution of one or more marketed

product to develop is, poor when tested by official compendial test procedure.

3. Drug particle size – to establish the evidence that the particle size may affect bioavailability.

4. The physical form of drug – to establish that certain polymorphs, solvates or complexes have poor

dissolution characteristics and hence bioavailability may be affected.





5. Presence of specific excipients – to establish evidence that specific excipients may alter dissolution or

absorption, hence bioavailability may also be affected.

6. The nature of tablet or capsule coating – to establish evidence that coating may interfere with the

disintegration or dissolution of the formulation.

Biopharmaceutic Classification System (BCS): The biopharmaceutic classification system was developed

primarily in the context of immediate release (IR) solid oral dosage forms. It is the scientific framework for

classifying drug substances based on their aqueous solubility and intestinal permeability. It is a drug development

tool that allows estimation of the contributions of three major factors namely dissolution, solubility and intestinal

permeability that affect oral drug absorption from immediate release solid oral dosage forms. The interest in this

classification system is largely because of its application in early drug development and then in the management

of product change through its life cycle. Goals of the BCS Guidance:

1. To improve the efficiency of drug development and the review process by recommending a strategy for

identifying expendable clinical bioequivalence tests.

2. To recommend a class of immediate-release (IR) solid oral dosage forms for which bioequivalence may

be assessed based on in vitro dissolution tests.

3. To recommend methods for classification according to dosage form dissolution, along with the solubility

and permeability characteristics of the drug substance.

Classification: According to BCS, drug substances are classified as:

Class I: High Solubility – High Permeability

Class II: Low Solubility – High Permeability

Class III: High Solubility – Low Permeability

Class IV: Low Solubility – Low Permeability

Combined with the dissolution, the BCS takes into account the three major factors governing

bioavailability parameters namely, dissolution, solubility and permeability. This classification is associated with

drug dissolution and absorption model, which identifies the key parameters controlling drug absorption as a set of

dimensionless numbers. Absorption number, defined as the ratio of the mean residence time to mean absorption

time. Dissolution number, defined as the ratio of mean residence time to mean dissolution time.

METHODS FOR ASSESSING BIOAVAILABILITY

Direct and indirect methods may be used to assess drug bioavailability. The in-vivo bioavailability of a

drug product is demonstrated by the rate and extent of drug absorption, as determined by comparison of

measured parameters, eg, concentration of the active drug ingredient in the blood, cumulative urinary

excretion rates, or pharmacological effects.

For drug products that are not intended to be absorbed into the bloodstream, bioavailability may be

assessed by measurements intended to reflect the rate and extent to which the active ingredient or active

moiety becomes available at the site of action.

The design of the bioavailability study depends on the objectives of the study, the ability to analyze the

drug (and metabolites) in biological fluids, the pharmacodynamics of the drug substance, the route of

drug administration, and the nature of the drug product.

Pharmacokinetic and/or pharmacodynamic parameters as well as clinical observations and in-vitro studies

may be used to determine drug bioavailability from a drug product.

Pharmacokinetic methods: These are indirect methods. Assumption is that pharmacokinetic profile reflects the

therapeutic effectiveness of a drug. Advantages include the results are accurate, reliable, reproducible.

a) Plasma / blood level time profile: Time for peak plasma (blood) concentration (t max) Peak plasma drug

concentration (Cmax) Area under the plasma drug concentration–time curve (AUC).

b) Urinary excretion studies: Cumulative amount of drug excreted in the urine (Du) Rate of drug excretion in the

urine (dD u/dt). Time for maximum urinary excretion (t).

Pharmacodynamic methods: Involves direct measurement.(measurement of pharmacologic or therapeutic end

point)

Disadvantages: High variability, difficult to measure, limited choices, less reliable, more subjective. Drug

response influenced by several physiological & environmental factors

Maximum pharmacodynamic effect (E max).

Time for maximum pharmacodynamic effect.





Area under the pharmacodynamic effect–time curve.

Onset time for pharmacodynamic effect.

They involve determination of bioavailability from:

a) Acute pharmacological response.

b) Therapeutic response.

In-vitro dissolution studies 1. Closed compartment apparatus

2. Open compartment apparatus

3. Dialysis systems.

Plasma drug concentration: Measurement of drug concentrations in blood, plasma, or serum after drug

administration is the most direct and objective way to determine systemic drug bioavailability. By appropriate

blood sampling, an accurate description of the plasma drug concentration–time profile of the therapeutically

active drug substance(s) can be obtained using a validated drug assay.

The time of peak plasma concentration, t max, corresponds to the time required to reach maximum drug

concentration after drug administration. At t max, peak drug absorption occurs and the rate of drug absorption

exactly equals the rate of drug elimination. Drug absorption still continues after t max is reached, but at a slower

rate. When comparing drug products, t max can be used as an approximate indication of drug absorption rate. The

value for t max will become smaller (indicating less time required to reach peak plasma concentration) as the

absorption rate for the drug becomes more rapid. Units for t max are units of time (eg, hours, minutes).

The peak plasma drug concentration, C max, represents the maximum plasma drug concentration

obtained after oral administration of drug. For many drugs, a relationship is found between the pharmacodynamic

drug effect and the plasma drug concentration. C max provides indications that the drug is sufficiently

systemically absorbed to provide a therapeutic response. In addition, C max provides warning of possibly toxic

levels of drug. The units of C max are concentration units (eg, mg/mL, ng/mL). Although not a unit for rate, C

max is often used in bioequivalence studies as a surrogate measure for the rate of drug bioavailability.

The area under the plasma level–time curve, AUC, is a measurement of the extent of drug bioavailability.

The AUC reflects the total amount of active drug that reaches the systemic circulation. The AUC is the area under

the drug plasma level–time curve from t = 0 to t = ∞, and is equal to the amount of unchanged drug reaching the

general circulation divided by the clearance.

where F = fraction of dose absorbed, D0 = dose, k = elimination rate constant, and VD = volume of distribution.

The AUC is independent of the route of administration and processes of drug elimination as long as the

elimination processes do not change. The AUC can be determined by a numerical integration procedure,

such as the trapezoidal rule method.

The units for AUC are concentration time (eg, μg hr/mL).

For many drugs, the AUC is directly proportional to dose. For example, if a single dose of a drug is

increased from 250 to 1000 mg, the AUC will also show a four fold increase.

In some cases, the AUC is not directly proportional to the administered dose for all dosage levels. For

example, as the dosage of drug is increased, one of the pathways for drug elimination may become

saturated. Drug elimination includes the processes of metabolism and excretion. Drug metabolism is an

enzyme-dependent process.

For drugs such as salicylate and phenytoin, continued increase of the dose causes saturation of one of the

enzyme pathways for drug metabolism and consequent prolongation of the elimination half-life.

The AUC thus increases disproportionally to the increase in dose, because a smaller amount of drug is

being eliminated (ie, more drug is retained). When the AUC is not directly proportional to the dose,

bioavailability of the drug is difficult to evaluate because drug kinetics may be dose dependent.





Plasma drug concentration time curve

Urinary drug excretion data: Urinary drug excretion data is an indirect method for estimating bioavailability.

The drug must be excreted in significant quantities as unchanged drug in the urine. In addition, timely urine

samples must be collected and the total amount of urinary drug excretion must be obtained.

The cumulative amount of drug excreted in the urine, D∞u is related directly to the total amount of drug

absorbed. Experimentally, urine samples are collected periodically after administration of a drug product. Each

urine specimen is analyzed for free drug using a specific assay.

The relationship between the cumulative amounts of drug excreted in the urine and the plasma level–time

curve shows when the drug is almost completely eliminated, the plasma concentration approaches zero and the

maximum amount of drug excreted in the urine, D∞u is obtained.

The rate of drug excretion, because most drugs are eliminated by a first-order rate process, the rate of

drug excretion is dependent on the first-order elimination rate constant k and the concentration of drug in the

plasma Cp. In the maximum rate of drug excretion, (dD u/dt) max, is at point B, whereas the minimum rate of

drug excretion is at points A and C. Thus, a graph comparing the rate of drug excretion with respect to time should

be similar in shape as the plasma level–time curve for that drug.

The total time for the drug to be excreted, in and, the slope of the curve segment A–B is related to the rate

of drug absorption, whereas point C is related to the total time required after drug administration for the drug to be

absorbed and completely excreted t = ∞. The t∞ is a useful parameter in bioequivalence studies that compare

several drug products.

Rate of excretion

Acute pharmacological response

In some cases, the quantitative measurement of a drug in plasma or urine lacks an assay with sufficient

accuracy and/or reproducibility. For locally acting, non systemically absorbed drug products, such as

topical corticosteroids, plasma drug concentrations may not reflect the bioavailability of the drug at the

site of action.

An acute pharmacodynamic effect, such as an effect on forced expiratory volume, FEV1 (inhaled

bronchodilators) or skin blanching (topical corticosteroids) can be used as an index of drug

bioavailability. In this case, the acute pharmacodynamic effect is measured over a period of time after

administration of the drug product.





Measurements of the pharmacodynamic effect should be made with sufficient frequency to permit a

reasonable estimate for a time period at least three times the half-life of the drug. This approach may be

particularly applicable to dosage forms that are not intended to deliver the active moiety to the

bloodstream for systemic distribution.

The use of an acute pharmacodynamic effect to determine bioavailability generally requires

demonstration of a dose–response curve.

Bioavailability is determined by characterization of the dose–response curve. For bioequivalence

determination, pharmacodynamic parameters including the total area under the acute pharmacodynamic

effect–time curve, peak pharmacodynamic effect, and time for peak pharmacodynamic effect are obtained

from the pharmacodynamic effect–time curve. The onset time and duration of the pharmacokinetic effect

may also be included in the analysis of the data.

The use of pharmacodynamic endpoints for the determination of bioavailability and bioequivalence is

much more variable than the measurement of plasma or urine drug concentrations.

Effects such as change in ECG or EEG readings, pupil diameter, etc are related to the time course of a

given drugs.

Bioavailability can be determined by construction of pharmacologic effect time curve as well as dose

response graph.

The drawback of this method is that, the response tends to more variable. Moreover, the observed

response may be due to an active metabolite whose concentration is not proportional to concentration of

parent drug responsible for the pharmacological effect.

Therapeutic response: Theoretically, this method is most definite among all.

It’s based on observing clinical response to a drug formulation given to a patient suffering from disease

for which the drug is intended to be used.

A major drawback is that quantification of observed response is unreliable for assessment of

bioavailability.

In- vitro dissolution study: Drug dissolution studies may under certain conditions give an indication of drug

bioavailability. Ideally, the in-vitro drug dissolution rate should correlate with in-vivo drug bioavailability (see

and on in-vivo–in-vitro correlation, IVIVC). Dissolution studies are often performed on several test formulations

of the same drug. The test formulation that demonstrates the most rapid rate of drug dissolution in vitro will

generally have the most rapid rate of drug bioavailability in vivo.

Closed compartment apparatus: Non sink condition

Open compartment apparatus: Perfect sink condition

Dialysis system: This method is useful for very poorly aqueous soluble drugs for which maintenance of sink

condition would require large volume of dissolution fluid.

Clinical observations: Well-controlled clinical trials in humans establish the safety and effectiveness of drug

products and may be used to determine bioavailability. However, the clinical trials approach is the least accurate,

least sensitive, and least reproducible of the general approaches for determining in-vivo bioavailability. The FDA

considers this approach only when analytical methods and pharmacodynamic methods are not available to permit

use of one of the approaches described above. Comparative clinical studies have been used to establish

bioequivalence for topical antifungal drug products (eg, ketoconazole) and for topical acne preparations. For

dosage forms intended to deliver the active moiety to the bloodstream for systemic distribution, this approach may

be considered acceptable only when analytical methods cannot be developed to permit use of one of the other

approaches.

Analytical methodology: The selected analytical method should be

Sufficiently sensitive to permit detection of low concentration of drug.

Reproducible

Must be specific for unmetabolized drug as well as capable of determining concentration of drug in

presence of metabolites, constituents of blood/ urine.

Stable Isotope Studies: This approach involves the simultaneous administration of test product and the reference

product, using each subject as his own control. The reference contains the drug, which has been synthesized to

contain a stable isotope such as such as 2H, 15N, 13C or 18Oin a position in a drug molecule that is not





susceptible to metabolism and does not result in kinetic differences due to presence of isotope. The sample is

collected and the comparisons are made of quantity of labeled and unlabeled drug in each sample, using

sophisticated detection systems involving mass spectroscopy.

A method for the calculation of bioavailability in slow release formulations in the presence of within-

individual variability: In the present study they propose a model-independent method based on the combination

of the area under the curve of serum drug levels and the mean residence time for evaluating the amount of

bioavailability when within-individual variability is present in the serum clearance of the drug, administered as a

slow release formulation (SRF), and this follows linear pharmacokinetic behavior.

The method assumes that the modifications in the area under the curve of the serum levels induced by the

within-individual variability in the kinetic behavior of the drug lead to a variation of the same proportions in the

mean residence time of the serum levels curve and that this parameter can be used as a correction factor in the

ratio of the areas under the curve of serum levels in bioavailability studies.

The method allows one to calculate the fraction of dose absorbed from the SRF without having to

measure the disposition clearance of the drug either when using the reference formulation or when the drug is

administered as a SRF. The method is easy to apply and has a minimum mathematical complexity. The validity of

the method was evaluated using simulated data with either no error or containing a random error of 10%.

Bioavailability Problems: There are a number of examples of drugs products which have exhibited

bioavailability problems in the past. These examples are all pre-1976 [Gibaldi, 1984]. This is an indication that

more attention is now being given to formulation development during drug development. Three chlorpropamide

formulations were tested and the peak plasma concentration after administration of one brand was less than half

the peak concentration after the other two formulations (figure.1.).

Figure.1. Plot of Cp versus Time

The text reports a number of bioavailability problems with digoxin. One example is particularly

interesting. Doctors in Israel noticed 15 cases of digoxin toxicity between Oct/Dec 1975 with almost no reports

for the same period the previous year. It was found that the local manufacturer had changed the formulation to

improve dissolution without telling the physicians. Urinary data suggested a two-fold increase in availability of

the new formulation.

Again there are a number of examples in the text. One report described an incidence of phenytoin

intoxication in Australia in 1968 and 1969. Apparently the tablet diluent was changed from calcium sulfate to

lactose. Later studies showed that the bioavailability was higher from the dosage form containing lactose. Other

drugs with problems in the past include Acetazolamide, Aminosalicylate, Ampicillin, Aspirin, Ascorbic Acid,

Chloramphenicol, Chlorothiazide, Diazepam, Furosemide, Iron, Levodopa + 10 (Gibaldi, 1984).

CONCLUSION

Today, various pharmaceutical companies are developing generic drug products. Bioequivalence study is

important for generic drug approval process. It is our hope that, this review will provide an easy quick overview

for Regulatory consideration required for bioequivalence study. This review covers major aspect of requirement

of bioequivalence study along with the regulatory specification.





REFERENCES

Asif M Tamboli, Pavan Todkar, Priti Zope and FJ Sayyad, An Overview on Bioequivalence: Regulatory

Consideration for Generic Drug Products Journal of Bioequivalence & Bioavailability, 2(4), 2010, 086-092.

Dalton JT and Yates CR, Bioavailability of Drugs and Bioequivalence, Encyclopedia of pharmaceutical

technology, Third Edition, Volume 1, Swarbrick J, Editor, 2007, Informa Healthcare. USA Inc, 164-176.

Davit BM, Conner DP, fabian-Firtsch B, Highly Variable Drugs: Observations from Bioequivalence Data

Submitted to the FDA for New Generic Drug Applications, AAPS J, 10, 2008, 148-156.

Karalis V, Macheras P, Van Peer A, Bioavailability and Bioequivalence: Focus on Physiological Factors and

Variability, Pharm Res, 25, 2008, 1956-1962.



Harish G et.al. Indian Journal of Research in Pharmacy and Biotechnology


THE EFFECT OF SUPERDISINTEGRANTS ON THE DISSOLUTION OF

CALCIUM CARBONATE FAST DISSOLVING TABLETS 1Mohammed Farhana,

1J.Preeti,

1Md Faizulla,

1Budda Chellibabu,

1Harish.G*,

2Rajnesh Kumar Singh

1Nimra College of Pharmacy, Jupudi,Vijayawada

2Micro Advance Research Centre, Bangalore


ABSTRACT

The objective of the present study is to design and evaluate the effect of disintegrating agents such as

Starch, Cross Caramellose Sodium, Sodium starch glycolate and crosprovidone Calcium carbonate

conventional tablets simultaneously. The nature of calcium carbonate which forms cake on standing

which affects the drug disintegration process there by inhibits the drug release from the Conventional

Tablet. Hence in the present study the effect of disintegrating agents at different concentrations is carried

out on both the drugs simultaneously and finding out the best concentration followed by stability studies

for a period of 3 months.

Key words-Calcium Carbonate, Disintegrating agent, fast dissolving Tablets

INTRODUCTION

Despite increasing interest in controlled-release drug delivery systems, the most common tablets are those

intended to be swallowed whole and to disintegrate and release their medicaments rapidly in the gastrointestinal

tract (GIT). The proper choice of disintegrant or superdisintegrant and its consistency of performance are of

critical importance to the formulation development of such tablets. Drug release from a solid dosage form can be

enhanced by addition of suitable disintegrants. In more recent years, increasing attention has been paid to

formulating fast dissolving and/or disintegrating tablets that are swallowed, but also orally disintegrating tablets

that are intended to dissolve and/or disintegrate rapidly in the mouthThe present study is an attempt to select best

possible combination of drug and disintegrating agent to formulate rapidly disintegrating tablet of calcium

carbonate conventional tablets which disintegrates faster thereby reducing the time of onset of action. Lactose is

selected as diluents, Starch, Sodium starch glycolate, CCS and crospovidone were selected as disintegrants. PVP

K 30M paste was used as a binder in all formulations, Magnesium stearate and Talc as a Lubricant, Aerosil as a

Glidant.


Calcium carbonate Procured from MicroLabs,Bangalore , Cross Caramellose Sodium, Sodium Starch

Glycolate Procured from Signet chemicals,Mumbai, Anhydrous lactose Procured from Jain

Enterprises,Chennai,Aerosil ,Talc from Nice Chemicals Ltd, Chennai

Table 1(a): Formulation Tablet of Calcium carbonate conventional tablet

FA- Starch, FB- Croscarmelose Sodium, FC-Sodium starch glycolate, FD & FE- Crospovidone

Formulation FA1 FA2 FA3 FB1 FB2 FB3 FC1 FC2 FC3 FD1 FD2 FE1 FE2

Calcium

Carbonate (mg)

250 250 250 250 250 250 250 250 250 250 250 500 500

Starch 50 100 150 - - - - - - - - - -

CCS - - - 10 20 30 - - - - - - -

SSG - - - - - - 40 50 60 - - - -

CP - - - - - - - - - 40 80 40 80

Lactose 570 520 470 610 600 590 580 570 560 580 540 340 300

PVP 50 50 50 50 50 50 50 50 50 50 50 20 20

Talc 50 50 50 50 50 50 50 50 50 50 50 50 50

Magnesium

Sterate

30 30 30 30 30 30 30 30 30 30 30 50 50

Total Weight 1000 1000 1000 100

0

1000 1000 1000 1000 1000 1000 1000 1000 1000






EVALUATION OF FORMULATED CALCIUM CARBONATE TABLETS

Evaluation of blend characteristics: The Calcium carbonate granules were prepared using wet granulation

method. Various formulations were made as shown in table: 1(a). The Formulated calcium carbonate granules

were evaluated for Pre-formulation parameters like angle of repose, bulk density, tapped density, Compressibility

index and Hausner’s Ratio.

Using FA (Starch) as Disintegrant: The angle of repose of formulated calcium carbonate tablet was in the range

of 20°-30°. Normally if the value falls between 20°-30°, it shows good flow property. The bulk density and

tapped density were found to be in the range of 0.37 to 0.38 g/cm3 and 0.44 to 0.45g/cm

3 respectively. A Hausner

ratio was within the range of 1.16 to 1.17, lesser than 1.25 is considered to be an indication of good flow property.

The compressibility index was within the range of 5-15 hence falls within the excellent range.

Using FB (Croscarmelose Sodium) as Disintegrant: The angle of repose of prepared calcium carbonate tablet

was in the range of 20°-30°. Normally if the value falls between 20°-30°, it shows good flow property. The bulk

density and tapped density were found to be in the range of 0.34 to 0.36 g/cm3 and 0.39 to 0.40g/cm

3 respectively.

The Hausner’s ratio was within the range of 1.07 to 1.18, lesser than 1.25 is considered to be an indication of

good flow property. The compressibility index was within the range of 5-15 hence falls within the excellent range.

Using FC (Sodium Starch Glycolate) as Disintegrant: The angle of repose of prepared calcium carbonate tablet


density and tapped density were found to be in the range of 0.35 to 0.36 g/cm3 and 0.39to 0.41g/cm

3 respectively.


good flow property. The compressibility index was within the range of 5-15 hence falls within the excellent range.

Using FD (Crospovidone- Internal) as Disintegrant: The angle of repose of prepared calcium carbonate tablet


density and tapped density were found to be in the range of 0.36 to 0.38 g/cm3 and 0.40 to 0.41g/cm

3 respectively.


good flow property. The compressibility index was within the range of 5-15 hence falls within the excellent

range.

Using FE (Crospovidone- Internal& External)) as Disintegrant: The angle of repose of prepared calcium

carbonate tablet was in the range of 20°-30°. Normally if the value falls between 20°-30°, it shows good flow

property. The bulk density and tapped density were found to be in the range of 0.36 to 0.37 g/cm3 and 0.40 to

0.41g/cm3

respectively. The Hausner ratio was within the range of 1.10 to 1.11, lesser than 1.25 is considered to

be an indication of good flow property. The compressibility index was within the range of 5-15 hence falls within

the excellent range.

Post- Compressional Characteristic: The post compressional characteristic for all the formulated batches was

found to be within the limits as per Indian pharmacopeia 2007. The hardness was found to be within the range of

3.5 to 5.5 Kg/cm2 in all the formulations indicating good mechanical strength with an ability indicating physical

and mechanical strength with an ability to withstand physical and mechanical stress conditions while handling. In

all the formulations, the friability value is less than 1% giving an indication that tablets formulated are

mechanically stable. All the tablet formulations passed the weight variation test. The weight of all the

formulations was found to be within the limits. The assay of all the formulations was found to be with in the 98%

to 100.5% acceptable limits. The results of disintegration time of all the tablets prepared by wet granulation found

to be varied with change in concentration of disintegrating agents from few seconds to several minutes.

Formulations FD 1 and FE1 disintegrated within 3min and found to be more effective. The disintegration time of

the tablets using different disintegrants decreases in the following order CP > Croscarmellose sodium > SSG >

Starch. It is observed that, when CP is used as disintegrant, tablets disintegrate rapidly with in less time compared

to other tablets prepared using croscarmellose sodium, starch and sodium starch glycolate disintegrants. Though

tablets prepared by intra and extra granulation method found to be more effective in comparison with formulation

prepared by only extra granulation. When concentration of Starch, SSG, CCS and BC is increased, the

disintegration time was reduced significantly.

STABILITY STUDIES

Drug molecules are of reactive naturally, the additives are considered to be inert substance but this may not

be true for all additives in a formulations. Hence, in developing any formulation, when additive are selected the

same must be super imposed on to drugs and with other additives present in the formulation, to see how





compatible they are with the other formulation ingredients. Real time study of ICH guidelines involves storage of

products at 30°C & 65% RH for the complete proposed shelf life period, and analyzing the product sample every

month in the first 3 months, every 3 months from 4th month onwards up to one year, every 6 month in the second

year of storage, afterwards once in a year till shelf life is completed. ICH guidelines also demands for storing

samples at 40 ° c and 75 % RH (stress condition or accelerated stability studys) for relatively short period of time

(3-6 months) which depends on claimed shelf life period as well as the zone (zone 1/2/3/4 of the world) to which

the product is meant to be exported. This later study (with stress conditions) is to mine the alternating climates

condition during the shelf life of the product. The stability parameter for all the formulation were evaluated after

15, 30, 45, 60, and 90 days for 40 °C at 75% RH and the values were been tabulated in table given in Table 11.

CONCLUSION Selecting appropriate formulation excipients and manufacturing technology can obtain the design feature

of fast disintegrating tablet. The disintegrants have the major function to oppose the efficiency of the tablet binder

and the physical forces that act under compression to form the tablet. The stronger the binder, the more effective

must be the disintegrating agents in order for the tablet to release its medication. Ideally, it should cause the tablet

to disrupt, not only into the granules from which it was compressed, but also into powder particles from which the

granulation was prepared. From this study, it is concluded that the disintegrants such as Starch, SSG, CCS was

compared with crospovidone disintegrants and in this study calcium carbonate tablet using crospovidone as

disintegrant prepared by intra and extra granulation method was found to be the most effective as they disintegrate

rapidly when compared to other disintegrants, and the percentage drug release also shows a higher dissolution

profile.

Table 1: Post-compressional characteristic of Calcium carbonate Tablets Using FA as disintegrant

Formulation Weight

variation(mg)

Thickness

(mm)

Diameter

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

FA1 Compiles 7.194 18.99 3.5-5.5 0.291 91.8263

FA2 Compiles 7.29 19.06 3.5-5.5 0.386 93.1792

FA3 Compiles 7.39 18.97 3.5-5.5 0.254 99.9458

Table 2: Post-compressional characteristic of Calcium carbonate Tablets Using FB as disintegrant

Formulation Weight variation

(mg)

Thickness

(mm)

Diameter

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

FB1 Compiles 7.31 18.91 3.5-5.5 0.355 99.2762

FB2 Compiles 7.46 19.03 3.5-5.5 0.318 98.4949

FB3 Compiles 7.27 19.11 3.5-5.5 0.347 99.5952

Table 3: Post-compressional characteristic of Calcium carbonate Tablets Using FC as disintegrant


(mg)

Thickness

(mm)

Diameter

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

FC1 Compiles 7.251 17.96 3.5-5.5 0.314 99.6604

FC2 Compiles 7.564 18.42 3.5-5.5 0.389 98.8846

FC3 Compiles 7.387 18.55 3.5-5.5 0.296 98.8863

Table 4: Post-compressional characteristic of Calcium carbonate Tablets Using FD as disintegrant


(mg)

Thickness

(mm)

Diameter

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

FD1 Compiles 7.27 19.11 3.5-5.5 0.214 98.4771

FD2 Compiles 7.39 19.27 3.5-5.5 0.296 99.2737

Table 5: Post-compressional characteristic of Calcium carbonate Tablets Using FE as disintegrant


(mg)

Thickness

(mm)

Diameter

(mm)

Hardness

(Kg/cm2)

Friability

(%)

Assay

(%)

FE1 Compiles 6.94 18.72 3.5-5.5 0.184 98.2401

FE2 Compiles 7.05 18.92 3.5-5.5 0.213 99.1880

http://www.pharmainfo.net/excipients





Table 6: Disintegration profile of Calcium carbonate tablets using FA as disintegrant Formulation With Disk Without Disk

I II I II I II

FA1 11min 43 sec 10 min 30 sec 10min 52sec 14min 32 sec 15min 11sec 15min 48 sec

FA2 8min 2sec 9min 33 sec 8min 18 sec 11min 14sec 12min 31 sec 11min 56 sec

FA3 4min 41 sec 5min 8 sec 4min 55sec 9min 23sec 9min 51 sec 8min 50sec

Table 7: Disintegration profile of Calcium carbonate tablets using FB as disintegrant


I II I II I II

FB1 9min 21sec 8min 55 sec 10min 05sec 11min 15 sec 11min 24 sec 10min 55min

FB2 7min 43sec 8min 11 sec 8 min 5sec 9min 22 sec 9min 17 sec 10 min 31 sec

FB3 5min 22 sec 5min 42sec 6min 31sec 6 min 4 sec 7min 41sec 7min 18sec

Tab 8: Disintegration profile of Calcium carbonate tablets using FC as disintegrant


I II I II I II

FC1 11min 41 sec 10min 21 sec 10min 54 sec 14min 11sec 14min 56sec 13min 34sec

FC2 8min 43sec 9min 21sec 9min 5sec 12min 37sec 14min 12sec 12min 44sec

FC3 4min 21sec 5min 32 sec 4min 13sec 6min 23sec 6min 47 sec 6min 43sec

Tab 9: Disintegration profile of Calcium carbonate tablets using FD as disintegrant Formulation With Disk Without Disk

I II I II I II

FD1 5 min 51 sec 6min 11 sec 5min 33sec 9min 46sec 8min 23 sec 9min 11sec

FD2 3min 11 sec 2 min 47 sec 2min 17sec 5min 25 sec 4min 21 sec 5 min 41 sec

Tab 10: Disintegration profile of Calcium carbonate tablets using FE as disintegrant Formulation With Disk Without Disk

I II I II I II

FE1 6min 19 sec 6min 5sec 5min 54sec 11min 19 sec 10min 47 sec 11min 14 sec

FE2 4min 45 sec 3min 52 sec 4min 32sec 9min 11sec 9min 42sec 8 min 4 sec

Stability studies:

Tab 11: Stability studies of calcium carbonate Optimized batch Characteristics 40

0C ± 2

0C, 75% ± 5%RH

Initial 15days 30days 45days 60days 75days 90days

Description White compiles compiles compiles compiles compiles compiles

Weight variation

(mg)

compiles compiles compiles compiles compiles compiles compiles

Thickness (mm) 6.31 6.27 6.24 6.28 6.23 6.22 6.24

Diameter (mm) 19.07 19.01 18.89 18.97 18.84 19.84 19.89

Hardness(kg/cm2) 3 3 3 3 3 3 3

Friability (%) 0.09 0.04 0.01 0.01 0.01 0.01 0.01

Assay (%) 98.32 98.30 99.74 98.74 98.56 98.25 98.73

Disintegration

(With disk)

4min

5sec

5min 19

sec

4min

34sec

4min

55sec

4min

5sec

4min 33sec 4min

56sec

Disintegration

(Without disk)

8min 15 sec 8min40

sec

9min23

sec

8min

54sec

8min19

sec

7min 55sec 7min

49sec





REFERENCES

Agdish Singh, Current Status of Tablet Disintegrants:A Review, Drug. Dev. Ind. Pharn, 18(3), 1992, 375-383.

Alekha K, Charles Gayser, Hector Fausett, Evaluation of Quick Disintegrating Calcium Carbonate Tablets,

AAPS Pharm.Sci.Tech, 1(3), 2000, 20.

Andries F Marais, Mingna Song, Melgardt M, Effect of compression force, humidity and disintegrant

concentration on the disintegration and dissolution of directly compressed furosemide tablets using

croscarmellose sodium as disintegrant, Tropical.J .Pharm Res, 2(1), 2003, 125-135.

Antony P J, Sanghavi NM, A New Binder for Pharmaceutical Dosage Forms,

Drug. Dev. Ind. Pharm, 23(4), 1997, 413-415.

Banker SG, Modern pharmaceutics, second edi, Marcel Dekker, newyork, 1990, 372-379.

Basak SC, Rao YS, Manavalan R, Rao PR, Development and In vitroevaluation of an oral Floating matrix tablet

formulation of ciprofloxacin, Ind.Pharm. Sci, 66, 2004, 313-316.

Bi al yen, Rapidly Disintegrable multiparticular Tablets, Chem.Pharma.Bull., 18(9), 1995, 1308-1310.

Bi Y, Sunada H, Yonezawa Y, Preparation and evaluation of a compressed tablet rapidly disintegrating in the oral

cavity, Chem. Pharm. Bull, 44, 1996, 2121-2127.



T K Gopal et.al. Indian Journal of Research in Pharmacy and Biotechnology


PHYTOCHEMICAL AND PHARMACOLOGICAL STUDIES ON WHOLE

PLANT OF ASYSTASIA GANGETICA T.k.Gopal*, Megha.G, D.Chamundeeswari, C.Umamaheswara Reddy,

Faculty of Pharmacy, Sri Ramachandra University, Chennai, India


ABSTRACT

Asystasia gangetica (Linn) T.Anderson belonging to the family Acanthaceae is an ornamental plant

and used as a vegetable in times of food scarcity. It is having many medicinal properties and is used in

folklore medicine for various ailments like asthma, rheumatism, swellings etc. The whole plant of

Asystasia gangetica was used for the study. The solvent methanol was used for the extraction as it was

reported to extract the maximum phytoconstituents. The preliminary phytochemical analysis indicated the

presence of compounds like Flavonoids, Alkaloids, Glycosides, Coumarins, Tannins, Steroids, Terpenoids

and Saponins. The nutritive value of the methanolic extract was evaluated by estimating the

Carbohydrate, Protein and Lipid content. The results justified its use as a vegetable in times of food

scarcity. The secondary metabolites content like Phenols, Tannin and Flavonol content was also

estimated.

The Methanolic Extract of the whole plant was subjected to fractionation by Column chromatography

with solvents of graded polarity. A Flavonoid Mixture Fraction was obtained in the Ethyl acetate:

Methanol (50: 50) fraction. This mixture was characterised by chemical tests, Thin Layer

Chromatography and High Performance Liquid Chromatography. The HPLC characterisation reported

the presence of 4 known Flavonoids and 2 unknown compounds. The four Flavonoids were Luteolin,

Quercetin, Kaempferol and Isorhamnetin. The invitro pharmacological activities were performed for the

methanolic extract of the whole plant of Asystasia gangetica.

The methanolic extract was selected for the pharmacological activities because it showed the

maximum anti-inflammatory effect as reported in literature. Protein Denaturation produces auto antigens

which may be a cause for the development of Rheumatoid arthritis. The study of Anti arthritic activity of

the methanolic extract showed a dose dependent inhibition of protein Denaturation and was comparable

to that of the standard. The methanolic extract also exhibited a dose dependent inhibition of ADP induced

platelet aggregation. The methanolic extract was also evaluated for its Anthelmintic activity. It showed a

dose dependent decrease in time taken for paralysis and death of worms. The methanolic extract and

Flavonoid Mixture fraction were subjected to tests on blood viscosity. The Flavonoid mixture fraction

exhibited a better decrement in blood viscosity than the methanolic extract.

KEY WORDS: Asystasia gangetica, Anti-Oxidant, Anti-Arthritic, In-vitro, Anthelmintic

INTRODUCTION

Plants play a pivotal role in health care. According to World Health Organisation (WHO), 80% of the

world’s population relies on traditional medicine, particularly plant drugs for primary health care. The practise of

traditional medicine is not new in India since it is the birth place of many traditional practices like Ayurveda,

Siddha and Unani. India is particularly well endowed with over 6000 medicinal plants and well recorded practical

knowledge of traditional medicine. The scientific mind will not be satisfied by mere claims no matter from

whatever source they originate, unless corroborated by experimental and clinical evidences. As it is evident that

plants are treasure house for many potent medicines, it is important to scientifically evaluate the traditional

practices as well as upgrade the existing knowledge and make it available to the general public.

The very important plant molecules like Digitoxin, Morphine, Vincristine, Quinine etc have been used as

prototypes for the discovery of newer synthetic molecules. The frequency of life threatening diseases and ailments

has increased worldwide and it is becoming an important cause of morbidity and mortality in developing

countries. Majority of the diseases like Atheroscelorosis, Arthritis, Diabetes, and Cancer occur due to free radical

generation. Plants contain naturally occurring anti-oxidants like Flavonoids, Tannins etc which can be utilised for

scavenging free radicals. Many of the plant Alkaloids affect the nervous system and hence have been used as

Anaesthetics, Psycho stimulants, Motor end Depressants etc.

Asystasia gangetica is an ornamental plant and has been used as a source of nutrition in times of food

scarcity. It is also used traditionally for many ailments and diseases. Already the traditional use of the leaves as

anti-asthmatic (Akah PA, 2003) was scientifically proven. But no work on its anti-arthritic, Anthelmintic and anti-






0

20

40

60

80

100

Pe

rce

nt

inh

ibit

ion

Concentration µg/ml

Methanolic Extract

platelet properties was performed. It was in this context that the multiple pharmacological activities of the whole

plant of Asystasia gangetica were investigated in the course of this study. Also an attempt was made to isolate the

possible bioactive molecule responsible for the pharmacological actions. Asystasia gangetica (Linn.) T.Anderson

is commonly known as Chinese violet. ‘Asystasia’ means inconsistency and it relates to the fact that the corolla is

slightly regular which is an unusual characteristic in the Acanthaceae family. The word ‘gangetica’ is derived

from Ganges River where it is presumed to occur. It is locally used as a potherb and leafy vegetable in times of

food scarcity. It is also promoted as cover plant in orchards as it checks soil erosion and prevents growth of

noxious weeds. It also attracts bees to orchards. It has high nutritive value and hence used as forage for cattle,

goats and sheep in south East Asia. It is also used as an ornamental plant.


a. Plant material: The whole plant was collected from Tirumala hills, Andhra Pradesh, in the month of

December 2009. The plant material was authenticated by National Institute of Herbal Science (PARC/2010/542),

West Tambaram, Chennai.

b.Preparation of the extract: The freshly collected whole plant was dried in shade and coarsely powdered with a

blender. 200 grams of the powder was subjected to continuous hot percolation using Soxhlet’s apparatus with the

solvent methanol for 48 hrs. The solvent was recovered by distillation in Rotary Vacuum Evaporator at 80oC. The

residue was stored in a desiccator and used in further studies.

Fig 1: Aerial parts of Asystasia gangetica Fig 2: Asystasia gangetica habit

c. Phytochemical Evaluation: Compounds like Flavonoids, Alkaloids, Tannins, and Glycosides etc. are

responsible for many pharmacological activities of a plant. Phytochemical evaluation gives the chemical nature of

the bioactive molecules responsible for pharmacological activity in a plant.

d. Pharmacological studies: The following Pharmacological studies has been carried out on the whole plant of

Asystasia gangetic such as determination of Anti-arthritic activity by inhibition of protein denaturation

(Mizushima et al., 1966 ), In - vitro determination of anti-platelet activity by inhibition of platelet aggregation

induced by ADP, In - vitro anthelmintic activity (Fabiyi, 1986) and Effect of plant extract on blood viscosity.


In- Vitro Pharmacological Activities

1. Anti-arthritic activity by inhibition of protein denaturation method: The percentage of inhibition of protein

Denaturation of the methanolic extract and the standard Diclofenac sodium are tabulated in Table.1 & represented

in fig 3. Table.1 Percent inhibition of Protein Denaturation by

Methanolic Extract & Diclofenac sodium

Fig 3: Anti-Arthritic activity of Asystasia gangetica

Concentration

μg/ml

Percentage Inhibition

of Methanolic extract

Percent Inhibition of

Diclofenac sodium

10 17.29 -

50 23.61 -

100 33.34 -

200 42.70 84.47

400 58.11 -

800 65.87 -

1000 78.94 -

The methanolic extract exhibited a dose dependent inhibition of Protein Denaturation for Anti Arthritic activity.

The maximum percent inhibition (78.94%) was exhibited by 1000 µg/ml of the methanolic extract.





2. Anti-platelet activity by inhibition of ADP induced platelet aggregation: The percentage transmittance of

the methanolic extract and the standard drug aspirin in different time intervals are tabulated in Table.2 &

represented in fig 4

Table.2 Percent transmittance of methanolic Extract & Aspirin for

Anti-Platelet Activity

Concentration

(µg/ml)

Percentage transmittance

0min 1min 2min 3min 4min 5min

100 2.311 2.433 2.453 2.546 2.743 2.764

200 3.764 3.001 3.437 3.813 3.760 3.941

400 4.111 4.024 4.512 4.589 4.708 4.800

500 5.452 5.675 5.876 5.783 5.863 5.944

ASPIRIN 6.769 6.654 6.540 6.502 6.471 6.411

Fig 4: Percent transmittance of Methanolic Extract & Aspirin for

Anti-Platelet Activity

The methanolic extract showed a dose dependent inhibition of aggregation for Anti Platelet activity and the

maximum inhibition was exhibited by 500 µg/ml of Methanolic extract.

3. In- Vitro Anthelmintic Activity

The average values of the time taken for paralysis and death of Pheretima posthuma are tabulated in Table.3 and

represented in fig 5

Table.3 Time taken for Paralysis & Death in Albendazole and Methanolic Extract treated groups

Group Solution Concentration

(mg⁄ml)

Time taken for

paralysis (min)

Time taken for

death (min)

I Control Solution - - -

I Albendazole

(Standard)

10 38±0.3 56±0.3

I Test Extract 10 45±0.2 54±0.5

50 37±0.6 43±0.3

100 25±0.4 30±0.8

200 19±0.9 24±0.4

Values are Mean± SEM of 6 observations.

0

2

4

6

8

100 200 400 500 ASPIRIN

Pe

rce

nt

Tran

smit

tan

ce


0 minute

1 minute

2 minutes

3 minutes

4 minutes

5 minutes





Fig 5: Time taken for Paralysis & Death in Albendazole and

Methanolic Extract treated groups The Methanolic extract showed a dose dependent decrease in time taken for paralysis and death of Pheretima

posthuma. The maximum dose of 500 mg/ml of methanolic extract showed the least time taken for paralysis and

death of the earth worms.

10 mg/ ml of Methanolic

Extract 50 mg/ ml of Methanolic Extract 100 mg/ ml of Methanolic

Extract

200 mg/ ml of Methanolic

Extract 10 mg/ml of Standard solution Control solution

Fig.6. In Vitro anthelmintic activity of various concentrations of methanolic extract of whole plant of

Asystasia gangetica

0

10

20

30

40

50

60

10 50 100 200

Tim

e in

min

ute

s

Concentration mg/ml

Methanolic Extract (Paralysis time in min) Methanolic extract (Death time in min) Albendazole (paralysis time)

Albendazole (death time)





4) Effect of methanolic extract and flavanoid mixture fraction on blood viscosity

The viscosity of Methanolic extract treated blood is tabulated in Table.4 and represented in fig 7.

Table.4 Effect of Methanolic Extract on Blood Viscosity in time span of 90 minutes

Concentration

(µg/ml)

0

minute

30

Minutes

60

minutes

90

minutes

Relative blood

viscosity

100 2.2 2.2 2.1 2.1

2.2 200 2.1 2 2 1.9

300 1.9 1.9 1.9 1.8

400 1.8 1.7 1.7 1.6

500 1.6 1.6 1.5 1.5

Fig: 7 Effect of Methanolic extract on Blood viscosity

The effect of various concentrations of Flavanoid fraction on blood viscosity at various time intervals is

tabulated in Table.5 and represented in fig 8.

Table.5 Effect of Flavonoid Mixture Fraction on Blood viscosity in time span of 90 minutes

Concentration

(µg/ml)

0minute 30minutes 60minutes 90minutes Relative blood

viscosity

100 2.2 2.1 1.9 1.9

2.2

200 1.9 1.8 1.8 1.7

300 1.7 1.7 1.6 1.6 400 1.6 1.5 1.5 1.5 500 1.5 1.5 1.4 1.4

Fig 8: Effect of Flavonoid Mixture Fraction on Blood viscosity

The methanolic extract showed a dose dependent decrease in Blood viscosity in a span of 90 minutes. The

Flavanoid Mixture fraction showed better decrease in viscosity than methanolic extract.

0

0.5

1

1.5

2

2.5

100 200 300 400 500

Vis

cosi

ty

concentration µg/ml

0 minute

30 minutes

60 minutes

90 minutes

Relative viscosity

0

0.5

1

1.5

2

2.5

100 200 300 400 500

Vis

cosi

ty


0 minute

30 minutes

60 minutes

90 minutes

Relative viscosity





CONCLUSION The study justified the use of the plant as a food source by quantifying the nutritive value. The study also

justified the use of plant as Anthelmintic and anti-Arthritic in folklore medicine. The methanolic extract also

exhibited inhibition of platelet aggregation and decrease in blood viscosity. Also this is the first report of the

flavonoids Luteolin, Quercetin, Kaempferol and Isorhamnetin being isolated from the Methanolic Extract of the

whole plant of Asystasia gangetica. The study also reports that the decrease in blood viscosity by the methanolic

extract was due to the Flavonoid mixture, which was isolated by column chromatography.

REFERENCES

Akah PA, Ezike AC, Nwafor SV, Okoli CO, Enwerem NM, Evaluation of the anti asthmatic property of Asystasia

gangetica leaf extracts. Journal of Ethnopharmacology, 89, 2003, 25-36.

Athnasiadau S, Interaction of anti-inflammatory drugs with serum proteins especially with some biologically

active proteins. J.pharma.pharmacol, 20, 1968, 69-73.

Athnasiadau S, I Kyriazakis, F Jackson, RL Coop, Direct Anthelmintic effects of condensed tannins towards

different gastrointestinal nematodes of sheep: in vitro and in vivo studies, Vet.Parasitol, 99, 2001, 205-219.

Brown JH, Inhibition of heat-induced denaturation of serum protein by mixtures of non-steroidal anti-

inflammatory agents, Proc.soc.exp.biol.med, 128, 1968, 225-228.

Chamundeswari D, In vitro antiplatlet activity-guided fractionation of aerial parts of Melthoria maderaspatana.

Indian Journal of Pharmaceutical sciences, 68(5), 2006, 668-670.

Connar AO, The in vitro anti-denaturation effects induced by natural products and non-steroidal compounds in

heat treated (immunogenic) bovine serum albumin is proposed as a screening assay for the detection of anti-

inflammatory compounds, without the use of animals, in the early stages of the drug discovery process, West

Indian med j, 57, 2008, 327-331.

Ezike AC, Akah PA, Okoli CO, Bronchospasmolytic activity of the extract and fractions of Asystasia gangetica

leaves. International Journal of Applied Research in Natural Products, 3, 2008, 8-13.

Hack B, Analytical method of determination of mineral& metallic elements. Hand book of Analytical practice.

Dolphin ands, John(Ed) N.Y.Incorp.press ltd, 2000, 126-149.

Harborne, J.B, Phytochemical methods, Chaponann & Hall, London, 1973, 1-271.

Iqbal, Z, Lateef M, Ashraf M, Jabbar A, Anthelmintic activity of Artemisia brevifolia in sheep. J.of

Ethnopharmacology, 93, 2004, 265-268

John S, Estimation of total Flavonoid content by two complementary colorimetric methods, Journal of food

&drug analysis, 10, 2002, 178-182.

Kokate, CK Texbook of Pharmacognosy, Nirali prakashnan, Publication, 1985 112-124.

Kritikar, K.R and B.D Basu, Indian Medicinal plants 2nd

E d., Basu, Allahabad, India, 1933, 231-236.

Kumar R, Journal of Experimental Pharmacology, 2, 2010, 29-36.

Lancet.S, Screening test for anti-rheumatic drugs, j.nat prod, 2, 1966, 443-445.





INVESTIGATION OF IN-VITRO ANTI-OXIDANT, ANTI-INFLAMMATORY

AND ANTI-ARTHRITIC ACTIVITY OF AERIAL PARTS OF SECURINEGA

LEUCOPYRUS (WILLD.) MUELL T.K.Gopal*, T.Sheela, D.Chamundeeswari, C.Umamaheswara Reddy

Faculty of Pharmacy, Sri Ramachandra University, Chennai, India

Corresponding author: E.Mail: [email protected]

ABSTRACT

Securinega leucopyrus (willd.) Muell. (Euphorbiaceae) is a thorny, large shrub or small tree. In

Ayurveda it has a lot of potential medicinal uses. The decoction of leaves are used to dress the

cancerous wounds. The juice or paste of the leaves along with tobacco used to destroy worms in

sores. Its leaf decoction useful externally in the treatment of piles. The present study is designed to

carry out Anti-oxidant, Anti-inflammatory, Anti arthritic activity by Invitro methods. The Securinega

leucopyrus was authenticated and the anatomical features were done which shows calcium oxalate

crystals, fibres, glandular trichomes, sclereids, vascular bundles (secondary xylem and phloem). The

microscopical evaluation also performed by determination of stomatal index and vein islet number.

The In-vitro anti-oxidant activity revealed that the Alcoholic extract showed more activity as

compared to Chloroform, Ethyl acetate, n-Hexane. In DPPH (1,1-diphenyl-2-picrylhydrazyl) radical

method Alcoholic extract showed more activity and in nitric oxide scavenging method Hydro

alcoholic, Alcoholic extracts showed more activity. Chloroform extract showed more activity in

hydroxy radical method. In Total anti-oxidant method Alcoholic extract showed more activity. The

Anti-inflammatory activity also performed by Human Red Blood Corpuscle (HRBC) Membrane

Stabilization Method. The results were tabulated and the values are represented in the figure. In this

Ethyl acetate extract showed more activity. The Anti-arthritic activity is performed by inhibition of

protein denaturation method. And the results are given in table and values are represented in graph.

This shows Alcoholic extract gave more activity. The above results shows that the presence of

phytoconstituents like Glycosides, Flavanoids, Alkaloids, steroids and Tannins may be responsible for

Anti-oxidant, Anti-inflammatory, Ant arthritic activities.

Key words: Securinega leucopyrus, Anti-oxidant, Anti-inflammatory, Anti-arthritic.

INTRODUCTION

Plants are used as medicine since time immemorial. Approximately one-third of the top selling drugs

in the world are derived from natural products and their derivatives. The traditional approach makes use of

material that has been found by trial and error over many years in different systems of medicine (Cotton,

1996). Plants are used for discovery of modern medicine in four basic ways, as a source of direct medicinal

agents, serve as a raw material base for elaboration of more complex semi-synthetic chemical compounds,

chemical structure derived from phyto constituents can be used as models for new synthetic compound (Anon,

1994), Plants can be used as taxonomic markers for discovery of new therapeutic compounds (Cox.P.A,

1990). According to the world health organization (WHO), (Murry et al.,) 80% of the world’s population has

faith in traditional medicine, particularly plant drugs for their primary healthcare.

India is a goldmine of well recorded and traditionally well practical knowledge of herbal medicine.

India officially recognizes over 3000 plants for their medicinal value. It is estimated generally that over 6000

plants in India are use in traditional, folk and herbal medicine, representing 75% of the medicinal needs of the

third world countries. The frequency of life threatening infections has increased worldwide and it is becoming

an important cause of morbidity and mortality in immuno-compromised patients in developing countries. Eg:

Cancer, Cardio vascular diseases, Rheumatoid arthritis, Cataracts, Alzheimer’s diseases and inflammatory

disorders. (Dev,1999). Several anti-oxidants of plants origin are experimentally proved and used as effective

protective against oxidative stress. They play an important role in major health problems like cancer. Free

radicals are produced in large amount during metabolic disease conditions like diabetes, hypertension,

atherosclerosis, urolithiasis, ulcers etc. This free radicals attack DNA, protein molecules, enzymes and cells

leading to change in genetic material and cell proliferation (Cancer).

Plants which contain carotenoids, flavonoids and Tannins can be utilized to scavenge the excess free

radicals from human body. An inflammatory disease like Rheumatoid arthritis is one of the most common

immuno inflammatory conditions affecting the population worldwide. About 1% of the worldwide adult

population is affected by Rheumatoid arthritis with above twice as many females suffers as male. In recent

years, attempts have been made to investigate the indigenous drug against Arthritis. Research in the field of

indigenous plant is a significant aspect of developing a safer anti arthritic principle through isolation,






characterization, identification and biological studies. (Pratt, 1992). Securinega leucopyrus (willd.) Muell,

belonging to the family Euphorbiaceae commonly known as Bushweed, Indian snow berry, is a thorny woody

shrub or a small tree distributed in different parts of India (Ayensu, 1996). Its leaves and fruits are edible. The

berry is sweet. The slender branches are reported to be utilized for preparing wicker-baskets and for thatching

(Kirtikar, 1918). The leaves of the plant contain germicidal properties.

In Chattisgarh, the decoction of leaves is used to dress the cancerous wounds. It is used in

combination with tobacco. The juice or paste of the leaves along with tobacco used to destroy worms in sores.

Its leaf decoction useful externally in the treatment of piles and it is used to wash the wounds of cattle. It is

used as popular veterinary medicine (Bakshu, 2001). The other uses are sweet, cooling, diuretic, aphrodisiac,

tonic and are useful in vitiated conditions of pitta, burning sensation, strangury, seminal weakness, general

debility, larvicide, paralysis, paresis, piscicide, insecticide. The anti-microbial properties of securinega

leucopyrus is already reported (Bakshu, 2001). The leaves are set as a disinfectant and its paste is used to

extract the extraneous materials from body tissues without surgery. Even though Securinega leucopyrus has

lot of potential medicinal uses, the study on this plant is very less. Considering the importance of this plant,

the present study was undertaken.

Plant collection: It consists of dried aerial parts of Securinega leucopyrus (willd.) Muell. (Euphorbiaceae) is a

thorny, large shrub or small tree. The plant Securinega leucopyrus were collected from Tribal Women’s

Welfare Trust’s Herbal Garden, Thandarai, Chengalpattu (Dist), Tamilnadu in the month of December and

authenticated at National Institute of Herbal Science, West Tambaram, Chennai (PARC/2009/289).

Fig 1: Morphology of Securinega Leucopyrus (willd.) Muell

METHODS AND MATERIALS

Collection and processing of plant material: Then the aerial part of plant material was cut into small

fragments and dried in shade until the fracture is uniform and smooth. The dried leaves, stem portions were

separately powdered by using a blender and sieved in sieve No 60 (Gahan, 1984). The coarse powder 500 gm

was subjected to maceration for 72 hours, followed by exhaustive maceration for 48 hours by using various

solvents of increasing polarity [n- hexane> Chloroform> Ethyl acetate>Alcohol> Water] by decanting and

drying the marc after each extraction. The solvents were recovered by distillation of the extracts at 750C to

800C. The extracts were dried under desiccator and the percentage yield was calculated. The residues obtained

were used for the screening of the phytochemical analysis. These n-Hexane, Chloroform, Ethyl acetate,

Ethanol and Hydroalcoholic extracts were used for the preliminary phytochemical investigation,

chromatographic analysis and its pharmacological evaluation (Sundaresan, 1991).

In-Vitro Pharmacological Evaluation: The aerial part of the plant of Securinega leucopyrus (willd.) Muell

had been stuied for its anti-oxidant activity by DPPH method (Yohozawa, 1998), nitric oxide scavenging

activity (Alderson, 2001), determination of total anti-oxidant activity (Prieto, 1999) scavenging of hydroxyl

radical deoxyribose method (Elizabeth, 1990), in-vitro anti-inflammatory activity by human red blood

corpuscle (HRBC) membrane stabilization method, in-vitro anti-arthritic activity by inhibition of protein

denaturation method.


Anti-oxidant activity by DPPH radical scavenging method: The chloroform and ethyl acetate, alcohol,

hydro alcohol extracts showed a dose dependent increase in anti-oxidant activity in DPPH method. About

82.5%, 88.42% were observed in Chloroform and Alcoholic extracts of Securinega leucopyrus. But in Hexane

extract the effect was decreased when dose increases. The presence of flavanoids, Alkaloids, Tannins and

Steroids in these extracts may be responsible for free radical scavenging activity. From the results it is made

clear that Securinega leucopyrus possess free radical scavenging property and the order was, Alcohol >

Chloroform > Ethyl acetate > Hydro alcohol >Hexane. DPPH is a relatively stable free radical. The assay is

the measurement of scavenging ability of Antioxidants towards the stable radical DPPH. The plant containing





flavanoids, Alkaloids, Tannins and Steroids reduces the radical to the corresponding hydrazine when it reacts

with a hydrogen donors in the anti-oxidant principle. In DPPH method results are highly reproducible. The

active constituent present in the plant donates an electron to reduce the DPPH radical to its corresponding

hydrazine. The chemical constituents present in this plant like Flavanoids, Tannins, Alkaloids and Steroids

may be responsible for this activity. They shows the anti-oxidant activity by inhibition of enzymes involved in

oxidation systems.

Anti -Oxidant Activity by Nitric Oxide Scavenging Method: All extracts of Securinega leucopyrus shown

a dose dependent increase in Nitric oxide scavenging property .About 85% inhibition was observed by

Hydroalcoholic extract. But in Hexane and Ethyl acetate extracts the effect was very less compared to

standard. The presence of flavonoids Tannins and steroids in these extracts may be responsible for Nitric

oxide scavenging activity. Nitric oxide is a radical produced in mammalian cells, involved in the regulation of

various physiological processes. In the present study the nitrite produced by the incubation of sodium nitro

prusside in standard phosphate buffer at 25◦

c was reduced by plant extract. This may be due to the anti-

oxidant principles present in the plant extract which compete with oxygen to react with nitric oxide, there by

inhibiting the generation of more deleterious products such as Nitric anhydride (N 2O3 ) and perhydroxy nitrite

(ONO ---

) (Chen et al 2001). This activity is due to the presence of flavonaids, Tannins and steroids. They

inhibit the free radicals by inhibition of enzymes involved in oxidation systems (5- Lipoxygenase,

cycloxygenase, mono oxygenase, xanthine oxidase.

Determination of total anti-oxidant activity: The Ethanolic, Hydro alcoholic extracts showed maximum

effect due to the presence of flavanoids, Reducing sugars, Alkaloids, Tannins and steroids. The values were

expressed as equivalents of Vitamin E. From the results it is made that Securinega leucopyrus possess free

radical scavenging activity through anti-oxidant property.

Anti-oxidant activity By Hydroxy radical scavenging method: All the extracts of Securinega leucopyrus

shown a dose dependant increase in nitric oxide scavenging property. About 87.11% inhibition was observed

by Alcoholic extract. The Chloroform and Hydroalcoholic extract showed 78.56%, 78.52% inhibition. But in

Ethyl acetate and Hexane extract the effect was very less compared to standard. The presence of flavanoids

Tannins and steroids in these extracts may be responsible for Hydroxy radical scavenging activity. Ferrous

salts can react with H2O2 and form hydroxyl radical via Fention’s reaction. The iron required for this reaction

is obtained either from the pool of iron or the heme containing protein. The hydroxyl radical (OH)-

thus

produced may attack the sugar of DNA deoxy causing ribose fragmentation , base loss, and DNA strand

breakage. The generation of (OH)- in fenton reaction is

due to the presence of iron ions. When the Fe

2+/Fe

3+

redox couple is bound by certain chelators, the OH● fragmentation is prevented, whereas the increased colour

formation in the absence of crude extracts were observed in deoxy rebose assay. In this, the extract act as a

chelator of iron ions , binding to them, & preventing the formation of free radicals, though the extracts not

directly involved in the OH● scavenging . The results indicates

that the extracts of plant plays a major role in

the inhibition of ribose fragmentation and hence the decreased color formation in the deoxy rebose assay. The

free radical scavenging property of the crude extracts of plant against DPPH, Nitric oxide, Hydroxy radicals

and the Total anti-oxidant activity is clearly understood from the results of this chapter. The phytochemical

screening of the extract revealed the presence of flavonoids, tannins which is responsible for anti-oxidant

property.

Anti-inflammatory activity by HRBC membrane stabilization method: The all extracts shows a dose

dependent increase in Anti-inflammatory activity in HRBC membrane stabilization method. About 89.66%,

87.42%, 72.32% inhibitions were observed in Ethyl acetate, Chloroform and Alcoholic extracts. But in

Hexane and Hydro alcoholic extracts the effect was very less when compared to standard Diclofenac sodium.

This anti-inflammatory activity may be due to the presence of flavonoids, Tannins and Alkaloids. Flavonoids

may produce their Anti-inflammatory effect by a multitude of ways to inhibit the inflammatory processes.

Formation and release of various mediators of inflammation like Histamine and Prostaglandin are affected by

Flavanoids. They inhibit the increased capillary permeability during inflammation. The adhesion of leucocyte

to endothelial surface and subsequent migration is influenced by flavanoids. They inhibit the prostaglandin

and Leucotriene C4 in human platelet. They were investigated for lipoxygenase inhibitory activity. They

inhibits cytokine release from cells.

Anti-Arthritic Activity by Inhibition of Protein Denaturation Method: The results of anti-arthritic activity

of all extracts of Securinega leucopyrus on inhibition of protein denaturation method as shown in table18 and

figure 5.6. The Ethanol, Chloroform, Hydro alcoholic extracts showed maximum activity when compared to

n-Hexane and Ethyl acetate extracts. The effect is represented as, Ethanol > Water > Chloroform > Ethyl

acetate > Hexane extracts. The effect may be due to Alkaloids, Tannins, Flavanoids, Triterpenes and Reducing





sugars. Denaturation of protein is one of the cause of Rheumatoid arthritis. Production of auto antigen in

certain arthritic disease may be due to denaturation of proteins. The mechanism of denaturation probably

involves alteration in electrostatic hydrogens and hydrophobic di sulphide bonding. The present study,

Securinega leucopyrus capable of controlling the production of auto antigens and thereby it inhibit the

denaturation of protein in Rheumatic diseases.

Tab 1: The Percent inhibition of different extracts of Securinega leucopyrus on DPPH radical

scavenging activity

Concentrations in μg/ml

Hexane

extract CHCL3

extract EtoAc

extract Alcoholic

extract Hydoalcholic

extract Percent Inhibition of

Standard (Vit. E) 10 19.50 25.25 23.28 24.45 20.36 - 50 26.51 29.91 26.74 28.43 20.13 32.5

100 47.91 55.23 50.46 51.58 40.01 62.88 200 39.66 68.58 62.85 58.34 52.41 81.02 400 39.51 74.62 71.22 75.66 65.32 95.24 800 37.87 74.56 75.95 76.3 62.45 - 1000 34.77 82.51 78.32 88.42 67.41 -

Fig. 2: Effect of securinega leucopyrus on DPPH method

Tab 2: Percentage inhibition of different extracts Securinega leucopyrus by Nitric Oxide Scavenging

method.


Hexane

extract CHCL3

extract EtoAc

extract Alcoholic

extract Hydrialcocolic

extract Percent Inhibition of

Standard (Vit. E) 10 15.43 18.43 18.34 20.35 25.35 - 50 18.34 20.65 21.45 24.17 26.13 28.40

100 22.51 28.47 20.36 35.11 38.53 47.78 200 38.62 46.44 29.4` 62.43 69.42 71.34 400 45.23 55.91 37.22 63.12 67.37 87.28 800 47.54 52.13 37.13 62.56 74,91 - 1000 54.33 71.44 45.78 77.12 85.38 -





Fig 3: Effect of Securinega Leucopyrus on Nitric Oxide Scavenging activity

Determination of Total Antioxidant Activity: In this investigation the absorbance (Y-axis) of different

extracts were extrapolated to the concentration (X-axis) of standard Vit – E.

Fig 4: Determination of total anti-oxidant activity

Tab 3: Absorbance of Test extracts plotted on the above graph is as follows

Extracts 200 µg/ml

Hexane 0.6215

Chloroform 1.2710

Ethyl acetate 1.0126

Ethanol 1.4205

Hydroalcohol 1.3010

Tab 4: Estimation of Total anti-oxidant activity

Concentrations of

extracts in μg/ml

Equivalent to Standard Vit – E in μg/ml

Hexane Chlorofom Ethylacetate Ethanol Hydroalcohol

200 36.5 68 61.5 83.5 72.7 The above table shows the extrapolated values. Hence the effect of 200µg/ml of extracts is equivalent to the

effect of µg/ml of Vit – E with respect to the extracts.





Tab 5: Percent inhibition of Securinega leucopyrus in Hydroxyl Radical Scavenging method


Hexane

extract CHCl3

extract EtoAc

extract Alcoholic

extract Hydro

alcocolic

extract

Percent Inhibition of

Standard (Vit. E)

10 21.91 28.54 29.44 28.22 18.62 - 50 34.56 38.55 28.41 37.45 22.45 40.34

100 33.42 51.91 38.42 35.66 28.64 62.99 200 45.82 66.34 48.54 47.44 34.25 85.59 400 60.44 75.66 66.69 59.23 43.88 95.01 800 65.71 75.52 67.23 61.03 50.52 -

1000 77.30 86.12 71.56 66.45 58.34 -

Fig 5: Effect of Securinega Leucopyrus on Hydroxy Radical Scavenging Activity

Tab 6: Percent HRBC Membrane Stabilization of different extracts of Securinega leucopyrus


Hexane

extract Chloroform

extract

Ethyl

acetate

extract

Alcohol

extract

Hydro

alcohol

extract

Percent

stabilization of

Diclofenac sodium 10 29.77 34.21 35.67 25.14 20.67 -

50 29.93 34.52 36.73 28’56 29.54 -

100 40.56 42.51 47.43 32.56 37.54 -

200 47.82 55.32 68.23 45.06 36.77 93.52

400 43.18 69.76 71.94 62.45 34.52 -

800 59.43 75.87 82.54 62.44 40.54 -

1000 65.88 87.42 89.66 74.32 56.33 -

Fig 6: Effect of Securinega leucopyrus on Anti-inflammatory Activity





Tab 7: Percent denaturation of protein of different extracts of Securinega leucopyrus


Hexane

extract CHCL3

extract Ethanolic

extract Alcohol

extract Hydro

alcohol

extract

Percent stabilization of

Diclofenac sodium

10 18.34 26.95 27.57 29.45 24.55 - 50 25.67 32.86 32.74 38.56 37.13 -

100 38.33 48.21 30.51 40.54 39.45 - 200 37.34 47.54 38.43 57.82 56.99 94.14 400 45.81 56.90 62.55 67.90 57.82 - 800 45.58 69.44 74.21 73.51 67.95 - 1000 52.80 78.52 76.4 87.11 78.56 -

Fig 7: Effect of securinega leucopyrus on Anti Arthritic Activity

CONCLUSION

From this study we can conclude that Securinega leucopyrus showed the presence of Alkaloids,

Sugars, Flavonoids and Tannins. This is identified by phytochemical analysis and confirmed by thin layer

chromatography. The present study on Securinega leucopyrus showed that the plant has moderate anti-oxidant

activity and it compared with standard vitamin E. The plant also showed anti-inflammatory and anti-arthritic

activity and it compared with standard Diclofinac sodium. Chloroform, Ethyl acetate, Ethanol and

Hydroalcohol fractions are having significant anti-inflammatory and antiarthritic activity may be due to the

presence of alkaloids, Phytostrols, Flavonoids, saponins, Tannins, and Lignans. The future work will be the

determination of Anti-oxidant, Anti-inflammatory and Anti arthritic activities by Invivo methods.

REFERENCES

Alderson WK, Cooper CE, Knowels RG, Nitric oxide systhesis, structure, function & inhibition, Journal of

Biochemistry, 2001, 357:593-615.

Anon K, Ethnobotany in search for new drugs. Ciba foundation symposium, John Wiley & sons, Newyork,

1994,188.

Ayensu ES, World medicinal plant resourses. In conservation for productive agriculture (VL chopra and TN

Khoshoo, edds) ICAR, 1996, New Delhi, India-11-42.

Bakshu LM, Ram AJ, Raju RBV, Fitoterapia, Volme-72, No-8, Dec 2001, 930-9330

Chen Y, Zhen R, Jia Z, Flavonoids as super oxide scavengers and anti-oxidants, Free radical Bio Med, 9,

1990, 19-26.

Cotton, 1996, The traditional approach makes use of material that has been found by trial and error over many

years in different countries and systems of medicine

Elizabeth K and Rao MNA, 1990, Oxygen radical scavenging activity of curcumin, Indian J Pharm, 58, 1990,

237.





Gahan, 1984, Plant Histo chemistry and phyto chemistry. An introduction academic press, Florida, U.S.A,

1984, 300.

Kirtikar KR, Basu B.D, Indian medicinal plants, volm-5, 1918, 97-98.

Murray CJL and Lopez AD, The global burden of Diseases.1996.

Pratt DE, natural anti-oxidants from plant material, in phenolic compounds in food & their effects on health II:

Anti-oxidants and cancer preservation (AVS symposium series 507) edited by M. Hang, (American chemical

society, Washington DC), 1992, 54.

Prieto P, Pineda M, Aguilar M. Spectrophotometric quantification of antioxidant capacity through the

formation of a phosphomolybdenum complex: specific application of vitamin E. Anal. Biochem, 269, 1999,

337-341.

Sundaresan V, De Britto AJ, Preliminary studies on some Phyto medicinal plants of thirunelveli hills.

Journal of economic and taxonomic Botany 23, 1991, 377-38.

Yohozawa T, Chen CP, Dong E, Tanaka T, Nonaka GT, Nishioka I, Studies on the inhibitory effect of

Tannins & flavonoids against radical. Bio chemical pharmacology, 56, 1998, 212-222.



Sampath Kumar et.al. Indian Journal of Research in Pharmacy and Biotechnology


TRANSDERMAL SONOPHORESIS TECHNIQUE- AN APPROACH FOR

CONTROLLED DRUG DELIVERY K.P.Sampath Kumar

1*, Debjit Bhowmik

2, M.Komala

2

1. Department of Pharmaceutical Sciences, Coimbatore medical college,

2. Karpagam University, Coimbatore

*Corresponding author: Email:[email protected]

ABSTRACT

Sonophoresis (phonophoresis) has been shown to increase skin permeability to various low and

high molecular weight drugs, including insulin and heparin. However, its therapeutic value is still being

evaluated. Some obstacles in transdermal sonophoresis can be overcome by combination with other

physical and chemical enhancement techniques. Use of ultrasound in therapeutics and drug delivery has

gained importance in recent years, evident by the increase in patents filed and new commercial devices

launched. The present review discusses new advancements in sonophoretic drug delivery in the last two

decades, and highlights important challenges still to be met to make this technology of more use in the

alleviation of diseases. new formulations tried in sonophoresis, synergistic effects with techniques such as

chemical enhancers, iontophoresis and electroporation, as well as the growing use of ultrasound in areas

such as cancer therapy, cardiovascular disorders, temporary modification of the blood-brain barrier for

delivery of imaging and therapeutic agents, hormone replacement therapy, sports medicine, gene therapy

and nanotechnology.

Keywords: Sonophoresis, Permeability, higher frequency, Patient compliances.

INTRODUCTION

Sonophoresis is a process that exponentially increases the absorption of semisolid topical compounds

(transdermal delivery) into the epidermis, dermis and skin appendages. Sonophoresis occurs because

ultrasound waves stimulate micro-vibrations within the skin epidermis and increase the overall kinetic energy of

molecules making up topical agents. It is widely used in hospitals to deliver drugs through the

skin. Pharmacists compound the drugs by mixing them with a coupling agent (gel, cream, ointment) that transfers

ultrasonic energy from the ultrasound transducer to the skin. The ultrasound probably enhances drug transport

by cavitation, microstreaming and heating. Sonophoresis is also used without drug delivery in physical therapy,

and as a complementary modality for iontophoresis.

Application of ultrasound to the skin increases its permeability (sonophoresis) and enables the delivery of

various substances into and through the skin. Ultrasound has been used extensively for medical diagnostics and to

a certain extent in medical therapy.The generation of ultrasound and mechanism of sonophoresis with particular

emphasis on the role of cavitation (both inside and outside the skin), thermal effects, convective transport, and

mechanical effects also included. Sonophoresis is a localized, non-invasive, convenient and rapid method of

delivering low molecular weight drugs as well as macromolecules into the skin. The ultrasound waves generate

tiny bubbles of water on skin surface; this causes the skin surface to lightly get worn out. This allows the drug to

pass through the skin surface efficiently. Sonophoresis occurs because ultrasound waves stimulate micro-

vibrations within the skin epidermis and increase the overall kinetic energy of molecules making up topical

agents.

Sonophoresis, or ultrasound, creates holes in the skin, and allows fluids to travel into or out of the body.

When sound is emitted at a particular frequency, the sound waves disrupt the lipid bilayers. This method can be

used for delivery of steroids, systemic drugs such as Insulin and antigens for vaccination. Ultrasound transdermal

drug delivery system in noninvasive way is used for Diabetics to control blood sugar level through short term and

long term delivery of Insulin. Noninvasive drug delivery (as capsule formulation) is used for acne, psoriasis.

These systems enhance activity of transdermal patches. The higher the frequency, the more dispersed the

transmission.

Advantages of Using Sonophoresis as a Physical Penetration Enhancer

Enhanced drug penetration (selected drugs) over passive transport.

Allows strict control of transdermal penetration rates.

Low risk of introducing infection as the skin remains intact

Reduction of dosing frequency and patient compliance


http://en.wikipedia.org/wiki/Transdermal

http://en.wikipedia.org/wiki/Epidermis_(skin)

http://en.wikipedia.org/wiki/Dermis

http://en.wikipedia.org/wiki/Kinetic_energy

http://en.wikipedia.org/wiki/Pharmacists

http://en.wikipedia.org/wiki/Cavitation

http://en.wikipedia.org/wiki/Iontophoresis





Improved control of the concentrations of drugs with small therapeutic indices.

Reduction of fluctuations in plasma levels of drugs

Avoids hepatic first pass elimination and gastrointestinal irritation.

Substitute’s oral administration when the route is unsuitable as in case of vomiting, diarrhea.

Permit both local and systemic effects and less risk of systemic absorption than injection.

Easy termination of drug delivery in case of toxicity, through termination of ultrasound.

Disadvantages of using sonophoresis as a physical penetration enhancer

Stratum corneum must be intact for effective drug penetration.

Can be time consuming to administer

Applications of Sonophoresis:

1. Ultrasound helps in treatment of wide varieties of sports injuries such as tennis elbow, tendon problems,

repairing damaged ligaments, muscle spasms, stiff joints, fractured bones and cartilage. Also used in

healing of wounds, damaged skin, skin rejuvenation, nerve stimulation, and improving the strength and

elasticity of scar tissues.

2. Ultrasound with Topical Anesthesia rapidly decreases pain of intravenous cannulation.

3. Low frequency ultrasonic gene delivery.

4. The dolphin therapy arouses a great interest in the whole world, since it causes analgesic effects, removal

of depression, and improvement of learning abilities of the children suffering from autism.

5. Sonophoresis is also being used in drug enhancement in granulomas and tumors.

6. Sonophoresis is being investigated as a way of extracting compounds such as glucose.

7. In the treatment of sick fish by University of Maryland‘s Center of Marine Biotechnology. The current

method uses intraperitoneal injections which are costly and highly labour intensive. In this experiment,

ultrasound was applied to water containing fish and compound of interest. The ultrasound waves

increases the permeability of compound into the tissues of the skin and gills. This method is highly cost

and labour effective.

8. Sonophoresis also used in treatment of glaucoma, corneal infection and nail delivery, to Increase the

permeability of drugs.

Future Trends: Vaccination In recent years, the potential for exploiting the skin for purposes of vaccination has

received a great deal of attention. One common strategy is to use an adjuvant, which is a compound used to

enhance the immune response to vaccine compounds Ultrasound can be used to enhance skin permeability to both

the adjuvant and the vaccine, and hence to facilitate their delivery to the target cells. Gene therapy Gene therapy is

a technique for correcting defective genes that are responsible for disease development, most commonly by

replacing an ‘abnormal’ disease-causing gene with the ‘normal’ gene The most obvious candidate diseases for

cutaneous gene therapy are the severe forms of particular geno dermatoses (monogenic skin disorders), such as

epidermolysis bullosa. Other applications might be healing of cutaneous wounds such as severe burns and skin

wounds of diabetic origin.

CONCLUSION: Sonophoresis is the enhancement of migration of drug molecules through the skin by ultrasonic

energy Sonophoresis occurs because ultrasound waves stimulate micro-vibrations within the skin epidermis and

increase the overall kinetic energy of molecules When sound is emitted at a particular frequency, the sound waves

disrupt the lipid bilayers The higher the frequency, the more dispersed the transmission.

REFERENCES

Allen LV, Popovich NG, Ansel HC, Ansel’s pharmaceutical dosage forms and drug delivery systems.

Transdermal drug delivery systems. 8th ed. India: Gopsons papers ltd.; 2006, 298-315.

James Swarbrick, Transdermal Delivery: Sonophoresis, Encyclopedia of pharmaceutical technology, 3 rd edition,

Volume-6, 2007, 3828-3842.

Mitragotri S, Blankschtein D, and Langer R. Transdermal drug delivery using lowfrequency sonophoresis.

Pharmaceutical research, 13, 1996, 411-420.

Mr. Ashish Pahade, Dr. Mrs. V.M.Jadhav, Dr. Mr. V.J.Kadam, Sonophoresis: an overview, International Journal

of Pharmaceutical Science, 3(2), 2010, 24-32





N.K.Jain, Sonophoresis: Biophysical of Transdermal Drug Delivery, Controlled and Novel Drug Delivery, 1 st

edition, 1997, 208-235.

Ogura M, Paliwal S, Mitragotri S, Low-frequency sonophoresis: Current status and future prospects. Advanced

Drug Delivery Reviews, 60, 2008, 1218–1223.

Pahade A, Jadhav VM, Kadam VJ, Sonophoresis: an overview. International Journal of Pharmaceutical Sciences

Review and Research, 3, 2010, 24-32.

Tang H, Wang CCJ, Blankschtein D, and Langer R, An Investigation of the Role of Cavitation in LowFrequency

Ultrasound-Mediated Transdermal Drug Transport, Pharmaceutical Research, 19, 2002, 1160-1169.

Terahara T, Mitragotri S, Kost J, Langer R, Dependence of low-frequency sonophoresis on ultrasound

parameters; distance of the horn and intensity. International Journal of Pharmaceutics, 235, 2002, 35-42.

Tezel A, Sens A, Tuchscherer J, and Mitragotri S. Frequency Dependence of Sonophoresis, Pharmaceutical

Research, 18, 2001, 1694-1700.



Navin Dixit et.al. Indian Journal of Research in Pharmacy and Biotechnology


A COMPREHENSIVE REVIEW OF ELADI VATI Navin Dixit*, Sheo Dutt Maurya , Bhanu P.S.Sagar

I.E.C.College of Eng & Technology, Greater Noida (U.P)

*Corresponding author: Email: [email protected]

ABSTRACT

Herbal medicines are the synthesis of therapeutic experiences of generations of practising physicians

of indigenous systems of medicine for over hundreds of years while nutraceuticals are nutritionally or

medicinally enhanced foods with health benefits of recent origin and marketed in developed countries.

The marketing of the Herbal drugs under the category of the Neutraceutical is unethical. Herbal

medicines are also in great demand in the developed world for primary health care because of their

efficacy, safety and lesser side effects. They also offer therapeutics for age-related disorders like memory

loss, osteoporosis, immune disorders, etc. for which no modern medicine is available. Eladi Vati is a pill

containing cardamom, cinnamon, long pepper, dates, raisins and sugar. It treats bronchitis, cough, cold

and other respiratory diseases.

Keywords: Eladi vati , pill, Herbal medicines, respiratory diseases.

INTRODUCTION

India despite its rich traditional knowledge, heritage of herbal medicines and large biodiversity has a

dismal share of the world market due to export of crude extracts and drugs. WHO too has not systematically

evaluated traditional medicines despite the fact that it is used for primary health care by about 80% of the world

population? HERBAL medicine is still the mainstay of about 75–80% of the world population, mainly in the

developing countries, for primary health care because of better cultural acceptability, better compatibility with the

human body and lesser side effects. They have stood the test of time for their safety, efficacy, cultural

acceptability and lesser side effects. The chemical constituents present in them are a part of the physiological

functions of living flora and hence they are believed to have better compatibility with the human body. A soothing

remedy with Ela (Chhoti elaichi / Lesser cardamom), Mulethi, Misri, Munakka and other demulcents that is

extremely useful in respiratory and gastric manifestations, such as dry cough, sore throat, nausea, excessive thirst

and pittaja disorders.

Indications: Eladi Vati is extremely useful in the following conditions

Sore throat, dry cough and cold.

Chronic bronchitis.

Hiccups, nausea and vomiting.

Loss of appetite, hyperacidity and gastritis.

Burning micturition and dysuria.

Actions: Pacifies aggravated pitta.

Soothes the throat.

Relieves excessive thirst.

Vati & Gutika: Medicines prepared in the form of tablet or pills are known as Vati and Gutika. These are made

of one or more drugs of plant, animal or mineral origin. It has simple composition, no side effect, and

contraindication. It has been using since long time.

Benefits: Helps to relieve cough, cold, fever, hiccups, vomiting, dizziness, hematemesis (blood vomiting), and

abdominal pain.

It relieves excessive thirst, spleen diseases and gout.

Side Effects: There are no known side effects of this medicine.

Over-dosage may cause slight burning sensation in abdomen.

It is better to avoid this tablet during pregnancy





MATERIALS USED IN FORMULATION OF ELADI VATI

Table 1: Materials used in formulation of eladi vati

Ingredients English Name Scientific Name Quantity

Ela Cardamom Elettaria cardamomum 6 g

Patra Cinnamon Cinnamomum tamala 6 g

Twak Long pepper fruit Cinnamomum zeylanicum 6 g

Pippali Piper longum 24 g

Sita Sugar 48 g

Madhuka Licorice Glycyrrhiza glabra 48 g

Kharjura Dates Phoenix dactylifera 48 g

Draksha Raisin Vitis vinifera 48 g

Madhu Honey 48 g

EVALUATION PARAMETER OF ELADI VATI

Eladi Gutika (EG), a traditional Ayurvedic polyherbal formulation is used as a remedy

for Kasa (Cough), Svasa (Asthma), Bhrama (Vertigo),Raktapitta (Bleedingdisorders), Jvara (Fever), Amvata (Rhe

umatism) etc. In the present work, an attempt has been made to develop pharmacognostic standards for EG. The

raw materials of EG were subjected to proximate analysis prior to preparation of EG. EG was prepared using raw

materials of pharmacopoeial quality. Powder microscopy of EG showed the presence of discerning anatomical

characters which were present in the raw materials. A simple, rapid, accurate and sensitive HPTLC method was

developed and validated for the quantitation of piperine from EG. Method was validated as per ICH guidelines

and applied for stability studies of EG stored for different storage periods at room temperature. A comparative

evaluation of EG prepared in-house was carried out with three available marketed samples in terms of their

respective piperine content.

Acute toxicity study of EG in Albino Swiss mice revealed that it is safe at the dose of 2 g/kg body weight

in animal. Evaluation of EG by these scientific methods can be used as quality control tool for the manufacturing

and processing of EG. Eladi Gutika (EG), is an Ayurvedic polyherbal formulation of seven ingredients. Eladi Vati

is an Ayurvedic tablet used in treating cough, cold, fever and vomiting. It is used mainly in respiratory and

gastric conditions. Eladi Vati is a soothing remedy, which is prepared using various ingredients like Mulethi,

Misri, Ela, Munakka and other demulcents. All these ingredients are useful for sore throat, nausea, dry cough,

excessive thirst and pittaja disorders. It has simple composition, no side effect and contraindication. Piperine, a

major alkaloid of Pippali (one of the ingredient of EG) is reported as a bioavailability enhancer, anti-

inflammatory, anticonvulsant and antiulcer agent. There are reports on extraction of piperine using various

extraction techniques and its estimation using analytical tools from single herbs and polyherbal formulations. But,

there are no methods reported for its estimation from complex matrix of EG.

CONCLUSION

Eladi Vati is an Ayurvedic tablet used in treating cough, cold, fever and vomiting. It is used mainly in

respiratory and gastric conditions. It helps to relieve cough, cold, fever, hiccups, vomiting, dizziness,

haematemesis and abdominal pain. It relieves excessive thirst, spleen diseases and gout. It is a natural aphrodisiac

and useful in all bleeding conditions.

REFERENCES

Ayurvedic Formulary of India (AFI), Part I. Government of India, Ministry of Health and Family Welfare. New

Delhi: Department of Health, Controller of Publications Civil Lines; 1989, 181-182.

Bajada S, Singla AK, Bedia KL, Liquid chromatographic method for determination of piperine in rat plasma:

Application to pharmacokinetics, J Chromatogr B, 776, 2002, 245-9.

Capasso R, Izzo AA, Borrelli F, Russo A, Sautebin L, Pinto A, Effect of piperine, the active ingredient of black

pepper on intestinal secretion in mice, Life Sci, 71, 2002, 2311-7.

Evans WC, Trease and Evans Pharmacognosy, 14 th ed, London: SW Saunders Company Ltd, 1996, 319.

Hamrapurkar PD, Jadhav K, Zine S, Quantitative estimation of piperine in Piper nigrum and Piper longum using

high performance thin layer chromatography, J Appl Pharm Sci, 1, 2011, 117-20.





Patel RK, Kanani RJ, Patel VR, Patel MG, Development and validation of HPTLC method for simultaneous

quantification of vasicine and piperine in Vasavaleha, Int J Pharm Res, 2, 2010, 14-7

Shailajan S, Singh A, Tiwari B. Quality control and standardization of an ayurvedic Taila formulation. Int J

Biomed Res Anal, 1, 2010, 78-81.

Shailajan S, Yeragi M, Purohit A, Optimized separation and quantification of eugenol from a traditional Unani

medicine Jawarish-e-Bisbasa using HPTLC, Int J Pharm Sci Rev Res, 9, 2011, 146-51.

Shialajan S, Menon S, Polymarker based standardization of an ayurvedic formulation Lavangadi Vati using high

performance thin layer chromatography, J Pharm Res, 4, 2011, 467-70.

Tapadiya G, Metku M, Deokate U, Khadabadi S, Saboo S, Sahu K, Quantitative estimation of piperine from

pharmaceutical dosage form by HPTLC, Asian J Pharm Cli Res, 2, 2009, 47-50.



Pulak Majumder and Susmita Majumder Indian Journal of Research in Pharmacy and Biotechnology


PREPARATION AND CHARACTERIZATION OF SOME HERBAL OINTMENT

FORMULATIONS WITH EVALUATION OF ANTIMICROBIAL PROPERTY *Pulak Majumder

1 and Susmita Majumder

2

1Dept. of Pharmacognosy, Rajiv Gandhi Institute of Pharmacy, Trikaripur, Kasaragod Dist, Kerala, India,

2Dept. of Biotechnology, Heritage Institute of Technology, Anandapur, Kolkata, India.


ABSTRACT

The Indian subcontinent is enriched by a variety of flora- both aromatic and medicinal plants. Herbal

drugs constitute a major part in all the traditional systems of medicine. There are approximately 1250 Indian

medicinal plants, which are used in formulating therapeutic preparation according to Ayurveda and other

traditional system of medicine. This work has been an approach to formulate a modern Ayurvedic formulation

with single and double drug combination. Both the formulations are found to be very efficacious in all the

parameters which was conducted for its characterization and also found enhance antimicrobial property. This

study may give a brief importance for modernization of Ayurvedic preparation and also the importance of some

herbs which can be distinct within some time period.

Key words: Peperomia pellucida, Cymbopogon citrate, Ointment, Antimicrobial property.

INTRODUCTION

According to survey report by WHO, about 25 per cent of prescribed human medicines are derived from

plants and 80 per cent people still depend on traditional system of medicines. The herbal wealth of India and the

knowledge of their medicinal properties have a long tradition, as referred in Rig veda and other ancient literature.

The topography of India in the tropical belt with its varied climatic zones made it a vast storehouse of medicinal

plants. The quality assessment of herbal formulations is of paramount importance in order to justify their

acceptability in modern system of medicine. One of the major problems faced by the herbal industry is the

unavailability of rigid quality control profiles for herbal materials and their formulations. Regulatory bodies have

laid down the standardization procedures and specifications for Ayurvedic preparations. The World Health

Organization (WHO) has appreciated the importance of medicinal plants for public health care in developing

nations and has evolved guidelines to support the member states in their efforts to formulate national policies on

traditional medicine and to study their potential usefulness including evaluation, safety, and efficacy

(Organisation Mondiale De La Sante, 1992).

Peperomia pellucida (Fig 1) is a commonly known as Shiny Bush or silver bush, fleshy tropical annual,

shallow-rooted herb, usually growing to a height of about 15 to 45 cm. It is characterized by fibrous roots,

succulent stems, shiny, heart-shaped, fleshy leaves and tiny, dot-like seeds attached to several fruiting spikes. It

has a mustard-like odor when crushed. P. pellucida plant possesses various pharmacological properties like

Analgesic (Aziba PI et al), Anti-inflammatory (De Fatima Arrigoni, 2004) antipyretic (Alam Khan, 2008),

chemotherapeutic, broad spectrum antibiotic, refrigerant, anticancer (Arrigoni-Blank Mde F, 2002) and

depressant of Central Nervous System (Khan, 2007) etc. There are popular descriptions of this plant to lower

cholesterol levels or used on proteinuria and as diuretic.

The Cymbopogon citrates (Fig 2) commonly known as lemongrass or Citroengrass, Fever Grass, a

perennial plant with brawny stalks and somewhat broad and scented leaves. This species grows in thick bunches

that often develop to a height of six feet (1.8 meters) and approximately four feet (1.2 meters) in breadth. The

leaves of the plant are similar to straps and are 0.5 inch to 1 inch (1.3 cm to 2.5 cm) in width and around three feet

(0.9 meter) in length, and possess stylish apexes. The plant bears leaves round the year and they are vivid bluish-

green and when mashed they emit an aroma akin to lemons. Apart from the herb’s aromatic, ornamental and

culinary uses, lemongrass also provides therapeutic benefits to cure grouchy conditions, nervous disorders, colds

and weariness. The essential oils extracted from lemongrass have a yellow or yellowish-brown hue and this liquid

is known to be antiseptic. Very often the oil is applied externally to treat disorders like athlete’s foot (tinea pedia).

Lemongrass is also used as a carminative to emit digestive gas, a digestive tonic, a febrifuge, analgesic as well as

an antifungal (Chiori, 1977), anti microbial (Kokate, 1971), astringent , aids in avoiding panic, melancholy,

nervousness, rheumatism and sprains (Carbajal, 1989), suppress coughs, and as a diuretic and sedative (Carlini,

1986). The stalks and leaves of the lemongrass are widely used in culinary; In addition, Cymbopogon citratus is

extensively used by the cosmetic industry in the manufacture of soaps as well as hair care products. In an earlier






study, medicinal plants which have been reported individually of its anti microbial activity along with different

other pharmacological properties on different experimental studies, but there are no such kind of existing suitable

pharmaceutical formulations available for the single or combinational form of those drugs.

This attempt has been made to formulate and evaluate the pharmaceutical doses form i.e ointment by first

time combining those herbals and also single drug ointment formulation. This approach may provide a suitable

and more effective utilization of those pharmaceutically impotent herbs.


Collection and Authentication of plant: Plants of Peperomia pellucida and Cymbopogon citrates were collected

from the road sides of Trikaripur forest areas, Kasaragod district of Kerala, India, in the month of May 2011 in a

quantity sufficient for all the experiments in a single batch and the plant materials were authenticated. One each

voucher specimen was submitted and preserved in the Department of Pharmacognosy, Rajiv Gandhi Institute of

Pharmacy, Trikaripur (Herbarium no. RGIP/10-12/003 & 004). The plant parts were washed under running tap

water, cut into small pieces of 2-3cm and shade dried (300C, 50 ± 5% relative humidity) for 15days. The shade

dried plant material was powdered using a dry grinder to get the coarse powder (sieve no. 10/44). The powder was

stored in air tight container for further use.

Fig 1: Peperomia pellucida Fig 2: Cymbopogon citrate

Preparation of plant extract and isolation of oil: The powder of Peperomia pellucida was subjected solvent

extraction. The powder material was refluxed with ethanol (90%) in a Soxhlet extractor for 18 hrs in batches of

50g each cycle. The marc was gently pressed and dried before completing the extracting.

The extracts obtained by the above techniques were concentrated in vacuum under reduced pressure using a rotary

flash evaporator. Fresh leaves of Cymbopogon citrates were subjected to Clevenger apparatus with sufficient

quantity of water with glycerin and boiled up to complete extraction of oil from the plant part.

Preliminary phytochemical screening: The preliminary phytochemical screening was carried out according to

the recommended standard procedures (Kokate, 1997) (Wallis, 1985) (Khandalwal, 2008).

Physical Evaluation of formulated ointments: Organoleptic parameters: Organoleptic parameters like color, odor and taste of all the formulations were carried

out.

Determination of pH: The pH value of a solution was determined by digital pH meter (Mettler Toledo). The pH

meter was operated according the manufacturer’s instructions. First the apparatus was calibrated using buffer of 4,

7 and 9 pH. The electrodes were immersed in the solution and the pH was measured.

Homogeneity:All the developed ointments were tested for homogeneity by visual inspection. They were tested

for their appearance.

Evaluation of Herbal Ointment and Polyherbal Ointment

Ointment formulations: Two types of drug formulations (ointments) were prepared by using standard ointment

base for standardization and evaluation microbial activity. For topical application 5g of each extract was

separately incorporated with 100g of simple ointment IP.

The formula for simple ointment I.P. is:

S.No Ingredients Quantity (gm)

1 White bees wax 20

2 Hard paraffin 30

3 Cetosteryl alcohol 50

4 White soft paraffin 900





Consistency or hardness of an ointment (Pharmacopeial Forum):

The consistency or hardness of an ointment was measured by Penetrometer. Prepare the test samples by one of the

following procedures:

Carefully and completely fill three containers, without forming air bubbles. Level if necessary to obtain a

flat surface. Store the samples at 25±0.5°C for 24 hrs, unless otherwise prescribed.

Store three samples at 25±0.5°C for 24hrs. Apply a suitable shear to the samples for 5 min. carefully and

completely fill three containers, without forming air bubbles, and level if necessary to obtain a flat

surface.

Melt three samples and carefully and completely fill three containers, without forming air bubbles. Store

the samples at 25±0.5°C for 24hrs, unless otherwise prescribed.

Place the test sample on the base of the Penetrometer. Verify that its surface is perpendicular to the vertical axis of

the penetrating object. Bring the temperature of the penetrating object to 25±0.5°C and then adjust its position

such that its tip just touches the surface of the sample. Release the penetrating object and hold it free for 5sec.

Clamp the penetrating object and measured the depth of penetration. Repeat the procedure for remaining two

containers.

Spreadability studies of ointment (Wood, 1963): Spreadability of the formulation was determined by an

apparatus suggested by Muttimer et al., which was suitably modified in the laboratory and used for the study. It

consists of a wooden block, which was provided by a pulley at one end. A rectangular ground glass plate was

fixed on this block. An excess of ointment (about 3 gm.) under study was placed on this ground plate. The

ointment was then sandwiched between this plate and another glass plate having the dimension of fixed ground

plate and provided with the hook. A 1 Kg. weight was placed on the top of the two plates for 5 minutes to expel

air and to provide a uniform film of the ointment between the plates. Excess of the ointment was scrapped off

from the edges. The top plate was then subjected to pull of 80 gms. With the help of string attached to the hook

and the time (in seconds) required by the top plate to cover a distance of 10 cm. be noted. A shorter interval

indicates better Spreadability.

Spreadability is measured as S = M × L /T

M= weight tide to upper slide

L= length of glass slides

T= Time

Tube Extrudability studies of ointment: It is a usual empirical test to measure the force required to extrude the

material from tube. The method applied for determination of applied shear in the region of the rheogram

corresponding to a shear rate exceeding the yield value and exhibiting consequent plug flow one such apparatus is

described by Wood et al. In the present study, the method adopted for evaluating ointment formulation for

extrudability was based upon the quantity in percentage of ointment and ointment extruded from tube on

application of finger pressure. More quantity extruded better was extrudability. The formulation under study was

filled in a clean, lacquered aluminum collapsible tube with a nozzle tip of 5 mm opening and applies the pressure

on tube by the help of finger. Tube extrudability was then determined by measuring the amount of ointment

extruded through the tip when a pressure was applied on tube.

PHARMACOLOGICAL STUDIES

Topical sensitivity Test: Both two ointments (Herbal and Polyherbal) were tested for their skin sensitivity tests

by applying to the elbow of the hand in selected human volunteers and observed the side effects if any, as a set of

parameters like skin inflammation, skin irritation reddening of skin (allergic reactions) etc.

Microbiological studies (Bauer, 1966): The antibacterial activity of various ointment formulations evaluated

against various strain of anaerobic and aerobic microorganism by standard cup plate method and the inhibition

zone diameters were measured with the help of zone reader. Mucus fungus, Lactobacillus, Escherichia coli

(aerobic organism) were used for testing of antibacterial activity. Nutrient agar media was used for aerobic

bacterial culture and incubated at temperature 37°C±2°C for 48 hrs.


The various physicochemical parameters utilized to evaluate the prepared ointment formulations are shown in

table 2.





Preparation of Ointments: Herbal ointment was prepared by incorporating 5% of Cymbopogon citrates oil in

simple ointment base (Fig. 3). Polyherbal ointment was prepared by equal amount of (5%) both the drug

Peperomia pellucida plant extract and Cymbopogon citrates, oil incorporated in simple ointment base (Fig.3).

Fig.3. Herbal and polyherbal ointment formulation

Preliminary Phytochemical Investigation: The isolation, purification and identification of active constituents

are chemical methods of evaluation. Qualitative chemical tests are also included under chemical evaluation.

Preliminary phytochemical screening which is performed to establish a chemical profile of a crude drug is a part

of chemical evaluation. All the three different plant parts extracts were screened for phytochemical investigation

by different phytochemical tests to check the presence or absence of a group of phytochemical constituents. These

phytochemical tests shown the presence of carbohydrates, alkaloids, tannins, steroids, triterpenoids, flavonoids

etc. are present in stem extract whereas in root extract contains all above except flavonoids as mentioned in Table

1. Leaf extract were also found to contain all the constituents including saponins.

Table 1: Preliminary phytochemical analysis of various parts of Peperomia pellucida.

Chemical Constituents Root Stem leaf

Carbohydrates + - -

Proteins - - -

Alkaloids +++ ++ +

Saponins - - +

Tannins ++ ++ +++

Flavonoids ++ ++ +++

Steroids + + +

Triterpenoids + + +

Glycoside +++ ++ ++

+ = Present, - = Absent.

Table-2 Physical Evaluation of formulated ointments

Evaluation parameters Herbal ointment

Polyherbal ointment

Description Colour yellow Dark brown

Odour Aromatic Characteristic

Consistency Soft semisolid Soft semisolid

Phase separation Nil Nil

pH (10℅w/v solution) 6.2 6.5

Hardness or consistency 149mm 198mm

Spreadability 47.2g/s 24.6g/s

The pH of the formulations lies in the normal pH range of human skin (6.8 ± 1). All the formulations did not

produce any skin irritation, i.e , erythema and edema for about a week when applied over the skin. These

formulations did not produce any skin irritation for about a week when applied over the skin.





Fig 4: Spread ability of ointment

From the data it is clearly evident all the physicochemical characteristics of both the formulation were found

satisfactory and there were no significance changes in evaluation parameters. The formulation (polyherbal

ointment) was found to be more satisfactory than Herbal ointment. Spreadability results (graph-1) of both the

ointments found satisfactory.

The antimicrobial activity of polyherbal ointment and Herbal ointment were determined (Fig. 4). The zones of

inhibition of various strains of aerobic and anaerobic organisms are depicted in table 4. The zone of inhibition for

both the ointment (after one month) found to be nearer to standard Gentamycin.

Table 3. Antimicrobial activity of Herbal Ointment and Polyherbal Ointment

Diameter of Zone of inhibition (cm)

Ointment E. coli Mucus fungus Lactobacillus

Herbal Ointment 1.25± 0.2 1.50±0.12 1.11±0.32

Polyherbal Ointment 1.85±0.42 1.93±0.31 1.71±0.13

Standard(Gentamycin) 2.21±0.23 2.00±0.11 2.11±0.23

Control 0.166±0.11 0.41±0.23 0.08±22

E. coli Mucus fungus Lactobacillus

Fig. 5. Antimicrobial activity of Herbal and Polyherbal Ointment formulations

CONCLUSION

A combinational therapy is the need of hour to treat eczema and pruritis. This can be achieved by

Clotrimazole and Ichthammol (an antifungal and antiseptic).In this study, ointment was formulated with different

bases like white soft paraffin, cetostearyl alcohol, hard paraffin, and light liquid paraffin. By combining these

drugs with appropriate ointment bases (as polyherbal formulation) a better therapy and patient compliance can be

achieved. Both the formulations may shows the batter way to use these drugs with more significant way and more

over WHO also emphasizes the herbal medicine for treatment. This study showed the convenient preparation and

more effective use of both the herbs with modernized formulations with comparatively old form of medicine. This

affords may leads a beginning of proper utilization and conservation of medicinally important herbs and cost

effective treatment in future.





ACKNOWLEDGEMENT

Authors are thankful to the principal, Rajiv Gandhi Institute of Pharmacy, Trikaripur and Heritage

Institute of Technology, Anandapur for their necessary facilities and constant support to successfully completion

of this research work.

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Shyam Bihari Sharma et.al Indian Journal of Research in Pharmacy and Biotechnology


THE EFFECTS OF AIR POLLUTION ON THE ENVIRONMENT AND

HUMAN HEALTH Shyam Bihari Sharma

1*, Suman Jain

1, Praveen Khirwadkar

2, Sunisha Kulkarni

1

1. School of Studies in Pharmaceutical Sciences, Jiwaji University, Gwalior, Madhya Pradesh, INDIA

2. Institute of pharmacy, Vikarm University, Ujjain (M.P)

*Corresponding Author: [email protected]

ABSTRACT

Pollution is the introduction of contaminants into the natural environment that cause adverse

change. Air pollution includes anything introduced by humans into the atmosphere with a damaging

effect. The main cause of air pollution is the burning of fossil fuels in cars, in planes and for the

production of electricity. Air pollution occurs when any chemicals or biological matter that can harm

humans or other living things is introduced into the atmosphere. Pollutants in the air include carbon

dioxide, carbon monoxide, sulphur dioxide and small particles that are a result of burning different

materials, especially coal. These pollutants not only harm individuals by causing disease, but also

harm the environment by adding to global warming. Pollution affects all humans, children more so

than adults because children spend more time outside and take in more air, and air pollution, when

breathing, especially when exerting them.

The main air pollutants affect the lungs most. Sulphur dioxide irritates eyes, nose and throat.

When inhaled, cause severe lung problems, such as asthma, bronchitis, emphysema and lung cancer.

Nitrogen dioxide damages lung tissue and can restrict airways and cause emphysema. It also leads to

formation of ozone, which can eat holes in lung tissue, aggravate asthma and leave people susceptible

to respiratory disease. Carbon monoxide, which is invisible and odourless, can lead to damage of the

heart and central nervous system, headaches, dizziness, convulsions and death. Air pollution affects

the whole earth ecosystem. Global warming, caused by carbon dioxide building up in the atmosphere,

traps heat in the earth's atmosphere, which can cause dramatic climate changes that shift the delicate

balance of ecosystems around the world. A number of chemicals that have been released into the air,

such as chlorofluorocarbons, have caused depletion of the protective ozone layer, which causes

harmful ultraviolet radiation to reach the earth. Air pollution may be prevented only if individuals

and businesses stop using toxic substances that cause air pollution in the first place. This would

require the cessation of all fossil fuel-burning processes, from industrial manufacturing to home use

of air conditioners.

Keywords: Pollution, Air pollution, Pollutants, Atmosphere, Carbon dioxide, Global warming.

INTRODUCTION

Air pollution is the introduction into the atmosphere of chemicals, particulates, or biological materials

that cause discomfort, disease, or death to humans, damage other living organisms such as food crops, or

damage the natural environment or environment. The atmosphere is a complex dynamic natural gaseous

system that is essential to support life on planet Earth. Stratosphericozone depletion due to air pollution has

long been recognized as a threat to human health as well as to the Earth's ecosystems. Indoor air pollution and

urban air quality are listed as two of the World’s Worst Toxic Pollution Problems in the 2008 Blacksmith

Institute World's Worst Polluted Places report.

Air pollution, contamination of the air by noxious gases and minute particles of solid and liquid

matter (particulates) in concentrations that endanger health. The major sources of air pollution are

transportation engines, power and heat generation, industrial processes, and the burning of solid waste.

The combustion of gasoline and other hydrocarbon fuels in automobiles, trucks, and jet airplanes

produces several primary pollutants: nitrogen oxides, gaseous hydrocarbons, and carbon monoxide, as well as

large quantities of particulates, chiefly lead. In the presence of sunlight, nitrogen oxides combine with

hydrocarbons to form a secondary class of pollutants, the photochemical oxidants, among them ozone and the

eye-stinging peroxyacetylnitrate (PAN). Nitrogen oxides also react with oxygen in the air to form nitrogen

dioxide, a foul-smelling brown gas. In urban areas like Los Angeles where transportation is the main cause of

air pollution, nitrogen dioxide tints the air, blending with other contaminants and the atmospheric water vapor

to produce brown smog. Although the use of catalytic converters has reduced smog-producing compounds in

motor vehicle exhaust emissions, studies have shown that in so doing the converters produce nitrous oxide,

which contributes substantially to global warming.

Pollutants: A substance in the air that can be harmful to humans and the environment is known as an air

pollutant. Pollutants can be in the form of solid particles, liquid droplets, or gases. In addition, they may be


http://en.wikipedia.org/wiki/Earth%27s_atmosphere

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http://en.wikipedia.org/wiki/Biomolecule

http://en.wikipedia.org/wiki/Natural_environment

http://en.wikipedia.org/wiki/Earth

http://en.wikipedia.org/wiki/Stratosphere

http://en.wikipedia.org/wiki/Stratosphere

http://en.wikipedia.org/wiki/Blacksmith_Institute

http://en.wikipedia.org/wiki/Blacksmith_Institute

http://www.infoplease.com/encyclopedia/science/solid-waste.html

http://www.infoplease.com/encyclopedia/science/gasoline.html

http://www.infoplease.com/encyclopedia/science/automobile.html

http://www.infoplease.com/encyclopedia/science/ozone.html

http://www.infoplease.com/encyclopedia/science/smog.html

http://www.infoplease.com/encyclopedia/science/global-warming.html





natural or man-made. Pollutants can be classified as primary or secondary. Usually, primary pollutants are

directly emitted from a process, such as ash from a volcanic eruption, the carbon monoxide gas from a motor

vehicle exhaust or sulphur dioxide released from factories. Secondary pollutants are not emitted directly.

Rather, they form in the air when primary pollutants react or interact. An important example of a secondary

pollutant is ground level ozone — one of the many secondary pollutants that make up photochemical smog.

Some pollutants may be both primary and secondary: that is, they are both emitted directly and formed from

other primary pollutants.

Major primary pollutants produced by human activity include

Sulphur oxides (SOx) - especially sulphur dioxide, a chemical compound with the formula SO2. SO2 is

produced by volcanoes and in various industrial processes. Since coal and petroleum often contain sulphur

compounds, their combustion generates sulfur dioxide. Further oxidation of SO2, usually in the presence of a

catalyst such as NO2, forms H2SO4, and thus acid rain.This is one of the causes for concern over the

environmental impact of the use of these fuels as power sources.

Nitrogen oxides (NOx) - especially nitrogen dioxide are emitted from high temperature combustion,

and are also produced naturally during thunderstorms by electric discharge. Can be seen as the brown haze

dome above or plume downwind of cities. Nitrogen dioxide is the chemical compound with the formula NO2.

It is one of the several nitrogen oxides. This reddish-brown toxic gas has a characteristic sharp, biting odor.

NO2 is one of the most prominent air pollutants.

Carbon monoxide (CO)- is a colourless, odorless, non-irritating but very poisonous gas. It is a product

by incomplete combustion of fuel such as natural gas, coal or wood. Vehicular exhaust is a major source of

carbon monoxide.

Volatile organic compounds - VOCs are an important outdoor air pollutant. In this field they are often

divided into the separate categories of methane (CH4) and non-methane (NMVOCs). Methane is an extremely

efficient greenhouse gas which contributes to enhancedglobal warming. Other hydrocarbon VOCs are also

significant greenhouse gases via their role in creating ozone and in prolonging the life of methane in the

atmosphere, although the effect varies depending on local air quality. Within the NMVOCs, the aromatic

compounds benzene, toluene and xylene are suspected carcinogens and may lead to leukemia through

prolonged exposure. 1,3-butadiene is another dangerous compound which is often associated with industrial

uses.

Particulates, alternatively referred to as particulate matter (PM), atmospheric particulate matter, or

fine particles, are tiny particles of solid or liquid suspended in a gas. In contrast, aerosol refers to particles and

the gas together. Sources of particulates can be man made or natural. Some particulates occur naturally,

originating from volcanoes, dust storms, forest and grassland fires, living vegetation, and sea spray. Human

activities, such as the burning of fossil fuels in vehicles, power plants and various industrial processes also

generate significant amounts of aerosols. Averaged over the globe, anthropogenic aerosols—those made by

human activities—currently account for about 10 percent of the total amount of aerosols in our atmosphere.

Increased levels of fine particles in the air are linked to health hazards such as heart disease,altered lung

function and lung cancer.

Persistent free radicals connected to airborne fine particles could cause cardiopulmonary disease.

Toxic metals, such as lead and quick silver, especially their compounds: Chlorofluorocarbons (CFCs) -

harmful to the ozone layer emitted from products currently banned from use.

Ammonia (NH3) - emitted from agricultural processes. Ammonia is a compound with the formula

NH3. It is normally encountered as a gas with a characteristic pungent odor. Ammonia contributes

significantly to the nutritional needs of terrestrial organisms by serving as a precursor to foodstuffs and

fertilizers. Ammonia, either directly or indirectly, is also a building block for the synthesis of many

pharmaceuticals. Although in wide use, ammonia is both caustic and hazardous.

Secondary pollutants include: Particulates created from gaseous primary pollutants and compounds in

photochemical smog. Smog is a kind of air pollution; the word "smog" is a portmanteau of smoke and fog.

Classic smog results from large amounts of coal burning in an area caused by a mixture of smoke and sulfur

dioxide. Modern smog does not usually come from coal but from vehicular and industrial emissions that are

acted on in the atmosphere by ultraviolet light from the sun to form secondary pollutants that also combine

with the primary emissions to form photochemical smog.

Ground level ozone (O3) formed from NOx and VOCs. Ozone (O3) is a key constituent of the

troposphere. It is also an important constituent of certain regions of the stratosphere commonly known as the

Ozone layer. Photochemical and chemical reactions involving it drive many of the chemical processes that

http://en.wikipedia.org/wiki/Carbon_monoxide

http://en.wikipedia.org/wiki/Ground_level_ozone

http://en.wikipedia.org/wiki/Smog

http://en.wikipedia.org/wiki/Sulphur_oxide

http://en.wikipedia.org/wiki/Acid_rain

http://en.wikipedia.org/wiki/Nitrogen_oxide

http://en.wikipedia.org/wiki/Nitrogen_dioxide

http://en.wikipedia.org/wiki/Thunderstorms

http://en.wikipedia.org/wiki/Electric_discharge

http://en.wikipedia.org/wiki/Haze

http://en.wikipedia.org/wiki/Plume_%28hydrodynamics%29


http://en.wikipedia.org/wiki/Incomplete_combustion

http://en.wikipedia.org/wiki/Volatile_organic_compounds

http://en.wikipedia.org/wiki/Global_warming

http://en.wikipedia.org/wiki/Particulates

http://en.wikipedia.org/wiki/Radical_%28chemistry%29#Persistent_radicals

http://en.wikipedia.org/wiki/Metal

http://en.wikipedia.org/wiki/Lead

http://en.wikipedia.org/wiki/Mercury_%28element%29

http://en.wikipedia.org/wiki/Chlorofluorocarbons

http://en.wikipedia.org/wiki/Ozone_layer

http://en.wikipedia.org/wiki/Ammonia

http://en.wikipedia.org/wiki/Smog

http://en.wikipedia.org/wiki/Ultraviolet

http://en.wikipedia.org/wiki/Ground_level_ozone





occur in the atmosphere by day and by night. At abnormally high concentrations brought about by human

activities (largely the combustion of fossil fuel), it is a pollutant, and a constituent of smog.

Peroxyacetyl nitrate (PAN) - similarly formed from NOx and VOCs.

Minor air pollutants include: A large number of minor hazardous air pollutants. Some of these are regulated

in USA under the Clean Air Act and in Europe under the Air Framework Directive.

A variety of persistent organic pollutants, which can attach to particulates: Persistent organic pollutants

(POPs) are organic compounds that are resistant to environmental degradation through chemical, biological,

and photolytic processes. Because of this, they have been observed to persist in the environment, to be capable

of long-range transport, bioaccumulate in human and animal tissue, biomagnify in food chains, and to have

potential significant impacts on human health and the environment.

Anthropogenic sources (human activity) mostly related to burning different kinds of fuel

"Stationary Sources" include smoke stacks of power plants, manufacturing facilities (factories) and

waste incinerators, as well as furnaces and other types of fuel-burning heating devices. In developing and poor

countries, traditional biomass burning is the major source of air pollutants; traditional biomass includes wood,

crop waste and dung.

"Mobile Sources" include motor vehicles, marine vessels, aircraft and the effect of sound etc.

Chemicals, dust and controlled burn practices in agriculture and forestry management. Controlled or

prescribed burning is a technique sometimes used in forest management, farming, prairie restoration or

greenhouse gas abatement. Fire is a natural part of both forest and grassland ecology and controlled fire can be

a tool for foresters. Controlled burning stimulates the germination of some desirable forest trees, thus

renewing the forest.

Fumes from paint, hair spray, varnish, aerosol sprays and other solvents

Waste deposition in landfills, which generate methane. Methane is highly flammable and may form

explosive mixtures with air. Methane is also an asphyxiant and may displace oxygen in an enclosed space.

Asphyxia or suffocation may result if the oxygen concentration is reduced to below 19.5% by displacement.

Military, such as nuclear weapons, toxic gases, germ warfare and rocketry

Natural sources Dust from natural sources, usually large areas of land with little or no vegetation

Methane, emitted by the digestion of food by animals, for example cattle

Radon gas from radioactive decay within the Earth's crust. Radon is a colorless, odorless, naturally

occurring, radioactive noble gas that is formed from the decay of radium. It is considered to be a health

hazard. Radon gas from natural sources can accumulate in buildings, especially in confined areas such as the

basement and it is the second most frequent cause of lung cancer, after cigarette smoking.

1. Smoke and carbon monoxide from wildfires

2. Vegetation, in some regions, emits environmentally significant amounts of VOCs on warmer days.

These VOCs react with primary anthropogenic pollutants—specifically, NOx, SO2, and anthropogenic

organic carbon compounds—to produce a seasonal haze of secondary pollutants.

3. Volcanic activity, which produce sulfur, chlorine, and ash particulates

Air pollutant emission factors are representative values that people attempt to relate the quantity of a

pollutant released to the ambient air with an activity associated with the release of that pollutant. These factors

are usually expressed as the weight of pollutant divided by a unit weight, volume, distance, or duration of the

activity emitting the pollutant (e.g., kilograms of particulate emitted per tonne of coal burned). Such factors

facilitate estimation of emissions from various sources of air pollution. In most cases, these factors are simply

averages of all available data of acceptable quality, and are generally assumed to be representative of long-

term averages.

There are 12 compounds in the list of POPs. Dioxins and furans are two of them and are intentionally

created by combustion of organics, like open burning of plastics. The POPs are also endocrine disruptor and

can mutate the human genes.

The United States Environmental Protection Agency has published a compilation of air pollutant

emission factors for a multitude of industrial sources. The United Kingdom, Australia, Canada and many other

countries have published similar compilations, as well as the European Environment Agency.

Health effects: Air pollution is a significant risk factor for multiple health conditions including respiratory

infections, heart disease, and lung cancer, according to the WHO. The health effects caused by air pollution

may include difficulty in breathing, wheezing, coughing and aggravation of existing respiratory and cardiac

conditions. These effects can result in increased medication use, increased doctor or emergency room visits,

more hospital admissions and premature death. The human health effects of poor air quality are far reaching,

http://en.wikipedia.org/wiki/Peroxyacetyl_nitrate

http://en.wikipedia.org/wiki/Category:Hazardous_air_pollutants

http://en.wikipedia.org/wiki/Clean_Air_Act_%28United_States%29

http://en.wikipedia.org/wiki/Persistent_organic_pollutant

http://en.wikipedia.org/wiki/Fuel

http://en.wikipedia.org/wiki/Power_plant

http://en.wikipedia.org/wiki/Roadway_air_dispersion_modeling

http://en.wikipedia.org/wiki/Chemicals

http://en.wikipedia.org/wiki/Controlled_burn

http://en.wikipedia.org/wiki/Paint

http://en.wikipedia.org/wiki/Hair_spray

http://en.wikipedia.org/wiki/Varnish

http://en.wikipedia.org/wiki/Aerosol_spray

http://en.wikipedia.org/wiki/Landfill

http://en.wikipedia.org/wiki/Methane

http://en.wikipedia.org/wiki/Asphyxiant

http://en.wikipedia.org/wiki/Nuclear_weapon

http://en.wikipedia.org/wiki/Toxic_gas

http://en.wikipedia.org/wiki/Germ_warfare

http://en.wikipedia.org/wiki/Rocket

http://en.wikipedia.org/wiki/Dust

http://en.wikipedia.org/wiki/Methane

http://en.wikipedia.org/wiki/Flatulence

http://en.wikipedia.org/wiki/Digestion

http://en.wikipedia.org/wiki/Animal

http://en.wikipedia.org/wiki/Cattle

http://en.wikipedia.org/wiki/Radon

http://en.wikipedia.org/wiki/Radioactive_decay

http://en.wikipedia.org/wiki/Earth%27s_crust

http://en.wikipedia.org/wiki/Cigarette

http://en.wikipedia.org/wiki/Smoke


http://en.wikipedia.org/wiki/Wildfires

http://en.wikipedia.org/wiki/Volcano

http://en.wikipedia.org/wiki/Sulfur

http://en.wikipedia.org/wiki/Chlorine

http://en.wikipedia.org/wiki/Tonne

http://en.wikipedia.org/wiki/United_States_Environmental_Protection_Agency

http://en.wikipedia.org/wiki/United_Kingdom

http://en.wikipedia.org/wiki/Australia

http://en.wikipedia.org/wiki/Canada

http://en.wikipedia.org/wiki/European_Environment_Agency





but principally affect the body's respiratory system and the cardiovascular system. Individual reactions to air

pollutants depend on the type of pollutant a person is exposed to, the degree of exposure, the individual's

health status and genetics.

The most common sources of air pollution include particulates, ozone, nitrogen dioxide, and sulfur

dioxide. Both indoor and outdoor air pollution have caused approximately 3.3 million deaths worldwide.

Children aged less than five years that live in developing countries are the most vulnerable population in terms

of total deaths attributable to indoor and outdoor air pollution.

The World Health Organization states that 2.4 million people die each year from causes directly

attributable to air pollution, with 1.5 million of these deaths attributable to indoor air pollution.

"Epidemiological studies suggest that more than 500,000 Americans die each year from cardiopulmonary

disease linked to breathing fine particle air pollution. . ." A study by the University of Birmingham has shown

a strong correlation between pneumonia related deaths and air pollution from motor vehicles. Worldwide

more deaths per year are linked to air pollution than to automobile accidents. A 2005 study by the European

Commission calculated that air pollution reduces life expectancy by an average of almost nine months across

the European Union. Causes of deaths include aggravated asthma, emphysema, lung and heart diseases, and

respiratory allergies. The US EPA estimates that a proposed set of changes in diesel engine technology could

result in 12,000 fewer premature mortalities, 15,000 fewer heart attacks, 6,000 fewer emergency room visits

by children with asthma, and 8,900 fewer respiratory-related hospital admissions each year in the United

States.

The US EPA estimates allowing a ground-level ozone concentration of 65 parts per billion, would

avert 1,700 to 5,100 premature deaths nationwide in 2020 compared with the current 75-ppb standard. The

agency projects the stricter standard would also prevent an additional 26,000 cases of aggravated asthma and

more than a million cases of missed work or school.

The worst short term civilian pollution crisis in India was the 1984 Bhopal Disaster. Leaked industrial

vapours from the Union Carbide factory, belonging to Union Carbide, Inc., U.S.A., killed more than 25,000

people outright and injured anywhere from 150,000 to 600,000. The United Kingdom suffered its worst air

pollution event when the December 4 Great Smog of 1952 formed over London. In six days more than 4,000

died, and 8,000 more died within the following months. An accidental leak of anthrax spores from a biological

warfare laboratory in the former USSR in 1979 near Sverdlovsk is believed to have been the cause of

hundreds of civilian deaths. The worst single incident of air pollution to occur in the US occurred in Donora,

Pennsylvania in late October, 1948, when 20 people died and over 7,000 were injured.

A new economic study of the health impacts and associated costs of air pollution in the Los Angeles

Basin and San Joaquin Valley of Southern California shows that more than 3800 people die prematurely

(approximately 14 years earlier than normal) each year because air pollution levels violate federal standards.

The number of annual premature deaths is considerably higher than the fatalities related to auto collisions in

the same area, which average fewer than 2,000 per year.

Diesel exhaust (DE) is a major contributor to combustion derived particulate matter air pollution. In

several human experimental studies, using a well validated exposure chamber setup, DE has been linked to

acute vascular dysfunction and increased thrombus formation. This serves as a plausible mechanistic link

between the previously described association between particulates air pollution and increased cardiovascular

morbidity and mortality.

Effects on cardiovascular health: A 2007 review of evidence found ambient air pollution exposure is a risk

factor correlating with increased total mortality from cardiovascular events (range: 12% to 14% per a 10

microg/m3 increase). Air pollution is also emerging as a risk factor for stroke, particularly in developing

countries where pollutant levels are highest. A 2007 study found that in women air pollution is associated not

with hemorrhagic but with ischemic stroke. Air pollution was also found to be associated with increased

incidence and mortality from coronary stroke in a cohort study in 2011.

Effects on cystic fibrosis

Main article: Cystic fibrosis

A study from around the years of 1999 to 2000, by the University of Washington, showed that patients near

and around particulates air pollution had an increased risk of pulmonary exacerbations and decrease in lung

function. Patients were examined before the study for amounts of specific pollutants like Pseudomonas

aeruginosa or Burkholderiacenocepacia as well as their socioeconomic standing. Participants involved in the

study were located in the United States in close proximity to an Environmental Protection Agency. During the

time of the study 117 deaths were associated with air pollution. Many patients in the study lived in or near

large metropolitan areas in order to be close to medical help. These same patients had higher level of

http://en.wikipedia.org/wiki/Epidemiological

http://en.wikipedia.org/wiki/Cardiopulmonary

http://en.wikipedia.org/wiki/University_of_Birmingham

http://en.wikipedia.org/wiki/Pneumonia


http://en.wikipedia.org/wiki/Diesel_engine

http://en.wikipedia.org/wiki/Myocardial_infarction

http://en.wikipedia.org/wiki/Emergency_room

http://en.wikipedia.org/wiki/Asthma

http://en.wikipedia.org/wiki/India

http://en.wikipedia.org/wiki/Bhopal_Disaster

http://en.wikipedia.org/wiki/Great_Smog

http://en.wikipedia.org/wiki/London

http://en.wikipedia.org/wiki/Anthrax

http://en.wikipedia.org/wiki/Biological_warfare

http://en.wikipedia.org/wiki/Biological_warfare

http://en.wikipedia.org/wiki/USSR

http://en.wikipedia.org/wiki/Yekaterinburg

http://en.wikipedia.org/wiki/Donora,_Pennsylvania

http://en.wikipedia.org/wiki/Donora,_Pennsylvania

http://en.wikipedia.org/wiki/Los_Angeles_Basin

http://en.wikipedia.org/wiki/Los_Angeles_Basin

http://en.wikipedia.org/wiki/San_Joaquin_Valley

http://en.wikipedia.org/wiki/Cystic_fibrosis

http://en.wikipedia.org/wiki/Pseudomonas_aeruginosa

http://en.wikipedia.org/wiki/Pseudomonas_aeruginosa

http://en.wikipedia.org/wiki/Burkholderia_cenocepacia






pollutants found in their system because of more emissions in larger cities. As cystic fibrosis patients already

suffer from decreased lung function, everyday pollutants such as smoke, emissions from automobiles, tobacco

smoke and improper use of indoor heating devices could further compromise lung function.

Effects on COPD and asthma

Main article: Chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) includes diseases such as chronic bronchitis and emphysema.

Researches have demonstrated increased risk of developing asthma and COPD from increased exposure to

traffic-related air pollution. Additionally, air pollution has been associated with increased hosptializations and

mortality from asthma and COPD.

A study conducted in 1960-1961 in the wake of the Great Smog of 1952 compared 293 London

residents with 477 residents of Gloucester, Peterborough, and Norwich, three towns with low reported death

rates from chronic bronchitis. All subjects were male postal truck drivers aged 40 to 59. Compared to the

subjects from the outlying towns, the London subjects exhibited more severe respiratory symptoms (including

cough, phlegm, and dyspnea), reduced lung function (FEV1 and peak flow rate), and increased sputum

production and purulence. The differences were more pronounced for subjects aged 50 to 59. The study

controlled for age and smoking habits, so concluded that air pollution was the most likely cause of the

observed differences.

It is believed that much like cystic fibrosis, by living in a more urban environment serious health

hazards become more apparent. Studies have shown that in urban areas patients suffer mucushypersecretion,

lower levels of lung function, and more self diagnosis of chronic bronchitis and emphysema.

Links to cancer

A review of evidence regarding whether ambient air pollution exposure is a risk factor for cancer in 2007

found solid data to conclude that long-term exposure to PM2.5 (fine particulates) increases the overall risk of

nonaccidental mortality by 6% per a 10 microg/m3 increase.PMID 19235364

Exposure to PM2.5 was also associated with an increased risk of mortality from lung cancer (range: 15% to

21% per a 10 microg/m3 increase) and total cardiovascular mortality (range: 12% to 14% per a 10 microg/m3

increase). PMID 19235364

The review further noted that living close to busy traffic appears to be associated with elevated risks

of these three outcomes (increase in lung cancer deaths, cardiovascular deaths, and overall nonaccidental

deaths. PMID 19235364

The reviewers also found suggestive evidence that exposure to PM2.5 is positively associated with

mortality from coronary heart diseases and exposure to SO2 increases mortality from lung cancer, but the data

was insufficient to provide solid conclusions.

In 2011, a large Danish epidemiological study found an increased risk of lung cancer for patients who

lived in areas with high nitrogen oxide concentrations. In this study, the association was higher for non-

smokers than smokers. An additional Danish study, also in 2011, likewise noted evidence of possible

associations between air pollution and other forms of cancer, including cervical cancer and brain cancer.

Effects on children

Around the world, children living in cities with high exposure to air pollutants are at increased risk of

developing asthma, pneumonia and other lower respiratory infections. Because children are outdoors more

and have higher minute ventilation they are more susceptible to the dangers of air pollution. Risks of low

initial birth weight are also heightened in such cities.

The World Health Organization reports that the greatest concentrations of particulates are found in

countries with low economic world power and high poverty and population growth rates. Examples of these

countries include Egypt, Sudan, Mongolia, and Indonesia. However even in the United States, despite the

passage of the Clean Air Act in 1970, in 2002 at least 146 million Americans were living in non-attainment

areas—regions in which the concentration of certain air pollutants exceeded federal standards. These

dangerous pollutants are known as the criteria pollutants, and include ozone, particulates, sulfur dioxide,

nitrogen dioxide, carbon monoxide, and lead.

Protective measures to ensure children's health are being taken in cities such as New Delhi, India

where buses now use compressed natural gas to help eliminate the "pea-soup" smog.

References

Davis, Devra. When Smoke Ran Like Water: Tales of Environmental Deception and the Battle Against

Pollution. Basic Books. ISBN 0-465-01521-2,2002.

Grossni, Mark. "Human cost of valley's dirty air: $6.3 billion". Sacramento Bee. 2008.

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http://en.wikipedia.org/wiki/Chronic_bronchitis

http://en.wikipedia.org/wiki/Emphysema

http://en.wikipedia.org/wiki/Great_Smog

http://en.wikipedia.org/wiki/FEV1

http://en.wikipedia.org/wiki/Cystic_fibrosis

http://en.wikipedia.org/wiki/Mucus

http://www.ncbi.nlm.nih.gov/pubmed/19235364?dopt=Abstract



http://en.wikipedia.org/wiki/Low_birth_weight

http://en.wikipedia.org/wiki/Low_birth_weight

http://en.wikipedia.org/wiki/World_Health_Organization

http://en.wikipedia.org/wiki/Egypt

http://en.wikipedia.org/wiki/Sudan

http://en.wikipedia.org/wiki/Mongolia

http://en.wikipedia.org/wiki/Indonesia

http://en.wikipedia.org/wiki/Clean_Air_Act_%28United_States%29

http://en.wikipedia.org/wiki/Non-Attainment_Area

http://en.wikipedia.org/wiki/Non-Attainment_Area

http://en.wikipedia.org/wiki/Criteria_pollutant

http://en.wikipedia.org/wiki/New_Delhi

http://en.wikipedia.org/wiki/Compressed_natural_gas

http://en.wikipedia.org/wiki/International_Standard_Book_Number

http://en.wikipedia.org/wiki/Special:BookSources/0-465-01521-2

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Sahagun, Louis "Pollution saps state's economy, study says". Los Angeles Times.2008

Kay, Jane. "Bad air costing state's economy billions". San Francisco Chronicle. 2008

Farrah J. Mateen& Robert D. Brook "Air pollution as an emerging global risk factor for stroke" JAMA,

305(12) ,2011,1240-1.

Miller K. A., Siscovick D. S., Sheppard L., Shepherd K., Sullivan J. H., Anderson G. L., Kaufman J. D.

"Long-term exposure to air pollution and incidence of cardiovascular events in women.".The New England

journal of medicine, 356 (5), 2007, 447–458.

Christopher H. Goss, Stacey A. Newsom, Jonathan S. Schildcrout, Lianne Sheppard and Joel D. Kaufman

"Effect of Ambient Air Pollution on Pulmonary Exacerbations and Lung Function in Cystic Fibrosis".

American Journal of Respiratory and Critical Care Medicine, 169 (7), 2004, 816–821.

Zoidis, John D. "The Impact of Air Pollution on COPD". RT: for Decision Makers in Respiratory Care, 1999.

Gehring, U., Wijga, A. H., Brauer, M., Fischer, P., de Jongste, J. C., Kerkhof, M., Brunekreef, B.Traffic-

related air pollution and the development of asthma and allergies during the first 8 years of life.American

journal of respiratory and critical care medicine, 181(6), 2010, 596-603.

Andersen, Z. J., Hvidberg, M., Jensen, S. S., Ketzel, M., Loft, S., Sorensen, M., Raaschou-Nielsen, O.

Chronic obstructive pulmonary disease and long-term exposure to traffic-related air pollution: a cohort

study.American journal of respiratory and critical care medicine, 183(4), 2011, 455-461.

Committee of the Environmental and Occupational Health Assembly of the American Thoracic Society,

Health effects of outdoor air pollution. American journal of respiratory and critical care medicine, 153(1),

1996, 3-50.

J. Sunyer, "Urban air pollution and Chronic Obstructive Pulmonary disease: a review". European Respiratory

Journal, 17(5), 2001, 1024–1033.

Raaschou-Nielsen, O, Andersen ZJ, Hvidberg M, Jensen SS, Ketzel M, Sorensen M, Tjonneland A, Lung

cancer incidence and long-term exposure to air pollution from traffic, Environmental health perspectives,

119(6), 2011, 860-865.

Raaschou-Nielsen, O., Andersen, Z. J., Hvidberg, M., Jensen, S. S., Ketzel, M., Sorensen, M., Tjonneland, A.,

Air pollution from traffic and cancer incidence: a Danish cohort study. a global access science source,

10,2011, 67.

Committee on Environmental Health, Ambient Air Pollution: Health Hazards to Children, Pediatrics, 114 (6),

2004, 1699–1707.

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http://www.sfgate.com/health/article/Bad-air-costing-state-s-economy-billions-3185388.php

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FORMULATION AND EVALUATION OF ORODISPERSIBLE TABLETS OF

CINNARIZINE BY SUPER DISINTEGRANTS ADDITION METHOD Praveen Khirwadkar

1, Kamlesh Dashora

2, Shyam Bihari Sharma

3*

1Institute of Pharmacy, Vikram University, Ujjain, Madhya Pradesh, India

2Institute of Pharmacy, Vikram University, Ujjain, Madhya Pradesh, India

3Jiwaji University, Gwalior, Madhya Pradesh, India


Abstract

The aim of the present work is to study the Preformulation parameters for Cinnarizine

orodipersible tablet. The objective of pre-formulation study is to generate information useful to the

formulator in developing stable and bioavailable dosage form. Administration of conventional tablets

of Cinnarizine has been reported to exhibit fluctuations in plasma drug levels, either in manifestation

of side effects or reduction in drug concentration at the receptor sites. Conventional Cinnarizine

tablets available in market are not suitable where quick onset of action is required. Besides, the

conventional tablets also show poor patient compliance particularly by the geriatric and pediatric

patients who experience difficulty in swallowing, and by those who are bed ridden or who are

traveling and do not have an easy access of water. Usage of excipients like low-substituted

hydroxypropyl cellulose, crospovidone helps for the faster disintegration and faster release of the

drug from the dosage form. The Preformulation studies were carried out in terms of test for

identification (physical appearance, melting point, and UV spectrophotometer), solubility profile, and

determination of partition coefficient and quantitative estimation of drug. All the observations and

results showed that the Cinnarizine could serve as suitable candidate for fast dissolving tablet that

may improve the bioavailability.

Keywords: Preformulation study, Orodispersible tablet, Patient compliance, Super disintegrants

INTRODUCTION

Tablets are solid preparations each containing a single dose of one or more active ingredients and are

obtained by compressing uniform volumes of particles. The objective of the design and manufacture of the

compressed tablet is to deliver orally the correct amount of drug in the desired location and to have its

chemical integrity protected to the point. Recent advances in Novel Drug Delivery System (NDDS) aims to

enhance safety and efficacy of drug molecule by formulating a convenient dosage form for administration and

to achieve better patient compliance. One such approach is “Mouth Dissolving Tablet”. This is an innovative

tablet technology where the dosage form containing active pharmaceutical ingredients disintegrates rapidly,

usually in a matter of seconds, without the need for water, providing optimal convenience to the patient.

Innovators and inventor companies have given these tablets various names such as orally disintegrating tablets

(ODT), mouth dissolving (MD), fast melting, fast dissolving or Orodisperse. The European Pharmacopoeia

defines Orodisperse as a tablet that can be placed in the mouth where it disperses rapidly before swallowing.

Researchers have formulated ODT for various categories of drugs, which are used for therapy in which rapid

peak plasma concentration is required to achieve desired pharmacological response. These include

neuroleptics, cardiovascular agents, analgesics, anti-allergic and drugs for erectile dysfunction. The concept of

Mouth Dissolving Drug Delivery System emerged from the desire to provide patient with conventional mean

of taking their medication. Difficulty in swallowing (Dysphasia) is a common problem of all age groups,

especially elderly and pediatrics, because of physiological changes associated with these groups of patients.

Other categories that experience problems using conventional oral dosage forms includes are the

mentally ill, uncooperative and nauseated patients, those with conditions of motion sickness, sudden episodes

of allergic attack or coughing. Sometimes it may be difficult to swallow conventional products due to

unavailability of water. These problems led to the development of novel type of solid oral dosage form called

“Mouth Dissolving Tablets”. This tablet disintegrates instantaneously when placed on tongue, releasing the

drug that dissolves or disperses in the saliva. Produce rapid onset of action.

In such a cases Bioavailability of

drug is significantly greater than those observed from conventional tablet dosage form.


Preformulation studies: Preformulation studies are the first step in the rational development of dosage form

of a drug substance. The objective of Preformulation studies are to develop a portfolio of information about

the drug substance, so that this information useful to develop formulation. Preformulation can be defined as

investigation of physical and chemical properties of drug substance alone and when combined with excipients.

Preformulation investigations are designed to identify those physicochemical properties and excipients that






may influence the formulation design, method of manufacture, and pharmacokinetic-biopharmaceutical

properties of the resulting product.

Organoleptic Characteristics: The color, odor, and taste of the drug were characterized and recorded using

descriptive terminology.

Drug Excipients Compatibility Study: IR Study were done for following drug, drug- excipients ratio

1. API alone (Cinnarizine)

2. API: Ac-Di-Sol®

→ 1: 1

3. API: Kollidone®

→ 1: 1

Super disintegrants addition method: Specified quantity of Cinnarizine, mannitol, Avicel 102, aspartame,

crospovidone, Ac-di-sol, talc and magnesium stearate were weighed accurately and passed through 60

#screen. All the materials were transferred to mortar and triturated till it mixed uniformly. The resulting

powdermixture was compressed into tablets using singlepunch tablet machine. Formulation of tablets wasre

presented in Table.1.

Table.1.Composition of Orodispersible Tablet of Cinnarizine

Formulation F1

(mg)

F2

(mg)

F3

(mg)

F4

(mg)

F5

(mg)

F6

(mg)

F7

(mg)

F8

(mg)

F9

(mg)

Cinnarizine 25 25 25 25 25 25 25 25 25

Manitol 30 30 30 30 30 30 30 30 30

crospovidone 8 12 16 8 12 16 8 12 16

Ac-di-Sol 2 2 2 4 4 4 6 6 6

Aerosil 2 2 2 2 2 2 2 2 2

Aspartame 3 3 3 3 3 3 3 3 3

Magnesium Stearate 1 1 1 1 1 1 1 1 1

Avicel 102Up to 200 200 200 200 200 200 200 200 200

RESULTS

Evaluation of the blend, evaluation of the tablet for post compression parameters, cumulative

percentage of drug release were done and noted in the tables 2 to 4. Stability studies as well as compatibility

studies were conducted and presented in table 5 and figures 1 to 3 respectively.

Table.2. Evaluation of the Powder Blend

Batch

Code

Bulk

Density

Tapped

Density

Angle Of

Repose

Percentage

Compressibility

Hausner’s

Ratio

F1 0.52 0.58 25.32 16.25 1.163

F2 0.53 0.62 26.23 18.23 1.200

F3 0.59 0.60 28.23 20.06 1.164

F4 0.54 0.78 23.56 14.52 1.170

F5 0.57 0.55 25.33 15.25 1.135

F6 0.59 0.77 29.36 17.23 1.152

F7 0.51 0.71 24.56 12.56 1.207

F8 0.53 0.69 30.21 18.31 1.212

F9 0.54 0.74 31.12 19.18 1.237

Table.3. Physical parameters of mouth dissolving Tablet

Batch

Code

Weight

Variation

Thickne

ss (Mm)

Hardness

(Kg/Cm2)

Friability

(%)

Disintegration

Time (Sec)

Wetting

Time (Sec)

Assay

(%)

F1 pass 2.56 3.4 0.73 30.6 ±1.25 64.0 ±1.35 98.14

F2 pass 2.57 2.5 0.76 43.7 ±2.46 65.8 ±0.35 99.02

F3 pass 2.60 2.5 0.79 56.4 ±2.42 79.0 ±0.85 100.51

F4 pass 2.63 3.2 0.74 29.6 ±1.22 32.4 ±1.15 98.91

F5 pass 2.65 3.0 0.78 30.6 ±1.25 65.0 ±1.35 100.04

F6 pass 2.66 2.5 0.80 33.0 ±1.00 35.0 ±0.95 99.86

B7 pass 2.51 3.0 0.69 23.3 ±0.58 29.1 ±1.05 98.92

F8 pass 2.52 2.5 0.65 35.5 ±0.50 41.7 ±1.45 101.05

F9 pass 2.54 2.5 0.66 36.5 ±0.50 34.12 ±1.45 100.3





Table.4.The following are the results of the dissolution studies of the formulated

Table.5. Stability parameters of formulation B7 stored at temperature 40C/75 % RH 40oC.

Parameters Control sample After 15 Days After 30 Days After 60 Days

Drug Content (%) 100.34 99.57 99.19 99.00

Disinintegration time (Sec) 25.3 26.2 26.8 25.90

Wetting Time (Sec) 29.1 30.2 30.9 30.25

Hardness(Kg/Cm2) 3.00 3.1 3.3 3.4

Fig. No 1: IR Spectrum of Pure Drug (Cinnarizine) Fig. No. 2 IR Spectrum of Cinnarizine + Ac-di-sol

Fig. No. 3 IR Spectrum of Cinnarizine + Crospovidone

CONCLUSION

Cinnarizine is a histamine H1-receptor antagonist is the most frequently prescribed drug in treatment

of motion sickness, vomiting, allergic reaction, vertigo and insomnia. Conventional Cinnarizine tablets

available in market are not suitable where quick onset of action is required. To overcome these problems,

there is a need to develop a rapidly disintegrating dosage form, particularly one that would rapidly disintegrate

in saliva and could be administered without water anywhere anytime. No such mouth dissolving tablet of

Cinnarizine is available in the market.

The present investigation was aimed to evaluate the possibility of using different parameters for the

development of fast dissolving of Atenolol. Preformulation studies were done using various parameters such

as identification test of Atenolol. The description and appearance, melting point and solubility were also

performed for further characterization & it was found that all results are satisfactory.

REFERENCES

Bhushan SY, Sambhaji SP, Anant RP, and Kakasaheb RM, New drug delivery system for elderly, Indian

drugs, 37, 2000, 312-318.

Time

(Min)

Cumulative percentage Drug Release

F1 F2 F3 F4 F5 F6 F7 F8 F9

0 0 0 0 0 0 0 0 0 0

2 63.75 60.82 55.00 74.97 71.47 69.06 75.77 73.64 72.65

4 75.09 65.75 62.97 84.39 81.97 79.91 88.09 84.34 80.88

6 81.79 79.04 75.65 92.68 89.75 88.85 93.08 90.85 89.58

8 92.19 86.34 87.78 96.65 95.32 94.90 98.08 97.79 94.46

10 99.3 96.22 97.32 99.6 98.08 97.61 99.89 98.75 97.90





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http://www.emc.medicines.org.uk/



Mohanraghupathy S et.al Indian Journal of Research in Pharmacy and Biotechnology


EFFECTIVE HYPOGLYCEMIC ACTION OF METFORMIN COMBINATIONS

AGAINST DEXAMETHASONE INDUCED DIABETES MELLITUS IN RATS Mohanraghupathy S*, Jayabharath N, Bhuvana tejaY, Hameera khanam B, Lavanya lahari B

Department of Pharmacology, Annamacharya College of Pharmacy, Kadapa, Andhra Pradesh


ABSTRACT

Diabetes mellitus is a chronic metabolic disorder occurs due to the damage of β-cells of islets

of Langerhans in pancreas. In general patients suffering for diabetes mellitus always use the medicines,

apart from the medicines they take insulin also. In our study we did the preclinical experimental study of

Metformin combinations on Albino Wistar rats such that which combination is effective for the patients

who are using purposefully using the insulin. Our studies shown that Metformin + Glipizide shown the

effective hypoglycemic results. For inducing diabetes mellitus we introduced Dexamethasone

intraperitoneally as it is a steroidal anti-inflammatory agent and it is having hypoglycemic action as

adverse effect.

Key Words: Diabetes mellitus, metformin, glimepiride, glipizide

INTRODUCTION Diabetes Mellitus: It is a metabolic disorder characterized by hyperglycemia, glycosuria, hyperlipidaemia,

negative nitrogen balance and sometimes ketonaemia. A widespread pathological change is thickening of

capillary basement membrane, increase in blood vessel wall matrix and cellular proliferation resulting in vascular

complications like lumen narrowing, early atherosclerosis, and sclerosis of glomerular capillaries, retinopathy,

neuropathy and peripheral vascular insufficiency.

Insulin-dependent diabetes mellitus (IDDM), juvenile onset diabetes mellitus: There is β cell destruction in

pancreatic islets, majority of cases are auto immune (type 1A) antibodies that destroy β cells are detectable in

blood, but some are idiopathic (type 1B) no β cell antibody is found. In all type 1 cases circulating insulin levels

are low or very low, and patients are more prone to ketosis. This type is less common and has a low degree of

genetic predisposition.

Noninsulin-dependent diabetes mellitus (NIDDM), maturity onset diabetes mellitus: There is no loss or

moderate reduction in β cell mass; insulin in circulation is low, normal or even high, no anti-β-cell antibody is

demonstrable; has a high degree of genetic predisposition; generally has a late onset (past middle age). Over 90%

cases are type II DM. causes may be:

1) Abnormality in gluco-receptor of β cells so that they respond at higher glucose concentration or relative β

cell deficiency.

2) Reduced sensitivity of peripheral tissues to insulin: reduction in number of insulin receptors, ‘down

regulation’ of insulin receptors. Many hypertensives are hyperinsuliaemic, but normoglycaemic; exhibit

insulin resistance associated with dyslipidaemia (metabolic syndrome). Hyperinsulinaemia has been

implicated in causing angiopathy.

3) Excess of hyperglycemic hormones (glucagon, etc.) /obesity: cause relative insulin deficiency-the β cells

lag behind.

4) The third main form, gestational diabetes occurs when pregnant women without a previous diagnosis of

diabetes develop a high blood glucose level. It may precede development of type 2 DM.


Metformin, Glimipride, Glipizide and Dexamethasone were purchased from the near community pharmacy.

Kits used for the estimation of albumin, total cholesterol, blood urea nitrogen were purchased from Erba

Diagnostics. Blood glucose levels were estimated with the help of Glucometer which was purchased from

community pharmacy. Male Wistar albino rats weighing 150-250gms, Manjunatha institute of animal house,

Bangalore.

Maintenance of animals: Animals were kept for 1 week to acclimatize laboratory conditions before starting the

experiment. The animal house temperature was always maintained within the range of approximately 22-290c.

The relative humidity of animal house was maintained within the range of 30-70% throughout the duration of

experiment. Sufficient diet and water was supplied every day. All the experimental procedures and protocols used

in the study were approved by institutional animal ethical committee (IAEC).


http://en.wikipedia.org/wiki/Gestational_diabetes





0

50

100

150

200

250

Normal Diabetic Metformin + Glimepride

Metformin + Glipizide

Body weight

Fasting blood glucose levels

Post prandial blood glucose level

Experimental protocol

Group – I: It consists of normal animals which were received no dug and only food was kept. This group consists

of four animals.

Group – II: It consists of four animals which were received inducing agent Dexamethasone of 0.75mg/kg body

weight with normal food.

Group – III: It consists of four animals which were received inducing agent Dexamethasone of 0.75mg/kg body

weight. And also these animals received the combination of hypoglycemic agents [Metformin (500mg/kg body

weight) + Glimepride (1mg/kg body weight)] with normal food.

Group – IV: It consists of four animals which were received inducing agent Dexamethasone of 0.75mg/kg body

weight. And also these animals received the combination of hypoglycemic agents [Metformin (500mg/kg body

weight) + Glipizide (5mg/kg body weight)] with normal food.

Biochemical parameters: After intraperitoneal administration of Dexamethasone in rats the Diabetes was

induced after thirty days. The dose was calculated by taking the body weight of rat. The blood glucose levels were

examined with help of glucometers. When the fasting blood glucose levels shown 150mg/dl in rats those animals

were considered as diabetic. And we examined the body weight additionally. The results of Body weight and

Blood glucose levels are given below:

Table 1: Blood glucose levels of the experimental animals

Fig 1: Blood glucose levels of the experimental animals

Tab 2: Estimation of Total protein, Cholesterol, Blood Urea Nitrogen, Serum Creatinine

Groups Total protein Cholesterol Blood Urea

Nitrogen

Serum

creatinine

Control 6.99 82.20 42.66 0.50

Diabetic 3.77 182.2 0.78 0.80

Metformin+Glimipride 5.02 98.8 0.62 0.62

Metformin + Glipizide 6.05 91.9 0.59 0.58

Groups Normal Diabetic Metformin +

Glimepride

Metformin +

Glipizide

Body weight (gm) 230 175 185 187

Fasting blood glucose levels (mg/dl) 90 147 150 160

Post prandial blood glucose level (mg/dl) 100 180 114 100





0

20

40

60

80

100

120

140

160

180

200

control diabetic met+gl met+gz

Total protein

Cholesterol

BUN

Serum creatinine

Fig 2: Estimation of Total protein, Cholesterol, Blood Urea Nitrogen, Serum Creatinine

CONCLUSION

The Metformin combination therapy on Dexamethasone induced diabetes mellitus on rats proved that

instead of administration of insulin to the patients with excess blood glucose levels oral therapy is always safe. It

also denotes that the metabolic activity of the organs as well as the different mechanism of action of medicines is

going to reduce the blood glucose level perfectly. Glipizide and Metformin combination showed the valuable

results on therapy when compared to Glimipride. The both drugs Glimipride and Glipizide are the secretogogues

enhancing the insulin levels as well as the Metformin is having the property of reducing the glucose level by

glucose reuptake by the cells. Finally we concluded that Metformin and Glipizide combination is the best.

DISCUSSION

Patients suffering with diabetes mellitus with blood glucose levels above 200mg/dl always have a

little pain on taking insulin through intramuscular route. Hypoglycemic agents with different combinations when

used through oral route are not only effective but also safe for the patients. The metabolic rate of the medicines

should always be examined properly because, in some patients the medicines after prolong use may not work due

to improper binding of the drug molecules at the receptor sites. This factor leads to increased blood glucose level.

Immediately the patient should not be advised to take insulin because, insulin is having adverse hypoglycemic

effect. Patients are advised to have the physical exercise like walking or working like gardening, sweeping, lifting

mild luggages etc because whatever the diet we are taking it is not only digested in the stomach but also should be

utilized in the form of energy in tissues through work.

REFERENCES

Akhila shetty J and Divya Choudhary, Effect of insulin plant (Costus igneus) leaves on Dexamethasone induced

hyperglycemia, JAR. 1, 2010, 100.

Department of health and families, Glucometers, RHA, 2005, 1-4.

Good Man and Gilman, Stephen N. Davis and Daryl K. Granner, Insulin and oral hypoglycemic agents and the

pharmacology of the endocrine pancreas, The pharmacological basis of therapeutics, 10th edition, 1679.

Harsh Mohan, Diabetic Nephropathy, Text book of pathology, 6th editon, 2010, 677.

Padmaja Udaykumar, Insulin and oral hypoglycaemics, Text book of Medical pharmacology, 2nd

edition, 2009,

486.

Vasanth Muthu Swamy, Lal Krishna, Murugan SS, CPCSEA guidelines for laboratory animal facility, 2009, 9-10



Suruchi Singh et.al Indian Journal of Research in Pharmacy and Biotechnology


A REVIEW ON MEDICINAL PLANTS HAVING ANTIOXIDANT POTENTIAL S.K Sharma, Lalit Singh, Suruchi Singh

*

Sunder Deep Pharmacy College, Ghaziabad, U.P, India.

*Corresponding author: E.mail: [email protected]

ABSTRACT

Natural compounds from plants and other life forms (bacteria, fungi, marine organisms)

represent a major source of molecules with medicinal properties. Among them, antioxidant substances

are of particular interest. The understanding of the central role that oxidative stress holds in the

progression of disorders as varied as: cardiovascular diseases, degenerative conditions, rheumatic

disorders, metabolic syndrome, and in aging, makes antioxidant capacity to a key-feature of modern,

multipotent remedies. A lot of medicinal plants, traditionally used for thousands of years, are present in a

group of herbal preparations of the Indian traditional health care system (Ayurveda) named Rasayana

proposed for their interesting antioxidant activities.

Keywords: Antioxidant, Ayurveda, Rasayana, Oxidative stress.

INTRODUCTION

Antioxidants are substances that may protect your cells against the effects of free radicals. Free radicals

are molecules produced when your body breaks down food, or by environmental exposures like tobacco smoke

and radiation. Free radicals can damage cells, and may play a role in heart disease, cancer and other diseases.

Studies suggest that a diet high in antioxidants from fruits and vegetables is associated with a lower risk of cancer,

cardiovascular disease, Parkinson's disease and Alzheimer's disease. A plant-based diet protects against chronic

oxidative stress-related diseases. Dietary plants contain variable chemical families and amounts of antioxidants. It

has been hypothesized that plant antioxidants may contribute to the beneficial health effects of dietary plants. Our

objective was to develop a comprehensive food database consisting of the total antioxidant content of typical

foods as well as other dietary items such as traditional medicine plants, herbs and spices and dietary supplements.

Since ancient times, the medicinal properties of the plant materials improve the quality and nutritional

value of plants has been investigated in the recent scientific form. While, flavonoids are a group of polyphenolic

developments throughout the world, due to their potent compounds with known properties, which include free

antioxidant activities. The antioxidants have been reported to have radical scavenging, inhibition of hydrolytic

and oxidative to prevent oxidative damage caused by free radical.

Antioxidants Potential Plants

Free radicals are atoms or molecules with singlet, i.e. unpaired electron which makes them highly

reactive. Oxidative free radicals are generated by metabolic reactions create a chain reaction leading to

membrane and other lipid peroxidation, DNA damage, etc. This has been implicated in atherosclorosis

(oxidated LDL is more atherogenic), cancers, neurodegenerative and inflammatory bowel diseases. Many

endogenous and dietary compounds like superoxide dismutase, ferritin, transferrin, reruloplasmin,

tocopherol, carotene and ascorbic acid have anti oxidant and free radical scavenging properties. Small

amounts of reactive oxygen species are continually formed in the body in the cell membrane and close to the cells

organelles. They act where they are generated. Hence, they can damage most cell structures including membrane

lipids, proteins, enzymes and nuclic acids.

The body has mechanisms to produce the small amounts of oxidants normally formed during

metabolic reaction. Reactive species such oxidants are formed in controlled amounts by neutrophil

leucocytes on exposure to microbes are beneficial to the body in that they participate in destroying the

microbes. Excess of oxidants, however, can be harmful to the body. Liver is also under constant threat of oxidants

and some of the free radical especially H2O2. Lipid peroxidation has been demostred as one of the important

feature after exposure to hepatotoxic substances and also is a measure of extent of hepatic damage. Several

herbs and herbal formulations are available for the scavenging activity. In addition to this there is a

global trend to revive the traditional systems of medicines and renewed interest in the natural remedies for

treating human ailments. Antioxidants have important preventive roles, not only on undesirable changes in the

flavor and nutritional quality of food, but also on tissue damage in various human diseases. Almost all organisms

are well protected against free radical damage by either enzymes or compounds, such as ascorbic acid, α-

tocopherol and gluthione.






When the mechanism of antioxidant protection unbalanced by the deterioration of different factors,

physiological functions can occur which result in diseases or accelerated aging. Consequently, it is important to

find compounds that prevent oxidation. Antioxidants have important preventive roles not only on undesirable

changes in the flavor and nutritional quality of food, but also on tissue damage in various human diseases. They

are effective in prevention of degenerative illnesses, such as different types of cancers, cardiovascular and

neurological diseases, cataracts and oxidative stress dysfunctions. Polyphenols are the most significant

compounds for the antioxidant properties of plant raw materials. Then antioxidant activity of polyphenols is

mainly due to their redox properties, which allow them to act as reducing agents, hydrogen donors, singlet oxygen

quenchers, metal chelators and reductants of ferryl hemoglobin. Medicinal plant parts are commonly rich in

phenolic compounds, such as flavonoids, phenolic acids, stilbenes, tannins, coumarins, lignans and lignins. These

compounds have multiple biological effects including antioxidant activity.

CONCLUSION

As antioxidant is a molecule capable of slowing or preventing the oxidation of other molecules. Oxidation

is achemical reaction that transfers electron from a substance to an oxidizing agent. Oxidation reactions can

produce free radicals, which start chain reactions that damage cells. Antioxidants are the substances that inhibit

oxidation and are capable of counteracting the damaging effects of oxidation in body tissue. They prevent damage

caused by free radicals. Free radicals are very unstable molecules with an unpaired electron and are important

intermediates in natural processes involving control of vascular tone, cytotoxicity and neurotransmission. Free

radicals cause many human diseases like cancer, Alzheimer’s disease, cardiac reperfusion abnormalities, kidney

disease and fibrosis etc. Antioxidants play many vital functions in a cell and have many beneficial effects when

present in foods.

Table 1. List of plants exhibit antioxidant characteristics and their chemical constituents

PLANT NAME PLANT PART MAIN CHEMICAL CONSTITUENTS

Withania somnifera

Ocimum sanctum

Piper nigrum

Arentium lappalo

Scutellaria barbata

Daucus carrota

Coleus ferscoli

Salvia sclarea

Eugenia caryophylla

Allium sativum

Zingiber officinalis

Ginkgo biloba

Vitis vinifera

Berries, leaves,

roots

Leaves, Seeds

Fruit

Root

Leaves,

Leaves, Seed, Root

Roots

Entire plant, seed

Inflorescence

Leaves, Bud

Leaves, Rhizome

Plant

Fruit, Seed

Ascorbic acid,α-tocopherol and reduced glutathione,

superoxide dismutase,ascorbate peroxidase, catalase,

peroxidase & polyphenol oxidase

Ascorbic acid,β-carotene, β-sitosterol, eugenol,Palmitic acid,

tannin

Ascorbic acid, β carotene,Lauric acid, myristic acid, palmitic

acid, piperine

Insulin, tannic acid

Gallic acid

Alanine, α tocopherol, ascorbic acid, camphene,eugenol,γ-

terpinene, histidine Antitoxin

Ferscolin

ˠ-terpinene, linalyl acetate, myrcene,

Palmitic acid, rosemarinic acid

Acetyl-eugenol, Ascorbic acid, β -carotene, β-sitosterol,

caryophyllene oxide, eugenol, isoeugenol

Alanine, Ascorbic acid, β-sitosterol,Caffeic acid, Kaemferol,

Methionine

6-Gingerol,alanine, Ascorbic acid, Histidine, Lauricacid,

Methionine, Myristic acid,Palmitic acid, Tryptophan

EGB 761,Ginkgogolide

Alanine, α-tocopherol, Ascorbic acid, β -carotene, β-

sitosterol, Histidine, OPC, Methionine, Palmitic acid,





Citrus aurantifolia

Cymbopogon citratus

Commiphora myrrha

Myristica fragrance

Olea europaea

Mentha piperata

Catharanthus roseus

Rosemarionus

officinalisL

Santalum album

Curcuma domestica

Acorus calamus

Alisma plantago-

aquatica L.

Allium ursinum L.

Cotinus coggygria Scop.

Angelica sylvestris L.

Anthriscus cerefolium

Anthriscus sylvestris

Carum carvi L.

Ery

ngium campestre L.

Sanicula europaea L.

Achillea millefolium s.l.

Arctium lappa L.

Artemisia absinthium L

Artemisia vulgaris L.

Bellis perennis L

Bidens tripartita L.

Carlina acaulis L.

Carthamus tinctorius L.

Cichorium intybus L.

Cirsium arvense (L.)

Scop

Fruit

Leaves

Resin, Sap

Seed, Leaf

Leaf

Leaf

Leaf

Entire Plant

Leaf, Oleoresin

Fruit, Wood

Rhizome

Rhizome

Flowering aerial

parts, roots

Leaf

Leaf

Root, Grains

Root, Flowering

aerial part

Flowering aerial

part

Fruits

Flowering aerial

part

Flowering aerial

part

Leaf, root

Flowering aerial

part

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Root

Flower

Flowering aerial

part, root

Leaf

Flowering aerial

part

Flowering aerial

parts

selenium

Alanine, α –pinene,ascorbic acid,β -Sitosterol, caffeic acid,

Eugenol, Linalylacetate, Palmitic acid, Tannin

Β-sitosterol,Myrcene,Selenium

Β-Sitosterol,campestrol,eugenol

Lauric acid,Myrcene, Palmitic acid

Α -tocopherol,apigenin, β -carotene, γ -

tocopherol,kaempferol, Luteolin

Menthol, Limonene

Vincristine, Vinblastine

Carsonic acid, Rosemaric acid,

Β –sitosterol , Caryophyllene oxide, eugenol,isoeugenol

Alanine, eugenol, β -sitosterol, Palmitic acid, phenol

Curcumin,tannins, phenolic acids

Only antioxidative fractions devoid of beta-asarone should

be used,

Triterpene (alisol B)

Flavonoids, sulfur-containing compounds

Flavones, aurones, chalcones

Flavonoids, coumarins

Flavonoids (apiin), lignans

Flavonoids (quercetin, apigenin)

Flavonoids, volatile oil

Flavonoids, triterpenes

Rosmarinic acid derivative

Flavonoids, tannins, volatile oil

Flavonoids

Flavonoids

Flavonoids

Flavonol glycosides

Flavonoids

Flavonoids

Flavonoids

Phenolic acids, flavonoids

Phenolic acids,acidic polysaccharides with unprecised

structure

Flavonoids

Flavonoids, volatile oils


Flavonoids


Flavonoids, polysaccharides (mucilages)

Flavonoids

Flavone 6-C-Glycosides





Conyza canadensis L.

Cronq.

Hieracium pilosella L

Matricaria recutita L.

Onopordum acanthium

Solidago virgaurea L.

Taraxacum officinale

agg.

Tussilago farfara L

Betula pendula Roth

Alliaria petiolata

Capsella bursa-pastoris

Nasturtium officinale

Humulus lupulus L.

Sambucus nigra L.

Sambucus ebulus L.

Viburnum lantana L.

Viburnum opulus L.

Evonymus europaeus L.

Cornus mas L.

Corylus avellana L.

Juniperus communis L.

Hippophae rhamnoides

Elaeagnus angustifolia .

Equisetum arvense L.

Calluna vulgaris (L.)

Vaccinium myrtillus L.

Anthyllis vulneraria L.

Genista tinctoria L.

Lotus corniculatus L.

Melilotus officinalis L.

Pallas

Ononis spinosa L.

Trifolium arvense L.

Flowers

Flowering aerial

parts

Flowering aerial

parts

Root, Flowering

aerial parts

Leaf

Leaf

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Glandulae

Flowers

Leaf

Branches

Branches

Grains

Fruits

Grains, Leaf

Fruits

Fruits

Leaf, Branch

Flowering aerial

parts

Flowering aerial

parts

Leaf, Fruit

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts

Bark, Flowers

Bark

Flavonoids, glucosinolates

Flavonoids, glucosinolates

Flavonoids

Flavonoids

Flavonoids

Flavonoids, procyanidins

Flavonoids, procyanidins

Flavonoids

Flavonoids, phenolic acids

Phenolic acids

Flavonoids

Flavonoids, carotenoids

Flavonoids

Flavonoids

Flavonoids

Anthocyans

Flavonoids,isoflavones (genistein)

Flavonoids, triterpenes

Flavonoids

Triterpenes

Isoflavones

Isoflavones

Isoflavones

Tannins, procyanidins, flavonoids

Tannins, procyanidins,

Flavonoids

Xanthones, phenolic acids

Tannins, gallic acid

Flavonoids, tannins

Flavonoids

Flavonoids

Tannins, flavonoids

Flavonoids, phenylpropanoids

(verbascoside)


Flavonoids

Flavonoids


Flavonoids

Flavonoids

Flavonoids


Flavonoids





Trifolium pratense L.

Trifolium repens L.

Quercus petraea L.

Quercus robur L.

Centaurium erythraea

L.

Erodium cicutarium L.

Geranium

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts, roots

Flowering aerial

parts

Flowering aerial

parts

Flowering aerial

parts



Flavonoids

Flavonoids, iridoids

Phenolic acids,flavonoids, carotenoids

Polysacharides, flavanoids

Polysaccharides (mucilages), flavonoids

Flavonoids, coumarins

phenylpropanoids (verbascoside)

Tannins,

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Beta vulgaris, Mal NJ Nutr, 16(3) 2010, 397-408.

TK Gopal, Harish G, D Chamundeeswari, C Umamaheswara Reddy, In-vitro Anti-Oxidant Activity of Roots of

Boerhaavia diffusa Linn, Research journal of Pharmaceutical, Biological and Chemical Sciences, 4, 2010, 782-

788.



Vijayakumar and Hindumathy Indian Journal of Research in Pharmacy and Biotechnology


INVITRO ANTI-INFLAMMATORY ACTIVITY OF STRYCHNOS POTATORUM

LINN SEED BY HRBC MEMBRANE STABILIZATION V.Vijayakumar*

1, Dr C.K.Hindumathy

2

1. Department of Pharmaceutical Chemistry, Vinayaka Missions College of Pharmacy.

2. Department of Biotechnology, V.M.K.V. Engineering College.

* Corresponding author: E-mail: [email protected]

ABSTRACT

In the present study, strychnos potatorum linn belonging to the family Loganiaceae (Strychnaceae)

was studied. It is a folkloric medicinal plant used to treat antibacterial, antifungal, anti diabetic,

antioxidant and anti-inflammatory. Since many flavonoids have remarkable anti-inflammatory activity the

present work aims at evaluating the anti inflammatory activity of seeds strychnos potatorum linn by

HRBC membrane stabilization method. The prevention of hypo tonicity induced HRBC membrane lysis

was taken a measure of anti inflammatory activity. The anti-inflammatory activity of hydroalcholic

extract was comparable to that of the standard drug Hydrocortisone. The percentage protection for the

hydroalcholic extract and hydrocortisone were 100 at µg/ml. the hydroalcholic extract of strychnos

potatorum linn seed has significant anti-inflammatory activity (54.95±0.74).

Key words: Anti-inflammatory, strychnos potatorum linn, HRBC membrane stabilization,

Hydroalcholic extract.

INTRODUCTION

Inflammation is a normal protective response to tissue injury that is caused by physical trauma, noxious

chemicals or microbiological agents. Inflammation is the result of concerted participation of more number of

proliferative factors (like Vasoactive, chemotatic) at different stages and there are many targets for anti

inflammatory activity. (Tripathi, K.D, 2004). The irrespective tissue injury is a type of inflammatory response

suppressed by glucocorticoids and this is the basic clinical uses and also it interferes with several steps in the

inflammatory response. Numbers of corticoids are only palliative; do not act on the inflammation instead of they

favor spread of infections capacity of defensive cells to kill microorganisms is impaired at the same time interfere

with healing and scar formation. The alternate use of corticoids is hazardous other than the corticosteroids the

NASIDs are also used to treat inflammation .the main mechanism of action of the NASIDs are the inhibition of

prostaglandin (PG) synthesis or preferential or selective COX-2 inhibition. Due to the inhibition of prostaglandin

(PG) synthesis it may produce toxic effects like bleeding, inhibition of platelet function, gastric mucosal damage,

asthma and anaphylactic reactions may cause some individuals. (Tripathi, K.D, 2004).the plant strychnos

potatorum linn commonly known as Nirmali and otherwise called clearing nut. The plant is distributed throughout

India, Ceylon, Burma, and Deccan. The present work aims at evaluating the anti inflammatory activity of

strychnos potatorum linn seed by HRBC membrane stabilization.


Plant material: The fresh seeds of strychnospotatorum Linn were collected from shervaroys hills, Salem district,

Tamilnadu, India. The collected seeds were identified and authenticated by the Botanist Dr. A. Balasubramanian

(consultant central siddha research) Executive Director ABS Botanical Garden, Salem, Tamilnadu.

Preparation of extract: The dried seeds are powdered using mixer grinder; 1000gm of the powdered seeds was

packed evenly in the soxhelet extractor and subjected to extraction with Hydroalchol (70% ethanol, 30%

water).After extraction, the solvent was distilled off and the extracts were concentrated on water bath to a dry

residue and kept in a desiccator.

Anti inflammatory activity: The HRBC membrane stabilization has been used as a method to study the anti-

inflammatory activity. (Gandidasan.R, 1991) Blood was collected from healthy volunteers. The collected blood

was mixed with equal volume of sterilized Alsever solution (2% dextrose, 0.8% sodium citrate, 0.05% citric acid

and 0.42 % sodium chloride in water).the blood was centrifuged at 3000 rpm and packed cells were washed with

isosaline (0.85 % , pH 7.4) and a 10% v/v suspension was made with isosaline.

The assay mixture contains the drug (at various concentrations as mentioned in the table), 1 ml phosphate

buffer (0.15 M, pH 7.4), and 2 ml of hypo saline (0.36%) and 0.5 ml of HRBC suspension. Hydrocortisone

sodium was used as the reference drug. Instead of hypo saline 2 ml of distilled water was used in the control. All

the assay mixtures were incubated at 370C for 30 minutes and centrifuged. The haemoglobin content in the






supernatant solution was estimated using spectrophotometer at 560 nm. The percentage of hemolysis was

calculated by assuming the hemolysis produced in the presence of distilled water as 100%. The percentage of

HRBC membrane stabilization or protection was calculated by using the following formula,


The lysosomal enzymes released during inflammation produce a various disorders. The extra cellular

activity of these enzymes is said to acute or chronic inflammation. The non steroidal drugs act either by inhibiting

these Lysosomal enzymes or by stabilizing the membrane. (Rajendran Vadivu, 2008) since HRBC membrane are

similar to Lysosomal membrane components the prevention of hypo tonicity induced HRBC membrane lysis is

taken as a measure of anti inflammatory activity of drugs. The results were reported in table 1. It was observed

from the table 1 and figure 1 that the hydroalcholic extract shows significant anti inflammatory activity at the

concentration 100 µg/ml, which is comparable to the standard drug hydrocortisone sodium. The anti inflammatory

activity of the extract were concentration dependent, with increasing concentration the activity is also increased.

The hydroalcholic extract has significant anti inflammatory activity.

Table.1. In-vitro anti inflammatory activity strychnos potatorum linn seed

Concentration in

( µg/ml )

Activity ( % protection ) Standard Hydrocortisone

sodium HAESP

Control - -

20 43.53±0.62 55.48±0.57

40 46.41±0.87 57.71±0.54

60 49.00±0.50 59.21±0.95

80 52.57±0.68 63.56±0.68

100 54.95±0.74 67.50±0.52

(Values are expressed as SEM of 3 readings)

Figure1. In-vitro anti inflammatory activity strychnos potatorum linn seed

CONCLUSION

The extract exhibited membrane stabilization effect by increasing hypo tonicity induced lysis of

erythrocyte membrane. The erythrocyte membrane is analogus to the Lysosomal membrane (Chou, 1977) and its

stabilization implies that the extract may as well stabilize Lysosomal membrane. Stabilization of Lysosomal

membrane is very important in limiting the inflammatory response by preventing the release of Lysosomal

substances of activated neutrophil such as bacterial enzymes and proteases which cause further tissue damages.

(Murugasan, 1981) From the investigation it was concluded that the hydroalcholic extract of strychnos potatorum

linn has significant membrane stabilization property and it was comparable to the standard drug hydrocortisone.

0

20

40

60

80

20 40 60 80 100

Act

ivit

y %

pro

tect

ion

concentration in µg /ml

Invitro anti inflammatory effect of strychnos

potatorum linn

HAESP

STANDARD DRUG





REFERENCES

Chou CT, The Anti inflammatory effect of Tripterygium Wilfordihook on adjuvant induced paw edema in rats

and inflammatory mediator’s release, Phytother Res, 11, 1997, 152-154

Gandidasan R, Thamaraichelvan A, Baburaj S, Anti inflammatory action of Lannea coromandelica by HRBC

membrane stabilization. Fitoterapia, 1991, voll LXII, 81-83.

Kirtikar K.R, Basu B.D, Indian Medicinal plants. International book distributors, Dehradum.1975; II edn; Vol III;

1971.

Murugasan, Vember, S, Damodharan, C, C. Studies on erythrocyte membrane IV, In-vitro hemolytic activities of

Oleander extract .Toxicol Lett, 8, 1981, 33-38.

Nirmala Devi K, Periyanayagam K, in-vitro Anti inflammatory activity of Plectrnthus Amboinicus (Lour) spring

by HRBC membrane stabilization, International journal of pharmaceutical studies and Research, 1(1), 2010; 26-

29.

Rajendran Vadivu, Lakshmi K.S.In vitro and in-vivo anti inflammatory activity of Leaves of Smplocos

cochinchinenisis (Lour) Moore ssp Laurina. Bangladesh JPharmacol, 3, 2008, 121-124.

Ram P Rastogi, Mehrotra B N, Compendium of Indian Medicinal plants, CDRI Lucknow and publication and

information Directorate, New Delhi, 2, 1970, 79-201.

Tripathi KD, Essentials of Medical Pharmacology, Jaypee brothers medical publishers (P) Ltd, New Delhi, 2004,

V edn; 167-181, 257-259.

Warrier P.S, Indian Medicinal plants, Arya Vaidhya Sala, Kottakkal, Orient Longmann limited. Hyderabad, Voll

IV, 315-317.



Ramanji Naik Indian Journal of Research in Pharmacy and Biotechnology


SYNTHESIS AND CHARACTERIZATION OF 1, 3, 4-OXADIAZOLE AND 1,3,4-

THIADIAZOLE Ramanji Naik *

Vaagdevi College of pharmacacy and research center, Nellore, AP


ABSTRACT

There are vast numbers of pharmacologically active heterocyclic compounds in regular clinical use.

The presence of heterocyclic structures in diverse types of compounds is strongly indicative of the

profound effects such structure exerts on physiologic activity, and recognition of this is abundantly

reflected in efforts to find useful synthetic drugs. The 1, 3, 4-oxadiazole and 1,3,4- thiadiazole nucleus has

emerged as one of the potential pharmacophore responsible for diverse pharmacological properties.

Literature is flooded with reports of a variety of biological activities of 2, 5-Disubstituted-1,3,4-

oxadiazoles and 1,3,4- thiadiazole. The search for better anticonvulsant drug and the importance of 2,5-

disubstituted 1,3,4-oxadiazoles and 1,3,4- thiadiazole as anticonvulsant pharmacophores, prompted us to

design, synthesize and evaluate a series of differently substituted 1,3,4-oxadiazoles and 1,3,4- thiadiazole

for their potential anticonvulsant activity by autodock software.

KEY WORDS: 1, 3, 4-oxadiazole and 1, 3, 4- thiadiazole , synthesis

INTRODUCTION

The search for new effective and safe drugs has led today’s researchers to improve the existing drugs by

increasing their potency, duration of action and decreasing their toxic side effects. Structural activity studies

shows that a variation in the ring system or incorporation of different biologically active ring systems or minor

group modifications extends distinct biologically active ring systems or minor group modifications extends

distinct pharmacological effects upon the drug molecules. The past quarter of a century has seen a great increase

in the technical applications of heterocyclic compounds, in a Pharmaceutical field due to their various synthetic

strategies. This attracted the researchers and enormous approaches were made from their side to evaluate its

activities.

MATERIAL AND METHODS

Isonicotinic Acid, Indole 3-Acetic Acid Procured from SISCO research lab, Quinoline 2-carboxylic acid,

Ethanol PROCURED FROM FISCHER LABS, Phenyl Isothiocyanate, Methanol, Potassium Iodide, Sodium

Hydroxide PROCURED FROM SISCO research lab

SCHEME

STEP I STEP II

RCOOH RCOOClRCONHNH2

NH2NH2 .H2O

(Ia-e) (IIa-e)

PCl5 , CCl4

, 2 hr

STEP III

RCONHNH2 +

N

C

S

RCONHNHC=S

NH

Conc H2SO4

I2/KI

Ethanolic NaOH

O

NN

HN

R

S

NN

HN

R

(IIa-e)

(IIIa-e)

(IVa-e)

(Va-e)

STEP IV






Table.1.Different substitution in compounds IVa-e and Va-e

Compound R Compound R

Iva

N

Va

N

IVb

NH

H2C

Vb

NH

H2C

IVc

N

Vc

N

IVd Cl

Vd

Cl

IVe H2N

Ve H2N

EXPERIMENT

Synthesis

General method of Synthesis of Acid chlorides (Ia-e): A mixture of substituted acid (0.03 mole) and

phosphorus pentachloride (0.05ml) in anhydrous carbon tetra chloride (20ml) was refluxed for 2 hour at 1000c.

Solvent was distilled off and the solid acid chloride thus obtained was used for further reaction.

General method of Synthesis of Acid hydrazides (IIa-e): In the acid chloride (0.03 mole), hydrazine hydrate

was added (0.1 mole) drop wise at 00c and the resultant mixture was stirred for 5 hours at room temperature. A

solid that separated out was washed with aqueous sodium carbonate (10%) and dried in vaccum. It was

recrystalized from methanol to the pure crystalline solid was obtained.

General method of Synthesis of substituted thiosemicarbazide (III-a-e): Substituted thiosemicarbazides (IIIa-

e)were synthesized by refluxing substituted acid hydrazides (0.03moles) with phenyl isothiocyanates (0.03moles)

in 20 mL of ethanol on a boiling water bath for 5-6 h. After completion of reaction, the reaction mixture was

concentrated and kept overnight at room temperature. The needle shaped crystals of thiosemicarbazides so

obtained were filtered.

Synthesis of compound (IVa-e): 2-(phenyl) amino-5-substituted-1,3,4-thiadiazoles (IVa) were synthesized by

cyclization of substituted thiosemicarbazides (0.004 mole) with sulfuric acid at 0-5OC. After completion of

reaction, the mixture was poured onto crushed ice; the solid so separated was filtered, washed with water and

recrystallization from methanol yielded the pure compound.

Synthesis of compound (Va): A solution of substituted thiosemicarbazide (0.004 mole) and NaOH (5 M, 2 ml) in

25ml of absolute ethanol was cooled with continuous stirring for half an hour. To this mixture, iodine in KI (5%)

was added drop wise, till the color of iodine persisted at room temperature and the mixture was refluxed for 2 h

on a water bath. After completion of reaction, the mixture was poured onto crushed ice and the solid so separated

was filtered and washed with water. The recrystallization from petroleum ether:diethyl ether mixture (8:2, v/v)

gave the pure compounds.


Synthesis: A series of five membered thiadiazole and oxadiazole heterocyclic compounds was synthesized from

substituted thiosemicarbazides were obtained by the reaction between various heterocyclic or aromatic acid

hydrazides and phenyl isothiocyanate. The acid hydrazides were prepared by chlorination of the corresponding

acids, followed by treatment with hydrazine hydrate in 0-5oC.





SUMMARY AND CONCLUSION

Much attention has been paid to 1, 3, 4-oxadiazole and thiadiazole derivatives in recent years, because of

their showing anti-inflammatory, hypoglycemic, anticonvulsant, antimicrobial, anticancer and other activities. A

series of five membered thiadiazole and oxadiazole heterocyclic compounds was synthesized from substituted

thiosemicarbazides were obtained by the reaction between various heterocyclic or aromatic acid hydrazides and

phenyl isothiocyanate. The acid hydrazides were prepared by chlorination of the corresponding acids, followed by

treatment with hydrazine hydrate in 0-5oC. Melting points were determined by open capillary tube method and are

uncorrected. Purity of the compounds was checked on thin layer chromatography (TLC) plates (Silica gel G) in

the solvent system toluene: ethyl acetate: formic acid (5:4:1, v/v/v). The spots were located under iodine vapors

and UV light. Docking studies for the synthesized compounds were carried out against androgen receptor for

bearing the PDB id- 3PMX by the use of molecular docking software’s such as PYMOLWIN and Auto DOCK

and found that all the compounds showed good binding interaction with the receptor. The study also showed that

ARG 182 and SER 14 is the target site for binding of the drug to show anti-convulsant activity. Compounds Ve

and IVe were found to be more active when compared to other synthesized derivatives. Further, Characterization

by IR, 1H NMR and Mass spectra and in vivo studies for the synthesized compounds has to be carried out to

confirm the pharmacological activity.

Table.2.Structure and IUPAC Name of the synthesised Oxadizoles and thiadiazole derivatives

ompound Code Structure of the Compound IUPAC name

IVa S

NN

HN

N

N-phenyl-5-(pyridin-3-yl)-1,3,4-

thiadiazol-2-amine

IVb

S

NN

HN

HN

CH2

5-((1H-indol-3-yl)methyl)-N-

phenyl-1,3,4-thiadiazol-2-amine

IVd S

NN

HN

Cl

5-(4-chlorophenyl)-N-phenyl-1,3,4-

thiadiazol-2-amine

IVe S

NN

HN

O2N

5-(4-nitrophenyl)-N-phenyl-1,3,4-

thiadiazol-2-amine

Va O

NN

HN

N

N-phenyl-5-(pyridin-3-yl)-1,3,4-

oxadiazol-2-amine

Vb

O

NN

HN

HN

CH2

5-((1H-indol-3-yl)methyl)-N-

phenyl-1,3,4-oxadiazol-2-amine

Vd O

NN

HN

Cl

5-(4-chlorophenyl)-N-phenyl-1,3,4-

oxadiazol-2-amine

Ve O

NN

HN

O2N

5-(4-nitrophenyl)-N-phenyl-1,3,4-

oxadiazol-2-amine





Table.3. Physico-chemical data of the synthesised Oxadiazole and thiadiazole Compound

code

Molecular

formula

Colour Appearance/

Nature

Melting

point

Solubility TLC Yield

Rf value

Iva C13H10N4S white crystalline 136-38 Soluble in chloroform

0.72 74.32

IVb C17H14N4S brown powder 142-44 Soluble in

chloroform 0.62 77.27

IVc C17H12N4S white powder 168-70 Soluble in chloroform

0.79 73.35

IVd C14H10ClN3S white crystalline 152-54 Soluble in


IVe C14H12N4S brown powder 138-40 Soluble in chloroform

0.69 85.72

Va C13H10N4O white crystalline 266-68 Soluble in


Vb C17H14N4O brown powder 278-80 Soluble in chloroform

0.54 73.56

Vc C17H12N4O white powder 280-82 Soluble in


Vd C14H10ClN3O white crystalline 238-40 Soluble in chloroform

0.58 86.33

Ve C14H12N4O brown powder 236-38 Soluble in


*Solvent used for TLC= Toluene: Ethyl acetate: Formic acid (5:4:1), *Detecting agent – Iodine

Table.4. Autodock result of Compound IVa

Conformation Binding energy Inhibitory

constant

Ref RMS Hydrogen bond (Residue

Name And Distance)

1 -9.58 94.87 n/a none

2 -4.83 289.23 n/a none

3 -5.36 117.71 n/a SER14:HN1:..N (1.833 A0)

MET19:HN:..N (2.034 A0)

4 -4.45 548.91 n/a none

5 -9.93 52.26 n/a LEU94:O:..S (2.966 A0)

6 -9.29 154.41 n/a none

7 -9.72 344.13 n/a THR174:HN:..N (2.029 A0)

LEU12:O:..S 8 -10.6 17.12 n/a ILE100:O:..N (2.923A

0)

9 -4.65 392.35 n/a none

10 -4.87 268.09 n/a THR174:HG1:..N (2.056 A0)

THR174:HN:..N (2.118 A0)

LEU12:O:..S

Table.5. Autodock result of Compound IVb


constant


Name And Distance)

1 -5.35 120.78 n/a none

2 -4.93 242.14 n/a none

3 -4.55 463.08 n/a none

4 -5.19 157.39 n/a SER14:HN2:..N (2.073 A⁰) 5 -5.35 119.0 n/a none

6 -4.4 594.47 n/a none

7 -4.67 378.96 n/a MET19:HN:..N (2.151 A⁰) 8 -5.23 161.23 n/a none

9 -5.61 122.32 n/a SER14:HN1:..N (1.921 A⁰) 10 -5.47 118.21 n/a none





Table.6. Autodock result of Compound IVc


constant

Ref

RMS

Hydrogen bond (Residue

Name And Distance)

1 -5.74 61.84 n/a none

2 -5.07 192.31 n/a none

3 -5.33 123.97 n/a none

4 -5.27 136.93 n/a LEU12:O:..S (2.616 A⁰)

5 -5.33 123.22 n/a none

6 -5.2 154.5 n/a LEU12:O:..N (2.928 A⁰)

7 -5.94 44.42 n/a SER14:HN2:..N (2.19 A⁰)

8 -5.11 131.53 n/a none

9 -5.39 58.89 n/a none

10 -5.31 133.97 n/a LEU12:O:..S (2.527 A⁰)

Table.7. Autodock result of Compound IVd


constant

Ref

RMS


Name And Distance)

1 -5.35 120.78 n/a none

2 -4.93 242.14 n/a none

3 -4.55 463.08 n/a none

4 -5.19 157.39 n/a SER14:HN2:..N (2.073 A⁰)

5 -5.35 119.0 n/a none

6 -4.4 594.47 n/a none

7 -4.67 378.96 n/a MET19:HN:..N (2.151 A⁰)

8 -5.23 161.23 n/a none

9 -5.61 122.32 n/a SER14:HN1:..N (1.921 A⁰)

10 -5.47 118.21 n/a none

Table.8. Autodock result of Compound IVe


constant

Ref

RMS


Name And Distance)

1 -5.23 146.38 25.61 LYS104:HZ3:..O 1.775 A⁰

2 -6.15 31.28 21.67 ARG182:HH12:..O 1.9 A⁰

3 -5.86 50.41 20.85 ASN214:HD22:..O 2.018 A⁰ LYS251:HZ3:..N 1.983 A⁰

4 -22.05 68.76 32.44 none

5 -5.87 49.39 22.5 ARG203:HE:..O 2.041 A⁰

6 -5.25 141.25 21.0 LYS183:HZ3:..O 1.783 A⁰

7 -5.42 106.12 23.8 ARG182:HH12:..O 1.765 A⁰

8 -21.18 297.62 30.15 none

9 -6.64 13.55 21.62 ARG182:HH12:..O 2.007A⁰

10 -6.28 24.95 21.63 SER14:HN2:..N 2.084 A⁰ ARG182:HH12:..O 1.727 A⁰





Table.9. Autodock result of Compound Va


constant

Ref

RMS


Name And Distance)

1 -5.57 82.19 n/a SER14:NH2:..N 2.232 A⁰ THY61:HH:..N 1.916 A⁰

2 -4.61 419.09 n/a THR174:HG1:..N 1.991 A⁰ THR174:HN:..N 2.139 A⁰

LEU12:O:..O

3 -4.49 513.37 n/a SER14:HN1:..O 2.131 A⁰

4 -5.12 176.89 n/a SER14:O:..N 2.563 A⁰

5 -4.39 607.58 n/a PHE102:HN:..N,N 2.158A⁰ ILE100:O:..N

6 -4.52 487.57 n/a none

7 -5.01 212.17 n/a THR174:HN:..N 2.121 A⁰

8 -4.73 407.29 n/a THR174:HN:..N 2.023 A⁰

9 -4.47 520.35 n/a SER14:HN1:..O 2.011 A⁰

10 -5.26 146.87 n/a SER14:NH2:..N 2.123 A⁰

Table.10. Autodock result of Compound Vb


constant


Name And Distance)

1 -5.55 84.85 n/a LEU12:HN:..N (1.965A0)

2 -5.15 168.00 n/a none

3 -4.76 321.79 n/a MET19:HN:..N (1.887A0)

4 -5.13 173.75 n/a none

5 -4.73 339.34 n/a ALA175:NH:..N(2.026 A0)

6 -5.09 185.26 n/a none

7 -4.69 355.29 n/a ASN83:HD22:..O (2.151 A⁰)

8 -4.87 197.21 n/a MET19:HN:..N (1.927A0)

9 -4.63 341.37 n/a none

10 -4.59 183.23 n/a none

Table.11. Autodock result of Compound Vd


constant

Ref

RMS


Name And Distance)

1 -5.01 211.24 40.2 none

2 -4.89 261.15 39.49 none

3 -4.74 335.84 40.44 none

4 -4.09 998.83 41.13 LEU52:HN:..O (2.035 A⁰) LYS50:O:..N

5 -5.01 211.62 39.76 none

6 -4.98 223.17 39.73 none

7 -3.53 2.58 48.51 ASN83:HD22:..O (2.151 A⁰)

8 -5.01 212.84 39.91 none

9 -5.01 213.61 40.01 none

10 -4.88 265.81 39.36 none





Table.12. Autodock result of Compound Ve


constant

Ref

RMS


Name And Distance)

1 -6.89 8.92 19.06 SER14:HN2:..N(2.08A0)

ARG182:HH12:..O(2.185A0)

ARG182:HH22:..O(1.942A0)

ALA175:NH:..N(2.125 A0)

2 -6.99 7.46 19.04 ARG182:HH12:..O(1.806A0)

ARG182:HH22:..O(1.775A0)

ALA175:NH:..N(2.184 A0)

3 -6.03 37.7 20.26 TYR199:HH:..O(2.249A0)

ARG203:HH22:..O(1.87A0)

4 -6.06 36.43 22.58 MET19:HN:..O(1.997A0)

5 -6.9 8.78 18.84 ARG182:HH22:..O(2.045A0)

6 -6.92 8.43 19.4 SER14:HN2:..N(2.126A0)

ARG182:HH12:..O(1.94A0)

ARG182:HH22:..O(1.865A0)

7 -6.49 17.39 19.06 SER14:HN2:..N(1.963A0)

ARG182:HH12:..O(1.801A0)

ARG182:HH22:..O(2.022A0)

8 -6.93 8.32 19.06 ARG182:HH12:..O(1.805A0)

ARG182:HH22:..O(1.884A0)

9 -6.51 16.87 20.2 LEU94:HN:..N(2.169A0)

ARG148:HE:..O(1.638A0)

ARG148:HH22:..O(1.908A0)

10 -5.94 44.18 20.2 ARG182:HH12:..O(2.18A0)

ARG182:HH22:..O(1.855A0)

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Wei B T and Zhang Y M, Li M N, Synthesis and bioactivity study of 2,5- bismercapto-1,3,4-thiadiazoles

heterocyclic derivatives. Ind J Chem, 46, 2007, 544-9.

Zarghi A, Hajimahdi Z, Mohebbi S, Rashidi H, Mozaffari S, Sarraf S, Faizi M, Tabatabaee SA, Shafiee A,

Design and synthesis of new 2-substituted-5-[2-(2-halobenzyloxy)phenyl]-1,3,4-oxadiazoles as

anticonvulsant agents, Chem. Pharm. Bull, 56(4), 2008, 509–12.



Pruthvi Raj et.al Indian Journal of Research in Pharmacy and Biotechnology


PREPARATION, CHARACTERIZATION AND EVALUATION OF

OLMESARTAN MEDOXOMIL-Β CYCLODEXTRIN COMPLEXES V. Prudhvi Raj*, Subhashis Debnath, Maleswari, M. Niranjan Babu

Department of Pharmaceutics, Seven Hills College of Pharmacy, Venkatramapuram, Tirupati – 517 561

* Corresponding author: [email protected]

ABSTRACT

Olmesartan medoxomil is a poor water soluble drug. There are numerous approaches available and

reported in literature to enhance the solubility of poor water soluble drugs among which cyclodextrin

complexation is predominant. β-cyclodextrin was used since this possess a special ability to complex

with drugs enabling them to increase solubility, reduce bitterness, enhance stability and decrease

tissue irritation upon dosing. Olmesartan medoxomil-β cyclodextrin complexes were prepared and

characterized by FT-IR and SEM, studies. The results showed the formation of true inclusion

complexes at molar ratio 1:5. In contrast crystalline drug was detectable in all other products.

The dissolution of Olmesartan medoxomil from all the prepared complexes has been carried out to

determine the most appropriate ratio that can be used for further development like tablet formulation

for oral delivery. The complexes prepared by physical mixture method in 1:5 ratio showed superior

dissolution profile when compared to complexes prepared in other ratios. From this research work

it can be concluded that β CD has potential to increase the solubility of Olmesartan medoxomil and

true complexes are formed in 1:5 ratio and because of the higher dissolution profile it can be used for

further formulations.

KEY WORDS: β-cyclodextrin, complex, inclusion complexes, dissolution

INTRODUCTION

Orally administered drugs completely absorb only when they show fair solubility in gastric

medium and such drugs shows good bioavailability. Recently more than 40% NCEs (new chemical entities)

developed in Pharmaceutical Industry are practically insoluble in water. These poorly water soluble drugs are

allied with slow drug absorption leading to inadequate and variable bioavailability and gastrointestinal

mucosal toxicity. Therefore, the improvement of drug solubility thereby its oral bio-availability remains

one of most challenging aspects of drug development process especially for oral drug delivery system.

There are numerous approaches available and reported in literature to enhance the solubility of poorly

water soluble drug. The techniques are chosen on the basis of certain aspects such as properties of drug under

consideration, nature of excipients to be selected and nature of intended dosage form. Among all the solubility

enhancement techniques inclusssion complex formation technique has been employed more precisely to

improve the aqueous solubility, dissolution rate, and bioavailability of poorly water soluble drugs.

Inclusion complexes are formed by the insertion of the nonpolar molecule or the nonpolar region of one

molecule (known as guest) into the cavity of another molecule or group of molecules (known as host). The most

commonly used host molecules are cyclodextrins. Olmesartan is one of the widely used selective AT1 subtype

angiotensin II receptor antagonist which is marketed under the names of Benicar in US, Olmetac in EU and

Canada, Golme in India. As many of the drugs even Olmesartan medoxomil is a poor water soluble drug. The

present work aims to investigate the potential of β cyclodextrin (β CD) complexes to increase the solubility

of Olmesartan Medoxomil.

MATERIAL & METHODS

Olmesartan medoxomil (MSN Laboratories, Hyderabad), β cyclodextrin (Merck limited, Germany), HCl (Sd fine

chemicals Ltd, Mumbai)

Phase solubility studies: Phase-solubility studies were carried out with double distilled water according to the

method described by Higuchi and Connors (1965). Excess amount of Olmesartan medoxomil (100 mg) was

added to 1ml stock solution with varying concentrations (3, 6,9,12, 15 and 18 mM) of β-CD. The suspensions

were then shaken at 37.0±0.5 ◦C for at least 2 days. After equilibrium attainment, the samples were filtered and

the concentration of Olmesartan medoxomil was determined spectrophotometrically on a UV-

spectrophotometer at 260 nm. Amount of drug present in the samples were calculated by using USP-Disso-v3

software. The complexation efficiency, which reflects the solubilizing power of CDs toward the drug, was

calculated from the tredline of the phase solubility diagram according to the equation





CE=Slope/1-slope=SoK1:1

Where so represents the solubility of the drug in the absence of cyclodextrin K1:1 represents the apparent

solubility constant = Slope/ So (1-slope)

Synthesis of inclusion complexes: Inclusion complexes of Olmesartan medoxomil with β cyclodextrin was

prepared by the Physical mixture method. The Olmesartan medoxomil - β cyclodextrin inclusion complexes in the

ratios 1:1, 1:2, 1:3, 1:4, 1:5 and 1:6 (molar ratios) were prepared by mechanical trituration by grinding the

accurately weighed amounts in a geometrical dilution method in the mortar and pestle for 30 min.

EVALUATION

Characterization of prepared complexes: Characterization studies of the Olmesartan medoxomil - β

cyclodextrin complexes by FT-IR and SEM were performed.

Invitro dissolution studies: Dissolution studies of samples weight equivalent to 20 mg were performed using

USP XXII apparatus at the stirring speed of 50 rpm and temperature maintained at 37±0.50C with 0.1N HCl as

dissolution medium. The samples of 5ml were withdrawn at regular intervals of 5,15,30,45,60 min. Every

time the samples are replaced with 5ml of the fresh dissolution medium. The filtrates of the samples were

analyzed at 260 nm. Percentage drug release and other parameters of the samples were calculated by using Disso

software PCP Disso V3 software. The dissolution in pH 6.8 phosphate buffer, pH 7.4 phosphate buffer has been

performed in the aforementioned procedure.


Phase Solubility Studies: The aqueous solubility of Olmesartan medoxomil at various concentrations of β CD

was studied. It was found that the solubility of drug increased with the increasing concentration of β CD.

The main reason might be the formation of inclusion complexes however other reasons like formation of

aggregations of cyclodextrin which promote the solubility of drugs might also have been involved. The

coefficient of determination (r2) value of the phase solubility diagram of Olmesartan medoxomil is found to

be 0.9878 (<0.990) therefore the diagram can be classified as Ap-type curve. This positive deviation suggests the

formation of higher order inclusion complexes with respect to β-cyclodextrin. The complexation efficiency of the

curve calculated was 0.04395. Phase solubility curve for the system was found to be of Higuchi’s AP type

phase solubility curve was presented in the Figure No. 1, Table No. 1.

Table No.1: Phase solubility studies

Molar conc. of

β-cyclodextrin (*10-3

)

Molar conc. of Olmesartan

medoxomil (*10-5

)

3 0.06

6 0.09

9 0.12

12 0.16

15 0.21

18 0.26

Figure No. 1: Phase solubility studies of olmesartan medoxomil





The practical yield obtained and the Percentage yield for drug- β-cyclodextrin complexes prepared in different

ratios was given in the table no. 2.

Table no.2: percentage yield of the prepared complexes of different ratios

Inclusion complexes ratio %yield

1:1 90.6

1:2 89.4

1:3 91.2

1:4 92.4

1:5 91.6

Evaluation of Inclusion Complexes:

Fourier Transform Infrared (FT-IR) Spectroscopy of Olmesartan medoxomil- β cyclodextrin complexes:

The FT-IR studies showed that the significant peaks of Olmesartan medoxomil are at showed the characteristic

peak at 3286.81which can be assigned to the O-H group and a peak at 1737.2 which can be assigned to the

presence of Ester carbonyl group. The absence of peaks at 3286.81, 1282.71 and 1552.75 and reduction of

intensities of peaks at 1707.06 and 1832.44 to considerable level in the complexes prepared in the ratio 1:5

confirms the formation of true inclusion complex with β-cyclodextrin in 1:5 ratio(Table no 3).

Table No.3: comparison of interpretation of IR spectrum of drug, β CD and complexes

Olmesartan

medoxomil

Peak

assignment

Drug: β CD

1:1 1:2 1:3 1:4 1:5

3286.81 O-H group 3306.10 3317.67 absent absent absent

1282.71 C-N stretching 1282.71 1282.71 absent absent absent

1707.06 Diaryl ketone 1707.06 1707.06 1707.06 1707.06 1707.06

1737.2 Ester carbonyl

group

1832.44 1832.44 1832.44 1832.44 1832.44

1552.75 C=C aromatic

stretching

1550.82 absent absent absent absent

Scanning Electron Microscopy (SEM) Studies: To study about the shape and surface characteristics SEM

studies were performed for the individual components and Olmesartan medoxomil- β cyclodextrin complexes.

Drug and β-CD inclusion complexes of drug showed significant difference in the microscopic structure. The

reduction in the crystallinity was observed in the inclusion complexes. Reduced size of the complexes is due to

the method of preparation used, physical mixture method for preparing complexes. Comparative SEM

photographs for Olmesartan medoxomil and Complexes were given in Figure No. 2. Except the complexes

prepared in 1:5 ratio, the SEM images of remaining complexes showed the presence of minute amounts of drug

(irregular shaped particles) which confirms the formation of the true inclusion complexes in 1:5 ratio which

supports the results obtained from FT-IR.





a. Pure Drug b. β CD

c. Drug : β CD (1:1) d. Drug : β CD (1:2)

e. Drug : β CD (1:3) f. Drug : β CD (1:4)

g. Drug : β CD (1:5)

Figure No. 2: Comparative SEM images of Drug-β CD complexes in different ratios

In-vitro dissolution studies: The results from dissolution studies carried in 0.1 N HCl and also with pH

6.8 phosphate buffer and pH 7.4 phosphate buffer showed that there is cyclodextrin complexes showed

improved dissolution profile with 1:5 complexes the showing the highest. The dissolution studies in 0.1N

HCl revealed that drug release for pure drug at 60 min is 33.75% while the complexes prepared in 1:5 ratio

exhibited drug release of 65.93% at 60 min. In 6.8 buffer for pure drug release at 60 min is 37.83% whereas the

complexes prepared in 1:5 ratio exhibited 76.49% of drug release at the end of 60 min. In 7.4 buffer for pure drug

release at 60 min is 38.99% whereas the complexes prepared in 1:5 ratio exhibited 74.83% of drug release

at the end of 60 min. The improved release of the drug can be attributed to the formation of the

inclusion complex with β CD. Invitro dissolution profile for both pure drug and formulation along with

standard deviations in 0.1 N HCl was given in Table No.4 [Figure No.3] for pH 6.8 phosphate buffer was

given in Table No.5 [Figure No.4] for pH 7.4 phosphate buffer was given in Table No.6 [Figure No.5] and the

comparison of dissolution profile for pure drug and complexes prepared in the ratio 1:5 was given in Table No.7.





Table no.4: Dissolution profile for pure drug and complexws in 0.1N HCL

Time

(min)

Average % drug release (n=2)

Pure

drug

1:1 1:2 1:3 1:4 1:5

0 0 0 0 0 0 0

5 30.35 34.73 39.25 41.11 44.96 45.62

15 34.11 38.11 40.93 44.79 49.06 52.78

30 34.69 40.98 44.74 47.69 52.78 57.32

45 35.55 40.01 45.25 49.28 57.66 60.95

60 33.75 41.56 46.96 51.27 58.55 65.93

Table no.5: Dissolution profile for pure drug and complexws in pH 6.8 phosphate buffer

Time

(min)


Pure

drug

1:1 1:2 1:3 1:4 1:5

0 0.00 0.00 0.00 0.00 0.00 0.00

5 34.00 39.25 44.65 46.90 51.70 53.50

15 38.24 43.07 46.55 51.06 56.34 61.59

30 38.90 46.31 50.85 54.34 60.55 66.73

45 39.86 45.21 51.43 56.14 62.68 70.85

60 37.83 46.96 53.37 58.40 67.07 76.49

Table no.6: Dissolution profile for pure drug and complex was in pH 7.4 phosphate buffer

Time

(min)


Pure

drug

1:1 1:2 1:3 1:4 1:5

0 0.00 0.00 0.00 0.00 0.00 0.00

5 34.45 39.10 44.95 47.50 51.55 53.35

15 35.39 42.92 46.40 51.06 56.18 61.44

30 36.64 46.15 50.71 54.34 60.39 66.43

45 37.59 45.06 51.58 56.14 62.38 70.70

60 38.99 46.20 52.62 57.50 66.02 74.83

Table No.7: In vitro dissolution profile for drug and olmesartan medoxomil-β cyclodextrin

inclusion complexes (1:5 ratio)

Time

0.1N HCL

pH 6.8 phosphate

buffer

pH 7.4 phosphate

buffer

Pure

drug

Complexes

(1:5 ratio)

Pure

drug

Complexes

(1:5 ratio)

Pure

drug

Complexes

(1:5 ratio)

0 0.00 0.00 0.00 0.00 0.00 0.00

5 30.35 45.62 34.00 53.50 34.45 53.35

15 34.11 52.78 38.24 61.59 35.39 61.44

30 34.69 57.32 38.90 66.73 36.64 66.43

45 35.55 60.95 39.86 70.85 37.59 70.70

60 33.75 65.93 37.83 76.49 38.99 74.83





Figure No. 3: Dissolution profile for pure drug and complex in 0.1 N HCl

Figure No. 4: Dissolution profile for pure drug and complex in pH 6.8 phosphate buffer

Figure No. 5: Dissolution profile for pure drug and complex in pH 7.4 phosphate buffer

SUMMARY AND CONCLUSION

Phase solubility curve for the system was found to be of Higuchi’s AP type phase solubility curve.

The coefficient of determination (r2) value of the phase solubility diagram of Olmesartan medoxomil is found to

be 0.9878 (<0.990). This positive deviation suggests the formation of higher order inclusion complexes with

respect to β-cyclodextrin. The complexation efficiency of the curve calculated was 0.04395. Results from

SEM studies confirmed the formation of true complexes of drug Olmesartan medoxomil with β CD in 1:5

ratio supporting the results from FT-IR. The results from dissolution studies carried in 0.1 N HCl and also with

pH 6.8 phosphate buffer and pH 7.4 phosphate buffer showed that cyclodextrin complexes showed





improved dissolution profile and complexes prepared in the ratio 1:5 is showing the highest dissolution profile

which exhibited drug release of 65.93% in 0.1N HCl and exhibited 76.49% of drug release in pH 6.8 phosphate

buffer and 74.83% of drug release in pH 7.4 phosphate buffer at the end of one hour.

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Vibhooti and Preethi Indian Journal of Research in Pharmacy and Biotechnology


WAFERS TECHNOLOGY – A NEWER APPROACAH TO SMART DRUG

DILEVERY SYSTEM Papola Vibhooti

*, Kothiyal Preeti

Shri Guru Ram Rai Institute of Technology & Sciences Dehradun, Uttarakhand, India

*Corresponding author: E-mail: [email protected]

ABSTRACT The lyophilized wafer developed throughout this review is an effective and versatile drug delivery

system for oromucosal application. This has been established from the extensive physicochemical and

physic mechanical profiling conducted. Through a screening and selection of polymers, HPC had the

lowest gelation characteristics and was therefore suitable for the development of the wafer system.

Suitable excipient and polymer combinations were established which allowed for the development of

rapidly disintegrating and prolonged release wafer systems. The wafer system containing HPC, lactose,

mannitol and glycine had the ability to disintegrate within 30 seconds. The modified wafer system,

consisting of pectin cross linked with zinc ions serving as the drug reservoir, and mucoadhesive polymer

combination of pectin, carmellose and gelatin, provided effective release of model drug diphenhydramine

hydrochloride over approximately six hours. The modification of this technology to provide a prolonged

release mucoadhesive system seems promising. It is envisaged that this system will be applicable to many

drugs requiring the extended release of bioactive material. Therefore, the lyophilized wafer matrices

developed in this study are highly effective in the rapid delivery of drugs, using the oral route as a site of

administration.

KEY WORDS: Glioblastoma, glycospingolipids, acylceramides, matrices and lyophilized.

INTRODUCTION

Therapeutic value and pharmacoeconomic value have in recent years become major issues in defining

health care priorities under the pressure of cost containment. The improvement in drug therapy is a consequence

of not only the development of new chemical entities but also the combination of active substances and a suitable

Delivery system. The treatment of an acute disease or chronic illness is mostly accomplished by delivery of one or

more drugs to the patient using various pharmaceutical dosage forms. Tablets, pills, capsules, suppositories,

creams, ointments, liquids, aerosols, and injections are in use as drug carriers for many decades (Shayne, 2008)

(Bhalla, 1999).

These Conventional types of drug delivery systems are known to provide a prompt release of the drug.

Therefore, to achieve as well as to maintain the drug concentration within the therapeutically effective range

needed for treatment, it is often necessary to take this type of drug several times a day, resulting in the significant

fluctuation in drug levels. For all categories of treatment, a major challenge is to define the optimal dose, time,

rate, and site of delivery. Recent developments in drug delivery techniques make it possible to control the rate of

drug delivery to sustain the duration of therapeutic activity and/or target the delivery of drug to a special organ or

tissue. Many investigations are still going on to apply the concepts of controlled delivery for a wide variety of

drugs (Chein, 1992).

The basic rationale for controlled drug delivery is to alter the pharmacokinetics and pharmacodynamics of

pharmacologically active moieties by using novel drug delivery systems or by modifying the molecular structure

and or physiological parameters inherent in a selected route of administration. It is desirable that the duration of

drug action become more a design property of a rate - controlled dosage form and less, or not at all, a property of

the drug molecules’ inherent kinetic properties (Brannon, 1997).

The rationale for development and use of novel drug delivery systems may include one or more of the

following arguments

Decrease the toxicity and occurrence of adverse drug reactions by controlling the level of drug and/or

metabolites in the blood at the target sites.

Improve drug utilization by applying a smaller drug dose in a controlled – release form to produce the

same clinical effect as a larger dose in a conventional dosage form.

Control the rate and site of release of a drug that acts locally so that the drug is released where the activity

is needed rather than at other sites where it may cause adverse reactions.






Provide a uniform blood concentration and/or provide a more predictable drug delivery.

Provide greater patient convenience and better patient compliance by significantly prolonging the interval

between administrations.

The oral mucosa provides the ideal application site for many active ingredients. Their diffusion into the

dense network of capillaries ensures direct access to the blood stream –and excellent patient compliance.

Wafer – an innovative oral dosage form: New oral thin films, so-called wafers, thus creating new possibilities

for action profiles and patient compliance. Wafers are paper-thin polymer films used as carriers for

pharmaceutical agents. The innovative dosage form is taken orally but does not require water or swallowing.

Effective absorption of active ingredient: The wafer quickly dissolves in the oral cavity, and the active

ingredient can be absorbed into the blood - stream via the oral mucosa. The active ingredient, once absorbed by

the oral mucosa, thus bypasses the liver’s first-pass effect, which improves bioavailability. Depending on the

selected wafer type, the active ingredient’s release may also be delayed. In this case, it is absorbed after

swallowing via the gastrointestinal tract.

Positive aspects with wafers (industrial point of view):

Attractive dosage form with new active ingredients.

Improvement of established products.

Access to new indications by means of a new absorption profile even for existing active ingredients.

Optimization of bioavailability.

Increase patient compliance.

Innovative technology for product.

Increase of product appeal through innovative format.

Exclusivity and cutting edge technology position in the market through a step forward.

Advantages of wafers:

No first – pass effect*

Conrolled release

Improved bio-availability, translates to lower doses

Reduction of side-effects

Reduced impact on the gastro intestinal tract*

Discrete and easy application (no additional intake of liquids required)

Excellent compliance, especially in children and seniors

*Applies to all wafer types except the flash-dispersal wafer whose active ingredient is swallowed and

subsequently absorbed into the gastro-intestinal tract.

Marketing availability till date: Ranbaxy Laboratories received import permission for marketing the US FDA

approved product Gliadel (polifeprosan 20 with carmustine implant) Wafer. The company has signed an exclusive

licensing agreement with BioPro Pharmaceutical, USA, to promote and market Gliadel Wafer in India. Gliadel

Wafer is for the treatment of newly diagnosed high-grade malignant gliomas and recurrent glioblastoma

multiforme. There is very limited data available on the incidence of brain tumours in India, according to unofficial

sources, the estimated prevalence of CNS tumours in India is two to five new cases per 1,00,000 per year.

Another source estimates the total number of cases to be around 21,000 per year. Glioblastoma multiforme

constitutes about 60-65 percent of these primary brain tumours.

Type of wafers

Flash dissolved wafers

Melt away wafers

Sustained release wafers

Flash dispersed wafers





Anatomic and Physiological Considerations: Four sites within the buccal cavity have been used for drug

administration. The four regions have varying permeability, which plays a role in the absorption of drugs across

the oral mucosa. As seen in the four key areas are the buccal cavity, the lingual area, the palate and gingival

region. The most commonly used sites for drug administration of the four mentioned above is the sublingual and

buccal route. Using the sublingual route, the medicament is placed under the tongue, usually in the form of a

rapidly dissolving tablet. The anatomic site for drug administration between the cheek and gingival is known as

the buccal mucosa. The oral mucosa is composed of three layers. The first layer is the stratified squamous

epithelium; underneath this layer lies the basement Membrane .The basement membrane overlies the lamina

propria and submucosa.

Fig 1 : Difference between sustained release and flash-dispersal wafers

Fig 2 : Difference between flash dissolve and melt – away wafers

The constitution of the epithelium within the different sites of the oral cavity shows dissimilarity. The

gingival and hard palate are exposed to mechanical stress during eating, hence the epidermis is keratinized in a

similar manner as the skin. The epithelium in the soft palate, buccal and sublingual area is not keratinized,

therefore not containing ceramides and acylceramides which are associated with providing a barrier function

(Danckwerts, 2003). The mucosa of the buccal and sublingual region have only small amounts of ceramide, and is

thus more permeable when compared to other regions of the oral cavity (Squier, 1991). The presence of

membrane coating granules (MCGs) accounts for the differences in permeability amongst the various regions of

the oral mucosa. When cells go through differentiation from basal to flattened keratinous cells, MCGs are formed.

At the apical cell surface, MCGs merge with the plasma membrane and their contents are discharged into the

intercellular spaces. This occurs mainly in the upper one-third of the epithelium. MCGs are present in both





keratinized and nonkeratinised epithelia, however their composition is different. On the other hand, non-

keratinised epithelium contains MCGs that are nonlamellar and include cholesterol, cholesterol esters and

glycospingolipids.

A layer of mucus is present on the surface of the epithelial layer of cells. This plays a major role in cell-

to-cell adhesion, oral lubrication, as well as mucoadhesion of mucoadhesive drug delivery systems .A major

feature in the environment of the oral cavity is the presence of saliva. The salivary glands produce saliva,

responsible for protecting the soft tissues from abrasion during the mastication of food Saliva plays an essential

role in facilitating the disintegration of quick-disintegrating drug delivery systems (Shojaei, 1998). The buccal

and sublingual regions are different from each other in terms of anatomy, permeability to drug, and their ability to

retain a drug delivery system for a desired duration. Although the buccal mucosa is less permeable than the

sublingual mucosa and does not yield a rapid onset of action as seen with sublingual delivery, mucosa of the

buccal area has an expanse of smooth and relatively immobile surface, which is suitable for placement of a

retentive system. For buccal drug delivery, adhesion to the oral mucosa permits not only the intimacy of contact

and the possibility of improved drug absorption, but also the ability to achieve an optimum residence time at the

site of administration (Rathborne, 1994). These characteristics make the buccal mucosa a more appropriate site

for prolonged systemic delivery of drugs. The sublingual route is however more suitable for delivery systems

formulated either as rapidly disintegrating matrices or soft gels. These systems create a highly significant drug

concentration in the sublingual region prior to systemic absorption across the mucosa.

Fig 3. : Mucousal region of mouth [10] Fig 4: Composition of layers of mucosal

epithelium

(a) Keratinised (b) Non kerantinised

Fig 5. : Absorption through oral

mucosal

Mode of Action: The wafer quickly dissolves in the oral cavity, and the active ingredient can be absorbed into the

blood - stream via the oral mucosa. The active ingredient, once absorbed by the oral mucosa, thus bypasses the

liver’s first-pass effect, which improves bioavailability. Depending on the selected wafer type, the active

ingredient’s release may also be delayed. In this case, it is absorbed after swallowing via the gastrointestinal tract.

Manufacturing of wafers: The active ingredient in wafer is integrated into a polymer matrix. The typical size of

an oral film is between 2cm2

to 10cm2 , with a thickness of 20 micrometre to 500 micrometre.Oral thin films can

be composed of a single-layered system.The active ingredient may be prsented within the wafer matix in either a

dissolved an emulsified or a dispersed state.If required, it can also be bound in a complex form,for example,to

enable taste masking.

Open Matrix-Type Wafers and Tablets: With the introduction of the Zydis®

system in the late 1970s, the

concept of quick disintegrating drug delivery systems gained much attention. It was the first of this class of

delivery systems to be manufactured on a large scale. It is a freeze-dried wafer made from various standard tablet

adjuvants. The wafer essentially works on the principle of forming an open network containing the active

ingredient. The Zydis®

manufacturing process. The freeze-dried tablet disintegrates within 2-3 seconds, releasing





the active ingredient. The drug either forms dispersion or dissolves in the saliva, which is then swallowed and

absorbed via the GIT.

Fig.6.Production of Zydis®

lyophilized wafer [10]

The WOWTab®

(With-Out-Water tablet) has been produce by Yamanouchi Pharmaceutical Co. Ltd.

(Tokyo, Japan). This tablet is manufactured using conventional granulating and compression. The rapid

disintegration is attributed to the blending of a low and high mold ability saccharide. The unique combination of

saccharides provides sufficient mechanical strength as well as quick tablet disintegration.Fuisz Technology Ltd.

(Chantily, Virginia, USA) developed the Flash Dose®

tablet, which can dissolve in the patient’s mouth in less than

10 seconds. This has been achieved by the use of Shearform™

technology. The process involves a unique blend of

sugars being placed in a fast spinning machine and subjected to flash heat. By this process, long cotton-like fibres

called ‘floss’ are produced. The ‘floss’ is then cured by subjecting it to specific environmental conditions that

induce crystallisation, at this stage crystallisation modifiers may also be added. The matrix is then blended with

coated or uncoated microspheres containing the active drug. The floss is compressed using standard tabletting

equipment (Virely, 1990).

Fig.7. Manufacturing process of Flash Dose

Of the various open matrix-type wafers on the market, the Zydis®

system remains the most popular; as a result

making lyophilisation the most frequently used process for the manufacture of these systems.

Preparation of wafers: For the formulation of a rapidly disintegrating wafer, a polymer with low gelation

characteristics is desired (Misra, 1999). The gelation potential of polymers is highly dependent on it’s’ solubility.

Materials and Methods: Polymers utilised in the study include: sodium alginate, hydroxypropylmethyl cellulose

(HPMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), pectin, polyethylene oxide (PEO),

polyvinyl alcohol (PVA) (MW 124,000 - 186,000). Additionally, lactose and polystyrene cylindrical moulds of

total volume 60.31mm³ (diameter 16mm and depth of 2.4mm) were utilised. Materials used in the preparation of

simulated saliva were: Potassium Phosphate Monobasic (KH2PO4), Disodium Hydrogen Phosphate (Na2HPO4),

Sodium Chloride (NaCl).

Preparation of wafers: Polymers suitable for oramucosal preparations were identified based on information

provided in literature. A polymer (1%w/v) and lactose as a bulking agent (6%w/v) was added to deionised water

and mixed for 45 minutes. 1.5mL of the various polymer solutions were pipetted into the cylindrical cavities pre-

oiled with mineral oil. The formulation was subjected to a freeze-phase in a freeze-dryer at -60°C for 2 hours. The





drying-phase was executed at a pressure of 25 mtorr for 24 hours. Wafers were stored in glass jars with 2g of

desiccant sachets.

ANALYSIS OF WAFERS Weight Uniformity: Weight uniformity was used to assess the reproducibility of wafer production process.

Individual wafers were weighed, and standard deviations calculated. All experimentation was conducted in

triplicate. The reproducibility of the production process was demonstrated by the low standard deviations (SD)

calculated from the mass for each of the various polymer systems. Shows the results obtained from the various

polymer wafer systems. Mean weight of wafers manufactured (N=3)

Polymer Mean (g) ± SD

HPC 0.126 ± 0.0017

HPMC 0.122 ± 0.0002

Pectin 0.134 ± 0.0055

PEO 0.119 ± 0.0045

PVA 0.118 ± 0.0011

Sodium alginate 0.109 ± 0.0007

Although the standard deviation of the samples is low, slightly higher values were observed for polymers such as

pectin and PEO. This may be attributed to the high viscosity of the initial solution, and therefore greater

variability in the production process.

Gelation of Matrices: The main objective of this study was to formulate a rapidly dissolving wafer system. Thus

the matrix formation characteristics required assessment and formed the basis for the selection of a suitable

polymer. Gelation of the dosage form would delay the disintegration and ultimately the release of active

substance. A novel method was developed in order to assess the matrix forming profiles of the wafers. Wafers

were weighed before being placed in a Petri dish (diameter 85mm, depth 10mm) containing 20mL of simulated

saliva (pH 7.1). The Petri dish was agitated for a period of 30 seconds on a Vortex Genie2 on the slowest setting.

The contents of the Petri dish were sieved through a stainless steel mesh (pore size 1mm). The mass of the

remaining residue was determined on a balance (and used to calculate the rate of matrix formation.The simulated

saliva solution comprised 2.38g Na2HPO4, 0.19g KH2PO4 and 8g NaCl in 1000mL of deionised water (Guo,

1994).

Determination Limits for Formulation Variables: The lower and upper limits were determined using a trial

and error method. Wafers of varying polymer and diluents concentrations (up to 30%w/v of each) were made and

inspected visually. Polymers such as sodium alginate, pectin and PEO tended to form a gel-like substance when

hydrated and agitated rather than undergo disintegration.

Sodium alginate produced the highest amount of residue, possibly due to its low water solubility. In sharp

contrast, the highly hydrophilic polymers such as HPC were completely disintegrated within 30 seconds into

small particles which were able to penetrate through the pores on the sieve. The mass of intact material after

sieving of the various dissolved wafers tested. Based on the results obtained, HPC was identified as the most

suitable polymer for the wafer system, because no residue was produced after 30 seconds of hydration and

agitation in simulated saliva. This may be attributed to the fact that HPC is highly soluble in polar solvents and

therefore undergoes disintegration rapidly without forming a gel residue, ensuring rapid matrix disintegration.

Development of the Manufacturing Process: To establish the suitability of a mould in terms of ease of the

system removal, well plates, blister packs and disposable polystyrene trays were assessed. To overcome problems

of wafers sticking to the mould, various lubricant systems were considered. Magnesium Stearate, Span 60, Maize

oil and mineral oil were evaluated for their anti-adhesive properties. It was also necessary to determine suitable

timeframes for the lyophilisation process.

Established Parameters of Formulation Variables

Concentration of HPC: Lower and upper limits were determined to be 1%w/v and 10%w/v respectively. The

upper limit of 10%w/v was set because wafers of higher polymer concentrations were difficult to remove from the





mould. Some wafers produced with polymer concentrations below 5%w/v collapsed. Less than 1%w/v of HPC

was not sufficient to form the wafer matrix.

Concentration of Diluent: The concentration of the diluent would affect both the solubility and textural

properties of the matrices. Lower and upper limits were determined to be 1%w/v and 5%w/v respectively.

Concentrations of lactose higher than 5%w/v caused the wafer to be powdery and extremely fragile.

Type of Mouldl: A major problem that was encountered was the removal of the wafers from the moulds without

disrupting the delicate structure. Polystyrene trays proved to be the most successful, with minimal deformation of

the final product as these moulds could be easily split down the middle to release the wafer.

Type of Lubricant: As mentioned above, removal of the wafers from the mould was problematic. Mineral oil

produced the greatest ease of removal of the product as compared to the other lubricants analyzed, imparting

minimal hydrophobicity and having no effect on the taste of the final product as opposed to other substances such

as maize oil.

Freeze-Drying Parameters: Although the wafers appeared to be dry after a period of 24 hours, ‘melting’ and

discoloration of the matrices occurred on storage. This was attributed to moisture present within the products,

indicating that the freeze drying process needed to be conducted for a longer period. In future processes, this was

increased to 48 hours.

Fig.8. Mass of intact wafer after gelation studies using various polymers (N=3)

Concluding Remarks: It was necessary to gain a firm understanding of the key factors involved in the successful

production of a lyophilised wafer system. HPC was selected as the most appropriate polymer of the seven that

were assessed. It was expected that the type of diluent used in the wafer matrix would affect the disintegration rate

of the wafers. Mannitol which is more quickly soluble than lactose will be included in the experimental design to

assess its influence of this inclusion on the disintegration rate. The diluents will either be used on their own or in a

1:1 combination. To solve the problem of wafers collapsing, Seager (1998) recommended that glycine be used as

a collapse protectant. Therefore, concentrations of up to 0.6%w/v will be included in subsequent formulations.The

selection of a suitable polymer, determination of future formulation parameters and creation of problem-free

manufacturing techniques formed the basis of this part of the study.

General Statistical Approach to Wafer Formulation: A Face Centered Central Composite design was

developed with 5 factors and 4 centre points (Table 5.1).The equation for the design was as follows:

Response = b0 + b1*s + b2*t + b3*u + b4*v + b5*w + b6*s*s + b7*t*t + b8*u*u + b9*v*v + b10*w*w +

b11*s*t + b12*s*u + b13*s*v + b14*s*w + b15*t*u + b16*t*v + b17*t*w + b18*u*v + b19*u*w + b20*v*w

Where: s = Polymer Concentration; t = Diluent Type; u = Diluent Amount; v = Glycine Concentration; and

w = Fill Volume.

The responses that are generally measured are:

Disintegration profiles;

Rate of influx of simulated saliva into the matrix;

Friability;





Matrix yield value;

Matrix tolerance;

Matrix absorption energy;

Matrix resilience; and

Brinell Hardness Number (BHN).

Evaluation of CCF Responses

ANOVA Test: An Analysis of Variance (ANOVA) was conducted on the input variables of the wafers to

determine which input variables had a significant effect on the recorded output properties of the wafers. The

ANOVA was carried out using Essential Regression and Experimental Design V2.207 (Tan, 1994). Only the

linear terms were used to regress the data, since we were only interested in the effect that each input variable had

on the measured output variables at a 95% confidence interval.

Disintegration Profiles: The definition of a fast melting (or disintegrating) tablet appeared in a compendia

publication for the first time in 1998. However, neither the US Pharmacopeia north European Pharmacopeia have

defined a specific disintegration test (Yong, 2001). As a result, a novel method was developed to assess and

compare the disintegration profiles of the 30 samples manufactured according to the CCF. Wafers were weighed

before being placed in a petri dish containing 20mL of simulated saliva. The dish was allowed to slowly agitate

on a vortex mixer for a period of 20 seconds. The contents of the dish were sieved through a stainless steel mesh

(pore size 1mm). Particles that were able to pass through the pores of the sieve were considered to be sufficiently

disintegrated, while those captured by the sieve were termed the ‘residue’. The residue represents the portion of

the wafer that was not sufficiently disintegrated. The residue was measured in both the hydrated and dry state. For

wafers that were eroded very rapidly, the agitation time was reduced to 10 seconds. Tests were conducted in

triplicate. Based on the measurements documented, the following information was calculated, providing a

comprehensive disintegration profile for each wafer formulation:

Normalised Percentage Matrix Disintegrated per second (%/s)

Similar to the disintegration profiles, the influx of simulated saliva is calculated as a rate, allowing the various

formulations to be compared on the unit percentage per second.

Friability: Rapidly disintegrating systems prepared by the process of lyophilisation are known for having the

characteristic disadvantage of poor physical resistance (Dobetti, 2001). Problems anticipated as a result of this

include: breakage of tablet edges during handling and the inability of the tablet to be ejected and removed from a

conventional blister alveolus. These features need to be taken into consideration when determining the packaging

of the product. Friabilty was measured using a Roche friabilator (Hoffman la Roche, Basel, Switzerland). The

wafers (N=3) were accurately weighed before being placed into the friabilator. A rotation time of 4 minutes at 25

rpm was used. Tablets were removed and loose particles brushed off the surface. Wafers were re-weighed and the

percentage weight loss was calculated.

Structural Analysis His study focuses on the characterisation of matrix resilience, energy of absorption, matrix

yield value and matrix tolerance, using the TAXTplus Texture Analyser fitted with a 5kg load cell.

Following method of structural analysis:





Energy of Absorption: The energy of absorption is an indirect indication of the porosity of the wafers. A highly

porous wafer will exhibit a greater value for the energy of absorption because energy is accommodated within the

voids in the matrix. The energy of absorption is calculated by determining the area under the curve (AUC) of a

profile illustrating force (N) and distance (m) Note that for the AUC, the units of Newton metre (Nm) are

equivalent to Joules.

Matrix Yield Value: This test is indicative of a surface phenomenon, providing information about the superficial,

surface structure of the wafer. The matrix yield value is determined by creating a gradient between anchors 1 and

2. Anchor 2 represents the first point of major inflection on the force-distance profile. This is indicative of

primary fracture of the wafer matrix which results in a reduction of force.

Matrix Tolerance: On further application of force, the residual intact matrix undergoes complete fracture (Figure

5.3). The matrix tolerance value is indicative of the overall strength of the wafer. The second anchor indicates the

point of maximum force. The gradient is between anchors 1 and 2 in the matrix tolerance value. This indicates the

point at which total collapse of the matrix occurs.

Matrix Resilience: Matrix resilience profiles provide us with an understanding of the deformation characteristics

and the ability of the wafer to withstand pressure. The calculation of matrix resilience is provided by the ratio of

the AUC between anchors 2 and 3 and between anchors 1 and 2.

BHN: The BHN is an indication of the force required to indent the surface of the wafer, and is thus a measure of

the hardness of the surface of the wafer. BHN is calculated using the following equation.

Where: D = Diameter of ball probe = 3.175mm, d = Depth of indentation = 0.25mm, F = Force

CONCLUSION

The lyophilized wafer developed throughout this review is an effective and versatile drug delivery system

for oramucosal application. This has been established from the extensive physicochemical and physic mechanical

profiling conducted. Through a screening and selection of polymers, HPC had the lowest gelation characteristics

and was therefore suitable for the development of the wafer system. Suitable excipient and polymer combinations

were established which allowed for the development of rapidly disintegrating and prolonged release wafer

systems. The wafer system containing HPC, lactose, mannitol and glycine had the ability to disintegrate within 30

seconds. The modified wafer system, consisting of pectin cross linked with zinc ions serving as the drug reservoir,

and mucoadhesive polymer combination of pectin, carmellose and gelatin, provided effective release of model

drug diphenhydramine hydrochloride over approximately six hours.

A successful, reproducible, manufacturing technique was established by the optimization of the

lyophilisation cycle, employing mineral oil as a lubricant and polystyrene moulds providing wafers of suitable

characteristics. Characteristics that were critical to the mechanistic functioning of the wafer, such as rate of matrix

disintegration, rate of simulated saliva influx and friability, were extensively elucidated to determine the effects of

the formulation variables using ANOVA technology. A low concentration of polymer was associated with a high

Disintegration rate, friability and influx of simulated saliva. As predicted, an increase in the amount of diluents

present increased both the disintegration rate and friability.

The ANOVA method was used to present a comprehensive profile of the physic mechanical properties

such as matrix yield value, matrix tolerance, matrix absorption energy, matrix resilience and Brinell hardness

number.A firm understanding of the effects of formulation variables on the responses formed the corner stone of

the optimization process. Although the DSC did not form a component of the optimization process, the

information provided was integral in the determination of the effect of lyophilisation on the native ingredients.

Through this analytical process, it was accepted that lyophilisation did not significantly alter the Tg. The aim of

this study, to consider formulation variables in the statistical optimisation of the lyophilised wafer system was

achieved.





Future Prospects and Challenges (Dobetti, 2001): Historically, drug delivery has taken the form of injection,

infusion, ingestion, and inhalation, with additional variations of each category. For example ingestion may be in

tablet, capsule or liquid form; inhalation may be via use of a dry powder inhaler, an MDI, or a nebulizer. The

challenge for both drug and drug delivery companies is to deliver both existing and emerging drug technologies in

a manner that improves the benefits to the patients, healthcare workers and the healthcare system. Areas that are

being targeted for improvements through device development includes

Improved efficacy

Reduced side effects

Continuous dosing (sustained release)

Reduced pain from administration

Increased ease of use

Increased use compliance

Improved mobility

Decreased involvement of healthcare workers

Improved safety for healthcare workers

Reduced environmental impact (elimination of CFC’s)

To provide these benefits, a number of approaches are being (or in some cases have been) developed. The

common thread running through the approaches is the concept of self-administered, targeted, sustained release

with increased bioavailability. Determining which of the emerging approaches best meets stakeholder needs is a

complex, multifaceted problem. Although ingestion is probably the most widely accepted form of delivery it

presents difficulties for a number of important classes of drugs. Many drug delivery scientists view oral delivery

as the ideal drug delivery method. In the case of proteins and peptides, historical oral delivery mechanisms can

only delivery bioavailabities of a few percent. In some cases, dose limiting toxicity levels are caused by lack of

selectivity. In addition, due to the well known fragility and hygroscopicity of lyophilized products, an appropriate

packaging system for the wafers need to be developed to ensure that the dosage form reaches the patient and is

administered intact.

The modification of this technology to provide a prolonged release mucoadhesive system seems

promising. It is envisaged that this system will be applicable to many drugs requiring the extended release of

bioactive material.

Therefore, the lyophilized wafer matrices developed in this study are highly effective in the rapid delivery of

drugs, using the oral route as a site of administration. The manufacturing process is simple and reproducible. A

number of unique opportunities are presented for the formulation of a controlled release drug delivery system.

Fig 9: Calculation of energy of absorption (i.e. AUC) Fig.10.Determination of matrix yield





Fig.11. Determination of matrix tolerance Fig 12 : Determination of matrix resilience

Fig.13. Force – Distance profile for the computation of the BHN

REFERENCE

Shayne COX GAD, Pharmaceuticals Manufacturing Handbook of Production and Process, A John Wiley & Son,

INC, Publication, 5(1), 2008, 348.

Bhalla HL, Drug delivery Research in India a challenges and opportunity, J Controlled Release, 62, 1999, 65-68.

Chein YW, Novel Drug Dilevery Systems , Marcel Dekker , New York 1992, 1-2

Brannon - Peppas L, Polymer in Controlled Drug Delievery, Biometirial, 11, 1997, 1-14.

Danckwerts MP, Intraoral drug delivery: A comparative review, Amer. J. Drug Del, 1, 2003, 149-224

Squier CA , The permeability of oral mucosa, Crit. Rev. Oral Biol. Med, 2, 1991, 13-32

Shojaei AH, Buccal mucosa as a route for systemic drug delivery: A review, J. Pharm. Sci, 1(1), 1998, 15-30

Rathborne M, Drummond B and Tucker I, Oral cavity as a site for Systemic drug delivery, Adv. Drug Deliv. Rev,

13, 1994, 1-22.

Virely P and Yarwood RJ, Zydis: a novel, fast dissolving dosage form, Manuf. Chem, 61, 1990, 36-37





Guo J.H. Investigating the surface properties and bioadhesion of buccal patches, J. Pharm. Pharmacol, 46, 1994,

647-650,

Tan Y.T.F, Peh KK and Al-Hanbali O, Inveatigation of interpolymer complexation between Carbopol and

various grades of polyvinylpyrrolidone and effects on adhesion strength and swelling properties, J. Pharm. Sci,

4(1), 2001, 14,

Yong CS., Jung J, Rhee J, Kim C and Choi H, Physicochemical characterization and evaluation of buccal

adhesive tablets containing omeprazole, Drug Dev. Ind. Pharm, 27, 2001, 447-455,



Samaresh Pal Roy et.al Indian Journal of Research in Pharmacy and Biotechnology


EVALUATION OF ANTI-ULCER EFFECTS OF ETHANOLIC EXTRACT OF

DELONIX REGIA FLOWER Samaresh Pal Roy*

1, Kamlesh Prajapati

1, Ramji Gupta

2, Dipanwita Bhadra

4, Nikunj Patel

1, Archana

Batiwala1, Gautam Sonara

1, Neerav Gheewala

1, T. Kannadasan

3

1. Department of Pharmacology, Shree Dhanvantary Pharmacy College, Kim, Surat, Gujarat, India

2. Shree Dhanvantary Pharmaceutical Analysis and Research Centre, Kim, Surat

3. Anna University of Technology, Coimbatore, Tamilnadu

4. PDL Dept. Alkem Laboratory Limited, Amulya, Daman

*Corresponding author: Email: [email protected], Mobile: +919377077712

ABSTRACT

Ulcer is one of the most common disorder in the world. The available antiulcer drug in the

market have different side effect. So the people move to herbal drug for a better result and lesser

side effect. Delonix regia is a plant from the family leguminosae is extensively available in the

word. The earlier researcher found that the plant contain the chemical constitutent like tannin,

saponin, flavonoid, so it have a good antioxident property. So, this study is planned recurred the

antiulcer activity by ethanolic extract of the different doses (100mg/kg, 200mg,kg & 400mg/kg) of

the flower of Delonix regia (EEDRF) on ethanol induced ulcer model in experimental rat whereas

lansoprazole (8mg/kg) was taken as an standard drug. It has been found that the extract shows

significant antiulcer activity in a dose dependent manner. The protection of ulcer may due to the

presence of antioxidant principles present in the plant.

Keywords: Delonix regia, Antiulcer, Ethanol

1. INTRODUCTION

Gastric ulcer, the most common disorder of GIT has multifunctional causes in its pathophysiology. The

pathophysiology of peptic ulcer has been centralized on an imbalance between aggressive and protective factors

in the stomach such as acid-pepsin secretion, mucosal barrier, mucus secretion, blood flow, cellular regeneration,

prostaglandins and epidermal growth factors. Although hospital admission for uncomplicated peptic ulcers in

developed countries had begun decrease, there was a striking rise in admission for ulcer haemorrhage and

perforation among elderly people. This increase has been attributed to the increased use of non-steroidal anti-

inflammatory drugs (NSAIDs), alcoholic beverages, cigarettes and Helicobacter pylori infections.

It is now considered to be one of the modern age epidemics affecting nearly 10% of world population

during last decade have offered new insights in the therapy and prevention of peptic ulceration. Plants provide an

alternative strategy in search for new drugs. There is a rich abundance of plants reputed in traditional medicine

known to possess antiulcer properties. It is likely that plants will continue to be a valuable source of new

molecules which may, after possible chemical manipulation, provide new and improved antiulcer drugs.

Borrelli and Izzo reveal the extensive variety of chemical compounds isolated from medicinal plants with

antiulcer activity. Literature search revealed that herbs rich in flavonoids show several biological activities

including antiulcerogenic activity. This is an important reason to investigate antiulcer effects in medicinal plants

with traditional use in gastric diseases.

Delonix regia is a plant from the family leguminosae, is extensively cultivated in most regions of the

world. The flowers of Delonix regia are commonly used as an antibacterial, analgesic, antiulcer, anti-

inflammatory and antimicrobial. Reports indicate that pharmacological activities of Delonix regia flowers include

anti-inflammatory and analgesic (Ahmad & Aqil., 2003), antimicrobial, broad spectrum antibacterial and

antifungal activities.Wijayasirivardena et al 2009 reported the plant possess antiinflammatory activity.

Muruganathan et al. 2011 reported that the bark of Delonix regia showed significant anti-inflammatory

and anti-arthritic activity. Pradeepa et al. 2012 reported the leaf extract of Delonix regia showed antinociceptive

activity. The plant has been claimed to be useful as antioxidant (Aquil et al., 2006), larvicidal (Chockalingam et

al., 1990), antibacterial, antifungal (Ahmed et al., 2003), anti-inflammatory, analgesic (Muruganandam et al.,

2000), nutritional (Grant et al., 1991), antimalerial (Ankrah et al., 2003), antiperiodic, febrifuge, emetic, CNS

depressant (Rastogi et al., 1993) and antirheumatic (Khare et al., 2007). Its aqueous and alcoholic extracts were

active against roundworm. The bark contains leucocyanidin, lupeol, tannin, -sitosterol and free OH-proline as






major amino acid. Flower anthers are a rich source of zeaxanthin. Leaves contain tannins, lupeol and -sitosterol

(Khare et al., 2007). D. regia seeds contain lectins (Pando et al., 2002).

However, there is no scientific proof justifying the traditional use of Delonix regia flowers in treatment of

ulcer. Hence the present investigation was taken up to evaluate its potential gastroprotective efficacy in ethanol

induced experimental models of ulcer in rats.

2. MATERIALS AND METHODS

2.1 Plant material: Flowers of Delonix regia were collected from the field of Surat and the plant was

authenticated by Dr. Vinoo Parabia, Department of Bioscience, Veer Narmad South Gujarat University, Surat. A

herbarium specimen is deposited in our college museum.

2.2 Preparation of 70% EEDRF: The flowers were shade dried at room temperature and pulverized. The

powder obtained was subjected to successive soxhlet extraction with 70% ethanol (hydro-alcoholic extract). The

extract were concentrated under reduced pressure and stored in desicator until further use and the percentage yield

of corresponding extracts were calculated.

2.3 Animals: Whister albino rats (150-200g) and mice (18-25 g) of either sex were used for the study, obtained

from AFC, Shree Dhanvantary Pharmacy College, kim. After one week of acclimatization the animals were used

for further experiments. Approval from the Institutional Animal Ethical Committee for usage of animal in the

experiment was obtained as per the Indian CPCSEA guidelines.

2.4. Acute Toxicity studies: The acute toxicity was determined on albino mice by fixed dose method of OECD

Guide line given by CPCSEA. Groups of 6 mice were administered test drug by oral route in the range of 2000-

3000 mg/kg and mortality was observed after 24 hr.

2.5 Ethanol induced ulcer model (Surana et al., 2007): Albino rats of either sex weighing between 120 – 200 gm

were selected and divided into 5 groups of 6 animals each.

Table.1.Grouping of animals for the conduction of the trial

Group I Control (1 ml/200 g of 99.80% ethanol p.o.)

Group II Standard (Lansoprazole 8 mg/kg i.p.)

Group III 70% ethanolic extract of DR flowers 100 mg/kg p.o.

Group IV 70% ehtanolic extract of DR flowers 200 mg/kg p.o.

Group V 70% ethanolic extract of DR flowers 400 mg/kg p.o.

The animals were fasted for 24 hours with free access to water. Animals were given different doses of DRF

extract and standard drug lansoprazole as mentioned above. Thirty minutes after the treatment 1ml/200 g of

99.80% alcohol was administered p.o. to each animal. Animals were sacrificed 1 hr. after alcohol administration,

stomachs were isolated and cut open along the greater curvature and pinned on a soft board. The ulcer index was

measured.

Table.2.Codes for showing extent of ulcer activity in animals

Code Extent of ulcer activity

0 Normal stomach

0.5 Red coloration

1 Spot ulcers

1.5 Hemaorrhagic streaks

2 Ulcer > 3 mm < 5mm

3 Ulcers > 5mm

The percentage protection was calculated using the formula

Where Ut = Ulcer index of treated group

Uc = Ulcer index of control group.





2.6. Statistical analysis: Results were expressed as mean of # SEM (n=6). Statistical analysis was

performed with one way ANOVA followed by Turkey-Kramer multiple comparisons test. P values less

than 0.05 was considered to be statistically significant (p<0.05). 3. RESULTS

3.1. Acute toxicity: The acute toxicity study showed No animal died even at 2000mg/kg and hence the extract

was treated as non-toxic and 1/25th, 1/10

th & 1/5

th of the 2000mg/kg was selected for further investigations are

mentioned below.

I. 100 mg/kg (1/25th of 2000 mg/kg)

II. 200 mg/kg (1/10th of 2000 mg/kg)

III. 400 mg/kg (1/5th of 2000 mg/kg).

3.2. Antiulcer Activity: The extract (100, 200 and 400 mg/kg) significantly inhibited the ulcerogenic effect of

ethanol in rats. The percentage protection of ulcer is increase according to the increase in concentration of extract.

The higher doses i.e 200mg & 400 mg/kg of the ethanolic extract of DF flower shows significant ulcer protection

whereas 100mg/kg dose does not shows any significant effect. The standard drug lansoprazole (8mg/kg) shows a

highly significant result. The ulcer score also reveals the antiulcer protection of the extract. The results are

summarized in Table.3.

4. DISCUSSION

In the present investigation different doses of ethanolic extract of DR flowers (EEDRF) was screened for

gastroprotective properties by employing ethanol induced ulcer models in rats. It has been found that the ethanolic

extract of Delonix regia has shown the antiulcer property in a dose dependent manner. Ethanol provoked gastric

mucosal lesions are caused by the direct toxic effects of ethanol through the reduction in mucus production,

gastric mucosal blood flow and bicarbonate secretion. Endogenous glutathione and prostaglandin levels are also

lowered by ethanol while the release of histamine, influx of calcium ions, generation of free radicals and

production of leukotriene’s are all increased (somchit et al., 2007). The product of the 5-lipoxygenase pathway

may also play a key role in the development of ulcer induced by irritant agents such as ethanol. It has been

reported that leukotriene antagonist and 5-lipoxygenase inhibitors are capable of inhibiting alcohol and NSAIDs

induced gastric ulceration in rats (surana et al., 2007). In the present study also the significant protection exhibited

by the test extract against alcohol induced gastric ulceration may be due to inhibition of 5-lipoxygenase pathway

or leukotriene antagonistic activity.

5. CONCLUSION

The results of the present study indicate that ethanolic extract of Delonix regia flowers possessed

significant antiulcer properties, thus supports the traditional use of DR flowers in treatment of gastrointestinal

disorders.

6. ACKNOWLEDGEMENTS The authors express their gratitude to the entire fraternity of Shree Dhanvantary Pharmacy college &

Shree Dhanvantary Pharmaceutical Analysis & Research Centre for the completion of this project work.

A : Control

B : Std. Lansoprazole (8 mg/kg)

C : 70% Ethanolic Ext. (100 mg/kg)

D : 70% Ethanolic Ext. (200 mg/kg)

E : 70% Ethanolic Ext. (400 mg/kg)

Figure.1. Antiulcer activity of ethanolic extract of DR flowers





Table.3.Effect of 70% alcoholic extract of flower of Delonix regia on Ethanol-induced ulcer (Mean ± SEM,

n = 6 in each group) in rats

** P < 0.01 and *** P < 0.001 (Vs. Control) respectively

Control Standard

(lansoprazole)

EEDRF 100 mg/kg EEDRF 200 mg/kg EEDRF 400 mg/kg

Figure.2. Photograph showing open stomach of alcohol induced gastric ulceration rats

Gr.

No.

Bo

dy

wei

gh

t

Treatment ULCER INDEX Tota

l

scor

e

Mea

n u

lcer

Ind

ex ±

SE

M

%

of

pro

tecti

on

No

rma

l

colo

ure

d

sto

ma

ch

Red

colo

ura

tio

n

Sp

ot

ulc

er Haemorr

h-agic

streaks

Ulcer

≥3 but

≤5

Ulcer

≥ 5

I 160 Control - 0.5 1.0 1.5 2.0 - 5.0 6.00

± 0.632

-

160 - 0.5 1.0 1.5 2.0 - 5.0

170 - 0.5 1.0 1.5 2.0 - 5.0

170 - 0.5 1.0 1.5 2.0 - 5.0

150 - 0.5 1.0 1.5 2.0 3.0 8.0

175 - 0.5 1.0 1.5 2.0 3.0 8.0

II 180 Lansoprazol

e

8 mg/kg

- 0.5 - - - - 0.5 1.00

± 0.447

***

83.33

180 - 0.5 - - - - 0.5

175 - 0.5 - - - - 0.5

170 - 0.5 1.0 - - - 1.5

175 - - - - - - -

170 - 0.5 1.0 1.5 - - 3.0

III 160 Alcoholic

extract 100

mg/kg

- 0.5 1.0 1.5 - - 3.0 3.916

± 0.757

34.73

170 - 0.5 1.0 1.5 2.0 - 5.0

170 - 0.5 1.0 1.5 2.0 - 5.0

180 - 0.5 1.0 1.5 2.0 - 5.0

180 - 0.5 - - - - 0.5

180 - 0.5 1.0 1.5 2.0 - 5.0

IV 180 Alcoholic

extract

200mg/kg

- 0.5 1.0 1.5 2.0 - 5.0 2.166

± 0.909

**

63.90

180 - 0.5 - - - - 0.5

175 - 0.5 1.0 1.5 2.0 - 5.0

170 - 0.5 1.0 - - - 1.5

170 - 0.5 - - - - 0.5

160 - 0.5 - - - - 0.5

V 160 Alcoholic

extract

400mg/kg

- 0.5 1.0 1.5 2.0 - 5.0 1.416

± 0.735

**

76.40

175 - 0.5 1.0 - - - 1.5

180 - 0.5 - - - - 0.5

170 - 0.5 - - - - 0.5

170 - 0.5 - - - - 0.5

165 - 0.5 - - - - 0.5





REFERENCES

Ahmad I, Aqil F, Broad spectrum antibacterial and antifungal activities and potency of crude alcoholic extract and

fractions of Delonix regia flowers, 2nd

World Congress on “Biotechnological Development of Herbal Medicine”,

NBRI, Lucknow, UP, India, 2003:74.

Asolkar LV, Kakkar KK, Chakre OJ, Second Supplement to Glossary of Indian Medicinal Plants with active

principles-Part-I, New Delhi: National Institute of science Communication (CSIR), 1992:264.

Bhave AL, Bhatt JD, Hemavathi KG, Antiulcer effect of Amlodipine and its interaction with H2 blocker and

proton pump inhibitor in pylorus ligated rats, Indian J Pharmacol, 38(6), 2006, 403-07.

Biswajit Majumdar, Sursi Guha Ray Chaudhuri, Arun Ray, Sandip K Bandyopadhyay. Effect of ethanol extract of

Piper betle Linn leaf on healing of NSAID–induced experimental ulcer–A novel role of free radical scavenging

action, Indian J Exp Biol, 41, 2003, 311-15.

Borrelli F, Izzo AA, The plant kingdom as a source of anti-ulcer remedies, Phytother Res, 14, 2000, 581-91.

Deshpande SS, Shah GB, Parmar NS, Antiulcer activity of Tephrosia purpurea in rats, Indian J Pharmcol 35,

2003, 168-72.

Edward F, Gilman and Dennis G, Watson. Delonix regia-Royal Poinciana, Fact Sheet ST-228, a series of

Environemtnal Horticulture Departmrnt, Florida Cooperative Extension Service, Institute of Food and Agriculture

Sciences, University of Florida, Nov 1993.

Etuk EU, Agaie BM, Onyeyill PA, Ottah CU, The effect of Rabeprazole and its isomers on aspirin and histamine-

induced ulcers in rats, Indian J Pharmacol, 38(5), 2006, 357-58.

Ganachari MS, Shiv kumar. Anti-ulcer properties of Ziziphus jujuba Lam leaves extract in rats, J Nat Rem, 4(2),

2004, 103-08.

Goel RK, Sairam K, Dorababu M, Prabha T, Rao V, Effect of standardized extract of Ocimum sanctum Linn. on

gastric mucosal offensive and defensive factors, Indian J Exp Bio, 43, 2005, 715-21.

Hemamalini K, Vimal kumar Varma M, Antiulcer activity of Indigofera aspalathoids on chemically induced

ulcer models in rats and guinea pigs, Adv Pharmacol Tox, 7, 2006, 25-29.

Higham J, Kang JY, Majeed A, Recent trends in admissions and mortality due to peptic ulcer in England :

increasing frequency of haemorrhage among older subjects, Gut, 50, 2002, 460-64.

Jana U, Bhattacharya D, Bandopadhyay S, Pandit S, Debnath PK, Sur TK. Antiulcer activity of Digitrall: A

polyherbal drug in rats, Indian J Pharmacol, 37(6), 2005, 406-07.

Jungalwala FB, Cama HR. Carotenoids in Delonix regia (Gul Mohr) flower, Biochem J, 1962; 85:1.

Karpagam Kumara Sundari S, Murugesh, Nallu M, Antiulcer activity of certain Phenyl tosylates, Adv Pharmacol

Toxicol, 7(1), 2006, 7-9.

Kumar A, Ram II. Anti-ulcer properties of methanolic extract of Benincasa hispida (Thunb.) Cogn. Indian Drugs,

39(1), 2002, 9-13.

Lima ZP, Severi JA, Pellizzon CH, Brito ARMS, Solis PN, Caceres A, Can the aqueous decoction of Mango

flowers be used as an antiulcer agent, Ethnopharmacol, 106, 2006, 29-37.





Muruganathan G, and Mohan S. Anti-inflammatory and Anti-Arthritic activities of Delonix elata bark extracts, Int

J Res Ayu Pharm, 2(6), 2011, 1819-1821.

Pandit S, Sur TK, Jana U, Bhattacharyya D, Debnath PK. Antiulcer effect of Shankha bhasma: A preliminary

study, Indian J Pharmacol, 32, 2000, 378-80.

Pradeepa K, Krishna V, Venkatesh, Kumar GK, Kumar SRS, Hoskeri JH, Antinociceptive activity of Delonix

elata leaf extract, Asia Pacific J of Tropical biomedicine, 2012, 229-S231.

Sairam K, Ch V Rao, Dora Babu M, Vijay Kumar K, Agarwal VK, Goel RK. Anti-ulcerogenic effect of

methanolic extract of Emblia officinalis: an experimental study, J Ethnopharmacol, 82, 2002, 1-9.

Shah JS, Anand IS, Patel SK, Patel HU, Thakkar VT, A comparative study of antacid and antiulcer activity of two

marketed herbal products in North Gujarat region, Indian Drugs, 43(9), 2006, 763-65.

Somchit MN, Siti Rahmah S, Zuraini A, Ahmad Bustamam A, Zakaria ZA, Somchit N, Gastroprotective activity

of Spirulina platensis in acetic acid and ethanol induced ulcers in rats, J Nat Rem,7(1), 2007, 37-42.

Sonowski RA. The changing spectrum of therapy of active peptic ulcer disease, Modern Medicine, 58, 1990, 50-

58.

Surana SJ, Tatiya AU, Jain AS and Ushir YV. Antiulcer activity of Eranthemum Roseum (VAHL) R.BR on

ethanol induced ulcer in albino rats.Int.J.Pharmacol.Biol.Sci, 1(1), 2007, 65-66

Surana SJ, Tatiya AU, Jain AS, Ushir YV, Antiulcer activity of Eranthemum roseum (VAHL) R.Br on ethanol

induced ulcer in albino rats, Int J Pharmacol Biol Sci, 1(1), 2007, 65-69.

Wijayasirivardena C, Chauhan NG, Sharma PP, Lahore SK, Shah MB, Anti-inflammatory activity of Delonix

elata (L) gamble, J Nat Rem, 9(2), 2009, 209-215.



Dr. Praveen Kumar Indian Journal of Research in Pharmacy and Biotechnology


A STUDY ON MEDICATION NON-ADHERENCE IN AMBULATORY

DIABETIC PATIENTS AND NEED FOR PHARMACIST INTERVENTION FOR

IMPROVING PATIENT ADHERENCE Dr. Praveen Kumar G

Assistant Professor, C.L. Baid Metha College of Pharmacy, Chennai, Tamil Nadu.


ABSTRACT

Diabetes mellitus is a chronic disease and the prevalence for all age-groups worldwide was estimated to

be 2.8% in 2000 and 4.4% in 2030. Medication non adherence is a pervasive medical problem that is common

among patients with chronic disease. This study was conducted for a period of 1 month. It includes 154 randomly

selected patients who were interviewed by using a questionnaire regarding socio-economic characteristics,

adherence rates, barriers that affect adherence to medication use (using Morisky self report scale) and counseled

by pharmacist for 20-30 minutes. Of the total, 97(62.9%) were adherent and 57(37.01%) were non adherent to

medications. In our study the commonly cited intentional non-adherence was found to be self-decision(35.08%)

and ommision of drugs because of experiencing Side effects (22.8%). Confusion in dosage frequency (17.5%),

forgetfulness (10.5%) and financial difficulty (8.7%) were the other contributing factors for non-adherence.

Efforts were taken to increase the medication adherence and self care through patient education by pharmacists.

Patient’s medication compliance is a multifactor behavior in which the role of patient’s attitude is very important.

Interventions are needed to increase medication adherence so that patients can realize the full benefit of prescribed

therapies.

Key words: Diabetes mellitus, non adherent, pharmacist

INTRODUCTION

Diabetes mellitus is a chronic disease and the prevalence for all age-groups worldwide was estimated to

be 2.8% in 2000 and 4.4% in 2030.In India the prevalence of diabetes and pre-diabetes among adults were 5.1%

and 13.5%. Medication non adherence is a pervasive medical problem that is common among patients with

chronic disease. Factors such as uncontrolled diet, sedentary lifestyle, inappropriate therapeutic regimens as well

as medication non adherence have been known to have significant impact on glycemic control and outcome of

type 2 diabetes treatment. In unadjusted analyses non adherent patients had higher hospitalization (23.2%) and

higher mortality (5.9%). With this background information this study has been proposed to evaluate the probable

reasons for patient’s non adherence to prescribed anti diabetic medications in ambulatory care type 2 diabetes

patients and to improve the adherence rate through patient education.

Objective:

To assess adherence levels and factors affecting adherence in diabetic patients.

To educate patients on self care and adherence to the lifestyle modifications (diet and exercise) as well as

pharmacological therapy.

Methodology: The study was conducted in the Sri Ramachandra Hospital for a period of 1 month. A total of 154

randomly selected patients who were attending the diabetes outpatient department were interviewed by using a

questionnaire regarding socio-economic characteristics, adherence rates,barriers that affect adherence to

medication use(using Morisky self report scale) and counseled by pharmacist for 20-30 minutes.


Among the 154 patients, 42 (27.2%) male and 112(72.7%) female were taking a mean of 5.3 medicines

per patient to control diabetes and related comorbidities. Of the total, 97(62.9%) were adherent and 57(37.01%)

were non adherent to medications. In our study the commonly cited intentional non-adherence was found to be

self-decision(35.08%) and ommision of drugs because of experiencing Side effects (22.8%). Confusion in dosage

frequency (17.5%), forgetfulness (10.5%) and financial difficulty (8.7%) were the other contributing factors for

non-adherence. All together 128patients (83.12%) were on anti-hypertensive and 73patients (47.4%) were on lipid

lowering medicines. Efforts were taken to increase the medication adherence and self care through patient

education by pharmacists.



Dr. Praveen Kumar Indian Journal of Research in Pharmacy and Biotechnology


Factors for

non-adherence

Gender No. of

patients

%

Self decision Male 4 7.01

Female 16 28.07

Side effects Male 3 5.26

Female 10 17.54

Confusion Male 2 3.5

Female 8 14.03

Forget Male 2 3.5

Female 4 7.01

Financial

difficulty

Male 1 1.75

Female 4 7.01

Others Male 1 1.75

Female 2 3.5

GRAPH3 SD1: alter dosing schedule for convenience,

SD2: omit drugs if feeling ill, SD3:don’t care to take

drugs, SD4: think drugs are not effective.

GRAPH.2.SD:selfdecision,SE:sideeffects, C:confusion,

F:forget, FD:financial difficutly, O:Others

CONCLUSION

Patient’s medication compliance is a multifactor behavior in which the role of patient’s attitude is very

important. Interventions are needed to increase medication adherence so that patients can realize the full benefit of

prescribed therapies.

REFERENCES

Adisa R, Alutundu MB, Fakeye TO, Factors contributing to nonadherence to oral hypoglycemic medications

among ambulatory type 2 diabetes patients in Southwestern Nigeria, Pharmacy Practice, 7(3), 2009, 163-169.



Gowthami B et.al Indian Journal of Research in Pharmacy and Biotechnology


RECENT TRENDS IN POSITIVE AND NEGATIVE ASPECTS OF FOOD ON

BIOAVALABILTY OF DRUGS Gowthami B

1*, Sk Nahida Fazilath

2, Sanaulla Md

2, K Prudhvi Raj

2, Dastagiriah G

2, Tabassum Sk

2

1.Vaagdevi College of Pharmacy and Research Center, Nellore

2.Nimra College of Pharmacy

Corresponding Author: Email: [email protected]

ABSTRACT

The therapeutic effectiveness of a drug depends upon the ability of the dosage form to deliver the

medicament to its site of action at a rate & amount sufficient to produce the desired pharmacologic

response. This attribute of the dosage form is referred to as Physiologic availability or Biologic

availability or simply availability. For most of the drugs, the pharmacologic response is directly related

to the plasma levels. Thus, the term, Bioavailability is defined as the rate & extent (amount) of

absorption of unchanged drug from its dosage form. Bioavailability refers to the difference between the

amounts of a substance, such as a drug, herb, or chemical, to which a person is exposed and the actual

dose of the substance the body receives. Bioavailability accounts for the difference between exposure and

dose. A drug's therapeutic action or a chemical's toxicity is determined by the dose received at the target

site in the body. The dose at the target site is determined by the amount of the substance absorbed by the

body, which depends on its bioavailability. If a substance is ingested, for example, its bioavailability is

determined by the amount that is absorbed by the intestinal tract. If a substance is inhaled, its

bioavailability is determined by the amount that is absorbed by the lungs. Understanding bioavailability

is critical to determining the amount of a drug to administer or the level of chemical exposure that is

likely to produce toxicity.

Key words: Bioavailability, Bioavailability studies, Absolute and Relative bioavailability.

INTRODUCTION

In comparison to drugs, there are significant differences in dietary supplements that impact the evaluation

of their bioavailability. These differences include the following: the fact that nutritional supplements provide

benefits that are variable and often qualitative in nature; the measurement of nutrient absorption lacks the

precision; nutritional supplements are consumed for prevention and well-being; nutritional supplements do not

exhibit characteristic dose-response curves; and dosing intervals of nutritional supplements, therefore, are not

critical in contrast to drug therapy. In addition, the lack of defined methodology and regulations surrounding the

consumption of dietary supplements hinders the application of bioavailability measures in comparison to drugs. In

clinical trials with dietary supplements, bioavailability primarily focuses on statistical descriptions of mean or

average AUC differences between treatment groups, while often failing to compare or discuss their standard

deviations or inter-individual variation. This failure leaves open the question of whether or not an individual in a

group is likely to experience the benefits described by the mean-difference comparisons. Further, even if this

issue were discussed, it would be difficult to communicate meaning of these inter-subject variances to consumers

and/or their physicians.

FACTORS INFLUENCING BIOAVAILABILITY

Before the therapeutic effect of an orally administered drug can be realized, the drug must be absorbed. The

systemic absorption of an orally administered drug in a solid dosage form is comprised of three distinct steps:

1. Disintegration of the drug product

2. Dissolution of the drug in the fluids at the absorption site

3. Transfer of drug molecule across the membrane lining the gastrointestinal tract into the systemic

circulation.

http://en.wikipedia.org/wiki/Dose-response_relationship





Bioavailability and oral Dosage Forms Bioavailability Factors related to the dosage form

Physicochemical

properties of the drug

Formulation and

manufacturing variables Particle size

Crystalline structure

Degree of hydration of

crystal

Salt or ester form

Amount of disintegrant

Amount of lubricant

Special coatings

Nature of diluent

Compression force

Bioavailability Factors related to the patient

Physiologic factors Interactions with other substances Variations in absorption power along GI tract

Variations in pH of GI fluids

Gastric emptying rate

Intestinal motility

Perfusion of GI tract

Presystemic and first-pass metabolism

Age, sex, weight

Disease states

Food

Fluid volume

Other drugs

Factors influencing Gastric Emptying Rate

Factor Influence on gastric emptying rate

Increased viscosity of stomach contents Decreased

Body position lying on left side Decreased

Emotional state stress depression

anxiety

Increased or Decreased

Increased

Activity, exercise Decreased

Type of meal fatty acids, fats carbohydrates

amino acids

Decreased

pH of stomach contents decreased

increased

Decreased

Increased

Disease states gastric ulcers Crohn’s disease

hypothyroidism

hyperthyroidism

Decreased

Decreased

Increased

Drugs atropine propantheline narcotic analgesics

amitriptyline

metoclopramide

Decreased

Decreased

Increased

OBJECTIVES OF BIOAVAILABILITY STUDIES

1. Primary stages of development of a suitable dosage form for a new drug entity.

2. Determination of influence of excipients, patient related factors and possible interaction with other drugs

on the efficiency of absorption.

3. Development of new formulations of the existing drugs.

4. Control of quality of a drug product during the early stages of marketing in order to determine the

influence of processing factors, storage and stability on drug absorption.

PURPOSE OF BIOAVAILABILITY STUDIES

1. Bioavailability studies are performed for both approved active drug ingredients and therapeutic moieties

not yet approved for marketing by the FDA.





2. New formulation of active drug ingredients must be approved by the FDA before marketing to ensure the

safe and effective for its labeled indications for use.

3. According to FDA the drug product must meet all applicable standards of identity, strength, quality, and

purity. To ensure all these standards are met, the FDA requires bioavailability and pharmacokinetic

studies and where necessary, bioequivalence studies for all drug products.

4. Bioavailability links in vivo performance of the drug product used in clinical trials to studies

demonstration evidence of safety and efficacy.

5. For unjacketed drug that donor have NDA approval by the FDA, inviter and in vivo bioequivalence

studies must be performed on the drug formulation proposed for marketing as a genetic drug product.

Essential pharmacokinetic parameters, including the rate and extent of systemic absorption, elimination

half life, and rates of excretion and metabolism, should be established after single and multiple dose

administration. Data from these in vivo studies are important to establish recommended dosage regimens

and to support the drug labeling.

6. Used to define the effect of changes in the physicochemical properties of the drug substance and the

effect of the dug product on the pharmacokinetics of the drug.

7. Bioequivalence studies are used to compare the bioavailability of the same drug from various drug

products.

8. Bioavailability and bioequivalence can also be considered as the performance measures of the drug

product invivo. If the drug products are bioequivalent and therapeutically equivalent then the clinical

efficacy and the safety profile of these drug products are assumed to be similar and may be substituted for

each other.

CONSIDERATION IN BIOAVAILABILITY STUDY DESIGN

Bioavailability: Absolute versus Relative

Absolute bioavailability: When the systemic availability of a drug administered orally is determined in

comparison to its intravenous administration, is called as absolute availability (denoted by F).

AUC = Area under the Curve

D = Dose of administered drug

Its determination is used to characterize a drug’s inherent absorption properties from extra

vascular site.

Intravenous dose is selected as a standard due to its 100% bioavailability.

If the drug is poorly water soluble, intramuscular dose can be taken as standard.

Relative bioavailability: - When the systemic availability of a drug after administration is compared with that of

standard of the same drug it’s referred to as relative bioavailability (Fr).

In contrast to absolute bioavailability, it is used to characterize absorption of drug from its formulation.

F & Fr are expressed in percentages.

Single dose versus multiple dose studies: These are very common.

They are easy, offer less exposure to drugs & are less tedious.

However, it is difficult to predict the steady-state characteristic of a drug & inter-subject variability with

such studies.

On the other hand, multiple Dose study is difficult to control (poor subject compliance), exposes the

subject to more drugs & is highly tedious & time consuming. Nevertheless, such a study has a several

advantages.

1. More accurately reflects the manner in which the drug should be used.





2. Easy to predict the peak & valley characteristic of the drug since the bioavailability is determined at

steady – state.

3. Requires collection of fewer blood samples.

4. The drug blood levels are higher due to cumulative effect which makes its determination possible even by

the less sensitive analytic methods.

5. Can be ethically performed in patients because of the therapeutic benefit to the patient.

6. Better evaluation of the performance of a controlled release formulation is possible.

7. Nonlinearity in pharmacokinetics, if present, can be easily detected.

In Multiple Dose study, one must ensure that the steady state has been reached. For this, the drug should

be administered for 5-6 elimination half-lives before collecting the blood samples.

Human Volunteers- Healthy subjects verses Patients: Ideally, the bioavailability study should be carried out in

patients for whom the drug is intended to be used because of the apparent advantages;

1. The patient will be beneficial from the study.

2. Reflects better therapeutic efficacy of a drug.

3. Drug absorption pattern in disease states can be evaluated.

4. Avoids the ethical requirements of administering drugs to the healthy subjects.

In multiple dose study, they prefer patients rather than healthy humans.

The drawbacks of using patients as volunteers are equally large- disease, other drugs, physiologic

changes etc. may modify the drug absorption pattern.

Strict study conditions such fasting state required to be followed by the subject is also difficult.

In short, establishing a standard set of conditions necessary for a bioavailability study is difficult with

patients as volunteers.

Therefore, such studies are generally carried out in healthy human volunteers (20-40 yrs), preferably

male adults with body weight within narrow range +/- 10 %, under restricted dietary & fixed activity

conditions.

The consent of volunteers must be obtained and they must be informed about the conditions to be

followed during the course of studies – to abstain from any other medication for at least 2 weeks and

to fast overnight prior to and for a minimum of 2-4 hours post dosing as well as possible hazards if

any.

METHODS FOR ASSESSING THE BIOAVAILABILITY 1) Pharmacokinetic methods: These are very widely used and based on the assumption that the pharmacokinetic

profile reflects the therapeutic effectiveness of a drug. Thus these are indirect methods.

Indirect methods

a) Plasma level – time studies

b) Urinary excretion studies

2) Pharmacodynamic methods: These methods involve direct measurement of drug effect on a physiologic

process as a function of time.

Direct methods

a) Acute pharmacologic response

b) Therapeutics response

1) Pharmacokinetic methods

a) Plasma level – time studies: It is the most reliable Method, & based on assumption that two dosage forms

exhibit super imposable plasma level-time profiles in a group of subjects should result in identical therapeutic

activity.

With single dose study: Requires collection of samples for a period of 2 to 3 biological half lives. & make a plot

of Conc. Vs time of sample collection.

i.v. dose: sampling start within 5 min. and subsequent samples taken at 15 min intervals.

The 3 parameters of plasma level-time studies which are considered important for determining bioavailability are:

1. Cmax: The peak plasma concentration that gives an indication whether the drug is sufficiently absorbed

systematically to provide a therapeutic response.

2. tmax: The peak time that gives an indication of the rate of absorption.





3. AUC: The area under the plasma level-time curve that gives a measure of the extent of absorption or the

amount of drug that reaches the systemic circulation.

The extent of Bioavailability can be determined from equations:

F = [AUC] oral Div / [AUC] iv Doral

Fr = [AUC] test Dstd / [AUC] std Dtest

Where D stands for dose administered and subscripts i.v and oral indicates the route of administration. Subscripts

test and std indicate the test and standard dose of the same drug to determine relative bioavailability.

With multiple dose study: The method involves drug administration for at least 5 biological half-lives with a

dosing interval equal to or greater than the biological half-life to reach the steady state.

The extent of bioavailability for multiple dose study is given as:

Fr = [AUC]test Dstd τtest/ [AUC]std Dtest τstd

Where [AUC] values are under the plasma level- time curve of one dosing interval in a multiple dosing regimen

after reaching the steady state and τ is the dosing interval.

The Bioavailability can be determined from peak plasma concentration at steady state Css, max according to

following equation:

Fr = (Css,max)test Dstd τtest / (Css,max)std Dtest τstd

b) Urinary excretion studies: Based on the principle that the urinary excretion of unchanged drug is directly

proportional to the plasma concentration of drug. E.g. Certain diuretics and sulfonamides, urinary antiseptics such

as nitrofurantion and hexamine. Concentrations of metabolites excreted in urine is never taken into account in

calculations since a drug undergo presystemic metabolism before being absorbed. Method involves sampling at

regular intervals for a time span equal to7 biological half-lives. Total emptying of the bladder is necessary to

avoid errors.

The three major parameters examined in urinary excretion studies are:

1) (dxu/dt)max: The maximum urinary excretion rate, obtained from the peak of plot between rate of

excretion versus midpoint time of urine collection. It is analogous to Cmax. Its value increases as rate of

and/or extent off absorption increases.

2) (tu)max: The time for maximum excretion rate, it is analogous to the tmax. Its value decreases as the

absorption increases.

3) Xu: The cumulative amount of drug excreted in the urine, it is related to the AUC and increases as the

extent of absorption increases.

The extent of bioavailability is calculated from equation:

F = (Xu∞)oral Div / (Xu∞)iv Doral

Fr = (Xu∞)test Dstd / (Xu∞)std Dtest

With the multiple dose study to steady state , the equation is:

Fr = (Xu,ss)test Dstd τtest / (Xu,ss)std Dtest τstd

Where, (Xu,ss) is the amount of drug excreted unchanged during a single dosing interval at steady state.

Bioavailability of few drugs can be also determined by assay of biologic fluids other than plasma and urine.

For e.g. Theophyllline salivary secretion

Cephalosporin CSF and bile

2) Pharmacodynamic methods:

a) Acute pharmacologic response: When bioavailability measurement by pharmacokinetic methods is

difficult,an acute pharmacological effect such as a change in ECG or EEG readings, pupil diameter,etc.is related

to the time course of a given drug. Thus bioavailability can be determined by construction of pharmacological

effect-time curve as well as dose-response graphs. The method requires measurement of responses for at least 3

biological half-lives of the drug in order to obtain a good estimate of AUC.

Disadvantages: The pharmacological response tends to be more variable and accurate correlation between

measured response and drug available from the formulation is difficult.

The observed response may be due to an active metabolite whose concentrations not proportional to the

concentration of parent drug responsible for the pharm1acological effect.

b) Therapeutic response: Based on observing the clinical response to a drug formulation given to patients

suffering from disease for which it is to be used.





Drawback: Quantitation of observed response is too improper to allow for reasonable assessment of relative

bioavailability between two dosage forms of the same drug.

EFFECT OF FOOD ON BIOAVAILABILITY

Drugs are frequently taken with food, and patients often use mealtimes to remind them to take their

medications. However, food can have a significant effect on the bioavailability of drugs. The influence of food

on drug absorption has been recognized for some time, and several reviews have been published on the influence

of food on drug bioavailability. Food may influence drug absorption indirectly, through physiological changes in

the GI tract produced by the food, and/or directly, through physical or chemical interactions between the drug

molecules and food components. When food is ingested, stomach emptying is delayed, gastric secretions are

increased, stomach pH is altered, and splanchnic blood flow may increase. These may all affect bioavailability of

drugs. Food may also interact directly with drugs, either chemically (e.g. chelation) or physically, by adsorbing

the drug or acting as a barrier to absorption. In general, gastrointestinal absorption of drugs is favored by an

empty stomach, but the nature of drug-food interactions is complex and unpredictable; drug absorption may be

reduced, delayed, enhanced or unaffected by the presence of food. Following Table summarizes some of the

studies that have indicated the effect of food on the bioavailability of a variety of drugs.

FOOD-DRUG INTERACTIONS

Medicines can treat and cure many health problems. However, they must be taken properly to ensure

that they are safe and effective. Medications should be extremely specific in their effects, have the same

predictable effect for all patients, never be affected by concomitant food or other medications, exhibit linear

potency, be totally non-toxic in any dosage and require only a single dose to affect a permanent cure. However,

this ideal drug is still to be discovered. Many medicines have powerful ingredients that interact with the human

body in different ways. Diet and lifestyle can sometimes have a significant impact on drugs. A drug interaction is

a situation in which a substance affects the activity of a drug, i.e. the effects are increased or decreased, or they

produce a new effect that neither produces on its own. Typically, interactions between drugs come to mind (drug-

drug interaction). However, interactions may also exist between drugs and foods (drug-food interactions), as well

as drugs and herbs (drug-herb interactions). These may occur out of accidental misuse or due to lack of

knowledge about the active ingredients involved in the relevant substances. Interactions between food and drugs

may inadvertently reduce or increase the drug effect. Some commonly used herbs, fruits as well as alcohol may

cause failure of the therapy up a point of to serious alterations of the patient`s health. The majority of clinically

relevant food-drug interactions are caused by food induced changes in the bioavailability of the drug. Major side-

effects of some diet (food) on drugs include alteration in absorption by fatty, high protein and fiber diets.

Bioavailability is an important pharmacokinetic parameter which is correlated with the clinical effect of most

drugs. However, in order to evaluate the clinical relevance of a food-drug interaction the impact of food intake on

the clinical effect of the drug has to be quantified as well.

The most important interactions are those associated with a high risk of treatment failure arising from

a significantly reduced bioavailability in the fed state. Such interactions are frequently caused by chelation with

components in food. In addition, the physiological response to food intake, in particular, gastric acid secretion,

may reduce or increase the bioavailability of certain drugs.

Drug interactions can alter the pharmacokinetics and/or pharmacodynamics of a drug. The

pharmacodynamic interaction may be additive, synergistic, or antagonistic effects of a drug. Drug interactions

(DIs) represent an important and widely under recognized source of medication errors. The gastrointestinal

absorption of drugs may be affected by the concurrent use of other agents that have a large surface area upon

which the drug can be absorbed bind or chelate, alter gastric pH, alter gastrointestinal motility, or affect transport

proteins such as P-glycoprotein. A reduction only in absorption rate of a drug is seldom clinically important,

whereas a reduction in the extent of absorption will be clinically important if it results in sub therapeutic serum

levels.

Factors such as nonspecific binding, atypical kinetics, poor effector solubility, and varying ratios of

accessory proteins may alter the kinetic behavior of an enzyme and subsequently confound the extrapolation of in

vitro data to the human situation. Coenzyme Q-10 (CoQ10) is very widely consumed by humans as a food

supplement because of its recognition by the public as an important nutrient in supporting human health. It

interferes with intestinal efflux transporter P-glycoprotein (P-gp) and as result food-drug interactions arise. The

interaction of natural products and drugs is a common hidden problem encountered in clinical practice. The





interactions between natural products and drugs are based on the same pharmacokinetic and pharmacodynamic

principles as drug-drug interactions. Several fruits and berries have recently been shown to contain agents that

affect drug-metabolizing enzymes. Grapefruit is the most well-known example, but also sevillian orange, pomelo

and star fruit contain agents that inhibit cytochrome P450 3A4 (CYP3A4), which is the most important enzyme in

drug metabolism.

The study of drug-drug, food-drug, and herb-drug interactions and of genetic factors affecting

pharmacokinetics and pharmacodynamics is expected to improve drug safety and will enable individualized drug

therapy. Drugs can show their efficacy only if administered in appropriate quantity with appropriate combination

of drugs and foods and at appropriate time.

In contrast to the easy access to information on drug-drug interactions, the information about food-

drug interaction is not always available conveniently. It is a difficult and complex problem to accurately

determine the effects of food and nutrients on a particular drug. This article aims to help the healthcare

professionals specially physicians and pharmacists and patients to become more knowledgeable about drug and

food interactions.

Electronic search of literatures was conducted over a period of two months and all original research

and review articles were included in this study. No literature was older than 20 years. The drugs were selected and

reviewed on the basis of their general utilization pattern and realizing the need for reporting their interaction with

different dietary supplements for better therapeutic use of these drugs within the recommended dose regimen.

Fruit Juices: Among all fruit juices, grape fruit juice (GFJ) possesses high interaction with almost all types of

drugs. The juice modifies the body`s way of metabolizing the medication, affecting the liver`s ability to work the

drug through a person`s system.

Taniguchi in 2007 reported a case of purpura associated with concomitant ingestion of cilostazol,

aspirin and grapefruit juice in 79 years old man. His purpura disappeared upon cessation of grapefruit juice,

although his medication was not altered. The most probable cause of his purpura is an increase in the blood level

of cilostazol because of the inhibition of cilostazol metabolism by components of grapefruit juice; Numerous

reports have documented drug interactions with GFJ that occur via inhibition of CYP3A enzymes. Furano

coumarins present in GFJ inhibit the intestinal CYP 3A4 and have been shown to increase the oral bioavailability

of medications that are CYP 3A4 substrates like Felodipine, midazolam, cyclosporine and raise their

concentrations above toxic levels.

GFJ is generally contraindicated to patients taking psychotropics and it is advised to inform patients

about described interaction. The in vitro data suggest that compounds present in grapefruit juice are able to inhibit

the P-gp activity modifying the disposition of drugs that are P-gp substrates such as talinolol. The overall

exposure of some drugs can be increased by more than fivefold when taken with GFJ and increase the risk of

adverse effects. With new anticonvulsants, serum iron and sodium need to be monitored. Additionally, users are

advised to avoid drinking grape fruit juice within 1-2 hr(s) of taking these anticonvulsants.

Furanocoumarines and active bioflavonoids present in GFJ are also inhibitors of OATP and when

ingested concomitantly, can reduce the oral bioavailability of the OATP substrate, fexofenadine. Overall, a series

of flavonoids present in GFJ are identified as esterase inhibitors, of which kaempferol and naringenin are shown

to mediate pharmacokinetic drug interaction with most of the calcium channel antagonist and the statin groups of

drugs such as enalapril and lovastatin due to their capability of esterase inhibition.

Cholesterol-lowering agent lovastatin should be taken with food to enhance gastrointestinal absorption

and bioavailability. The absorption of rosuvastatin, another anti-hyper lipidemic agent, was significantly

decreased in the fed state compared with the fasting state, which suggests that rosuvastatin should be administered

on an empty stomach. Simvastatin, ezetimibe, pravastatin and fluvastatin may be taken without regards to food.

However, high fiber diets may lower the efficacy of these drugs. Concomitant administration of statins with food

may alter statin pharmacokinetics or pharmacodynamics, increasing the risk of adverse reactions such as

myopathy or rhabdomyolysis or reducing their pharmacological action. Consumption of pectin or oat bran

together with Lovastatin reduces absorption of the drug, while alcohol intake does not appear to affect the efficacy

and safety of Fluvastatin treatment.

Warfarin: Warfarin is commonly used to treat or prevent thromboembolic events. Patients taking warfarin are at

particular risk of interactions with dietary supplements, yet approximately 30% use herbal or natural product

supplements on a regular basis. There is a possible interaction between warfarin and a highprotein diet. The





potential for increased dietary protein intake to raise serum albumin levels and/or cytochrome P450 activity has

been postulated as mechanisms for the resulting decrease in international normalized ratio (INRs). Some

vegetables (broccoli, Brussels sprouts, kale, parsley, spinach, and others) are high in vitamin K. Eating large

quantities or making sudden changes in the amounts eaten of these vegetables, interferes with the effectiveness

and safety of warfarin therapy.

Eating charbroiled food may decrease warfarin activity, while eating cooked onions may increase

warfarin activity. Soy foods have been reported both to increase and to decrease warfarin activity. The

significance of these last three interactions remains unclear. The combination of warfarin administration and

cranberry juice ingestion appeared to be associated with an elevated INR without bleeding in elderly patient.

A number of studies have been documented on the interaction of warfarin and cranberry juice.26-30

Cranberry juice is a flavonoid, which has been shown to induce, inhibit, or act as a substrate for the biosynthesis

of several cytochrome P-450 (CYP) isoenzymes. Specifically, cranberry juice may inhibit the activity of

CYP2C9, the primary isoenzyme involved in the metabolism of S-warfarin. It was suggested that cranberry juice

increased the International Normalized Ratio (INR) of patients taking warfarin, but neither clearly identified

cranberry juice as the sole cause of INR elevation. If warfarin sodium is ingested with leafy green vegetables, the

hypoprothrombinemic effect of warfarin may be decreased and thromboembolic complications may develop.

Monoamnine oxidases: Antidepressant activity of monoamine oxidase inhibitors (MAOIs) was initially noted in

the 1950s. Although older monoamine oxidase inhibitors (MAOIs) are effective in the treatment of depressive

disorders, they are under-utilized in clinical practice due to main concerns about interaction with tyramine-

containing food (matured cheese, red vine, ripped bananas, yogurt, shrimp paste and salami) or so called cheese

reaction, since they are capable of producing hypertensive crisis in patients taking MAOIs. The first-generation

MAOIs such as phenelzine and isocarboxazid were largely nonselective inhibitors of both subtypes of MAO,

MAO (A) and MAO (B). These medications carried with them dietary restrictions. Tyramine is an indirectly

acting sympathomimetic agent, is degraded by MAO but in the presence of MAOIs, it escapes degradation and

reaches the systemic circulation where it is taken up by the adrenergic neuron, leading to a hypertensive crisis.

However, MAOIs have been well established as an effective intervention for people with treatment resistant

depression, and transdermal formulations may provide a valuable therapeutic option and eliminate the drug-food

interaction.

Antihypertensive Drugs: Patients placed on anti hypertensive drugs will benefit from concomitant moderate

sodium restricted diets. Propranolol serum levels may be increased if taken with rich protein food. A change in

diet from high carbohydrates/low protein to low carbohydrate/ high protein may result in increased oral clearance.

Smoking may decrease its plasma levels of by increasing its metabolism.

The intestinal absorption of celiprolol (beta-blocker) is inhibited when it is taken with orange juice.

Hesperidin, present in orange juice, is responsible for the decreased absorption of celiprolol. The absorption of

ACEs inhibitors is increased when taken on an empty stomach. While GFJ increases the bioavailability of

felodipine (Ca2 channel blocker).

Licorice extract, a common ingredient of dietary supplement contains glycyrrhizin and glycyrrhetinic

acid. It is a potent inhibitor of 11- bet- hydroxyl steroid dehydrogenase, it increases excess of cortisol to

mineralocorticoid receptors causing sodium retention and potassium depletion, so it may interfere with various

medicines including antihypertensive and antiarrhythmic agents. A high intake of liquorice can cause

hypermineralocorticoidism with sodium retention and potassium loss, oedema, increased blood pressure and

depression of the renin-angiotensin-aldosterone system. Studies showed that a daily consumption of glycyrrhizic

acid of 95 mg or more caused an increase in blood pressure. A practical guideline for an acceptable daily intake of

glycyrrhizic acid seems to be 9.5 mg a day. This means no more than 10-30g liquorice and no more than half a

cup of liquorice tea a day.

Antibiotics: Antibiotics are widely prescribed in medical practice. Many of them induce or are subject to

interactions that may diminish their anti-infectious efficiency or elicit toxic effects. Food intake can influence the

effectiveness of an antibiotic. Avoid coadministration of antibiotics with milk products which are rich sources of

divalent ions, such as calcium and magnesium that complex with some antibiotics and prevent their absorption.

The intake of dairy products, however, needs to be monitored and encouraged with appropriate consideration of

specific antibiotics involved.





A number of studies give evidence that fluoroquinolones forming slightly soluble complex with metal

ions of food show reduced bioavailability. Casein and calcium present in milk decrease the absorption of

ciprofloxacin. The effect of interaction of five fruit juices on the dissolution and absorption profiles of

ciprofloxacin tablets were determined. It was found that the absorption of ciprofloxacin (500 mg) tablets can be

reduced by concomitant ingestion of the GFJ. Therefore, to avoid drug therapeutic failures and subsequent

bacterial resistance as a result of sub-therapeutic level of the drug in the systemic circulation, ingestion of the

juice with ciprofloxacin should be discouraged.

Azithromycin absorption is decreased when taken with food, resulting in a 43% reduction in

bioavailability.

Tetracycline should be taken one hour before or two hours after meals, and not taken with milk

because it binds calcium and iron, forming insoluble chelates, and influencing its bioavailability. The effect of

milk added to coffee or black tea on the bioavailability of tetracycline was evaluated in healthy individuals.

Results showed that even a little quantity of milk containing extremely small amounts of calcium severely impair

the absorption of the drug, so that the presence of this metal ion should be carefully controlled in order to avoid

decreasing the available tetracycline.

Food-drug interactions may reduce the bioavailability of drugs taken after meals (negative food

effects). However, enteric-coated tablets that start to disintegrate when they reach the middle-to lower region of

the small intestine could reduce negative food effects. Results indicated that food-drug interactions were avoided

by separating the main absorption site of drugs from that of food components.

Analgesics and Antipyretics: Analgesics and antipyretics are used to treat mild to moderate pain and fever. For

rapid relief, acetaminophen should be taken in an empty stomach because food may slow the body absorption of

acetaminophen. Co-administration of acetaminophen with pectin delays its absorption and onset. NSAIDs like

ibuprofen, naproxen, ketoprofen and others can cause stomach irritation and thus they should be taken with food

or milk. Avoid or limit the use of alcohol because chronic alcohol use can increase the risk of liver damage or

stomach bleeding. The absorption of ibuprofen and oxycodone when given in the combination tablet was affected

by the concomitant ingestion of food. The C max and AUC0-alpha of ibuprofen were significantly increased after

single and multiple doses of Coca-Cola, thereby indicating increased extent of absorption of ibuprofen. The daily

dosage and frequency of ibuprofen must be reduced when administered with Coca-Cola.

Food intake did not appear to affect the extent of absorption (i.e, total exposure) of oral Diclofenac

potassium soft gelatin capsule at doses.

Bronchiodilators: Bronchodilators like theophylline, albuterol, and epinephrine possess different effects with

food. The effect of food on theophylline medications can vary widely. High-fat meals may increase the amount of

theophylline in the body, while high carbohydrate meals may decrease it. Avoid alcohol if taking theophylline

medications because it can increase the risk of side effects such as nausea, vomiting, headache and irritability.

Avoid eating or drinking large amounts of foods and beverages that contain caffeine (e.g., chocolate,

colas, coffee, and tea) since theophylline is a xanthine derivative and these substances also contain xanthine.

Hence consuming large amounts of these substances while taking theophylline, increases the risk of drug toxicity.

Additionally, both oral bronchodilators and caffeine stimulate the central nervous system. Patients may be advised

not to consume GFJ when taking theophylline, since it increases the bioavailability and monitoring of plasma

theophylline levels in patients consuming GFJ might be helpful in better management of patient care.

Antihistamines: Fexofenadine, loratadine, rupatadine, cimetidine cetirizine, are all antihistamines. It is best to

take prescription antihistamines on an empty stomach to increase their effectiveness. Rupatadine is commonly

used for the management of diseases with allergic inflammatory conditions.

A study indicates that concomitant intake of food with a single 20 mg oral dose of rupatadine exhibits

a significant increase in rupatadine bioavailability. Cimetidine is given with food to assist the maintenance of a

therapeutic blood concentration. A fraction of cimetidine is absorbed in the presence of food, allowing the

remaining drug to be dissolved once the gut is cleared. Thus, therapeutic levels are maintained throughout the

dosing interval. A study was conducted on a latest molecule esomeprazole (acid-reducer) and it was observe that

its bioavailability was reduced when taken within 15 min before eating a high-fat meal vs. that while fasting.

Antitubercular Drugs: Anti-tubercular drugs like isoniazid have been associated with tyramine and histamine

interactions. Inhibition of monoamine oxidase and histaminase by isoniazid can cause significant drug food

interactions. Food greatly decreases isoniazid bioavailability. Oleanolic acid, a triterpenoid exists widely in food,





medicinal herbs and other plants, has antimycobacterial activity against the Mycobacterium tuberculosis, when

administered with isoniazid, it exerts synergistic effect. High fat meals decrease the serum concentration of

cycloserine, a bacteriostatic anti-tubercular drug and results in incomplete eradication of bacteria.

Antidiabetics: Glimepiride is an antidiabetic and a new generation sulfonylurea derivative should be administered

with breakfast or the first main meal of the day. It has absolute bioavailability and the absence of food interaction

guarantee highly reproducible pharmacokinetics. Immediate release glipizide should be taken 30 minutes before

meals. However, extended release tablets should be taken with breakfast. The maximum effectiveness of

acarbose, an alpha-glucosidase inhibitor is attained when the drug is taken immediately at the start of each meal

(not half an hour before or after), because it delays the carbohydrate absorption by inhibiting the enzyme alpha-

glucosidase.

Thyroxine: Recent evidence pointed out the role of gastric acid secretion on the subsequent intestinal absorption

of thyroxine in relation with the timing of food ingestion as well as with pH impairment associated to frequent

gastric disorders like Helicobacter pylori infection and gastric atrophy. Levothyroxine is a derivative of thyroxine.

Grapefruit juice may slightly delay the absorption of levothyroxine, but it seems to have only a minor effect on its

bioavailability. Accordingly, the clinical relevance of the grapefruit juice-levothyroxine interaction is likely to be

small.

Drug interactions may be theoretical or clinically relevant. A summary table is given to highlight some

significant food-drug interactions. Some may be taken advantage of, to the benefit of patients, but more

commonly drug interactions result in unnecessary adverse events. Fortunately, undesirable drug interactions can

be prevented. Becoming more familiar with potential drug interactions can help clinicians predict and explain a

patient`s response to medications. Significant food effects complicate development of new drugs, especially when

clinical plans require control and/or monitoring of food intake in relation to dosing. The prediction of whether a

drug or drug product will show human food effect is challenging.

Antitumor Drugs: Mercaptopurine is a purine analog used for acute lymphoblastic leukemia and chronic

myelogenous leukemias. Since it is inactivated by xanthine oxidase (XO), concurrent intake of substances

containing XO may potentially reduce bioavailability of mercaptopurine. Cow’s milk is known to contain a high

level of XO. This interaction may be clinically significant. Therefore most patients should try to separate the

timing of taking mercaptopurine and drinking milk.

Tamoxifen is a successful anti-tumor agent. If taken with sesame seeds, it negatively interferes with

tamoxifen in inducing regression of established MCF-7 tumor size but beneficially interacts with tamoxifen on

bone in ovariectomized athymic mice.

Pharmacokinetic Interactions

Drug Absorption Interactions: Food may affect drug absorption in the GI tract by altering gastric pH, secretion,

gastrointestinal motility and transit time. This may result in a change in the rate of absorption or extent of drug

absorption or both. For example, azithromycin absorption is decreased when it is taken with food, resulting in a

43% reduction in bioavailability. Sustained-release theophylline products when taken with high-fat foods may

cause a sudden release (dose dumping) of theophylline, resulting in increased theophylline concentrations and

possible toxicity. Children are more prone to this interaction than adults.

In other cases, the components of the food, such as calcium or iron, may form complexes with the drug

that are less easily absorbed. Examples include tetracycline, sodium fluoride and ciprofloxacin .The absorption of

alendronate is impaired by food, calcium and almost everything, including orange juice and coffee. It should be

taken with plain water and nothing else should be consumed for at least 30 minutes. In many cases, the actual

mechanism by which food interferes with absorption is not known. Delayed absorption does not necessarily

reduce the total overall exposure to the drug; the area under the curve (AUC) may be equivalent regardless of how

the drug is taken.

A reduced rate of absorption may sometimes be useful in reducing the side effects of a drug, as in the

cases of ibuprofen, without reducing bioavailability. The bioavailability of some drugs may be enhanced by food.

For example, an acid environment is necessary for the absorption of ketoconazole. The absorption of griseofulvin

is increased by fat in a meal. Fenofibrate, mebendazole, isotretinoin, tamsulosin, carbamazpine and labetalol are

examples of drugs that will be better absorbed when taken with food.

Improved absorption of a drug may or may not have a significant effect on the drug’s efficacy. Patients

taking digoxin should avoid taking bran fiber, pectin-containing foods such as apples or pears, or fibercontaining,





bulk-forming laxatives at the same time, since these agents may bind to the digoxin, decreasing its absorption.

This interaction could result in decreased serum concentrations of digoxin and therapeutic effectiveness. It is

advisable to take some medications with food to reduce gastrointestinal irritation and possible nausea.

Examples of these medications include potassium supplements, ferrous sulfate, nonsteroidal anti-

inflammatory drugs, estrogen, prednisone, tacrine, terfenadine and nitrofurantoin.

Cholesterol-lowering agent lovastatin should be taken with food to enhance gastrointestinal absorption

and bioavailability. Simvastatin, pravastatin and fluvastatin may be taken without regard to food.

Drug Metabolism Interactions: Food may alter the hepatic metabolism of some drugs. It has been reported that

when administered with the antihypertensive drug felodipine, concentrated grapefruit juice caused an increase in

the bioavailability of felodipine. The mean felodipine bioavailability with grapefruit juice was 284% (range

164%–469%) that of water. This resulted in lower diastolic blood pressures and increased heart rate in the male

study volunteers. Adverse effects such as headaches, facial flushing and lightheadedness were more common after

ingestion of 250 ml grapefruit juice (125 ml frozen grapefruit concentrate plus 125 ml of water).

The bioavailability of nifedipine with grapefruit juice was 134% (range 108%–169%) of that with

water. Orange juice did not have these effects. It is postulated that flavonoid compounds in grapefruit juice

concentrate inhibit cytochrome P-450 metabolism of felodipine and nifedipine. This interaction could increase

both the efficacy and toxicity of these calcium channel blockers. There is potential clinical significance because

citrus juices are frequently consumed at breakfast, when many medications are also taken. Patients should be

advised of this possible interaction. First-pass hepatic metabolism of propranolol and metoprolol may be

decreased when either medication is taken with food, thereby enhancing bioavailability. Drug levels and

therapeutic efficacy may be increased due to this interaction.

Monoamine oxidase (MAO) inhibitors are known to interact with foods containing tyramine.

Tyramine is normally inactivated by the enzyme monoamine oxidase and this prevents tyramine from

accumulating in the body. Monoamine oxidase inhibitors cause increased levels of tyramine which can lead to a

hypertensive crisis. Patients taking monoamine oxidase inhibitors should avoid foods high in tyramine such as

aged cheeses, pickled fish, yeast extracts, red wine, some types of beer (including nonalcoholic beer), fava beans

and fermented products. High protein foods that have been aged, fermented, pickled, smoked or bacterially

contaminated are unsafe for patients taking MAO inhibitors. Foods considered safe when used fresh and in

moderation include sour cream, yogurt, meat extracts, chopped liver, dry sausage and alcoholic beverages.

Drug Excretion Interactions: Foods may alter the urinary pH, which can affect the activity of certain drugs. The

half-lives of some medications can be significantly changed by alterations in urinary pH. Therefore, the half-life

of acidic drugs will be extended in acidic urine because the drug is in its unionized form. However, the half-life of

an acidic drug in alkaline urine is reduced because the drug is in its ionized form. Foods such as milk, vegetables

and citrus fruits can alkalinize the urine. Meats, fish, cheese and eggs can acidify the urine. Foods may alter the

renal excretion of some medications.

Lithium and sodium compete for tubular reabsorption in the kidney. A high-salt diet causes more

lithium to be excreted, whereas a low-salt diet causes decreased renal excretion of lithium and an increase in

serum lithium levels.

Pharmacodynamic Interactions: Foods may interact with medications by altering their pharmacologic actions.

Diets high in vitamin K may cause antagonism of warfarin and decreased therapeutic efficacy of the

anticoagulant. Foods rich in vitamin K include green leafy vegetables (kale, turnip greens, spinach, broccoli and

brussels sprouts), cauliflower, chick peas, green tea, pork liver and beef liver.

Alcoholic beverages may increase the central nervous system depressant effects of medications such as

benzodiazepines, antihistamines, antidepressants, antipsychotic, muscle relaxants, narcotics or any drug with

sedative actions.

An example of a food potentiating the effect of a medication is coffee, as caffeine has additive effects

on theophylline. It has been reported that caffeine increased serum theophylline levels by 20%–30% and increased

the half-life of theophylline by decreasing clearance. Patients may complain of nervousness, tremor or insomnia.

Caffeine has some bronchodilatory effects, which may enhance the effects of theophylline. A lower dosage of

theophylline may be necessary for those patients who consume excessive quantities of coffee (more than 6 cups

daily).

ROLE OF PHARMACIST IN PREVENTION OF DRUG-FOOD INTERACTIONS





Pharmacists in every practice setting need to be vigilant in monitoring for potential drug-food

interactions and advising patients regarding foods or beverages to avoid when taking certain medications. It is

imperative for pharmacists to keep upto- date on potential drug-food interactions of medications, especially

today’s new drugs, so that they may counsel properly. In providing drug information to patients, pharmacists

often discuss potential side effects and how the medication should be taken. It is important to provide information

to patients on when to take their medications in relation to food intake.

Consequences of drug-food interactions may include delayed, decreased or enhanced absorption of the

drug. Food may also affect the bioavailability, metabolism and excretion of certain medications. The patient may

experience an adverse side effect or toxicity or may not receive the full therapeutic benefit of the medication. The

Joint Commission on the Accreditation of Healthcare Organizations (JCAHO) requires that a patient’s medication

profile include potential drug-food interactions, that the pharmacist call the prescriber whenever the potential for a

medication-food interaction exists and document the communication and follow-up action on the prescription or

order form, and that patients be given instructions and counseling regarding the potential for drug-food

interactions before their hospital discharge.

Elderly patients may be at a greater risk for drug-food interactions because they typically consume

more medications for their chronic medical conditions. A study of drug-nutrient interactions in long-term care

facilities found a significant relationship between the number of medications a resident consumed and the number

of drug-nutrient interactions for which a resident was at risk.

Counseling and Guidance about Drug- Food Interactions: The following information can be given to the

patients while dispensing the medicine.

Read the prescription label on the container. If you do not understand something or think you need

more information, ask your physician or pharmacist.

Read directions, warnings and interaction precautions printed on all medication labels and package

inserts. Even over-the-counter medications can cause problems.

Take medication with a full glass of water.

Do not stir medication into your food or take capsules apart (unless directed by your physician). This

may affect the efficacy of medication.

Do not take vitamin pills at the same time you take medication. Vitamins and minerals can interact

with some drugs.

Do not mix medication into hot drinks because the heat from the drink may destroy the effectiveness

of the drug.

Never take medication with alcoholic drinks.

Be sure to tell your physician and pharmacist about all medications you are taking, both prescription

and nonprescription.

Check with the pharmacist on how food can affect specific medications taken with the food.

Precautions to be taken

Medications need to be taken at different times relative to meals.

Consult a physician when health problems persist.

During pregnancy and nursing always consult a physician or pharmacist before taking any medication.

Drugs taken by the mother may affect the infant.

Check with a doctor or pharmacist for the proper way and time to take medication.

CONCLUSION

Interaction between foods and drugs can have profound influence on the success of drug treatment and

on the side effect profiles of many drugs. The clinical significance of drug-food interactions can be variable.

Some foods greatly affect drug therapy, resulting in serious side effects, toxicity or therapeutic failure. In some

instances, the interaction may have a beneficial effect by increasing drug efficacy or diminishing potential side

effects.

The interactions are not always detrimental to therapy, but can in some cases be used to improve drug

absorption or to minimize adverse effects. These interactions have received more attention recently, especially

drug interactions with grapefruit juice. As new drug approvals occur with ever increasing speed, there is less

information available about their adverse effects and interactions when the drugs reach the market. Pharmacists in

every practice setting need to be vigilant in monitoring for potential drug-food interactions and advising patients





regarding foods or beverages to avoid when taking certain medications. It is imperative for pharmacists to keep

up-to date on potential drug-food interactions of medications, especially today’s new drugs, so that they may

counsel properly to the patients.

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Sahithi B et.al Indian Journal of Research in Pharmacy and Biotechnology


A REVIEW ON COLLAGEN BASED DRUG DELIVERY SYSTEMS Sahithi B

*, Ansari Sk, Hameeda Sk, Sahithya G, Durga Prasad M, Yogitha Lakshmi

Nimra college of pharmacy, Vijayawada

Corresponding author: Email:[email protected]

ABSTRACT

Due to its biodegradability, biocompatibility, weak antigenicity and well-known safety profile,

collagen becomes a very useful carrier for the delivery of various kinds of drugs and agents like growth

factors, collagen possess some very unique properties as compare to other drug carriers that’s why a

numerous number of researches are in pipeline on this biomaterial. The main application of collagen is

collagen shields which are used in ophthalmology. However, fundamental awareness regarding collagen

biochemistry and the manufacturing knowledge in combination with understanding of the physico-

chemical properties is essential for fruitful application of collagen for drug delivery systems. The purpose

of this review article is to summarize information available on collagen dosage forms for drug delivery as

well as to communicate an outline regarding current preparation of collagen available in market includes

- collagen sponges for burns/wounds, mini-pellets and tablets, gel preparations in combination with

liposomes for sustained delivery of drug, formulations for transdermal drug delivery, and Nano spheres

for gene delivery, collagen matrices for cell culture.

Key words: Collagen, Drug delivery system, Biomaterial, Ophthalmology.

INTRODUCTION In the beginning of 1970s and 1980s the research on collagen was initiated by interested scientists and

commercial research laboratories expanding medical applications of biomaterials and connective tissue research.

In present years biotechnology has given a support to research the collagen material useful for drug delivery.

Basically Collagen is a protein which is widely used in medical field. Collagen plays a significant role in the

formation of organs and tissues, and is involved in different functional expressions of cells. Many natural

polymers and their synthetic analogues are used as a biomaterial, but the characteristics of collagen as a

biomaterial are different from those of synthetic polymers mainly in its mode of interaction inside the body. The

important features of collagen are its biocompatibility, biodegradability and weak antigenicity [Maeda et al.,

1999].In the body as compared with other natural polymers like gelatin and albumin. Collagen possesses good

abilityto penetrate a lipid-free interface. The primary reason for the usefulness of collagen in biomedical

application is that collagen can form fibers with extra strength and stability through its self-aggregation and cross-

linking.

Collagen: Basically collagen is a naturally existing protein present in the animal body, fibrous in nature, and

especially found in the connective tissue and flesh of mammals. Approximately 25%-35% of total body protein is

comprised of collagen, in the form of elongated fibrils; collagen is abundantly present in fibrous tissue like bone,

cartilage, tendons, blood vessels, ligament, skin, cornea, inter-vertebral disc and the gut. The synthesis of collagen

in the body is made by fibroblast cells. Collagens possess good tensile strength, and found both outside and inside

the body cells. In combination with elastic, collagen provides support to body tissues and organs, basically

collagen offers firmness and strength and elastic provides flexibility to body tissues. In fact gelatin which is used

in food and pharmaceutical industries is collagen that has been hydrolysed irreversibly.

Structure of collagen: Basically collagen possesses a triple helix structure, which generally made up of two

homologous chains (α-1) and one supplementary chain that varies slightly in its chemical composition (α-2).

These chains are polypeptide in nature and coiled around one another in a cable form. Each has a distinct turn in

the reverse direction, these chains are connected together chiefly by hydrogen bonds between nearby CO and NH

groups. The weight of collagen molecule is 300 kda and its structure is rope shaped and having a length of 300

NM and a width of 1.5 NM. The major content of glycine and amino acid residue is affecting the helix formation;

in each of three chains of collagen molecule the amino acids are regularly arranged. The sequence of amino acids

follows the pattern glycine-proline-X or glycine-X-hydroxyproline where X is the amino acid other than glycine,

proline or hydroxyproline; glycines constitute about 1/3 of total sequence and proline or hydroxyproline

accounting for the 1/6 of the sequence. This whole structure is joined with the help of hydrogen bonds and linking

peptide bonds.

Characteristics possessed by collagen:

Stretch-ability under stress condition collagen stretch rather than break





Strength

Biochemical compatibility

These three amino acid monomers are strongly fused and they look like single monomer

Several hydrogen bonds are present in collagen, on applying stress they can be wrecked and re-joined

after removal of pressure;

Collagen is biodegradable

Collagen show good absorption in-vivo

Collagen possesses weak antigenicity

Isolation and Purification of Collagen: Even though the mammalian body retains a plenty amount of collagen,

those tissues rich in fibrous possess collagen such as skin and tendons, are commonly used as preliminary

materials to produce collagen for use in transplants, wound dressings, or drug delivery systems. In addition

procaine, bovine and sheep collagen varieties derived from many different sources including marine sources,

human placenta, and recombinant human collagen from transgenic animals must be labelled. Autologous collagen

material deals additional gut alternative mucosa which is consumed in the building of surgical sutures. Collagen is

insoluble in organic solvents. Water-soluble collagen denotes only a minor fraction of total collagen and the

quantity depends on the age of the animal and kind of tissue extracted. In certain tissues, especially the skin of

young animals, cross linking is sufficiently little to extract a few percent in suitable conditions. Still, collagen

molecules present inside fibril masses can be separated and brought into aqueous solution. Though, the nature of

the crosslink dominant in different tissues decides the particular solvent to be used and the resulting yields.

Basically four types of collagen can be isolated and purified for the implementation in pharmaceutical industries

for the delivery of drugs are following such as

Natural salt soluble collagen

Alkali and enzyme treated collagen

acid soluble collagen

Insoluble collagen

Process to Isolate Neutral Salt Soluble Collagen:

Note- Most tissues have minute or no salt-extractable collagen. In demand to increase the yield for research

purposes animals can be put on the diet contain b-aminopropionitrile, an inhibitor of peptidyllysyl oxidase, yet

this method is in adequate for larger commercial scale.





Process to Isolate Acid Soluble Collagen:

Why Collagen can be used as a biomaterial for drug delivery:

Collagen is biodegradable and simply absorbed in the body

Collagen is a part of the body that’s why it is non-antigenic

Collagen is a non- toxic biopolymer

Collagen shows better biocompatibility

Collagen can be framed in a number of different forms

Shows synergism with other bioactive compounds

Haemostatic in nature and encourage blood coagulation

Compatible with synthetic polymers

By utilizing its functional group collagen can be easily modified to produce desired materials

Biodegradability of collagen can be controlled by cross-linking

Existing in plenty and simply purified from living organisms (constitutes more than 32% of vertebrate

tissues)

Biological plastic due to great ductile strength and negligible express ability.

The in vivo absorption of collagen is controlled by the use of cross-linking agents, such as:

Glutaraldehyde

Chromium tanning

Formaldehyde

Poly epoxycompounds

Acylazide

Carbodiimides

Hexamethylenediisocyanate

Physical treatment, such as ultra-violet/gamma-ray irradiation and dehydrothermal treatments have been

efficiently used for the introduction of cross links to the collagen matrix [10]. The use of collagen as a drug





delivery system is very comprehensive and diverse. It can be extracted into an aqueous solution and moulded

into various forms of delivery systems.

The applications of collagen as drug delivery systems are:

Used in the formation of microspheres for drug delivery

In formulation of nanoparticles for gene delivery

Collagen is used in the manufacturing of collagen Sponges for burns/wounds;

Development of tablets and pellets for the delivery of proteins;

Collagen is used for gel formulation and combined with liposomes for sustained delivery of drugs

In the treatment of cancer collagen is used as aqueous injection

Collagen is used in ophthalmology as collagen shields

Collagen is used as controlling material for the delivery of drugs in transdermal patches

As films for the delivery of human growth hormone, immune-stimulants, tetracycline, growth factors

For the delivery of glucocorticosteroids, microparticles of collagen are used.

Collagen delivery systems having a smooth release control can be attained by balancing the configuration

of the collagen matrix or attaching other proteins, such as Austin, fibronectin or by fusion of collagen with other

polymers, such as collagen/liposome and collagen/silicone.

Some specific type of drug delivery systems constructed on Collagen base:

Nanoparticles/Nano spheres/Microspheres: In the collagen fold configuration, the crystallites suspended in the

gel aggregates seem as a multiple chain system; this property is used to formulate aggregates as colloidal drug

delivery carriers. The construction of nanosphereis determined by a mixture of electronic and electrostatic forces

with sodium sulphate engaged as a liquefying reagent to facilitate greater charge-charge relations among plasmid

DNA and collagen. The stability of the produced collagen nanoparticles is depending on the molecular weight of

collagen, and temperature and pH is greatly affecting the molecular weight profile of the collagen solution , and

these are also further affect the non-covalent interactions liable for the molecular structure of collagen. The

nanoparticles and nanospheres based on biodegradable collagen; are enabled and enhanced uptake of exogenous

compounds such as anti-HIV in a number of cells, especially macrophages, that is an advantage of collagen based

nanoparticles as systemic delivery carriers; and they are also thermally stable and easily sterilized. Some other

drugs like steroids, cytotoxic drugs like Campthocin can be easily delivered in systemic circulation with the help

of collagen nanoparticles. Collagen based nanoparticles can be readily used in sustained and delayed release

formulation for steroids and antibiotics because of their: –

Large surface area

Smaller size

Great absorptive capability

Capacity to diffusing in water to form a colloidal solution

o For example: Dermal delivery of retinol enhanced in collagen nanoparticles. Retinol in collagen

nanoparticle was stable shows a quicker transportation of incorporated drug through the skin.

Fabrication methods for Collagen nanoparticles: There are basically four type of methods for the

manufacturing the protein based nanoparticles namely-

Emulsification

Desolvation

Coacervation

Spray drying

o And some additional methods are, Jet milling technique, fluidization and solvent precipitation

method, Interfacial polymerization etc.

Processes involved:

Emulsification: In this process, A collagen aqueous phase containing a hydrophilic surfactant and water, and an

organic phase containing a lipophilic surfactant, oil and water miscible solvent is mixed with rapid agitation by a

mechanical homogenizer at room temperature to form a homogeneous emulsion. Then the above emulsion will be

mixed in preheated oil (120) drop by drop resulting formation of collagen nanoparticles.





Aqueous phase

(Collagen solution)

Organic phase (oil)

Emulsification

O/w

emulsion

Heated oil

Collagen

Nanoparticles

Fig.1. Emulsification method for nanoparticles manufacturing

Desolvation : The process of Desolvation includes the addition of alcohol or natural salt as desolvation factor to

the collagen solution, which alters the tertiary structure of collagen, when the critical level of desolvation attained

the formation of collagen mass, starts lastly glutaraldehyde will be added as a cross-linking material, and then

nanoparticles is formed. This process was firstly employed by Marty and co-workers.

Fig2. Desolvation method for nanoparticles manufacturing

Coacervation: This method is similar to desolvation method, the difference is only in various parameters like-

temperature, molar ratio of organic solvent and protein, rate of solvent addition, concentration of cross-linker

used, pH, speed of homogenizer etc.

Spray drying: Basically spherical collagen nanoparticle is fabricated by this process. This process include the

spraying of dilute solution of collagen leads to the formation of hollow spheres using elevated temperature;





increased temperature can lead denaturation of collagen triple –helical structure. So collagen solution is sprayed

into liquid nitrogen to prevent denaturation. After that the fabricated nanospheres are successively frozen,

tempered, lyophilized, cross-linked, and sterilized.

Collagen based pellets/tablet: Collagen pellets are extensively used in Japan. These pellets are also known as

monolithic devices .They are tiny rods of approximately 1mm in diameter and 15 mm in length manufactured

from collagen by cutting, moulding, and drying. These devices are cylindrical in structure, and they can be

administered via injection using a syringe with a plugar. Pellets are very suitable for the local delivery of

lysozyme and minocycline in the treatment of clinical symptoms of periodontitis. These pellet are also proven to

be effective in vivo for the delivery of interleukin-2 (half-life =360 min for subcutaneously injected IL-2 mini-

pellets as compare to, 15 min and 8 min respectively for subcutaneous and intravenous injections of an aqueous

preparation). It is also proved that single subcutaneous injection of a mini-pellet cause a prolonged retention of

IL-2 and maximal concentration in the serum. The similar result was also observed for interferon. In 1989 Lucas

and his co-workers was designed a collagen pellet type controlled release delivery vehicle which is made up of

type-1 collagen for water soluble bone forming proteins.

Collagen films: Collagen films are basically sterilized films of collagen having thickness of 0.01-0.5 mm;

incorporated with drugs like steroids, antibiotics, hormones (obtained from rDNA technology E.g. human growth

hormone, rhBMP-2 etc.) intended for local action. They are simply manufactured by air- drying of casted collagen

mainly used as a barrier membrane, the drug incorporation in collagen films done by covalent bonding, hydrogen

bonding and by simple entrapment. Bradley used collagen films cross-linked by chromium tanning, formaldehyde

to sustain the release of medroxyprogesterone acetate. In this investigation cross-linking was done on films

already incorporated with drugs, endangering the Pharmacokinetic and pharmacodynamics activity of the drug.

This threat can be overcome with collagen materials which are cross-linked with glutaraldehyde prior to protein

incorporation. Collagen films also can be used as a drug carrier for antibiotics. E.g. when collagen films

containing tetracycline were implanted in rabbits suffering from periodontal disease; tetracycline was remained in

plasma for more than seven days and its activity was sustained for more than ten days. After four and seven weeks

the clinical symptoms of disease were decreased significantly. A European patent application was filed which

contain the research information regarding sustained release delivery of platelet derived growth factor; was

improved by multiple and single layer collagen films and it was found that this specific preparation had got a

release factor up to 100 h and thereby it leads to an enhanced wound healing in vivo. In the case of liver cancer or

tissue infection collagen film implants are very useful. When collagen film applied to eye it was totally

hydrolysed within 5-6 hr in the case of corneal tissue infection. Collagen sheets of micro-fibrous collagen was

used as local delivery carrier in the treatment of cancer, locally implanted collagen sheets which incorporated with

anticancer agents such as methotrexate and ectopocide needs low plasma concentration management. The

formation of bone tissues was enhanced when a collagen matrix and film were used as a carrier for gene delivery.

This shows that collagen based gene delivery system is very efficient as implant.

Collagen shields: Collagen shields is also known as collagen corneal shield, they are newly developed,

potentially versatile ophthalmic lens, which is made up of collagen, since collagen is a natural, commonly

available protein involved in the support and protection of vital structures, many researchers have tried to use

peripheral collagen to protect the surface of the eye in a variety of diseased states, like traumatic and non-

traumatic states after surgery; after corneal transplantation, radial keratomy. Generally collagen shields are

manufactured from bovine or procaine collagen, there are three kind of collagen shield available in market having

dissolving time of 12, 24, 72 hours. Bausch & Lomb Pharmaceuticals, a division of Bausch & Lomb, Inc.

acquired the rights to develop and market these collagen contact lenses, now known as Bio Cora collagen shields.

After much research, Bausch & Lomb has been able to produce the shields in a reproducible manner and in a

variety of shapes and thickness. Some other marketed preparations are (proshieldo, MediLenso, Fort Worth,

Chiron, TX, Irvine). These shields are able to enhance the penetration of corticosteroid, sub-conjunctival

antibiotics in eye. They are act as a short term bandage and allow sufficient oxygen transmission for essential

metabolism occurring in eye cornea.For the corneal surface lubrication these shields dissolve in collagen solution

that minimize lids rubbing. Mainly water soluble antibiotics and steroids are used in combination with collagen

shields for example- Vancomycin, Trimethoprim, Amphotericin-B,Gentamycin Polymyxin-B sulfate,

Tobramycin, steroids, pilocarpine, Application of collagen shields on cornea is demonstrated in figure-3.





Fig 3: Application of collagen shield on cornea

Marketed preparation of collagen shields:

Biocora®

ProshieldO®

MediLenso®

Irvine®

Chiron®

Collagen sponge: Collagen sponge are manufactured from pure bovine collagen obtained from bovine skin,

bovine collagen is firstly put into a solution having pH 3.0 and then stabilize into physical form of a sponge layer.

And then this sponge layer is combined with fibronectin, elastin or glycosaminoglycans to achieve a fluid

building capacity and elasticity. They are also manufactured by freeze-drying of alkali or acid, swollen collagen

containing 0.1- 5% dry matter content. Collagen sponges can be cross-linked with glutaraldehyde and

copolymerised with other synthetic as well as natural polymers, for example collagen sponges copolymerized

with PHEMA (polyhydroxyethylmethacrylate) are more hydrophilic in nature, retain wetness for longer period

and also possesses more tensile strength.

Collagen sponges were basically developed as hemostyptics and wound dressings but in present time they

are also used for antibiotics, steroids and growth factor delivery for wound healing and for bone forming implants.

The application of collagen sponges for delivery of topical agents (table 1)

Table.1.Application of collagen sponges for various topical agents

Collagen sponges are found very useful in dressing for leg ulcers, decubitus ulcer, donor sites, pressure

sores. The major benefits of collagen sponges includes their ability to absorb enormous quantity of tissue exudates

and smooth adherence to the wet wounds with preservation of micro climate as well as shielding against

secondary bacterial infection and mechanical harm, in addition collagen sponges promote inflammatory cells

activity to porous scaffolds and cellular growth. Thus collagen sponges can be considered asactive dressings,

which aid in the healing process.

Marketed preparations of collagen sponges: These are the some of the marketed formulation of the Collage

sponges available in the market such as Collarx®, Collatamp® G, Collatamp® Eg, Sulmycin® Implant,

Garamycin® Schwamm, Duracol®, Duracoll®, Gentacol®, Gentacoll®, Garacol®, Garacoll®, And Cronocol® -

Gentamicin Surgical Implants

Collagen hydrogels/gels: Collagen hydrogels/gels are processed by cross-linking of collagen with chemicals like

poly epoxy compounds, carbodiimides, polyphenolic compounds, aldehydes, and acyl azide compounds which

leads to the formation of bonds between molecules and fibrils. Collagen hydrogels possess a unique property of

soaking and swelling on hydration with biological fluids and they are also capable to maintain their integrity after

soaking.

Drugs/ agents Uses

Gentamycin

In septic focus in

abdomen

Growth factor

(rhBMP-2)

In bone formation

and wound healing





These hydrogels are very patients compliant because ease of application, high bio-adhesion and compatible with

large varieties of drugs and agents.Collagen gels are excessively used as injectables the most common form are-

1. Non-fibrillar viscous solution in aqueous medium

2. Fibersinjectables suspensions

For ophthalmic purpose these suspensions can be mixed with drugs and administered, these preparations are

patented, when inject; initially remains in liquid state and then after some time convert into gel.Shows a great

potential for sustained and controlled delivery of medicaments.

CONCLUSION AND FUTURE PERSPECTIVES

Collagen has various advantages as a biomaterial and is widely used as carrier systems for delivery of

drug, protein and gene. The examples described in this paper signify selected applications of collagen in the

biomedical field. The effective demonstration of usefulness of human skin substitutes made of collagen has leads

to the development of bioengineering tissues, such as blood vessels and ligaments. Although many applications of

collagen as a drug vehicle discussed in the paper, it should be noted the information regarding collagen is very

less as compare to synthetic polymers in literature because, the pure type-1 collagen is very costly, variability in

different forms, complex handling processes, and risk of Bovine spongiform encephalopathy (BSE). Beside them

collagen possess some very extraordinary properties which make it a very useful biomaterial for drug delivery

includes its biocompatibility, absorbability on biological membranes, no antigenicity, low toxicity, synergism

with other bioactive compounds etc. these advantage will carry the future development of this biomaterial.

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