“FORMULATION AND DEVELOPMENT OF DENTAL GEL CONTAINING CLOVE OIL FOR THE TREATMENT OF

PERIODONTAL DISEASES”

A Dissertation submitted to the

JAWAHARLAL NEHRU TECHNOLOGICAL UNIVERSITY ANANTHAPUR

In partial fulfillment of the requirements for the degree of

BACHELOR OF PHARMACY

Submitted by

SK. SANA BANU (12P21R0050)

K.CHANDANA (12P21R0014)

Y.CHARAN KUMAR (12P21R0015)

P.SATHEESH (12P21R0052)

P.SUBRAMANYAM (12P21R0062)

G.MANIKANTA (12P21R0035)

Under the Guidance of

V.VIJAY KUMAR M.Pharm., (Ph.D)

Associate Professor,

DEPARTMENT OF PHARMACEUTICS.

RAO’S COLLEGE OF PHARMACY

Chemudugunta (P.O.), Venkatachalam (M), Nellore, Andhra Pradesh.

2012 – 2016.

ABSTRACT :Aim The study was aimed to develop and evaluate dental gel containing clove oil as the chief constituent for the treatment of periodontitis. Methods It has a wide spectrum of antibacterial activity against a number of periodontal pathogens, hence it is selected for the treatment of periodontitis. Clove oil gel is formulated by using carbopol 934 as gelling agent, clove oil as medicinal agent, polyethylene glycol co-solvent, methyl paraben and propyl paraben as preservative and required quantity of distilled water as vehicle.

Results The clove oil was evaluated for physical parameters like acid value, ester value, specific gravity and refractive index and it shown satisfactory results. The prepared gel was evaluated for various properties such as antimicrobial activity, pH, spreadability, extrudability, drug content etc. In-Vitro experiments demonstrated that the formulation F3 is a suitable dosage form for the treatment of periodontitis. Clove oil showed the zone of inhibition of about 22.05±0.04 mm.

Conclusion On the basis of the result obtained in this present study we conclude that the gel formulations of clove oil F3 showed good physicochemical properties as well as good drug content compared to other formulations.

Key words Clove oil, Carbopol 934, Periodontitis, Anti microbial activity.

Ref: Vijaya Kumar Voleti*, Sana Banu Shaik, Chandana Konduru, Sathish Peyam, Charan Kumar Yaramsetti, Subramanyam Pasala, Shanmuga Pandiyan Pitchaimuthu. Formulation and development of dental gel containing clove oil for the treatment of human periodontal diseases. JCP. 2016;3(1): 1-7

2nd Indo-Korean Conference, Sullurpet, 30 th MARCH 2016 Theme: Herbals And Pharmaceuticals: Pivotal Issues & Concerns Venue: Gokula Krishna College of Pharmacy (Affiliated to JNTUA, Ananthapuramu, Approved by AICTE & PCI-New Delhi) SPSR Nellore, A.P., India – 524121

Acceptance of Poster/Oral Presentation21/03/2016Dear DelegateWe are pleased to inform you that your paper entitled “Formulation And Devfelopment Of Dental Gel Containing Clove Oil For The Treatment Of Human Periodontal Diseases ” has been accepted for presentation as “ Poster No:PPCE239” in Poster Session at the 2ndIndo – Korean conference-HPPIC2K16.As a presenter, you must be a registered delegate for the HPPIC2K16. However only the abstract of those presenters, who are registered by 25 March 2016, will be published in the final abstract book. Selected papers of the seminar might be published in “JOURNAL OF COMPREHENSIVE PHARMACY”. Those authors, who wish to publish their manuscript in the journal, should submit their full length papers on or before 25.03.2016.The full length manuscript should be in accordance with the instructions of Journal Of Comprehensive pharmacy”.You should have your registration badge at the poster venue in order to present the poster/oral.For any Queries Contact:

Kindly bring a copy of this letter and photo ID (College ID) for identification at the poster /oral presentation and also have to submit this letter to the registration committee during conference.The area provided for poster presentation will be about 1 meter (100cm), width by 1 meter height.We look forward to meet you at 2ndIndo – Korean conference, Sullurpet.Kind regardsChairman, Registration Committee-IKCSullurpet, SPSR Nellore. Dr .Pavan kumar Balagani Principal, Convenor, HPPIC2K16, Gokula Krishna college of pharmacy, sullurpet, SPSR Nellore, AP Email:nchp2k16@gmail.com

Mrs.M.Soujanya For registration: 9494795974

Mr. Deepak kumar For general information: 9652275737

Mr.SK Afsar For scientific session: 98855398761

Introduction Herbal gels:• Medicinal plants have been used as traditional treatments for numerous

human diseases for thousands of years and in many parts of the world. In rural areas of the developing countries, they continue to be used as the primary source of medicine. About 80% of the people in developing countries use traditional medicines for their health care [1]

Benefits of herbal drugs:• Herbal drugs have long era of use and better patient tolerance as well as

public acceptance.• Herbal drugs acts as a renewable source, which is our only hope for

sustainable supplies of cheaper medicines for the worlds growing population.

• The cultivation and processing of medicinal herbs and herbal products is environment-friendly.

• Throughout the world, herbal medicine has provided many of the most useful and vast variety of drugs to the modern medical science[2].

Pharmaceutical Gels• Defintion:

Gels are semisolid organic or inorganic colloids rich in liquid, consisting of hydrated threads or granules of the dispersed phase intimately associated with the dispersion medium

• Some gelling agents (carbomers) require a "neutralizer" or a pH adjusting chemical to create the gel after the gelling agent has been wetted in the dispersing medium[3].

• Gelling agents are used concentrations of 0.5% to 10%, depending on the agent.

• It is easier to add the active drug before the gel is formed if the drug doesn't interfere with the gel formation[4].

• Only Carbopol® 934P, methylcellulose, hydroxypropyl methylcellulose, and sodium carboxymethyl cellulose are recommended for oral administration.

Periodontal diseases• Periodontal disease is one of the most important concerns for dentists and

patients[5]. It is recognized as a major public health problem throughout the world and is the most common cause of tooth loss in

• The periodontium is the specialized tissues that both surround and support the teeth, maintaining them in the maxillary and mandibular bones. A variety of triggering factors like bacterial causes, dyscrasias, avitaminosis etc cause inflamed gums leading to gingivitis.

• In the United States 50% of adults have gingivitis affecting at least 3-4 teeth; two-thirds of the population has sub gingival calculus, and about a one-third have periodontitis [6] . Periodontal treatment aims to cure inflamed tissue, reduce the number of pathogenic bacteria and eliminate the diseased pockets.

• Mechanical therapy, chemotherapy and systemic administration of antibiotics are some of the clinical methods being utilized currently[7]

S.NO. Author Names Abstract References1. Reenu Yadav Formulation and characterization of antimicrobial oral gel

from some herbal extracts for treatment of periodontal diseases.

IJPPR. 2016, 5(2)

2. R.Bhramaramba Formulation and Evaluation of herbal gel containing Terminalia chebula Retz., leaves extract.

(SAJP). 2015, 4(3): 172-176.

3. Biresh Sarkar Formulation and Evaluation of herbal gel containing extract of Cedrus deodara.

(IJPCS). 2015,4(1)

4. Singh Rampal Formulation, optimizatioon and evaluation of aceclofenac transdermal gel

(JPSI). 2015,4(5)

5. T. Muthu Lakshmi Formulation and evaluation of herbal gel containing dalbergia sissoo roxb. bark extract.

(JPRCP).4(1):53-57

6. Dr Rohit Jain Herbs in periodontology – local drug delivery (WJPR).2014,3(2):1831-18407. Varsha B. Bagade Study of antimicrobial activity of herbal formulation (IJPLS).2013, 4(11)8. Deepak p pawar Formulation and evaluation of herbal gel containing

lantana camara leaves extract.(AJPCR).2013,6(3)

9. CharuM. Marya Investigation of In vitro inhibitory effect of clove essential oil and its two active principles on tooth decalcification by applying juice.

(IJD).2012

10. L. Nuñez Investigation of microbicide activity of clove essential oil (Eugenia caryophyllata) .

(BJM).2012,43(4)

11. Ganesh Misal Formulation and evaluation of Poly herbal gel. (IJNPR).2012,3(4)12. Manisha singh Formulation and evaluation Herbal Gel Containing

Ethanolic Extract of Ipomoea Fistulosa.(IJSR). 2012, 3(7)

13. Ilhami Gu¨ lc¸ Investigation of antioxidant activity of clove oil – A powerful antioxidant source.

(AJP). 2012, 5: 489-499

14. Euge´ nia Pinto Antifungal activity of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and dermatophyte species.

(JMM). 2009, 1454-1462

Literature Review

The study was aimed to develop and evaluate dental gel containing clove oil as the chief constituent for the treatment of periodontitis.

The objectives of the research work under taken are as follows:1. To perform clove oil

characterization.2. To formulate dental gel of

clove oil using gelling agents and other ingredients.

Plant ProfileSynonym: • Caryophyllum; clove flower; clove bud;

laungeBiological source:• Cloves consists of dried flower buds of

Eugenia caryophyllus (Myrtaceae). It should contain not less than 15% (v/v) of clove oil[8].

Chemical constituents:• 15-20% of volatile oil; 10-13% of tannin

(gallotannic acid), chromone and eugenin [9]. • The volatile oil contains eugenol (about 70 to

90%), eugenol acetate, methylamylketone, caryophyllenes and small quantities of ester and alcohols [10].

Uses:• Dental analgesic, carminative,• Stimulant, flavouring agent, an aromatic and

antiseptic

Taxonomical classification of clove plant (Syzygium aromaticum) Taxonomic HierarchyKingdom Plantae - plantes, planta, vegetal, plants

Subkingdom Viridiplantae

Infrakingdom Streptophyta – land plants

Superdivision Embryophyta

Division Tracheophyta – vascular plants, tracheophytes

Subdivision Spermatophytina – spermatophytes, seed plants,

phanerogames

Class Magnoliopsida

Superorder Rosanae

Order Myrtales

Family Myrtaceae – myrtles, myrtacees

Genus Syzigium P.Br.ex Gaertn

Species Syzygium aromaticum (L) Merr. & L.M.Perry – clove

Excipient profileCarbopol[2]

Nonproprietary Names:• BP: Carbomers• PhEur: Carbomers• USP-NF: CarbomerStructural Formula:

PEG[2]

Nonproprietary Names:BP: MacrogolsJP: Macrogol 400 Macrogol 1500 Macrogol 4000 Macrogol 6000 Macrogol 20000PhEur: MacrogolsUSP-NF: Polyethylene GlycolStructural Formula:

Uses:Ointment base; plasticizer; solvent; suppository base; tablet and capsule lubricant.

Use Concentration (%)

Emulsifying agent 0.1-0.5

Gelling agent 0.5-2.0

Suspending agent 0.5-1.0

Tablet binder 0.75-3.0

S.NO MATERIALS SUPPLIERS FUNCTION

1. Clove oil S.D.Fine chemicals Pvt. Ltd, Mumbai

Active pharmaceutical ingredients

2. Carbopol934 Roquette, Mumbai Gelling agent

3. Polyethylene glycol Isp Chemical Inc, Hyderabad Cosolvent

4. Glycerin Bayer Chemicals, Pune, Maharashtra

Drug solubiliser

5. Methyl paraben Merck Index chemicals, Hyderabad

Preservative

6. Propyl paraben Merck Index chemicals, Hyderabad

Preservative

7. Aspartame Nutrasweet, Vadodara, Gujarat Sweetening agent

8. Distilled water Bayer Chemicals, Pune, Maharashtra

Vehicle

Materials and Methods

Physico chemical characteristics of Clove oilAcid valueAcid value = potassium hydroxide consumed × 5.611 5.611

Weight (g) of the sample Saponification value:• Saponification value = mg of KOH consumed by 1 g clove oil• Weight of KOH = Normality of KOH × Equivalent weight× volume of KOH in litres• Volume of KOH consumed by 1 g of oil = [blank- test] Ester value • Ester value = Saponification value – Acid valueSolubility:• Clove oil is freely soluble in ethanolColour: • The colour of the formulation was checked out against a white background.Odour:• The odour of the gels was checked by mixing the gel in water and taking the smell.

Formulation of clove oil gel

Soaking •Soak carbopol 934 in water

Neutralization

•Neutralize with triethanolamine to pH 6.4

Addition of preservative •Addition of methyl and propyl paraben

Additon of co solvent and API

•Addition of propylene glycol and clove oil in another test tube

Addition of sweetner •Finally aspartame is added

Stirring •Stirring is done until a homogenous product is formed

Composition of gel formulations

INGREDIENTS F1 (gm) F2 (gm) F3 (gm) F4 (gm) F5 (gm)

Clove oil (ml) 0.75 0.75 0.75 0.75 0.75

Carbopol (g) 0.3 0.4 0.5 0.6 1

Polyethylene glycol

(ml)15 15 15 15 15

Glycerine (ml) 5 5 5 5 5

Methylparaben(g) 0.18 0.18 0.18 0.18 0.18

Propylparaben(g) 0.02 0.02 0.02 0.02 0.02

Aspartame (g) 0.4 0.4 0.4 0.4 0.4

Distilled water q.s q.s q.s q.s q.s

Evaluation of gel formulation

Appearance :• All the formulations of clove oil gel were pale yellow in colour. Consistency:• The consistency was checked by applying on skin.Greasiness:• The greasiness was assisted by the application on to the skin. Determination of pH:• The pH of gel was determined using digital pH meter by dipping the glass electrode completely into the gel

system [11]. Determination of viscosity:• Viscosities of the formulated gels was determined using Brooke field viscometer, spindle no. 7 and spindle

speed 60 rpm at 25◦C was used for gels, the corresponding dial reading on the viscometer was noted[12]. Determination of spreadability:• Spreadability was determined using following formula, S=M.L/T• Where S is the spreadability in grams.cm/sec, M is the mass in grams, T is the time in seconds. Determination of extrudability:• It was determined by sing a tube filled with the gel, having a tip of 5mm opening and by measuring the

amount of gel that extruded through the tip when a pressure was applied on the tube was noted down [13].

Determination of antimicrobial activity:Agar cup plate method was used for screening of antimicrobial activity of clove oil gel. Different concentrations of clove oil gel were placed aseptically in cups of agar plate which was previously inoculated with culture[14]. The plates were left at ambient temperature for 30 mins prior to incubation at 37◦C for 24 hrs. The broad spectrum antibiotic i.e., tetracycline was used as positive control for obtaining comparative results[15]. Plates were observed after 24-48 hrs incubation for the appearance of the zone of inhibition. Antimicrobial activity was evaluated by measuring the diameter of zones of inhibition (millimetres) of microbial growth.

Results

S.NO. PARAMETERS CLOVE OIL PROCURED

CLOVE OIL STANDARD

1. Colour Pale yellow Pale yellow

2. Odour Aromatic Aromatic

3. Acid value 3.66 3.84

4. Ester value 37.21 38.22

5. Solubility in ethanol Freely soluble Freely soluble

6. Density 1.02 g/ml 1.05g/ml

7. Refractive index 1.492 1.532

Table : 5 Physicochemical characteristics of clove oil

1

2

3

Fig: Antimicrobial activity on S.salivarius1 – F3 (Gel Formulation)2 – Clove oil3 – Tetracycline

5

Formulations

Appearance

pH Spreadability (g-cm/sec)

Extrudability %

Homogeneity Drug Content

F1 Pale yellow 6.6 18.20 92.14 Good 95.00

F2 Pale yellow 6.7 18.14 93.15 Good 95.20

F3 Pale yellow 6.7 17.49 94.10 Very good 95.40

F4 Pale yellow 6.6 16.72 90.23 Good 93.62

F5 Pale yellow 6.4 15.59 89.10 Very good 89.80

Table:6 Characteristics of gel formulations

Zone o

f inhibition (S

.saliva

rius)

Zone o

f inhibition (S

.sangu

is)

Zone o

f inhibition(L.

acidophilu

s)16.5

17.5

18.5

19.5

F3

Fig: 3 Antimicrobial activity on S.salivarius

F1 f2 F3 F4 F50

5

10

15

20

25

Zone of inhibition(S.salivarius)

Zone

of i

nhib

ition

(mm

)

Gel formulations

Fig: 4 Zone of inhibition of Streptococcus salivarius

F1 F2 F3 F4 F50

5

10

15

20

25Zone of inhibition (S.sanguis)

zone

of i

nhib

ition

(mm

)

Gel formulations

Fig:5 Zone of inhibition of Streptococcus sanguis

F1 F2 F3 F4 F50

5

10

15

20

25

Zone of inhibition (L.acidophilus)

zone

of i

nhib

ition

(mm

)

Gel formulations

Fig: 6 Zone of inhibition of Lactobacillus acidophilus

Discussions• The procured clove oil was characterized for the following parameters: Acid value : 3.66 Ester value : 37.43 Saponification value: 41.09 Density : 1.02gm/ml Refractive index: 1.492• The formulations were developed by using clove oil of same concentration and carbopol 934 at

different concentrations.All the formulations were pale yellow in colour and had characteristic odour of clove oil. The pH of all formulations ranged from 6.4-6.7, which was well within the normal pH range of buccal cavity 6-7. The spreadability of the gels was found to be in the range of 15.59-18.20 g-cm/sec, confirming that these gels may spread smoothly and uniformly. The formulations were glossy and translucent. The homogeneity and tube extrudability of all formulations was good.

• The drug content of the formulations was ranged from 89.8%-95.40% Table-6.The formulation F3 was found to have maximum drug content.

• The gel formulations of clove oil F3 showed good physicochemical properties as well as good drug content compared to other formulations.(Table -5,6). Hence, theses formulations were further selected for anti microbial studies. The results of anti microbial studies showed that gel formulation of clove oil F3 showed a maximum zone of inhibition on Streptococcus salivarius.

Conclusion • The clove oil was found to have anti microbial activity

against Streptococcus salivarius, Streptococcus sanguis, Lactobacillus acidophilus. The formulations developed from clove showed significant results so it can be further used commercially to develop dental gels after conducting clinical trials on human beings. Nevertheless further research is still needed in order to determine if they efficiently could substitute the synthetic antibiotics are used in combinations.

References 1. Euge nia Pinto; Lui s Vale-Silva; Carlos Cavaleiro and Li gia Salgueiro. Antifungal activity

of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and dermatophyte species. Journal of medical microbiology. 2009: 1454-1462.

2. Author. Handbook of pharmaceutical excipients. Pharmaceutical press. 2009.3. Chaieb, K. et al. (2007) The Chemical Composition and Biological Activity of Clove

Essential Oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): A Short Review. Phytotherapy Research, 21, pp. 501-506.

4. Gupta A. K., Tandon Neeraj, Dr. Sharma Madhu (2011). Quality Standards of Indian Medicinalplants, Indian Council of Medical Research, New Delhi, Volume 3

5. Ates D.A. and Erdogrul O.T. (2003) Antimicrobial activities of various medicinal and commercial plant extracts. Turkish Journal of Biology, 27: 157-162.

6. Suwipa U., Tanomjit S., Pechnoi S., Supreedee S., Prenee R. and Ithrat A. (2005) Study on antioxidant and antimicrobial activities of turmeric clear liquid soap for wound treatment of HIV patients. Journal of Science Education and Technology.27: 569-578.

7. Das K, Dang R, Machale UM, Fatepuri S. Formulation and evaluation of herbal gel containing Stevia leaves extract, The Pharma Review 2010, 8(44): 112-118.

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Aspergillus flavus. KMITL Sci. Tech J 2007; 7: 1-7.9. Satpathy B, Sahoo M, Sahoo P, Patra SR, Formulation and evaluation of gel containing essential

oils of Piper betle against skin infection pathogens. Int . J. Res Phar. Sci 2011; 2(3): 373-378.10. Hanoch Julianus Sohilait(2015). Chemical Composition of the Essential Oils in Eugenia

caryophylata, Thunb from Amboina Island, Science Journal of Chemistry ; 3(6): 9511. Ashley FP, Skinner A, Jackson P, Woods A, Wilson RF. The effect of a 0.1% cetylpyridinium

chloride mouthrinse on plaque and gingivitis in adult subjects. Br Dent J. 1984; 157:191-196. 12. Moran J, Addy M, Kohut B, Hovliaras CA, Newcombe RG. Efficacy of mouthrinses in

inhibiting the development of supragingival plaque over a 4-day period of no oral hygiene. J Periodontol;1994; 65:904-90

13. Lobene RR, Kashket S, Soparkar PM, Shloss J, Sabine ZM. The effect of cetylpridinium chloride on human plaque bacteria and gingivitis. Pharmacol Ther Dent. 1979; 4:33-47

14. Pawar Vinita A*, Bhagat Trupti B, Toshniwal Mitesh R, Mokashi Nitin D, Khandelwal K.R; Formulation and evaluation of dental gel containing essential oil of coriander against oral pathogens; Pawar Vinita A et al. Int. Res. J. Pharm. 2013, 4 (10).

15. Anurag Sharma*; Sumeet Dwivedi; Ganesh P. Mishra; Hemant Joshi; Formulation and Evaluation of Herbal Gel containing Extracts of Albezia Lebbeck linn; Am. J. PharmTech Res. 2012; 2(4)