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Experimental

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Experimental Work

Suresh Gyan Vihar University, Jaipur Page 105

5. MATERIALS AND EQUIPMENTS

5.1 Materials

Chemicals: : Lornoxicam, Flurbiprofen, Aceclofenac, Piroxicam, pH 7.4

buffer, potassium dihydrogen phosphate, sodium hydroxide, disodium

hydrogen phosphate, sodium hydroxide, n-octanol, Pluronic F-127, lecithin,

isopropyl myristate, Sodium sorbate, Sodium Benzoate, distilled water,

ethanol, methanol, and acetone.

Glassware: Beaker, volumetric flask, glass rod, conical flask, funnel, pipette,

measuring cylinder, separating funnel, slides, test tubes, capillary.

Equipment: UV double beam, stage micrometer, microscope, ocular lense,

Thiels melting point apparatus, hot air oven, pH meter, electronic balance,

dessicator.

5.2 Preformulation studies

1. Determination of solubility

a. Qualitative solubility

Qualitative solubility analysis of drugs were done by dissolving 5 mg of drug

in 5 ml of distilled water and different solvents such as HCl (0.1N), NaOH (0.05

N), Saline phosphate buffer (pH 7.4), Phosphate buffer(pH 9), Phosphate buffer

(pH 4), phosphate buffer (pH 2), ethanol, methanol, acetone and chloroform were

used to determine the solubility of drug.

b. Quantitative Solubility

Quantitative solubility analysis of drugs were done by 5 ml each solvent and

drug in gm(s) into the solvent till saturation of solvent. Different solvents were

used for the solubility determination like distilled water, Saline phosphate buffer

(pH 7), Phosphate buffer(pH 3.6), HCl (0.1N) and NaOH (0.05N). This is done to

determine the capacity of the solvent for dissolving the drug in it. The

concentration of drug is measured by UV spectrophotometer at 376 nm.

Experimental Work


2. Partition Coefficient

The 10 mg of drug was dissolved in 10 ml of distilled water and 10 ml of

carbon tetra chloride in separating funnel & shaken well for 3-4 hours and then

allowed to stand for at least 1 hour for phase separation. After that the water phases

were separated out and the concentration of drug was measured

spectrophotometrically after suitable dilution at 376 nm.

Po/w = Coil/Cwater

3. Melting Point

In this the presealed capillary was filled by the small amount of drug and the

capillary was placed in Melting Point Apparatus.The temperature when the drug starts

to melt and the temperature when drug complete melting was noted.

4. Particle Size

I) Calibration of eyepiece: Use standard stage micrometer to calibrate the eyepiece

micrometer and calculate for the least count (1 eye piece division)

Least Count = No. Of Stage Division / No. Of Eye Piece Div x 10.

II) Mounting of The sample: Transfer a small portion of the given sample on clean

slide and disperse it uniformly and place the slide on the stage of microscope.

III) Measurement of particle size: Focus the slide in low magnification (10x).

observe the particles than shift to high power (45x) and focus the slide. Measure the

size of each particle in terms of eyepiece divisions,a total of 100 particles should be

considered,tabulate the particles in terms of division of eyepiece and no. of particles

(frequency) obtained above,classify the diameter into size ranges and average

frequency of particles in terms of no. distribution.

5. Drug- excipient interaction study

A small amount of drug substance with excipients (like plronic F-127,

Lecithin, Sodium sorbate, Sodium benzoate, Isopropyl myristate, PEG 400,

Experimental Work


triethanolamine, carbopol 934, oleic acid, ethanol and propylene glycol) that is,

physical mixture of the drug and excipients (in 1:1 ratio were prepared to have

maximum likelihood interaction) were placed in a vial, and rubber stopper was placed

on the vial and sealed properly. A storage period of 2 weeks at 60°C, and the same

sample was retained for 2 months at 40°C. After storage the sample were observed

physically for liquefaction, caking, odour or gas formation, & discolouration.

6. Determination of λ max

A. Lornoxicam

The identification of drug was done by UV spectrophotometric method

reported by Nemutlu et al (2005). The small amount of drug is dissolved in 0.05 N

NaOH and scanned in UV range 200-600 nm in UV Double beam spectrophotometer.

The highest peak was determined, which is the λmax for the Lornoxicam .The spectral

data from this scan was used for the preparation of standard curve of Lornoxicam.

B. Flurbiprofen

100 mg of Flurbiprofen was dissolved in 100ml of ethanol. One ml of the

prepared stock solution was further diluted to 100 ml and finally scanned for

maximum absorbance using U.V. spectrophotometer in the range from 230 to 360 nm.

Average of triplicate readings was taken.

C. Aceclofenac

Aceclofenac was dissolved in PBS to obtain a 1000mcg/mL solution. This

solution was subjected to scanning between 200 – 400 nm and absorption maximum

was determined. The effect of dilution on absorption maxima was studied by diluting

the above solution to 20mcg/mL and scanned from 200 – 400nm.

D. Piroxicam

The identification of piroxicam done by UV spectrophotometer method. The small

amount of drug dissolve in 0.1N Hydrochloric acid and scanned in UV. The highest

peak is λmax for the piroxicam. From the spectra 333 nm ( λmax of piroxicam) was

obtained.

Experimental Work


2) Preparation of standard curve

A. Lornoxicam

a. Standard curve of lornoxicam in 0.05 N NaOH

Standard stock solution of Lornoxicam was prepared by dissolving 100 mg

drug in 100 ml 0.05 N NaOH (i.e.1000μg/ml). Aliquot of this solution are further

prepared by taking 10 ml of above solution and diluting it upto 100 ml (i.e. 100

µg/ml). From the above solution further dilutions are prepared in range of 5-35 µg/ml.

Absorbances were taken on UV spectrophotometer at 376 nm against 0.05 N NaOH as

a blank. From these absorbance’s, the standard curve is plotted. Standard curve

equation and regression value is obtained. The absorptivity coefficient of drug at

desired wavelengths was determined.

b. Standard curve of lornoxicam in 7.4 PBS

Standard stock solution of Lornoxicam was prepared by dissolving 10 mg drug

in 10 ml 7.4 PBS (i.e.1000μg/ml) containing 12mg of Tromethamine. This solution

was then sonicated till complete dissolution of drug. Aliquot of this solution are

further prepared by taking 10 ml of above solution and diluting it upto 100 ml (i.e. 100

µg/ml). From the above solution further dilutions are prepared in range of 4-24 µg/ml.

Absorbances were taken in UV spectrophotometer at 376 nm against 7.4 PBS as a

blank.From these absorbance’s, the standard curve is plotted. Standard curve equation

and regression value is obtained. The absorptivity coefficient of drug at desired

wavelengths was determined.

B. Flurbiprofen

a. Standard curve of Flurbiprofen in ethanol

100 mg of Flurbiprofen was accurately weighed and dissolved in ethanol in a

100 ml volumetric flask and the volume was made upto the mark with ethanol. The

above prepared solution of Flurbiprofen was subsequently diluted with ethanol to get

2, 4, 6, 8, 10, 12 μg per ml of the final solution. Then the absorbance was measured by

spectrophotometer at 248nm using ethanol as blank. Average of triplicate readings was

taken.

b. Standard curve of flurbiprofen in 7.4 PBS

Standard stock solution of Flurbiprofen was prepared by dissolving 10 mg drug

in 10 ml 7.4 PBS (i.e.1000μg/ml). Aliquot of 10 ml solution are further taking 10 ml of

Experimental Work


diluted upto 100 ml (i.e. 100 µg/ml). From the above solution are prepared in range of

10-90 µg/ml. Absorbances were taken on UV spectrophotometer at 248 nm against 7.4

PBS as a blank. From these absorbance’s, the standard curve is plotted. Standard

curve equation and regression value is obtained.

C. Aceclofenac

a. Standard curve of Aceclofenac in PBS

A stock solution containing 1 mg/mL of pure drug was prepared by dissolving

50mg of Aceclofenac in sufficient PBS to produce 50 mL solution in a volumetric

flask.Aliquot of 10 ml diluted upto 100 ml (i.e. 100 µg/ml). From the above solution

are prepared in range of 10-90 µg/ml. Absorbances were taken on UV

spectrophotometer at 273 nm against 7.4 PBS as a blank. From these absorbance’s,

the standard curve is plotted. Standard curve equation and regression value is obtained.

D. Piroxicam

a. Standard curve in 0.1N HCl

Standard stock solution of piroxicam was prepared by dissolving 100 mg drug

in 100 ml 0.1N HCL (i.e. 1000 μg/ml) . From the stock solution 10 ml was taken and

diluted upto 100 ml (i.e. 100 in 0.1N HCL. Again 0.2, 0.4, 0.6, 0.8, 1, 1.2, 1.4, 1.6, 1.8

and 2 ml solution was taken from this solution and dilute upto 10 ml with 0.1N HCL

to get the desired concentration range (2-20 μg/ml). The absorbance of drug was

measured at λmax 333 nm on UV spectrophotometer.

5.3 Results of preformulation studies

A. Lornoxicam

Identification of drug

The highest peak was found at 376 nm. Zero order derivative UV

spectrophotometric methods were used for the analysis of lornoxicam and the highest

peak was found at 367 nm. The drug samples were prepared with 0.05 N NaOH and

scanned with UV [Fig 5.1]. The values compared with standard drug volume, which is

reported in Merck Index.

1) Organoleptic Properties

Organoleptic properties of the drug sample were found to be as given in table

below-5.1

Experimental Work


Table 5.1 Organoleptic properties of Lornoxicam

Organoleptic properties Results

Colour Orange to Yellow powder

Crystallinity Amorphous in nature

Taste Slightly bitter in taste

Odour Odourless

2)Partition Coefficient

Partition Coefficient of Lornoxicam was found to be 1.7. The above value of

partition coefficient is nearby to the value of partition coefficient reported in Merck

index for Lornoxicam. The partition coefficient shows that the drug is lipophilic in

nature which makes it suitable for transdermal delivery via Pluronic lecithin

organogel.

3)Melting Point

Melting Point of Lornoxicam was found in range of 225-2280c which falls

under the melting point range specified in Merck index. This shows that the drug is

pure. The drug starts melting at 225º c and completes melting at 228º c which indicates

amorphous nature of drug.

4)Solubility Properties

Qualitative Solubility: The results of qualitative solubility of the drug in different

solvents are given below in the table 5.2.

Experimental Work


Table 5.2 Qualitative Solubility of drug in different solvents

Solvents(5ml) Solubility Properties of the drug(5mg)

Distilled Water +

0.1N HCl +++

3.6 pH Buffer ++

7.4 pH Buffer +++

9.2 pH Buffer +++

Ethanol ++

Methanol ++

Chloroform ++

Acetone +++

Hexane +

0.05 N NaOH ++++

+ Insoluble

++ Poorly soluble

+++ Slightly soluble

++++ Freely soluble

The results were showed that Lornoxicam is insoluble in distilled water &

hexane and very less solubility in organic solvents like ethanol, methanol, chloroform

& acetone but the drug was freely soluble in alkaline solvents like 7.4 pH buffer and

9.2 pH buffer. The drug showed high solubility in 0.05 N NaOH, which indicates the

acidic nature of the drug.

Quantitative Solubility: The results of quantitative solubility of the drug are given

below in the table 5.3.

Experimental Work


Table 5.3 Quantitative Solubility of drug in different solvents

Solvent Concentration of drug in solvent

0.05 N NaOH 6.306 mg of drug was present in 1ml of 0.05 N NaOH

0.1N HCl 0.664 mg of drug was present in 1 ml of 0.1N HCl

3.6 pH Buffer 0.732 mg of drug was present in 1 ml of 3.6 pH buffer



The observations showed that the solubility of Lornoxicam increases with the increase

of pH from 3.0 to 9.0, which indicates that the ionization of drug increases with the

elevating pH.

5.) Standard Curve

Standard curve of the drug in 0.05 N NaOH & 7.4 PBS was prepared by

method reported by Nemutlu et al (2005). The absorbances were taken out at 376 nm.

a. Standard curve of lornoxicam in 0.05 N NaOH

Absorbance’s of the drug at 376 nm in 0.05 N NaOH are given below in table

5.4.

Table 5.4 Absorbances of Lornoxicam at 376 nm in 0.05 N NaOH

S.

No.

Concentration (µg/ml) Absorbance

1 5 0.076

2 10 0.147

3 15 0.216

4 20 0.298

5 25 0.367

6 30 0.438

7 35 0.513

Experimental Work


Standard curve of Lornoxicam in 0.05 N NaOH at 376 nm is shown below in fig. 5.1

Fig 5.1: Standard Curve of Lornoxicam in 0.05 N NaOH

b. Standard curve of lornoxicam in PBS 7.4:

Absorbances of the drug at 376 nm in PBS 7.4 are given below in table 5.5-

Table 5.5 Absorbances of Lornoxicam at 376 nm in PBS 7.4

S. No. Concentration

(µg/ml)

Absorbance

1 0 0

2 4 0.1672

3 8 0.3354

4 12 0.4885

5 16 0.6515

6 20 0.8225

7 24 0.9663

Experimental Work

Suresh Gyan Vihar University, Jaipur 4

Fig 5.2 Standard Curve of Lornoxicam in 7.4 PBS

Fig. 5.1 & 5.2 showed linearity in the range of 5-35 µg/ml & 4-24 µg/ml with

regression coefficient of 0.9997 & 0.9996 respectively. It shows that the drug follows

Beer’s Lambert Law in these ranges.

7) Particle Size

The results of the Microscopic evaluation for the measurement of particle size

of the drug particles are given below in table 5.6.

Table 5.6 Particle size distribution of Lornoxicam

S. No. Size Range Mid Point

(M. P.)

No. of Particles

(N)

M. P. ×

N

M. P. × N × L. C.

(d)

1. 0-1 0.5 06 03 5.82

2. 1-2 1.5 09 13.5 12.15

3. 2-3 2.5 11 27.5 53.35

4. 3-4 3.5 27 94.5 183.33

5. 4-5 4.5 23 103.5 200.79

6. 5-6 5.5 24 132 256.06

∑n=100 ∑d=714.5

Experimental Work


Least Count (L. C.) = 1.94

Particle size was found to be 7.145 µm. Particle size distribution pattern

depicted in fig. 5.4 shows that drug particles are distributed in a range of 1-6 µm and

maximum number of particles are present in size range of 4-6 µm. This distribution

pattern also indicates that the drug is amorphous in nature.

From the above data particle size distribution graph is plotted which is shown in fig.

5.3.

Fig. 5.3 Particle Size Distribution of Drug (Lornoxicam)

8)Drug- excipient compatibility studies

Drug-excipient compatibility is performed by method described in merck

index. This study is done to find out compatibility between the drug and excipients.

From the results given in table 5.6, it can be concluded that there is no interaction

between excipients and drug. The drug and excipient are compatible with each other

and can be used for formulation of Gel.

The results of Drug-Excipient Compatibility studies are shown in table 5.7.

Experimental Work


Table 5.7 Drug-Excipient Compatibility Observations

S.No. Additives (50 mg each)

with drug

Observation at

60°C for 2 weeks

Observation at 40°C

for 2 month

Remarks

1. Drug ( lornoxicam) No interaction No interaction Accepted

2. Drug + pluronic F-127 No interaction No interaction Accepted

3. Drug + lecithin No interaction No interaction Accepted

4. Drug + isopropyl myristate No interaction No interaction Accepted

5. Drug + PEG 400 No interaction No interaction Accepted

6. Drug + Sodium sorbate No interaction No interaction Accepted

7. Drug + Sodium Benzoate No interaction No interaction Accepted

8. Drug + Triethanolamine No interaction No interaction Accepted

9. Drug + carbopol 934 No interaction No interaction Accepted

10. Drug + Oleic acid No interaction No interaction Accepted

11. Drug + Ethanol No interaction No interaction Accepted

12. Drug + propylene glycol No interaction No interaction Accepted

A. Flurbiprofen

Identification of drug: The λmax of Flurbiprofen was obtained by using double beam

uv visible spectrophotometer in the range of 230-360 nm..The maximum peak was

observed at 248 nm which is same as reported in I.P.

1) Organoleptic properties:

On the basis of organoleptic properties it was found that flurbiprofen was white,

crystalline powder, bitter taste and almost odorless shown in table 5.8.

Table 5.8: Organoleptic properties of flurbiprofen

S.No. Parameter Description

I) Color White Or Almost White.

II) Crystallinity Crystalline Powder.

III) Taste Bitter

IV) Odors Odorless Or Almost Odorless.

Experimental Work


2) Solubility determination: It was found that flurbiprofen was soluble in most of the

organic solvent and insoluble in water as shown in table 5.9.

Table 5.9 Solubility of Flurbiprofen


I) Qualitative Insoluble In Water.

Freely Soluble In-Ethanol(95 %),

Methanol ,Chloroform,

Acetone, Ph 7.4 Buffer

Quantitative solubility: The results of Quantitative solubility of the drug are given

below in the following table 5.10.

Table 5.10: Quantitative Solubility of drug in different solvents


Ethanol(95%) 7.412 mg of drug was present in 1ml of ethanol

Methanol 8.365 mg of drug was present in 1 ml of Methanol


Distilled water 0.03 mg of drug was present in 1 ml of distilled water

3) Partition co-efficient: The partition coefficient of Flurbiprofen in chloroform/water

was found to be 3.89. The obtained value of partition coefficient is suitable for

transdermal drug delivery.

4) Particle Size

The results of the microscopic evaluation for the measurement of particle size of the

drug particles are given below in table 5.11.

Experimental Work


Table 5.11: Particle size distribution of Flurbiprofen


(M. P.)

No. of Particles

(n)

M. P.* N M. P. * N *

L. C.(d)

1. 0-1 0.5 06 03 5.82

2. 1-2 1.5 09 13.5 12.15

3. 2-3 2.5 11 27.5 53.35

4. 3-4 3.5 27 94.5 183.33

5. 4-5 4.5 23 103.5 200.79

6. 5-6 5.5 24 132 256.06

∑n=100 ∑d=714.5

Particle size of Flurbiprofen = ∑d/∑n

=714.5/100

=7.145µm


5.4.

Fig. 5.4 Particle Size Distribution of Drug

Experimental Work


Particle size was found to be 7.145 µm. & distribution pattern depicted in fig. 5.4 .The

drug particles are distributed in a range of 1-6 µm and maximum number of particles

are present in size range of 4-6 µm.

5) Melting point:

The melting point of flurbiprofen was obtained by Thiels melting point apparatus. The

melting point was observed from 110-1120C which is approximately same as I.P.1996.

6) Standard curve in ethanol:

Standard curve of flurbiprofen in ethanol: the standard curve of flurbiprofen was

prepared in ethanol as shown in table 5.12 and fig 5.5

Table 5.12 Data for standard curve of flurbiprofen in ethanol

S.No. Concentration (μg / ml) Absorbance

1. 0 0

2. 2 0.098

3. 4 0.185

4. 6 0.368

5. 8 0.467

6. 10 0.599

7. 12 0.743

8. 14 0.879

Experimental Work


Fig 5.5. Standard curve of flurbiprofen in ethanol

h). Standard curve of flurbiprofen in 7.4 PBS

Standard curve of flurbiprofen in 7.4 PBS: the standard curve of flurbiprofen was

prepared in 7.4 PBS as shown in table 5.13 and fig 5.6

Table 5.13: Absorbance of Flurbiprofen at 248 nm in PBS 7.4

S. No. Concentration (µg/ml) Absorbance

1 0 0

2 10 0.149

3 20 0.248

4 30 0.350

5 40 0.451

6 50 0.562

7 60 0.628

8 70 0.738

9 80 0.837

10 90 0.902

Experimental Work


Fig 5.6. Standard curve of flurbiprofen in Pbs 7.4

7) Drug- Excipient compatibility studies


S.No. Additives (50 mg each)with drug

Observation at60°C for 2 weeks

Observation at40°C for 2 month

Remarks

1. Drug (flurbiprofen) No change No change Accepted

2. Drug + pluronic F-127 No change No change Accepted

3. Drug + lecithin No change No change Accepted

4. Drug + isopropyl myristate No change No change Accepted

5. Drug + PEG 400 No change No change Accepted

6. Drug + Sodium sorbate No change No change Accepted

7. Drug + Sodium Benzoate No change No change Accepted

8. Drug + Oleic acid No change No change Accepted

9. Drug + Ethanol No change No change Accepted

10. Drug + propylene glycol No change No change Accepted

Experimental Work


From the results given in table 5.11. it is concluded that there is no interaction between

excipients and drug. The drug and excipient are compatible with each other and can be

used for formulation of gel.

B. Aceclofenac

Identification of drug

The highest peak was found at 273 nm. The UV scan of the drug sample fig 5.7

showed highest peak at 276 nm which is nearby to the standard value reported in the

Merck index.

1)Organoleptic properties

Organoleptic properties of the drug sample was found to be as given in table

below.

Table 5.15: Organoleptic properties of Aceclofenac

Organoleptic properties Results

Colour Pale Yellow powder

Crystallinity Amorphous in nature

Taste Slightly bitter in taste

Odour Odourless

2)Partition coefficient

Partition Coefficient of Aceclofenac was found to be 1.3. The above value of

partition coefficient is nearby to the value of partition coefficient reported in Merck

index for Aceclofenac. The partition coefficient shows that the drug is lipophilic in

nature which makes it suitable for transdermal delivery via Pluronic lecithin

organogel.

Experimental Work


3)Melting point

Melting Point of Aceclofenac was found in range of 273-2780c which falls

under the melting point range specified in Merck index. This shows that the drug is

pure. The drug starts melting at 275º c and completes melting at 278º c which indicates

amorphous nature of drug.

4)Solubility Properties

Qualitative Solubility: The results of qualitative solubility of the drug in

different solvents are given below in the table 5.16.

Table 5.16 Qualitative Solubility of drug in different solvents

Solvents(5ml) Solubility Properties of the drug(5mg)

Distilled Water ++

0.1N HCl +++

3.6 pH Buffer +

7.4 pH Buffer ++++

9.2 pH Buffer +

Ethanol +++

Methanol ++

Chloroform ++

Acetone +

Hexane +

0.05 N NaOH ++

+ Insoluble

++ Poorly soluble

+++ Slightly soluble

++++ Freely soluble

The results showed that Aceclofenac is insoluble in acetone & hexane and very

less solubility in organic solvents like, methanol, chloroform. The drug showed high

solubility in 0.1 n HCL and ethanol, which indicates the acidic nature of the drug.

Experimental Work


Quantitative solubility: The results of Quantitative solubility of the drug are given

below in the table 5.17.

Table 5.17: Quantitative Solubility of drug in different solvents


0.05 N NaOH 1.324mg of drug was present in 1ml of 0.05 N NaOH

0.1N HCl 3.745 mg of drug was present in 1 ml of 0.1N HCl




5)Standard curve

Standard curve of the drug in 7.4 PBS was prepared. The absorbances were

taken out at 273 nm.

Standard curve of Aceclofenac in PBS

Absorbance’s of the drug at 273 nm in 0.05 N NaOH are given below in table 5.18

Table 5.18: Absorbances of Aceclofenac at 273 nm in PBS

S. No. Concentration (µg/ml) Absorbance

1 5 0.081

2 10 0.141

3 15 0.221

4 20 0.289

5 25 0.361

6 30 0.429

7 35 0.520

Experimental Work


Standard curve of Aceclofenac in PBS at 273 nm is shown below in fig. 5.7

Fig 5.7: Standard Curve of Aceclofenac in 0.05 N NaOH

Fig. 5.9 showed linearity in the range of 5-35 µg/ml & 4-24 µg/ml with

regression coefficient of 0.9993. This shows that the drug follows Beer’s Lambert Law

in these ranges.

6) Particle size



Table 5.19 Particle size distribution of Aceclofenac


(M. P.)

No. of Particles

(N)

M. P. ×

N

M. P. × N × L. C.

(d)

1. 0-1 0.5 05 2.5 4.625

2. 1-2 1.5 09 13.5 24.975

3. 2-3 2.5 14 35 64.75

4. 3-4 3.5 26 91 168.35

5. 4-5 4.5 21 94.5 174.85

6. 5-6 5.5 25 137.5 254.375

∑n=100 ∑d=691.925

Least Count (L. C.) = 1.85

Experimental Work


Particle size was found to be 6.91 µm. Particle size distribution pattern

depicted in fig. 7.4 shows that drug particles are distributed in a range of 1-5 µm and

maximum number of particles are present in size range of 4-7 µm. This distribution

pattern also indicates that the drug is amorphous in nature.

Fig. 5.8 Particle size distribution of drug (Aceclofenac)

6)Drug- excipient compatibility studies

Drug-excipient compatibility is performed by method described in merck

index. This study is done to find out compatibility between the drug and excipients.

From the results given in table 5.20 it can be concluded that there is no interaction

between excipients and drug. The drug and excipient are compatible with each other

and can be used for formulation of Gel. The results of Drug-Excipient Compatibility

studies are shown in table 5.20.

Experimental Work




with drug

Observation at

60°C for 2 weeks

Observation at

40°C for 2 month

Remarks

1. Drug ( Aceclofenac) No interaction No interaction Accepted

2. Drug + pluronic F-127 No interaction No interaction Accepted

3. Drug + lecithin No interaction No interaction Accepted

4. Drug + isopropyl myristate No interaction No interaction Accepted

5. Drug + PEG 400 No interaction No interaction Accepted

6. Drug + Sodium sorbate No interaction No interaction Accepted

7. Drug + Sodium Benzoate No interaction No interaction Accepted

8. Drug + Triethanolamine No interaction No interaction Accepted

9. Drug + carbopol 934 No interaction No interaction Accepted

10. Drug + Oleic acid No interaction No interaction Accepted

11. Drug + Ethanol No interaction No interaction Accepted

12. Drug + propylene glycol No interaction No interaction Accepted

D. Piroxicam

Identification of drug: The λmax of Piroxicam was obtained by using uv visible

spectrophotometer in the range of 230-400 nm.The maximum peak was observed at

354 nm which is same as reported in I.P-1996.

1) Organoleptic properties:

On the basis of organoleptic properties it was found that Piroxicam was white

powder, bitter in taste and almost odorless shown in table 5.21.

Experimental Work


Table 5.21: Organoleptic properties of Piroxicam


I) Color Off white to light yellow powder

II) Crystallinity Crystalline Powder.

III) Taste Sligtly Bitter in taste

IV) Odors Odorless

2) Solubility Determination Qualitative Solubility-

The value of qualitative solubility of piroxicam are shown below

Table 5.22: Qualitative solubility of piroxicam

Solvents (5 ml) Solubility properties of drug

Distilled water +

Acetone + + +

Methanol + + +

Ethanol + +

Simulated salivary fluid (6.8 pH) ++

0.1N HCL + +

Chloroform + +

Where-

+ : Poorly soluble, + + : Slightly soluble, + + + : soluble

Qualitative solubility studies of drug shown in table 5.22 depicted that the drug is

more soluble in organic solvents as compare to hydrophilic solvents so it can be

concluded that drug is lipophilic in nature.

Experimental Work


Quantitative solubility:

The result of quantitative solubility of piroxicam are given below-

Table 5.23: Quantitative Solubility of Piroxicam

Solvents (2 ml) Solubility (mg/ml)

Distilled water 0.027mg/ml

Acetone 22.4 mg/ml

Methanol 7.56 mg/ml

Ethanol 5.97 mg/ml

PBS (6.8 pH) 6.19 mg/ml

0.1N HCL 1.70 mg/ml

Chloroform 10.4 mg/ml

3) Partition co-efficient: The partition coefficient of Piroxicam in chloroform/water

was found to be 2.8-3. The obtained value of partition coefficient is suitable for

transdermal drug delivery.

4) Particle size



Table 5.24: Particle size distribution of Piroxicam

S. No. Size range Mid point No. of particles

(N)

M.P.*No. of

particle*L.C. = d

1 0-1 0.5 59 36.8

2 1-2 1.5 19 35.6

3 1-3 2.5 16 50

4 1-4 3.5 5 21.8

5 1-5 4.5 1 5.6

Total Ʃn = 100 Ʃd= 150.00

Particle size of Piroxicam was found 1.5 micrometer.

Experimental Work



5.9

Fig. 5.9: Particle size distribution of drug

Particle size was found to be 1.5 µm. Particle size distribution pattern depicted in fig.

5.8 shows that drug particles are distributed in a range of 1-6 µm and maximum

number of particles are present in size range of 0-2 µm.

5) Melting point:

The melting point of Piroxicam was obtained by Thiels melting point apparatus. The

melting point was observed from 197-2000C which is approximately same as I.P.1996.

6) Standard curve in 0.1 N HCL

Standard curve calibration of piroxicam was prepared in 0.1N HCl at 333 nm and 354

nm on UV spectrophotometer. The linearity range was found between 2 – 20μg /ml

which obeys Lambert beers low.

Experimental Work


Table 5.25: Standard curve of Piroxicam

S.No. Concentration. (μg /ml) Absorbance at

333nm in 0.1N

HCl.

Absorbance at

354nm in

SSF(6.8)

0 0 0 0

1 2 0.133 0.073

2 4 0.249 0.135

3 6 0.368 0.214

4 8 0.510 0.281

5 10 0.630 0.349

6 12 0.755 0.417

7 14 0.876 0.484

8 16 1.024 0.549

9 18 1.159 0.622

10 20 1.250 0.686

Fig 5.10: Standard curve calibration of Piroxicam

Experimental Work


7) Drug-Excipient Compatibility studies

This study is done to find out compatibility between the drug and excipients. The

results of Drug-Excipient Compatibility studies are shown in table 5.26.

Table 5.26: Drug-Excipient Compatibility Observations


with drug

Observation at

60°C for 2 weeks

Observation at

40°C for 2 month

Remarks

1. Drug (Piroxicam) No change No change Accepted

2. Drug + pluronic F-127 No change No change Accepted

3. Drug + lecithin No change No change Accepted

4. Drug + isopropyl myristate No change No change Accepted

5. Drug + PEG 400 No change No change Accepted

6. Drug + Sodium sorbate No change No change Accepted

7. Drug + Sodium Benzoate No change No change Accepted

8. Drug + Oleic acid No change No change Accepted

9. Drug + Ethanol No change No change Accepted

10. Drug + propylene glycol No change No change Accepted

From the results given in table 5.26, it is concluded that there is no interaction between

excipients and drug. The drug and excipient are compatible with each other and can be

used for formulation of gel.

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