formulasi

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Dhanashri et al. World Journal of Pharmacy and Pharmaceutical Sciences

DEVELOPMENT AND VALIDATION OF HERBAL ANTISEPTIC

TOPICAL FORMULATION

Dhanashri B. Nagulwar*1, Pravin K. Bhoyar 2, Jagdish R. Baheti 3, Dinesh M. Biyani 5,

Dharmendra R. Mundhada4 , Prasad P. Kathade 2

1. Department of Pharmaceutics, Sharad Pawar College of Pharmacy, Dist: Nagpur,

Maharashtra, India.

2. Siddhivinayak College of Pharmacy,Warora, Dist-Chandrapur, Maharashtra, India.

3. Shriman Suresh Dada Jain College of Pharmacy, Chandwad, Nasik, Maharashtra, India

4. Department of Pharmaceutics, Agnihotri College of Pharmacy, Wardha, Maharashtra,

India

5. Department of Pharmaceutical Sciences, Birla Institute of Technology, Mesra, Dist- Ranchi, Jharkhand, India.

ABSTRACT

For skin infections, topical treatments applied directly to the skin are

safest. Present work was decided to carry out antimicrobial activity

and antiseptic effect of formulated cream. Oil of J. regia oil of K.

galanga, methanolic extract of C. tora seeds and T. arjuna bark were

successfully extracted and phytochemically screened. Thin layer

chromatography of above extracts showed that it contains different

active constituents. The result of phytochemical screening reveals that

the major constituents of extracts were coumarins, flavonoids, steroids

and triterpenoids. The in- vitro antimicrobial activity showed that the

oil, methanolic extract and formulated cream posses prominent

activity compared with that of standard. Zone of inhibition were found

to be prominent in concentration 5g / ml of oil and 155ug / ml for C.

tora seeds extract. In antimicrobial activity of cream, the formulation

containing maximum percent of oils i.e. 5% and 155ug/ml of C. Tora

methanolic extract showed highest zone of inhibition. Validation of cream was done and was

found within the limits. The pH of cream was found to be in range of 6.5 7.0 which is good

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Article Received on 02 June 2012, Revised on 28 June 2012, Accepted on 13 July 2012

*Correspondence for

Author:

* Dhanashri B. Nagulwar, Department of Pharmaceutics,

Sharad Pawar College of

Pharmacy, Dist: Nagpur,

Maharashtra, India.

[email protected]

om

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for skin pH. The consistency of cream was in the range of 230 to 290 1/10mm. The

formulated cream was used for antiseptic action. Patient trial was carried out and successful

results were seen. For this purpose cream was applied topically. In human study, trials were

conducted on 10 patients suffering from burn infection, tinea pedis and acne. Signs of the

infection were healed in the period of 2 week.

Key Words: Skin, C. tora, tinea pedis, acne.

INTRODUCTION

India has an ancient heritage of traditional medicine. Materia medica of India provides lots of

information on traditional aspects of therapeutically important natural products. The

evaluation of these drugs is mainly based on phytochemical, pharmacological and allied

approaches including various techniques like chromatography and microscopy. [1] Herbals are

being used, in medicine from time immemorial because they have fitted the immediate

personal need, they are accessible and inexpensive. It is widely accepted that the use of

herbal medicine is well established and safe. [2]

For skin infections, topical treatments applied directly to the skin are safest. Drugs taken

orally affect both disease and normal tissues, thus increasing the chance of side effects.

Conventional skin disease treatments such as the drugs ketoconazole, ciclopirox, naftifine

and tolnaftate can irritate the skin, causing stinging, itching, redness, drying or allergic

reactions. Therefore treatment of many herbs can be used safely for superficial skin

infections.

Herbal medicine, now a days are gaining importance for treating many diseases due to their

significant effect and lesser side effects as compared to allopathic medicines.[3] Plants like

Clove, Creosote, Garlic, Chamomile, Goldenseal, Tea tree oil, Echinacea, Sassafras has

antiseptic and wound healing action. From literature survey, it was revealed that

phytochemical analysis of C. tora, j. regia, T. arjuna, k galangal confirmed the presence of

flavonoids, steroids, triterpenoids in methanolic extract and these extract exhibited highest

antimicrobial activity against S. aureus, B. subtilis, P. bacteria. Plants can be used for

antiseptic action therefore decided to use of these plants for further study.

From market survey, it was indicated that no cream is available containing combination of

these plants therefore decided to formulate antiseptic herbal cream for microbial infection.

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In present project, methnolic extract of C. tora seeds and n-hexane extract (oil) of j. regia

and k. galanga will be used for antiseptic action of cream. extracted oil are soluble in oily

phase and C. tora methanolic extract is soluble in aq. phase therefore it is expected that

extract should be compatible with cream base therefore decided to use o/w cream base for

development of herbal antiseptic topical formulation. o/w creams also cause less dryness and

burning, nonstaining and water miscible. Creams are well absorbed and tolerated by many

people. [4]

In addition to this combination of plant extracts formulation will be enriched with Aloe-vera

extract as emollient for the skin. Therefore these formulations will be expected to exhibit

better efficacy as compared in the marketed antiseptic formulation.

In present work main objective is development and validation of Herbal antiseptic cream, in

vitro evaluation of antimicrobial activity, and in vivo study, assessment of antiseptic activity

of developed cream on patients through various skin infections will be expected to exhibit

better antiseptic effect as compared in the marketed antiseptic formulation.

MATERIAL AND METHODS

Collection and authentication of plant material

The T. arjuna bark, J. rgia seeds, K. galanga rhizome, C. tora seeds were collected from the

Shri Shail Medi Pharma of Nagpur district. The plant specimen was dried and its herbarium

sheet was prepared and it is available in Department of Botany, Nagpur University, Nagpur.

Procurement of Material and Extraction

These parts of the above mentioned plants subjected to size reduction to get coarse powder.

Such powdered material was charged into the Soxhlet apparatus and extraction was carried

out using n-hexane as solvent for J. regia seeds and for K. galanga rhizomes whereas

petroleum ether (60-80o) and methanol (GR) for T.arjuna bark and C. tora seeds. Various

Phytochemical screening tests are carried out such as tests for sterol like salkowaski test,

liebermanns test, liebermann-burchard test.[5] Tests for alkaloids like dragendorffs reagent,

mayers reagent, wagners reagent, hagers reagent. Test for saponins and test for tannins like

ferric chloride test, lead acetate test, potassium dichromate test, gelatin solution test. Test for

flavonoids (Shinoda test) and Test for proteins like biuret test, xanthoproteic test, millon's test

(Mercury nitrate solution) and test for fixed oils are carried out.

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Thin Layer Chromatography of different extracts [6-7]

Chromatographic pattern of active extract of T. arjuna bark, C. tora seeds j. regia seeds and

K. galanga rhizome was studied by thin layer chromatography to find out the number of

phytoconstituents present in them. The adsorbent material used for the thin layer

chromatography was silica gel G.

Analysis of oil [8]

The various physical constants like acid value, saponification value and ester value were

determined by using standard methods of Indian Pharmacopoeia (1996).

Acid value

Oil sample (2 g) was accurately weighed and dissolved in 50ml of mixture of ethanol (95%)

and ether (1:1), which was previously neutralized with 0.1M potassium hydroxide solution.

The flask was connected with a reflux condenser and warmed slowly with frequent shaking

until the sample was dissolved; 1ml of phenolphthalein solution was added and titrated with

0.1M potassium hydroxide until the solution remained faint pink after shaking for 30sec. The

acid value was calculated from the expression.

Acid Value = 5.61 n / m

Where,

n = ml of KOH required.

w = weight in gram of sample.

Saponification value

Accurately weighed 2g of the oil was added in 250ml flask of borosilicate glass fitted with

reflux condenser. 25ml of 0.5M ethanolic potassium hydroxide was added and boiled under

reflux on water bath for 30min, Phenolphthalein solution (1 ml) was added and titrated with

0.5M hydrochloric acid (ml) and repeated the same procedure for blank (bml).

Saponification value = 28.05

Where,

w = weight in g of the sample

Ester value

Acid value and saponification value were determined as per the Indian pharmacopoeia

procedures and the ester value was calculated from the expression,

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Ester Value = Saponification Value Acid Value

Anti-microbial activity [9-11]

The antimicrobial activity of K. galanga oil, J. regia oil , methanolic exrtract of C. tora seeds

and T. arjuna bark were studied against Gram-positive S. aureus and B. substilis, P. bacteria

and C. albicans and A. niger fungi by agar diffusion method. For antibacterial activity

streptomycin while for antifungal activity griseofulvin was used as the standard.

Screening of antimicrobial activity was carried out using the cup plate agar diffusion method.

This method depends on the diffusion of drug through the solidified agar layer of a petridish

to an extent such that growth of the inoculated microorganism prevented entirely in a circular

area zone around the cup containing the solution of the compound under test. One loopful

of the stock culture was inoculated at 10 ml of agar slant previously in sterilized test tubes,

and incubated at 37oC for 24 h and 20oC for 48 h respectively for bacteria and fungi. About

3ml of distilled water was added to the test tube and a suspension of the culture was obtained

by shaking for few minutes. The solutions were made by dissolving the test oil compound in

minimum amount of DMSO (dimethyl sulphooxide), and solid compound in minimum

amount distilled water, volume was made with sterilized water to produce a concentration of

100g/ml.

Respective sterile medium was melted on water bath and kept at 45oC. In each sterile

petridish 25ml of molten medium was added and 107/ml of sub cultured organism under

study was inoculated.

Formulation of cream [12-13]

For the convenient application of oil and other methanolic extract and by considering the

antimicrobial activity, cream was selected as a suitable dosage for topical application. Creams

are nonstaining and water miscible. Creams are well absorbed and tolerated by many people.

Cream base formula was selected for the development of herbal antiseptic cream as shown in

table 1.

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Table 1: formula for development of cream base

Sr. No Ingredients % w/ w

Oil phase

1. Stearic acid 26.56%

2. Cetyl alcohol 1.0%

3. Propyl paraben 0.01%

4. Butylated hydroxyl toluene 0.02%

Water phase

1. Propylene glycol 5.0 %

2. Glycerine 5.0 %

3. Triethanol amine 1. 0 %

5. Methyl paraben 0.01%

6. Water 61.42%

Method of preparation

Oil soluble components were dissolved in oil phase and heated to 75C. Other water soluble

components were dissolved in aqueous phase and heated to 75C. After heating, oil phase

was added in aqueous phase with continuous stirring till a thick viscous cream was obtained.

Formulation development of herbal creams of botanical extract.

Topical herbal creams of mentioned botanical extracts were prepared as per the following

way on trial and error basis.

Batch I- In this batch, a single botanical extract (C. tora seeds methanolic extract ) was

added which was compatible with above base.

Batch II- In this batch, cassia tora seeds methanolic extract and Juglans regia oil were added

which were also compatible and more emollient than the pervious.

Batch III- In this batch, all mentioned extracts were added, it was noted that all extracts were

compatible with the base formula and cream texture and color was more acceptable than the

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previous batch. Cream formulated in trial batch III was taken as the master formula and

subjected for further evaluation study. Formulas for each trial batch cream as shown in table

2.

Table 2: Formula for each batch

Ingredients Batch I Batch II Batch III

Stearic acid 26.56% 26.56% 26.56%

Cetyl alcohol 1% 1% 1%

Butylated hydroxy toluene

0.02% 0.02% 0.02%

Propyl paraben 0.01% 0.01% 0.01%

J. regia oil - 5.0% 5.0%

K. galangal oil - - 5.0%

Glycerine 5% 5% 5%

Propyleneglycol 5% 5% 5%

Triethanol amine 1% 1% 1%

Methyl paraben 0.01% 0.01% 0.01%

C. tora seeds methanol extract

1.55%

1.55%

1.55%

Aloe vera extract - - 2.0%

Water 59.85% 54.15% 54.15%

Evaluation of cream

As per the requirements for skin creams specified in Indian standards (6608: 2004), following

parameters were used for evaluation of cream.

Thermal stability

For thermal stability testing, a humidity chamber and clear glass container of around 30ml

capacities with screw cap were used. With the help of spatula cream was inserted in the glass

containers and tapped to settle to the bottom and plug was inserted and tightens the cap. The

filled bottle was kept inside the incubator at 45 10C for 48h. On removal from the

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incubator, it was noted that no oil separation or any other phase separation was not observed,

formulated cream was stable at 450C.

Determination of pH of formulated Cream

The digital pH meter was calibrated using buffer solution of pH 4.01, 7.0 and 9.2. Cream was

taken in a beaker and the pH of the cream was determined.

Determination of total fatty substance content

About 2g of the accurately weighed formulated cream was taken into a conical flask, Dilute

Hydrochloric acid (25ml) was added and a reflux condenser was fitted into the flask and the

solution was boiled until it perfectly cleared. Then contents of the flask were poured into a

300ml-separating funnel and was cooled to room temperature. In portion of 10ml the conical

flask was rinsed with 50 ml of petroleum ether and poured into the separating funnel,

separating funnel was then shaked well and left until the layers were separated. An aqueous

phase was separated and all ether extract was then washed with water. This petroleum ether

extract was then filtered through a filter paper and dried the material in the flask at a

temperature 90 20C. of to constant mass.

Formula:

Total fatty substance = 100 M1/ M2

Where,

M1= mass (g) of the residue

M2 = mass (g) of the cream

Determination of residue

About 5g of the cream was taken in a weighed, clean and dry squat form weighing bottle and

dried to constant mass at 105 10 C. Cooled in a desiccator and weighted.

Formula: Residue = 100 M1/ M2

Where,

M1 = mass (gm) of the residue

M2 = mass (gm) of the cream

Test for lead

Standard lead solution was prepared by using 1.600gm of the lead nitrate taken in water and

the solution was made to 1000ml. solution (10ml) was Pipette out and diluted to 1000ml with

water. One mm of this solution contains 0.01mg of lead (Pb). About 2.00gm of cream was

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taken in a crucible and heated on a hot plate and then taken in a muffle furnace to ignite it at

6000 C to constant mass. Dilute hydrochloric acid 3ml (5N) was added and warmed and

volume was made to 100ml. solution was then filtered. In the second Nessler' s cylinder, 2 ml

of dilute acetic acid (1N) was added, volume was made with water to 25ml. standard

hydrogen sulphide(10ml) solution was added to each Nesslers cylinder and volume was

made with water (50ml) mixed and allowed to stand for 10min and the colour produced in

two Nesslers cylinders was compared. The colored produced with hydrogen sulphide was

matched against that obtained with standard lead solution.

Spreadability

Spreadability of cream was measured with the glass slide apparatus, excess of cream was

placed between two slides and 1 kg weight was placed on slide for 5 min. to compress the

sample to uniform thickness, time in seconds to separate two slides was taken as measure of

spreadability.

S = w l / t

where,

S = spreadability (g cm/sec)

w = weight on upper slide (g)

l = length of Slide (cm)

t = time taken in sec (sec)

Homogeneity

The developed cream was tested for homogeneity by visual inspection, after the cream have

been set in the container, spread on the glass slide for the appearance, tested for the presence

of any lumps, flocculates or aggregates.

Skin irritation test

The skin irritation was carried out on human volunteers. For formulated cream, five

volunteers were selected and 1.0g of formulated cream was applied on an area of two square

inch to the back of the hand. The volunteers were observed for lesions or irritation.

Consistency

Consistency of the formulation was determined by penetrometer model no. 2. Associated

instrument Pvt. Ltd. Culcutta.

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Microbial evaluation [14]

Microbial evaluation of herbal formulations is essential to check the limits of microbial

contamination and extents of pathogencity. This evaluation has direct correlation with the

quality of products. For the evaluation of total microbial count details of different count

media were used, nutrient agar medium used for the growth of bacteria and Potato dextrose

agar medium was used for the growth of fungi. [15]

In aseptic conditions cream equivalent to one gram was dissolved in 10ml of sterile water and

was serially diluted. The medium and apparatus required for experimental were sterilized in

an autoclave at 1210C for 15min. In aseptic conditions, 1ml of test sample was transferred to

petridish containing melted agar medium at about 420C and mixed well by rotating the

petridish. It was allowed to solidify and then incubated at 370C for 24h for detection of

bacteria. After incubation period, colonies were counted.

Formula:

No. of colonies dilution factor

No. of microorganisms =

Volume of sample

Stability testing of formulated cream.

For assessing the stability of formulated creams following parameters were taken into

consideration like Thermal stability testing, pH, Total fatty substance content, Total residue,

General test for lead, Consistency, Spreadability. [16] These studies are essential to ensure that

product is stable over its designated shelf life. The stability study was carried out for three

months as per ICH norms, at three different temperatures such as at room temperature, 450C

and 8 to 100C. Results of stability testing of formulated cream are given in table.

Patients study

Realising the importance of patient acceptability the formulation was filled into 20gm

capacity container and dispensed to the patients suffering from acne, Tinea pedis, burn, in the

dermatology department of government Ayurvedic hospital Nagpur. The study was

supervised under the Dr. Kabra and Dr. Deepak Madavi and others doctors.

Each Polyherbal cream (containing combination of methanolic extract of Cassia tora seeds,

Juglans regia oil, Kaemferia galanga oil) was prescribed to the patients suffering from skin

diseases. A retrospective analysis of their record revels that the cream was well tolerated by

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the patients. Consent form was signed by the patients before prescribing cream. The

treatment was given for two weeks to the patients suffering from burn infection, Tinea pedis

and acne.

Patient study for Burn infection

Three patients suffering from burn infection were taken for patients study. Damage in

superficial burns was not deeper than papillary epidermis.[17] Clinical features were oedema

redness, bleb formation and pain due to preserved nerve endings. Compliance of patients was

good against formulated cream than the marketed preparation.

Patient study for acne

Acne is an inflammatory skin disorder of the skins sebaceous glands and hair follicles and

occurs when the cells lining the sebum canals are injured or when follicles or pores are

blocked or clogged by sebum (oil) and dead cells mixed together.

Patient of either sex, age between 21- 45 yrs. were selected, two types of pigmentation were

observed having black coloured and white coloured pigmentation, while giving the treatment

it was observed that pimple infection did not spread further, cream act effectively on P.

bacteria, sebum irritation decreases, pores did not clog further.

Patients study for Tinea pedis

Tinea pedis is the fungal infection of fact mostly involving in between the toes. It is usually

associated with prolonged exposure to moisture. It involved, skin appears blanched with

cracks and associated itching. Treatment involves the application of cream base, formulated

cream and marketed preparation (skina cream).

RESULT AND DISCUSSION

All the extracts of plant material were subjected to preliminary phytochemical screening for

the detection of various plant constituents. Phytochemical screening of methanolic extract of

T. arjuna bark, C. tora seeds showed presence of sterols, saponins, tannins, flavonoids, sugar,

proteins and coumarin. Hexane extracts of J. regia seeds and K. galanga rhizome showed

positive test for sterols, flavonoid and coumarin hence both the extracts were used for further

study.

Chromatographic pattern of above mentioned plant extracts were studied by thin layer

chromatography to find out the number of constituents present in them. Stationary phase used

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was Silica gel G and for mobile phase different combinations of the solvents were tried for

the development of the solvent system for different extracts.

Mobile phase with Benzene: Ethyl acetate (8:2) showed maximum resolution and

reproductive results. After spraying with 50 % H2SO4 and in iodine vapour gives 3 spots with

Rf values 0.34, 0.61, 0.87. of petrolium ether extract T. arjuna bark and gives 3 spots with Rf

values 0.34, 0.54, 0.74 C. of petrolium ether extract of C. tora seeds.

Mobile phase with Butanol; Acetic acid; Water (6:2:2) showed maximum resolution and

reproductive results. After spraying with 50 % H2SO4 and in iodine vapour gives 2 spots with

Rf values 0.48, 0.76 methanol extract of. T. arjuna bark and 6 spots with Rf values 0.32, 0.45,

0.57, 0.74, 0.81, 0.96 of methanol extract of C. tora seeds.

Mobile phase with Benzene: Ethyl acetate: Acetone (8:1:0.5) showed maximum resolution.

After spraying with 10 % vanniline sulphuric acid and in iodine vapour gives 4 spots with Rf

values 0.32, 0.68, 0.82.0.94 of J. Regia oil and 6 spots with Rf values 0.38, 0.46, 0.52, 0.64,

0.77, 0.84 of K. galanga oil.

As shown in Table 11, acid value, saponification value and ester value of J. regia seeds oil

were found to be 15.54, 134.5, and 118.7 and of K. galanga oil were found to be 1.2, 105.4

and 121.7 respectively as shown in table 3.

Table 3: Evaluation parameters of J. regia and K. galanga oils

It was indicated that methanolic extract of C. tora seeds, K. galanga oil and J. regia oil had

maximum antimicrobial activity whereas methanolic extract of T. arjuna bark had not

maximum antimicrobial activity as compared to that of other plants extract so T. arjuna bark

extract was not used in formulation of cream as shown in table 4, 5, 6 and 7.

Parameter J. regia Oil

K. galanga oil

Acid value 15.54 1.2

Saponification Value 134.5 105.4

Ester value 118.7 121.7

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Table 4: Antibacterial activity of K. galanga oil.

Table 5: Antibacterial activity of J. regia oil.

Microorganism Zone of Inhibition in mm

J. regia oil Streptomycin

(5g/ml) 1g/ml 2g/ml 3g/ml 4g/ml 5g/ml

P. bacteria 9 11.2 12.5 13.3 14.1 18.2

B. subtilis 10.2 11.1 12.2 13.2 16.7 21.1

S. aureus 7 9 13 15 18.7 23.2

Table 6: Antibacterial activity of methanolic extract of C. tora seeds and T. arjuna bark extract

Extracts Zone Of Inhibition in mm

B. Subtilis S. aureus P. bacteria C. tora seeds

methanolic extract (155ug/ml)

14.3 16.4 13.2

T. arjuna methanolic extract

(170ug/ml) 3 2 -

Streptomycin 5ug/ml 18.2 21.1 23.2

Table 7: Antimicrobial activity of all combined extracts

Microorganism Zone of Inhibition in mm K. galangal Streptomycin

(5g/ml) 1g/ml 2g/ml 3g/ml 4g/ml 5g/ml P. bacteria 4 7 10 13 14 18.2 B. subtilis 7 10 14 15 17 21.1 S. aureus 7 9 13 17 19 23.2

Microorganism

Zone of Inhibition in mm K. galanga

oil +

5 g/ml

J. regia oil +

5g/ml

C. tora seeds + methanolic

extract 155 ug/ml

Streptomycin (5g/ml)

P. bacteria 16.7

19.5

21.9

18.2

B. subtilis 21.1

S. aureus 23.2

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Cream formulated in trial batch III was taken as the master formula and subjected for further

evaluation study. It was indicated that evaluation parameter of formulated cream are within

the limits as per the requirements for skin creams specified in Indian standards as shown in

table 8.

Table 8: Evaluation of Cream

The results for determination of total plate count of bacteria and fungi are shown in following

table 9 and 10.

Sr. No Parameter Cream base

Cream base +

C. tora methanolic

extract

Cream base + C. tora

methanolic extract +

J. regia oil

Cream base +

C. tora methanolic

extract +

J. regia oil +

K. galanga oil

1 Thermal stability testing No phase separation No phase separation

No phase separation

No phase separation

2 pH 6.4 6.6 6.7 6.8

3 Total fatty substance content, (%/gm) 26.32%/gm 27.56%/gm 32.97%/gm 37.78%/gm

4 General test for lead Passes the test Passes the test Passes the test Passes the test

5 Total residue, (%/gm) 37.26%/gm 38.81%/gm 42.57%/gm 46.94%/gm

6 Homogeneity No lumps No lumps No lumps No lumps

7 Spreadability 10.32gm.cm/sec 10.30gm.cm/se

c 12.59gm.cm/sec 13.01gm.cm/sec

8 Skin irritation test No irritation No irritation No irritation No irritation

9

Consistency at room temperature

237 233 239 243

At 450C 1/10 mm 276 273 179 283

At 8-10 0C 1/10 mm 151 149 159 165

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Table 9: Total number of viable bacteria present in formulated cream.

Table 10: Total number of viable fungi present in formulated cream.

Formulated cream

Types of microorganism

Fungi (C.F.U./ml)

Cream base Cream base + C. tora methanolic extract

Cream base + C. tora methanolic extract + J. regia oil

Cream base + C. tora methanolic extract + J. regia oil + K. galanga oil

3.29 102 2.42 102 2.21 102 1.15 102

From stability study of cream, it was indicated that cream was stable at three different

temperatures such as at room temperature 450C and 8 to 100C.

From ANOVA, it was indicated that there were no significant difference between the

consistency, pH, spreadability, total fatty substance content, total residue content values of

the different batches of formulated cream. Calculated F ratio were less than table F value,

therefore accepted hypothesis that there were no difference between the batches at 5% level

of significance.

Three batches of cream were formulated, F1, F2 and F3 and their antimicrobial activity was

carried out and the results of zone of inhibition are as shown in table 11, 12 and figure 1 and

2.

Formulated cream

Types of microorganism Bacteria (C.F.U./ml)

Cream base

Cream base +

C. tora methanolic extract

Cream base +


+ J. regia oil

Cream base +


+ J. regia oil

+ K. galanga oil

3.12 102 2.57 102 2.39 102 1.21102

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Table11: Antibacterial activity of cream

Microorganism

Zone of Inhibition in mm Formulated cream Streptomycin

(5g/ml) F1 F2 F3 P. bacteria 17.2 17.4 17.4 18.2 B. subtilis 19.7 19.7 19.7 21.1 S. aurbeus 22.2 22.3 22.3 23.2

Figure 1: Antibacterial activity of cream

Table 11: Antifungal activity of cream

Microorg

anism Zone of Inhibition in mm

Formulated cream 5ug/ml grisevofulvin

(5g/ml) F1 F2 F3

C. albicans

23.4 23.5 23.7 25.2

A. niger 20.6 20.9 20.9 21.7

Thus, maximum zone of inhibition is found in cream containing all polyherbal extract which

is more or less equal to that of standard.

Figure 2: Antifungal activity of of formulated cream

0

5

10

15

20

25

1 2 3

F1F2F3Sreptomycin

zone of inhibition in mm

Antibacterial activit

1 . B. subtilis2. S. aureus3. P. bacteria

0

10

20

30

1 2

F1F2F3grisevofulvin

zone of inhibition

in mm

Bacterial

Antifungal activity

1.B. subtilis2. S. aureus3. P. bacteria

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From patients study it was indicated that all the parameters and signs of the disease infection

were healed in the period of 2 week.

Following photo shows the effect of base, cream and marketed preparation on patients

suffering from Tinea pedis, burn injury and acne as shown in figure 3, 4, 5, 6 and 7.

Figure 3: Effect of marketed cream on tinea pedis patients.

Before treatment after one week after two week

Figure 4: Effect of formulated cream on tinea pedis patients


Figure 5: Effect of marketed cream on acne patients


Figure 6: Effect of formulated cream on acne patients

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Figure 7: Effect of formulated cream on burn patients.

REFERENECES

1. Kumar V, Parmar NS. Herbs: A Potential Source for the Development of New

Phytomedicinals. The Pharma Review. 2003; 1: 56.

2. Mukherjee PK. Quality Control of Herbal Drugs. 1st ed., New Delhi; Business Horizon

Pharmaceutical Publishers: 2002.

3. World Health Organization. Research Guidelines for Evaluating the Safety and Efficacy

of Herbal Medicines; 1993.p. 397.

4. Khan MR, Khihana M, Omoloso AD. Antimicrobial activity of Michelia champaca.

Fitoteripia, 2003; 73: 744.

5. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy. 1st ed., Mumbai; Nirali Prakashan:

1990.

6. Stalhl E. Thin Layer Chromatography. 2nd ed., Berlin; Springer- Verlay Berlin

Before treatment After one week After two week

Before treatment After one week After two week

Before treatment After one week After two week After two week

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Publisher:2005.

7.Wagne H, Bladt S. Plant Drug Analysis a TLC Atlas. 2nd ed.,

New Delhi; CBS Publisher and Distributors: 1995.

8. The Indian Pharmacopoeia. Govt. of India, Ministry of Health and Family Welfare, The

Controller of Publication, New Delhi 1996; A- 52.

9. Copland A, Nahar AT, Tomlinson CM. Antibacterial and free radical scavenging activity

of the seeds of Argimonia eupatoria. Fitoteripia, 2003; 73: 133.

10.Khan MR, Khihana M, Omoloso AD. Antimicrobial activity of Michelia Champaca.

Fitoteripia, 2003; 70: 659.

11. Pistelli L, Betroli AT, Lepori DH. Antimicrobial and antifungal activity of crude extracts

and isolated saponins from Astragalus verrucosus. Fitoteripia, 2002; 72: 340-4.

12.Pharmacopoeial standards for Ayurvedic Formulation. Central council

for research in Airbed and Sided; 1987.p. 65.

13. Ansel CA, Howard BG. Introduction to Pharmaceutical Dosage Form. 5th ed., New

Delhi; B.I. Publication: 1999.

14. The Indian Pharmacopoeia. Govt. of India, Ministry of Health and Family Welfare, The

Controller of Publication, New Delhi 1999; A- 57.

15. Bhaskaran S. Physical Pharmaceutics. 1st ed., New Delhi; Birla Publication Pvt. Ltd:

2007.

16. Vadas EB. Stability of Pharmaceutical Products. 20th ed., London; Lippincott Williams

and Wilkins publishers: 2002.

17. Goodman and Gilmans, The Pharmacology Basis of Therapeutics, 10th ed., London;

McGraw-Hill Publication: 2001.

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