1.Formulation & Development of Anti-acne cream using Garlic clovesByMiss Sneha S. Tathe(m.tech-II) Department of cosmetic technology,Kamla Nehru mahavidyalaya, Nagpur-09, 2011-2012

2. Acne Acne (acne vulgaris) is a disorderof hair-bearing skin in which hairfollicles develop obstructing hornyplugs (comedones) as a result ofwhich inflammation later developsaround the obstructed follicles,causing tissue destruction & scarformation. It is developed at puberty (teenage)when the sebaceous glands aremost active. 3. Causes of acne Hyperactive sebaceous glands (over active lipidsecretion )Hyperkeratosis (accelerated keratinization) at hairinfundibulum. Effect of bacteria ,The number of propionibacteriumacne found in the hair duct & on the skin.fig:p.acne 4. Development & formation of acneSteps Overactive secretion of sebum withhyperkeratosis causes the hair pores tobecome narrow & blocked to formcomedo. ( Fig. C) Comedo damages & resulting ininflammation of the tissuessurrounding the opening to form redpapules. (Fig.D) Horny material &sebum overflowsinto dermis to form pustules. (fig.E) Leukocytes attack the bacteria to from pus. Granulation often occur & scar remain. 5. Classification of acne Non-inflammatory acne I. Closed comedonesII. Open comedones Inflammatory acne- I. Papules- small pink bulb appear on the skin.II. Pustules- lesion with central visible core contain pus.III. Nodules- fill with deeper pus & heal with scarring. 6. About Active (Garlic Cloves) Synonyms Allium Biological source - Allium sativum Linn., Family - Liliaceae Part used - bulb(cloves) Description - color is white to pink, characteristic & aromaticodour.aromatic & pungent in taste.1.5 to 2.5cm Specific gravity - 1.046 The intact clove of garlic contain the odorless sulphur containing aminoacid derivative (+)-s-allyl-cysteine sulfoxide.Bulbs contain 29% carbohydrates, about 56% proteins (albumin), 0.1%fat, mucilage, and 0.06 to 0.1% volatile oil, phosphorous, iron andcopper and contains allyl propyl disulphide, diallyl disulphide, alliinand allicin. garlic contains sulphur compounds. This sulphur compound is presentin Alliin, Allicin , Organosulfur compounds , Vitamins, Minerals,Proteins and Amino Acids of the garlic . 7. Chemical structure of AllicinAllicin :- 2-propene-1-sulphinotioic acid-s-2-propenylester 8. Properties & uses of garlic bulb Garlic bulb is bitter, acrid, carminative, antiseptic, stimulant. It is expectorant,and disinfectant in the treatment of pulmonaryconditions. It is used in kapha , vata , constipation, bronchitis asthma. It is used in leucoderma ,leprosy and in dental caries. Allicin has an anti-bacterial activity It is used as an anti--cholesterol, antifungal and tonic. Garlic gives anti-inflammatory property. Garlic has a high sulfur content, which is beneficial for strong, suppleand smooth skin, hair and nails Garlic used as an antioxidant property. 9. Need of work Acne is the common skin problem which usually start in adolescence resolves by the mid 20s. Nearly 85% of people develop the skin condition of acne between the ages of 12-25 years. Acneresolves slowly at the age of 30-40 years. Acne often causes embarrassment to the sufferer. These are not heal the threatening disorders, but itdefiantly cast an impact on once personal self image. It must therefore be considered as a physiological state rather than a disease and as such falls verymuch in the cosmetic rather than the clinical field. It has been found in herbal beauty aids, can be perfectly applied to the demands of moderncosmetology both in terms of efficiency and safety , treatment with plant extracts have also shown thathuman body responds well to natural substances. Many herbs have been used in the treatment of acne for centuries and there is great benefit of usingherbs for this purpose. But there still lacks a proper scientific evidence to support the claims madeabout various properties of many herbs. It is reported that garlic cloves possess anti-bacterial properties. So it can be used in treating skindiseases. More over it can be regularly used herb and close to nature, which would give more benefits ofprotecting skin. Hence, the present project work is an humble attempt aimed at scientific evaluation of anti-acneproperty of garlic clove and also to formulate and develop anti-acne cream with garlic extract for thetreatment of acne. 10. Plan of work 11. EXPERIMENTATIONAND RESULTS 12. Extraction of Plant MaterialPreparation of raw garlic extract: 100gms of skinned raw garlic cloves were weighed and crushed in the mixer for 1min. together with 100 ml of water, then allowed standing for 30min. at 250c. after filtration through whatmann no. 4 filter paper to get raw garlic extract .Deodorization of raw garlic extract Raw garlic extract was kept at a temperature of 40c in a separating funnel for 5 days . 10ml Soybean oil was mixed into above raw garlic extract and mixture was prepared. This Mixture was kept in separating funnel at a temperature of 40c for period of 60 days. Semi-transparent brown liquid started to separate at the bottom of thefunnel and gradually increased in amount. Liquid garlic extract obtained which was completely free from odor. Liquid garlic extract was collected for further study. Analysis of Extract Sr. No.Test Result1)pH5.802) Color Semi-transparent brown3) Odorcharacteristic 13. Phytochemical Screening of ExtractSR. NO. TESTOBSERVATIONRESULTSA} Tests for carbohydrates1)Molishs testViolet color ring was observed +2)Fehlings test Brick red ppt. was observed+B} Tests for proteins1)Biuret testViolet color was observed+Millions test Brick red ppt. was observed+2)3)Xanthoprotein test Orange color ppt. was observed.+Sulphur test Black color obtained.+4)C}Tests for amino acids1)Ninhydrin test Purple color obtained+2)Tyrosine testDark red color observed+3)Cysteine testBlack ppt. formed. +D} Test for saponin glycosides1)Foam testPersistent foam was observed+E} Test for enzymes1) Yellowish brown color was not+Oxidase test2) changed. +Peroxidase test3) Pink color was not changed.+Catalase test Oxygen gas was evolved.F}Test for mucilage + Extract was showed swelling.G}Test for fixed oils and fats1)Spot testOil stains on the paper were appeared+2)Saponification testNeutralization was obtained. +H}Test for volatile oil _ Red color was not obtained. 14. Formulation & Development of Anti-Acne CreamBASE PRODUCTSR. NO. Ingredients Trial Trial Trial 100 50 (g) 1 2 3(g) With 3%With 5%With 10%(%) (%) (%)(Garlic (Garlic (Garlicextract) Extract)Extract)A} Oil Phase1. Stearic acid108 7 7 3.53.5 3.52.Glyceryl33 3 3 1.51.5 1.5 monostearate3.Isopropyl1 1 1 1 0.50.5 0.5myristate4. Bees wax4 4 2 2 1 115.Propyl paraben0.150.150.150.150.0750.075 0.075B} Water Phase6.Triethanolamine1 0.60.450.450.2250.225 0.2257. Glycerin5 5 5 5 2.52.5 2.58.Water 75.778.181.25 81.25 37.6335.63 30.639.Methyl paraben0.150.150.150.150.15 0.150.15C}Active Phase10.Raw Garlic - - - -3 510 extract 15. Observation for base formulations ObservationTrial 1Trial 2 Trial 3 AppearanceThe cream was veryThe cream was oily The cream was pearly thick with the poor but has good spreading white, with good spreading capacity. capacity.spreading , easy Skinabsorbencyandemollientproperties;hence this basewasselected for furtherexperimentation. pH6.42 6.426.42From the above observation the Trial 3 formulation was selectedfor further study and trial 1 and trial 2 were rejected. 16. Analysis of Finished Product o/w Anti-acne CreamI. Physical methodII.Chemical methodIII. Microbiological method1} Physical Methodsphysical parameters of the creamsPhysical Cream BaseCream with 2% Cream with 5% Cream with10% parameters (Garlic extract) (Garlic extract) (Garlic extract) Appearance Smooth creamySmooth creamySmooth creamy Smooth creamy Color Pearly whitePearly whitePearly whitePearly whiteOdor PleasantPleasantPleasantPleasant ConsistencySemisolid Semisolid SemisolidSemisolidSpredabilityVery good Very good Very goodVery good Oily/tacky feel NoNoNo NoSkin absorbencyEasy EasyEasy Easy 17. A} Determination of pH pH of the creamParameterCream Base Cream with2% Cream with 5% Cream with 10%Garlic extract Garlic extractGarlic extract pH 6.426.33 6.246.12B} Determination of thermal stability Thermal stability of the creams (450c) for 48 hoursparameter Cream BaseCream with2%Cream withCream with Garlic extract5% Garlic10% Garlicextract extract ThermalNo SeparationNo Separation No Separation No Separation stabilityof Phasesof Phases of Phases of Phases 18. 2} Chemical Methods A} Determination of TFMTotal fatty matter of the creamsSamplesCream Base Cream with 2% Cream with 5% Cream with 10%(g)Garlic extractGarlic extractGarlic extractTFM (% by mass)12.71 12.66 12.60 12.52 B} Determination of specific gravity of cream Specific gravity of the cream SamplesCream Base Cream with 2%Cream with 5% Cream with 10% (g) Garlic extract Garlic extract Garlic extract Specific1.05541.0542 1.05231.0506 Gravity (gm/ml) 19. Antimicrobial Study of active and product 3) Microbiological MethodResults of Zone of Inhibition of Garlic extract and cream with 2% garlic extract, cream with 5% garlicextract & cream with 10% garlic extract against S. aureus5%2% 10% Garlic extractMicro-organisms Garlic ExtractFormulationCream with 2% Cream with 5% Cream with 10% Garlic extractGarlic extract Garlic extract Staphylococcus 28mm11mm21mm 23mm aureus 20. Results of Zone of Inhibition of Garlic extract and cream with 2% garlic extract, cream with5% garlic extract & cream with 10% garlic extract against E. coliGarlic extract2%5% 10% Micro- Garlic ExtractFormulation organisms Cream with 2% Cream with 5% Cream with 10%Garlic extractGarlic extractGarlic extract Escherichia coli 37mm 12mm 16mm 25mm 21. Results of Zone of Inhibition of Garlic extract and cream with 2% garlic extract,cream with 5% garlic extract & cream with 10% garlic extract againstS.epidermidisGarlic extract 5%10% 2%Micro-organismsGarlic Formulation Extract Cream with Cream with 5% Cream with 10% 2% Garlic Garlic extractGarlic extract extractStaphylococcus epidermidis 24mm 13mm20mm22mm 22. Results of Zone of Inhibition of Garlic extract and cream with 2% garlic extract,cream with 5% garlic extract & cream with 10% garlic extract against P. acne Garlic extract 2% 5%10%Micro-organismsGarlicFormulation ExtractCream with 2% Cream with 5% Cream with 10% Garlic extractGarlic extractGarlic extractPropionibacterium 22mm10mm15mm20mmacneResult: - From the above observations it was decided to select creamwith 10% garlic extract for subjective evaluation. 23. Accelerated Stability Studies Determination of color changeThere was no color change was found in cream base and cream at 50 0c,roomtemperature & at 40c with 2% garlic extract ,cream with 5% garlic extract and creamwith 10% garlic extract thus it can be concluded that the sample of anti-acne cream isstable. Determination of Odor ChangeThere was no odor change was found in cream base and cream at 50 0c,roomtemperature & at 40c with 2% garlic extract ,cream with 5% garlic extract and creamwith 10% garlic extract thus it can be concluded that the sample of anti-acne cream isstable. Freeze Thaw TestThere was no color, odor, consistency change found in formulations .There wasno change in the color, odor, consistency of the formulations, thus it can beconcluded that the sample of the cream is stable at all temperature conditions. 24. Determination of pH Change at Different Temperature 6.5pH of product kept at 50 0c base pH 6 5.5 2%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 605% Time in days10%pH of product kept at room temp.6.5basepH65.5 2%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 5% Time in days 10% 6.6 pH of product kept at 4 0c 6.4base pH 6.2 62% 5.85%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 6010% Time in daysThere was only minor difference in the pH of the product at the different conditions of thetemperature found as compared with the initial. In all temperature conditions the pH wasalmost same. since not much changes found as compared to initial, thus it can be concludedthat the sample of the anti-acne cream is stable in all temperature conditions in term of pH. 25. Determination of Viscosity Change at Different TemperatureViscosity of product kept at 50 0c300000 viscosity in Cp200000base1000002% 05%Times in week1 week 2 week 3 week 4 week5 week 6 week7 week 8 week10% Time in weeks Viscosity of product kept at room temp. 400000Vscisity in base 200000 Cp 2% 5% 0 Times in week1 week2 week 3 Time in weeks week 4 week 5 week 6 week 7 week 8 week 10%Viscosity of product kept at 4 0c 400000Vscosity in Cp 200000 base0 2%Times in 1 week 2 week 3 week4 week 5 week6 week7 week 8 week5% week10%Time in weeksThere was only minor difference in the viscosity of the product at the different conditions of thetemperature found as compared with the initial. These changes have been recorded in the table.Since not much significant change was found in the viscosity of the product as compared to theinitial, thus it can be concluded that the sample of anti-acne cream is stable in all temperatureconditions in term of viscosity. 26. Determination of Globule Size at Different TemperatureGlobule size of product kept at 50 0c3gGlobule size2Base in m 2%1 5%0 10%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60time in daysGlobule size of product kept at room temp.3Globule size in2 Base m1 2%5%0 10%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 Globule size of product kept at 4 0c3 Globule size in m2Base12%0 5%2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56 58 60 10% Time in daysThe average particle size of the sample products were found to be slight increasing with time. There wasminor difference in the particle size of the product at the different temperature found as compared withinitial. These changes have been recorded in the table. Since not much significant change was found in theparticle size of the product as compared to the initial, thus it can be concluded that the sample of anti-acnecream is stable in all temperature conditions in term of particle size. 27. Subjective evaluationIt was decided to evaluate the effectiveness of anti-acne cream using garlic extracton 20 volunteers suffering from acne condition of age group between 18-30 years Procedure: Photographs of skin of all the subjects were taken. All the subjects were asked to apply cream with 10 % garlic extract on forearm for2 hours to check any skin sensitivity. All the subjects were instructed to apply cream with 10 % garlic extract on theirskin twice a day for one month. All the subjects were given a evaluation sheet and then they were asked to evaluatethe product for the above functional parameters on the basis of notations given inthat sheet. -ve : no reduction in sebum, no reduction in acne , no skin improvement. +ve : reduction in sebum, reduction in acne, skin improved. NI : no irritancy SE : spread easily. G Gender, A - Age 28. Evaluation of the Anti-acne cream with 10 % of garlic extract SubjectsParameter for the evaluation of productSr. No.GASkin ConsumerReduction spreading Reduction irritation improveEasy ofappeal sebumin acne typeSkin Skinin 1Female18Oily SE NI +ve +ve +veP 2Female19 NormalSE NI +ve +ve +veP 3Female20Oily SE NI +ve +ve +veP 4Female23Oily SE NI +ve +ve +veP 5Female26Oily SE NI +ve +ve +veP 6Female19DrySE NI +ve +ve +veP 7Female24oily SE NI +ve +ve +veP 8Female22 combination SE NI +ve +ve +veP 9Female15 combination SE NI +ve +ve +veP10Female17 combination SE NI +ve +ve +veP11Female22 NormalSE NI +ve +ve +veP12Female17 combination SE NI +ve +ve +veP13Female23Oily SE NI +ve +ve +veP14Female25Oily SE NI +ve +ve +veP15 Male 20 NormalSE NI +ve +ve +veP16Female20 combination SE NI +ve +ve +veP17Female25 NormalSE NI +ve +ve +veP18Female23 NormalSE NI +ve +ve +veP19 Male 19Oily SE NI +ve +ve +veP20 Male 23Oily SE NI +ve +ve +veP 29. Photographic Presentation of Product(with 10% active) Photograph of Volunteers before and after 1 monthCase I Case - IIICase - IIIBefore useAfter use Before useAfter use Case - II Before useAfter useFormulation containing garlic extract 10% was found to be effective in severeacne and several related problems. From consumer point of view the formulation were effective in reducing thesebum to control the infection and also to prevent the scaring and pimple marks. 30. 6. DISCUSSION & CONCLUSION Acne is a most common and more or less for every individual, affecting more than85% of teenagers of both sexes. It is significant disorder of skin during theadolescence. The study was planned to exploit the properties of garlic extract. It wasobserved during the literature survey that garlic extract has antiseptic, antibacterialand anti-inflammation properties. So it was decided to incorporate in the cream.For this purpose Garlic cloves were extracted. The properties of garlic were studiedfor various strains such as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli ,Propionibacterium acne which are responsible for acne. The resultwas indicated the fact that the extract is an effective bactericide. The cream baseformulation were selected and garlic extract was incorporated in it. Stability of the formulation is the important aspect for their shelf life and consumeracceptance. The stability study of the cream was carried out and the parametersstudied were color, odor, pH, and particle size. Accelerated stability study and the cream evaluation of the product indicate that theformulations are quite stable. The extract of garlic clove gave white color to cream. It was accepted by consumer because of its anti-bacterial activity against acne. The10% anti-acne cream was appreciated by volunteers. It reduced acne in volunteersafter application. The result of the study indicates that garlic extract is definitely having anti-acneproperty and it can be incorporated in cream base product. 31. 7. FUTURE SCOPEGarlic extract can be studied for anti-dandruffand antifungal properties. Garlic extract can be studied insect repellentproperty and can be used in the incense stickfor their insect repellent property. Garlic extract can be studied for antibacterialproperty and can be used in foot spray fortheir healing property. The antioxidant activity in the extract can befound out by the use of advanced techniquessuch as GC-MS, HPLC, NMR and UV. More stability parameters & toxicity studiescan be carried out further. 32. 8.REFERENCES Harsh Mohan; Textbook of pathology; Jaypee Brothers medical publication; 11th edition; 768. Michael S. Roberts, Kenneth A.Walters ; Dermal Absorption and Toxicity Assessment ; Informa HealthCare publication;2nd edition; vol.91;2007; 1. Takeo Mistui; New cosmetic science; Elsevier publication; 1st edition ;1997; 13,14,15. Barbara Daly ; daily Beauty Magazine ; 11,12. Doris.J.Bay; 100 Questions & Answers about acne; higher education and professionalpublication;2005;v,2. http://www.google.co.in/imgres?q=skin+structure&hl=en&gbv=2&biw=800&bih=466&tbm=isch&tbnid=lJL0qMEoXpLgM:&imgrefurl=http://cosbiology.pbworks.com/w/page/11556260/LESSON%2520601%2520%25E2%2580%2593%2520Skin%2520Structure%2520and%2520Function&docid=Zm3fUQGGa7jSAM&imgurl=http://cosbiology.pbworks.com/f/1248965341/module%2525206%252520%25252001.png&w=800&h=609&ei=4mKRT7-7KcuIrAecmiBQ&zoom=1&iact=hc&vpx=370&vpy=136&dur=4570&hovh=196&hovw=257&tx=103&ty=218&sig=113426323437045387229&page=3&tbnh=125&tbnw=164&start=20&ndsp=12&ved=1t:429,r:2,s:20,i:186 P. N. Bahl; Practice Of Dermatology ; 5th edition; 1982;23. Ronald Marks; Roxburghs common skin diseases; ELBS publication; 16th edition; 1995; 147. M. S. Balsam & E.Sagarin; Cosmetic Science & Technology ; 2nd edition; vol.3; 1957; 163.Takeo Mistui; New cosmetic science; Elsevier publication; 1st edition ;1997; 29-30. Takeo Mistui; New cosmetic science; Elsevier publication; 1st edition ;1997; 30-31. http://188.120.254.79/img/care_and_prevention/acne-stages_2_3790.jpg V.kumar; A.K. Abbas; N.fausto; J.C. Aster; Robbins & Carton pathologic basis of disease; Elsevierpublication ; 8th edition;2010;1197. Mark Lees Skin Care Beyond the Basics; 3rd edition; 2007; 217. 33. Mark Lees Skin Care Beyond the Basics; 3rd edition; 2007; 218. Vinod D. Rangari; Pharmacognosy & Phytochemistry; career publication; 1st edition; 2003;184,185. Soap, Detergent & Toiletries Review latest in soap, detergents, cosmetics, personal care ingredient andtoiletries magazine; vol. 41; 2010; 31. Orient Longman; Indian medicinal plants; Orient Longman pvt.; volume 1;1994; 93. G.K.Kokate, A.K.Purohit,S.B. Gokhale; pharmacognosy; Nirali prakashan;32nd edition; 9.184.Vinod D. Rangari; Pharmacognosy & Phytochemistry; career publication; 1st edition; 2003;185. G.K.Kokate, A.K.Purohit,S.B. Gokhale; pharmacognosy; Nirali prakashan;32nd edition; 9.184. K.M.Nadkarni; Indian Material Medica; popular prakashan pvt.ltd.; volume-I ; 1997; 66. http://hmtjournals.com/vol2_2/04.pdf Atta Ur-Rahaman; bioactive natural product chemistry; Elsevier publication; partD;vol.23;467. Atta Ur-Rahaman; bioactive natural product chemistry; Elsevier publication; partD; vol.23;468. www.nutritiondata.scif.com/facts/vegetables-and-vegetable-products/2446/2Soap, Detergent & Toiletries Review latest in soap, detergents, cosmetics, personal care ingredient andtoiletries magazine; vol. 41; 2010; 31,32. http://en.wikipedia.org/wiki/File:Cysteine-to-allicin-2D-skeletal.png Orient Longman; Indian medicinal plants; Orient Longman pvt.; volume 1; 1994; 93. G.K.Kokate, A.K.Purohit,S.B. Gokhale; pharmacognosy; Nirali prakashan;32nd edition; 9.184. Dr. K.M.Nadkarni; Indian Material Medica; popular prakashan pvt.ltd.; volume-I; 1997;66-69. http://www.phytochemicals.info/research/garlic-anti-inflammatory.Soap, Detergent & Toiletries Review latest in soap, detergents, cosmetics, personal care ingredient andtoiletries magazine; vol. 41; 2010; 35.Ferejdoon Shahidi, Chi-Tang-Ho; Phytochemical & PhytopharmaceuticalsAocs books and specialpublication; 2000; 63.http://www.freepatentsonline.com/4377600.html Khandelwal K.R.; Practical Pharmacognosy Techniques &Experiments; Nirali prakashan; 9th edition;1991;149. Khandelwal K.R.; Practical Pharmacognosy Techniques &Experiments; Nirali prakashan; 9th edition;1991;150. th 34. Khandelwal K.R.; Practical Pharmacognosy Techniques &Experiments; Nirali prakashan; 9th edition1991;153,154. C.K.Kokate; Practical pharmacognosy;Vallabh prakashan; 1990; 29. C.K.Kokate; Practical pharmacognosy;Vallabh prakashan; 1990; 28. G.K.Kokate, A.K.Purohit,S.B. Gokhale; pharmacognosy; Nirali prakashan;32nd edition; 8.13 .Textbook of Michael & Irene Ash ; Handbook Of Green chemicals;Synapse Information ResourcesInc.; 2nd edition; 888. Simon Quellen Field; Why theres Anti-Freeze in your toothpaste: The Chemistry of House HoldIngredients; Library of Congress Cataloging-in publication; 1st edition; 133. Paul Beringe; Remington: The Science And Practice Of Pharmacy; 21st edition ; 2006; 1086. G.K.Kokate, A.K.Purohit,S.B. Gokhale; pharmacognosy; Nirali prakashan;32nd edition; 9.166-9.167. Michael & Irene Ash ; Handbook Of Green chemicals;Synapse Information Resources Inc.; 2ndedition; 855. Michael & Irene Ash ; Handbook Of Green chemicals;Synapse Information Resources Inc.; 2ndedition; 910. Richard Mayers; The 100 Most Important Chemical Compounds; Greenwood publication ; 2007; 138. George W.A. Milne; Commercially Important Chemicals; 2005; 411. B.M.Mithal ; Textbook of Pharmaceutical Formulation;2nd edition; Vallabh Prakashan; 1980;90 Indian Standard: Skin creams Specifications; IS 6608:2004; 2nd Revision Bureau of IndianStandards,New Delhi, 2004; 3. Indian Standard: Skin creams Specifications; IS 6608:2004; 2nd Revision Bureau of IndianStandards,New Delhi, 2004; 3. Indian Standard: Skin creams Specifications; IS 6608:2004; 2nd Revision Bureau of IndianStandards,New Delhi, 2004; 3-4. L. Lachmann; The Theory & practice of Industrial Pharmacy; 4th edition; 1993; 527. th 35. Thank you