Analytical cosmetics:BIS specification and analytical methods for shampoo, skin-cream and toothpaste

Analytical cosmetics:BIS specification and analytical methods for shampoo, skin-cream and toothpaste, Indian standards, Schedule S and Schedule Q, Drugs and cosmetic act, standards for heavy metals in cosmetics, standards for coloring agents in cosmetics, annexures, list of approved colours and dyes as per schedule Q Part 1, list of colours permitted to be used in soaps, types of toothpaste, general requirements for ECO mark and BIS mark, BIS certification marking

1. ANALYTICAL COSMETICS: BIS SPECIFICATION AND ANALYTICAL METHODS FOR SHAMPOO, SKIN-CREAM AND TOOTHPASTE Presented by:- STEFFI THOMAS Assistant Professor SOP,LNCTU Bhopal

2. BIS -stands for Bureau of Indian Standards The quality and safety by Central Drugs Standard Control Organization (CDSCO) (Regulatory control over the quality of drugs, cosmetics and medical devices) As per Drug and Cosmetics Act 1940 and rules 1945, cosmetic means ‘ Any article intended to be rubbed, poured, sprinkled or sprayed on, or introduced into, or otherwise applied to the human body or any part thereof for cleansing, beautifying, promoting attractiveness or altering the appearance, and includes any article intended for use as a component of cosmetic ’. INTRODUCTION

3. Cosmetic products in India are regulated under the D & C act 1940 and rules 1945 and labelling declaration by BIS. BIS sets standards for cosmetics for the products which are under Schedule S of Drugs and Cosmetics rules 1945. BIS is the national standards body of India working under the aegis of Ministry of Consumer Affairs, Food and Public Distribution, Government of India. Cont..

4. According to IS 6608:200 , if all the raw materials requiring a test for heavy metals have been so tested and comply with the requirements, then the manufacturer may not test the finished cosmetic for heavy metals and arsenic. There should be a declaration for heavy metals and hexachlorophene (anti-bacterial) content for registration of cosmetic products. The test report should include result of lead, arsenic, mercury , other heavy metals and if any microbiological test. Undertaking from the manufacturer stating compliance of all the materials/pigments used, heavy metals (with the specified limits) and Hexachlorophene content in products with BIS and D & C rules 1945. Standards for heavy metals in cosmetics

5. While manufacturing skin creams and lipstick if dyes colour (pigments like lakes) are used then they shall comply with IS 4707 (Part I) It should be according to Schedule Q (which describes the list of dyes, colours and pigments permitted to be used in cosmetics and soaps) and CDSCO. Standards for coloring agent in cosmetics

6. Schedule Q is divided into 2 parts:- PART I :-It consists of the list of dyes, colours and pigments permitted to be used in cosmetics and soap as per amended by the BIS from time to time. PART II :-It consists of list of raw materials not permitted and not safe to be used in cosmetics Cont..

7. As per Schedule Q synthetic organic colours and natural organic colours used in cosmetics that are permitted for use shall not contain more than- - 2 ppm of Arsenic (calculated as Arsenic Trioxide) - 20 ppm of Lead (calculated as Lead) - 100 ppm of heavy metals other than Lead Cont..

8. Guinea green B Tartrazine Flaming red Amaranth Orange G Deep red (Maroon) Indigo Eosin TS Deep maroon ( Fanchon Maroon) Food yellow 18 Natural orange Lake red D Acid fast green List of approved colours and dyes as per Schedule Q Part I

9. Phthalocyanine blue Citrus red No. 2 Pigment yellow 3 Oil red no. 1/Solvent red 24/Oil red 3R Aqueous green paste Pigment orange 5 List of colours permitted to be used in soaps

10. Other ingredients must comply with the provisions of IS 4707 (Part II) Rule 134 of D & C rules prohibited on the use of cosmetics containing dyes, colours and pigments other than those specified by the BIS (IS 4707 Part I as amended) and Schedule Q According to Rule 145 of D & C Rules, the use of lead and arsenic compounds in cosmetics for the purpose of colouring is prohibited. The import of cosmetics in which a lead or arsenic compound has been used for the colouring purpose is not allowed under Rule 135. Cosmetics containing mercury compounds are not manufactured and imported under Rule 145 D and 135 D

11. IS 6356:2001. This standard prescribes the requirement and the methods of sampling and test for toothpaste. DENTIFRICE - It is any substance or combination of substances specially prepared for the public for cleaning the accessible surfaces of teeth. TOOTHPASTE - It is defined as a dentifrice in the form of a smooth, semi-solid, homogeneous mass containing acceptable ingredients such as abrasives/polishing agents, surfactants, humectants, binding agent and other appropriate substances for oral health maintenance. The product can be opaque, transparent or combination thereof, colored or white, packed in a suitable container from which it can be extruded in the form of a continuous mass. I.ANALYTICAL METHOD FOR TOOTHPASTE

12. TYPES :-Type 1-Non-fluoridated Type 2-Fluoridated SPECIFICATIONS 1. Composition :-The toothpaste shall not contain mono or disaccharides, e.g. sucrose or other readily fermentable carbohydrates. All the raw materials used shall conform to respective IS wherever they exist. 2. Dispensing :-The paste shall extrude from the collapsible tube or any other suitable container in which it is packed, at 27±2ºC in the form of continuous mass with the application of normal force, without the application of excessive force which would cause injury to the tube or container. It shall be possible to extrude the bulk of contents from the tube or container from the crimped end of the tube by rolling the tube gradually.

13. 3. Stability :-The toothpaste shall not show any physical sign of deterioration during normal conditions of storage and use. When subjected to a temperature of 45±2ºC for a period of 28 days the toothpaste should meet the requirements of the standard. 4. Packaging Material Inertness :-The collapsible tube or any other suitable container used for packaging should not corrode, deteriorate or cause contamination of the toothpaste during normal condition of storage and use. There should be no sign of corrosion, chemical attack or other damage.

14. 5. Acceptance test :-Toothpaste should comply with the requirements given in Table 1, when tested according to the methods given in Annexure B to Annexure G. 6. Shelf Life :-It should be declared by he manufacturer for all types of toothpaste. Manufacturing date (month and year) should be mentioned on tube and carton. The expiry date or “Best before use” shall be mentioned on the tube and carton. During the shelf life the product will meet the requirement of the standard.

15. 7.Additional requirements for the ECO-mark (Optional) (a) GENERAL REQUIREMENTS The product shall conform to the requirements for quality, safety and performance prescribed under the general requirements All the ingredients that go into the formulation of cosmetics shall comply with the provisions of IS 4707 (Part 1 and 2). The product shall meet other specific requirements as given in the standard. The product package shall display a list of key ingredients in descending order of quantity present.

16. The product should not be manufactured from any carcinogenic ingredients. The manufacturer should produce to BIS environmental clearance from the concerned state pollution control board as per the provisions of the Water (Prevention and Control of Pollution) Cess Act, 1977 and the Air (Prevention and Control of Pollution) Act, 1981 along with the authorization, if required under the Environment (Protection) Act, 1986 and the Rules made there under, while applying for ECO-mark. Additionally, provisions of the D & C Act, 1940 and the rules there under shall be complied with.

17. (b) SPECIFIC REQUIREMENTS Heavy metals like Lead and Arsenic shall not exceed 20 and 2 ppm respectively when tested by the respective method prescribed in IS. 8. Abrasivity (Type test) :-The toothpaste shall not exceed the limits of dentin abrasivity that of 2.5 times when tested as per the procedure given in Annex H. It may be noted that type test is recommended to be done on the formulation only once to pass the above criterion. This test need not be done for each and every batch. However, this test is a must if the abrasive system is changed in the formulation. It is not required for the new formulation as long as abrasive components are not changed.

18. 9. PACKING AND MARKING :- Packing :-Toothpaste shall be packed in collapsible tube or in any other suitable containers like sachets, pumps or other suitable dispensing systems. When packed in containers, the containers shall be properly sealed and have a leak-proof cap or closure. The containers, if necessary, may further be packed in cartons or any other suitable packaging material. The material for product packaging shall meet the parameters evolved under the scheme of labelling environment friendly packaging/packaging materials.

19. Marking :-The tubes and the cartons should be legibly marked with the following information:- Name and type of toothpaste Name and address of the manufacturer Net mass or volume of the material in the tube Batch number, in code or otherwise Month and year of manufacture Fluoride ion content in ppm for Type 2 toothpaste Expiry date or “Best use before…..” Foaming/Non-foaming List of key ingredients *Note:-this is exempted in case of pack sizes of 30g/60 ml or less

20. 10. BIS certification marking :- (a)The use of the standard mark is governed by the provisions of the Bureau of Indian Standards Act, 1986. and the Rules and regulations made thereunder . The details of the condition under which the license for the use of the standard mark may be granted to manufacturers or producers may be obtained from the Bureau of Indian Standards. (b)If the product is covered under ECO-Mark (optional), it shall be suitably marked with ECO-Mark logo besides Standard mark. The label may clearly specify that ECO-Mark is applicable to the contents or the package or both, as the case may be.

21. 11. Sampling :- Representative test samples of the material shall be drawn as prescribed in IS 3958. Number of tests and criteria for conformity-the tests for abrasivity , stabililty and container’s inertness shall be type tests and shall be performed for product approval whereas tests for dispensing, fineness, pH, heavy metals, arsenic, foaming power, fluoride content and microbial counts shall be carried out on each batch for acceptance of the product.

22. 12.Quality of Reagent :-unless specified otherwise, pure chemicals and distilled water (IS 1070) shall be employed in tests. *Note- ‘Pure chemicals’ mean chemicals that do not contain impurities., which affect the result of analysis.

23. S.NO. CHARACTERISTIC REQUIREMENT FOR NON-FLUORIDATED REQUIREMENT FOR FLUORIDATED METHOD OF TEST, ANNEXURE 1. Fineness (a)Particles retained on 150 μ IS sieve, % by mass, Max (b) Particles retained on 75 μ IS sieve, % by mass, Max 10 2.5 1 2.5 B - 2. pH of aqueous suspension 5.5-10.5 5.5-10.5 C TABLE 1 REQUIREMENTS FOR TOOTHPASTE

24. S.NO. CHARACTERISTIC REQUIREMENT FOR NON-FLUORIDATED REQUIREMENT FOR FLUORIDATED METHOD OF TEST, ANNEXURE 3. Heavy metals (Lead), ppm , Max 20 20 D 4. Arsenic, ppm , Max 2 2 E 5. Foaming power, ml, Min 50 50 F 6. Available fluoride ion, ppm , Max 50 1000 G 7. Microbial counts (a)Total viable counts per gram, Max (b)Gram negative pathogens per gram, Max 1000 Absent 1000 Absent IS 14648 IS 14648

25. Raw materials used in the toothpaste formulation fall into the following categories Polishing agents Surfactants Humectant Binding agent Others as per IS 4707 (Part 1 and Part 2) ANNEXURE A (INGREDIENTS CONVENTIONALLY USED IN THE MANUFACTURE OF TOOTHPASTE)

26. Squeeze the toothpaste and feel the presence of the particles/agglomerates/granules. Subject the toothpaste suspension to an ultrasonic treatment and pass through fineness test. Ultrasonification loosens out the agglomerates/granules into the constituent materials. ANNEXURE B (DETERMINATION OF FINENESS)

27. It is determined by using pH meter ANNEXURE C (DETERMINATION OF pH)

28. The colour produced with thioacetamide reagent in test solution is matched against the obtained with standard Lead solution ANNEXURE E (DETERMINATION OF ARSENIC) ANNEXURE D (DETERMINATION OF HEAVY METALS)

29. Strict attention should be paid to all the details of the procedure in order to ensure concordant results A suspension of the material in water is taken in a graduated cylinder and it is shaked 12 times under prescribed conditions The volume of the foam formed is observed after keeping the cylinder for 5 minutes. ANNEXURE F (DETERMINATION OF FOAMING POWER)

30. Water soluble species are converted to fluoride ion by acid hydrolysis. The fluoride ion activity is then determined potentiometrically with the help of fluoride ion sensitive electrode. ANNEXURE G (DETERMINATION OF FLUORIDE ION)

31. This annexure identifies the procedure for determination of dentifrice abrasivity using the RDA laboratory method. RDA-Relative Dentin Abrasivity (it is a scale that assigns the dentifrice a abrasivity value relative to standard reference abrasive) Abrasives include fluorides such as Sodium Fluoride, Stannous Fluoride or carbonates such as Calcium carbonate, Magnesium carbonate or oxides such as Aluminium oxide ANNEXURE J (LIST OF ADJUNCT INDIAN STANDARDS) ANNEXURE H (ABRASIVITY (RDA) MEASUREMENT TEST)

32. IS 7884:2004 (specification for surfactant based shampoo) II.ANALYTICAL METHODS FOR SHAMPOO S.NO. REQUIREMENTS DETAILS 1 Description Shampoo shall be in liquid, emulsion or a paste form. It may be coloured and perfumed 2 Physical characteristics When visually examined it should be free from any sediments. If it is in the form of any emulsion, it should be stable. When it is in the form of a paste, it should be free from agglomerated particles 3 Ingredients Raw materials should conform to the requirements prescribed in IS

33. S.NO. REQUIREMENTS DETAILS 3.1 Dyes It should comply with the provisions of IS 4707 (Part I), subject to the provision of Schedule Q of the Drugs and Cosmetics act, issued by Government of India 3.2 Other ingredients Shall comply with the provisions of IS 4707 (Part 2) 3.3 List of ingredients used conventionally in formulation of shampoo Annexure A 3.4 Compliance with Table 1 when tested in accordance with the method prescribed

34. 3.5. Additional requirements for ECO mark :- 3.5.1.The product shall conform to the requirements for quality, safety and performance 3.5.2.All the ingredients that goes into the formulation of cosmetics shall comply with the provisions of IS 4707 (Part I and Part II) 3.5.3.The product package shall display a list of key ingredients in descending order of the quantity present. 3.5.4.The product shall not be manufactured from any carcinogenic ingredients 3.5.5.The manufacturer shall produce to BIS environmental consent clearance from the concerned State Pollution Control Board as per the provisions of the Water (Prevention and Control of Pollution) Cess Act, 1977 and the Air (Prevention and Control of Pollution) act, 1981 along with authorization, if required under the Environment (Protection) act, 1986 and the rules made there under shall also comply with.

35. 3.6. Specific Requirements :- 3.6.1.Product shall be dermatologically safe when tested as per IS 4011 3.6.2.Biodegradable agents wherever used in cosmetic formulations shall be as per their limit finalized for synthetic detergents for ECO mark by the technical committee 3.6.3.Heavy metals calculated as Lead and Arsenic shall not exceed 20 and 2 ppm respectively when tested by the respective method prescribed in relevant Indian standards. 3.6.4.The material for product packaging shall meet the parameters evolved under the scheme of labelling environment friendly packaging/packaging materials.

36. 4. Packing and Marking :- 4.1.Shampoo shall be packed in glass or plastic containers or any other suitable containers 4.2.The container shall be legibly marked with the following information: Name of the material Manufacturer’s name and/or his recognized trademark, if any; Net content in volume for liquids and emulsions and in mass for pastes Month and year of manufacturing/packing Batch or lot number in code or otherwise ‘Best before use…’ (month and year to be declared by the manufacturer)

37. 4.3.BIS Certification marking- the container may also be marked with the standard mark. 4.3.1.The use of the standard mark is governed by the provisions of BIS act, 1986 and the rules and regulations made there under. The details of conditions under which the license for the use of Standard mark may be granted to manufacturers or producers may be obtained from the BIS 4.3.2.If the product is covered under ECO mark (optional), it shall be suitably marked with the ECO mark logo besides the standard mark. The label may clearly specify that ECO mark is applicable to the contents or the package or both, as case may be. If the product package or both, as case may be. If the product package is not separately covered under ECO mark scheme, it shall be clearly mentioned on the product that ECO mark label is applicable to contents only.

38. 5. SAMPLING 5.1.Representative samples of the material shall be drawn as prescribed in IS 3958 5.2.Tests for all the requirements shall be carried out on a composite sample 5.3.The shampoo shall be taken t have conformed to this standard if the composite sample passes all the tests.

39. 6. Quality of reagents Unless specified otherwise, pure chemicals and distilled water (see IS 1070) shall be employed in tests. ‘Pure chemicals’ shall mean chemicals that don’t contain impurities which affect the results of analysis.

40. ANNEXURE A (List of raw materials used in the formulation of surfactant based shampoo) A.1. DETERGENTS Sodium or potassium or ethanolamine salts of lauryl sulphonic acid Lauryl ether sulphates Sulphated monoglycerides Sodium alkyl sulpho -acetate Alkyl benzene polyoxyethyl sulphonates Sodium n- lauryl sarcosinate Sodium alpha olefin sulphonates Other synthetic detergents

41. A.2. FOAM STABILIZERS Ethanolamides or isopropanolamides of fatty acids Amine oxides Cocobetaines Cocoamidopropyl betaines A.3. CHELATING AGENTS Sodium polyphosphates Sodium salt of Ethylenediamine tetra acetic acid (EDTA)

42. A.4. SOLUBILIZING AGENTS Urea Aliphatic alcohol Sodium toluene sulphonate Sodium xylene sulphonate A.5. PRESERVATIVES Alcohols Formaldehyde Esters of para-hydroxy benzoic acid Sorbic acid Imidazolidinyl urea

43. A.6. OPACIFYING AGENTS Higher fatty alcohols Ethylene/propylene glycol stearates Mono and di-stearates of glycerol Zinc, calcium and magnesium salts of fatty acids PEG- Distearate 6000 Polyacrylates A.7. INORGANIC SALTS Sodium chloride Sodium sulphate Sodium phosphate Ammonium sulphate Ammonium phosphate Ammonium chloride

44. A.8. OTHER GROUPS OF INGREDIENTS Perfumes Dyes Conditioning agent Quarternary compounds Vitamins Vegetable oils Silicones Proteins Sunscreens A.9. EMOLLIENT Lanolin and its derivatives A.10. THICKENING AGENT Sodium carboxymethyl cellulose Methyl cellulose Methyl isopropyl cellulose Guar gum

45. ANNEXURE B - (Determination of non-volatile alcohol soluble matter) B-1 GENERAL This method determines the amount of non-volatile alcohol matter in surfactant based shampoos. B-2 REAGENTS B-2.1 Ethyl alcohol -Neutral, conforming to IS 321 B-2.2 Methyl red indicator solution -Dissolve 0.1 g of methyl red in 300 ml of ethyl alcohol and 200 ml of water B-2.3 Potassium chromate solution -10% solution B-2.4 Nitric acid dilute -1:4 (w/v) B-2.5 Silver nitrate solution -0.1 M

46. B-3 PROCEDURE B-3.1 Weigh accurately about 10 g of the sample into a 150 ml beaker. Evaporate on a steam-bath to almost complete dryness. Digest with 50 ml of 96% ethanol by heating on a steam bath for 2 min. Filter the hot alcoholic solution through a sintered glass filter funnel fitted to a Buchner flask to which suction is applied. Wash the beaker and the residue in the sintered glass funnel 5 times with 30 ml portions of hot ethanol.

47. B-3.2 Transfer the filtrate in the buchner flask to a weighed wide mouthed flat-bottomed flask. Evaporate nearly to dryness on a water bath and drive off the remaining alcohol by directing a gentle steam of dry air into the flask whilst continuously rotating the latter in the water bath. Heat the flask in an air over at a temperature of 105ºC until constant mass. Calculate mass percent of residue obtained Mass (% residue γ )=Mass of residue obtained ×100 Mass(g) of the material taken for test

48. B-3.3 Dissolve the residue in 50 ml of distilled water and add to it 2 drops of methyl red indicator solution. If the solution is yellow in colour , neutralize it by adding dilute nitric acid drop by drop to get pink colour . Titrate the solution with AgNO 3 solution using 2.5 ml of Potassium chromate solution as indicator, till a brown colour is obtained. Carry out a blank determination using the same quantity of all reagents except the sample.

49. B-3.4 Calculate the chloride content in shampoo in terms of molecular mass of NaCl in % by formula:- NaCl (in % X)= V×0.584 M Where , V=Volume (in ml) of standard AgNO 3 solution required for the material minus volume (in ml), of standard AgNO 3 solution required for the blank. M= Mass (in g) of the material taken for test.

50. B-3.5 Calculation To calculate % non-volatile alcohol soluble matter, subtract the mass percent of NaCl as determined in B-3.4 (X) from the mass percent of the residue (Y) obtained in B-3.2. % Non-volatile alcohol soluble matter=Y-X

51. ANNEXURE C (Determination of pH) pH meter equipped with glass rod. Determination of pH to be done at 27ºC

52. ANNEXURE D (Determination of foam height) Requirement for shampoo that is surfactant based:- S.NO CHARACTERISTICS REQUIREMENT METHOD OF TEST (REFER TO ANNEXURE) 1. Non-volatile alcohol soluble matter (% by mass) 10 B 2. pH 4-9 C 3. Foam height for 2 % solution, Min 150 mm D

53. ANNEXURE E (Ideal properties of shampoo) Ease of application Rinsing Easy wet combing Manageability Lustre Fragrance Low level of irritation Well preserved Good stability Economical

54. IS 6608:2004 (Indian standards skin-cream specification) IS 4707 (Part 1 and Part 2) REQUIREMENTS (1)Description:- Skin cream shall be in the form of a thick emulsion or unctuous mass (having greasy/oily/soapy feel) with a pleasant odour . It shall be white or pigmented or of uniform colour . III.ANALYTICAL METHODS FOR SKIN-CREAM

55. (2)Ingredients :-Unless specified otherwise, all the raw materials used in the manufacture of skin creams shall conform to the requirements prescribed in the relevant Indian Standards where such standards exist. The dyes, colours (pigments, lakes etc.) if used in the manufacture of skin creams shall comply with IS 4707 (Part 1) subject to the provision of Schedule Q of Drugs and Cosmetics Act, issued by Government of India. Other ingredients shall comply with the provisions of IS 4707 (Part 2)

56. (3)The material shall also comply with the requirements given in Table 1 when tested as prescribed in Table 1 Table 1:Requirements for skin-cream S.NO. CHARACTERISTICS REQUIREMENT METHOD OF TEST (REFER TO ANNEXURE) 1. Thermal stability To pass the test A 2. pH* 4-9 B 3. Total fatty substance content (% by mass), Min. 5 C 4. Total residue (% by mass), Min. 10 D 5. Heavy metals** (as Pb ), ppm , Max. 20 E 6. Arsenic**, ppm , Max. 2 F 7. Microbial content/limit (a)Total viable count (b)Gram – ve pathogen NMT 1000 Less than 10 IS 14648 IS 14648

57. *for creams based on beeswax and borax, pH shall be between 5-10 **if all the raw materials requiring test for heavy metals and arsenic have been so tested and comply with the requirements, then the manufacture may not test the finished cosmetic for heavy metals and arsenic.

58. Requirements for quality, safety and performance prescribed under this section All the ingredients that go into formulation of cosmetics shall comply with the provisions of IS 4707 (Part 1 and Part 2). The product shall also meet specific requirements as given in the standard The product package shall display a list of key ingredients in descending order of quantity present. The product shall not be manufactured from any carcinogenic ingredients. The manufacturer shall produce to BIS environmental consent clearance from the concerned State Pollution Control Board as per the provisions of the Water (Prevention and Control of Pollution) Cess Act, 1977 and the Air (Prevention and Control of Pollution) act, 1981 along with authorization, if required under the Environment (Protection) act, 1986 and the rules made there under shall also comply with. SPECIFIC REQUIREMENTS (ADDITIONAL REQUIREMENTS FOR ECO MARK)

59. SPECIFIC REQUIREMENTS Product shall be dermatologically safe when tested as per IS 4011 Heavy metals calculated as Lead ( Pb )-NMT 20 ppm and Arsenic 2 ppm , when tested by the respective method prescribed in Indian Standards.

60. PACKING AND MARKING Packing :-Material should be packed in suitable well-closed containers Marking :- The container shall be legibly marked with the following information:- (a) Name of the material (b)Manufacturer’s name and/or his recognized trademark, if any; (c)Net content (d)Month and year of manufacturing/packing (e)Batch or lot number in code or otherwise (f)‘Best before use…’ (month and year to be declared by the manufacturer) (g)List of key ingredients and NOTE-This is exempted in case of pack sizes of 30 g/60 ml or less (h)Any other information required by statutory authorities.

61. SAMPLING Representative samples of the material shall be drawn as prescribed in IS 3958 Tests for all the characteristic shall be carried out on the composite samples as per methods referred in table 1 The material shall be taken to have conformed to the standard if the composite sample passes all the tests.

62. QUALITY OF REAGENTS Unless specified otherwise, pure chemicals and distilled water (see IS 1070) shall be employed in tests. ‘Pure chemicals’ shall mean chemicals that don’t contain impurities which affect the results of analysis.

63. ANNEXURE A (Test for thermal stability) A humidity chamber/incubator controlled at 60-70% relative humidity and 45±1ºC. The sample shall be taken to have passed the test, if on removal from the incubator shows no oil separation or any other phase separation

64. ANNEXURE B (Determination of pH ) A digital pH meter equipped with a glass electrode can be used.

65. ANNEXURE C (Determination of total fatty substance content) The emulsion is broken with dilute mineral acid and the fatty matter is extracted with petroleum ether. It is weighed after removal of the solvent.

66. ANNEXURE D (Determination of residue) Weigh accurately about 5g of the material in a weighed, clean and dry squat form weighing bottle and dry to constant mass at 105±1ºC. Cool in a desiccator and weigh.

67. ANNEXURE E (Test for heavy metals) The colour produced with hydrogen sulphide solution is matched against that obtained with standard lead solution.

68. ANNEXURE F (Determination of Arsenic) Arsenic present in a solution of the material is reduced to arsine, which is made to react with mercuric bromide paper. The stain produced is compared with the standard stain.

69. THANKS