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GB/T 7918.3-1987 PDF English

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GB/T 7918.3-1987: Standard methods of microbiological examination for cosmetics. Fecal coliforms
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GB/T 7918.3-1987: Standard methods of microbiological examination for cosmetics. Fecal coliforms

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GB NATIONAL STANDARD OF THE PEOPLE’S REPUBLIC OF CHINA UDC 668.58.576.85.07 GB 7918.3-87 Standard methods of microbiological examination for cosmetics - Fecal coliforms Issued on: MAY 28, 1987 Implemented on: OCTOBER 01, 1987 Issued by. Ministry of Health of PRC

Table of Contents

1 Method summary... 3 2 Culture media and reagents... 3 3 Instruments... 6 4 Operation steps... 7 5 Test result report... 7 Additional information... 8 Standard methods of microbiological examination for cosmetics - Fecal coliforms Fecal coliform bacteria originate from the feces of humans and warm-blooded animals. The detection of fecal coliform bacteria indicates that the cosmetics have been contaminated by feces and may contain other intestinal pathogens or parasites. Therefore, fecal coliform bacteria are listed as important health indicator bacteria.

1 Method summary

According to the biological characteristics of fecal coliform bacteria, for example, Gram-negative non-spore-forming bacteria can ferment lactose to produce acid and gas when cultured at 44 °C for 24 ~ 48 h, it can produce typical colonies on selective culture media, decompose tryptophan to produce indigo matrix.

2 Culture media and reagents

2.1 Lactose bile salt culture medium Ingredients. Peptone. 20 g Porcine bile salt. 5 g Lactose. 5 g 0.4% bromocresol purple aqueous solution. 2.5 ml Distilled water. 1000 ml Preparation method. Dissolve peptone, bile salt, lactose in distilled water. Adjust pH to 7.4.Add indicator. Mix well. Dispense into test tubes (add a small inverted tube to each test tube). Sterilize it at 115 °C (10 lb) for 20 min. 2.2 Double concentration lactose bile salt medium According to the above lactose bile salt medium composition, the amount of distilled UDC 668.58.576.85.07 GB 7918.3-87 water remains unchanged, whilst the amount of other ingredients is doubled. 2.3 Eosin methylene blue (EMB) agar Ingredients. Peptone. 10 g Lactose. 10 g Dipotassium hydrogen phosphate. 2 g Agar. 20 g 2% eosin aqueous solution. 20 ml 0.5% methylene blue aqueous solution. 13 ml Distilled water. 1000 ml Preparation method. First add agar to 900 ml distilled water. Heat to dissolve. Then add dipotassium hydrogen phosphate peptone. Mix well. Dissolve it. Then make up to 1000 ml with distilled water. Correct the pH value to 7.2 ~ 7.4.Dispense it into flasks. Sterilize it at 121 °C (15 lb) for 15 min under high pressure for use. Add lactose and heat to melt agar before use. Cool to about 60 °C. Add sterilized eosin-methylene blue solution aseptically. Shake well. Pour into a plate for later use. 2.4 Peptone water (for indigo matrix test) Ingredients. Peptone (or tryptone). 20 g Sodium chloride. 5 g Distilled water. 1000 ml Preparation method. Heat and melt the above ingredients. Adjust the pH value to 7.0 ~ 7.2.Divide into small test tubes. Sterilize at 121 °C (15 lb) for 15 min. 2.5 Indigo matrix reagent Covank reagent. Dissolve 5 g of p-dimethylaminobenzaldehyde in 75 ml of amyl alcohol. Then slowly add 25 ml of concentrated hydrochloric acid. Test method. Inoculate bacteria in peptone water and culture at 44 °C for 24 h. Add 0.3 ~ 0.5 ml of Covank reagent along the tube wall. Shake the test tube gently. Positive ones show deep rose red in the reagent layer. 2.6.2.1 Fix the smear on the flame. Add crystal violet staining solution. Stain for 1 min. Use water to wash it. 2.6.2.2 Add Gram iodine solution. Let it action for 1 min. Use water to wash it. 2.6.2.3 Add 95% alcohol to decolorize for about 30 seconds. Or add alcohol to the entire smear. Immediately pour it off. Then add alcohol to the entire smear. Decolorize for 10 seconds. Use water to wash it. 2.6.2.4 Add counterstain solution dropwise. Counterstain for 1 minute. Use water to wash it. Wait to dry. Examine under a microscope. 2.6.3 Staining results Gram-positive bacteria are purple, whilst Gram-negative bacteria are red. Note. If a 1.10 dilute carbolic acid fuchsin stain solution is used as the counterstain solution, the counterstaining time only takes 10 seconds.

3 Instruments

3.1 Constant temperature water bath or water-proof constant temperature box. 44 °C. 3.2 Thermometer. 3.3 Microscope. 3.4 Glass slide. 3.5 Inoculation loop. 3.6 Electric furnace. 3.7 Conical flask. 3.8 Test tube. 3.9 Small inverted tube. 3.10 pH meter or pH test paper. 3.11 High pressure sterilizer. 3.12 Sterilized pipette. 3.13 Sterilized plate. ......
Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al.


      

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