Chinese Pharmacopoeia- 1110 Biological Tests- 1101 Sterility Test(1)
2021-06-11
1110 Biological tests 1101 Sterility test Sterility test is a method for determining whether sterile drugs, biological products, medical instruments, raw materials, accessories and other varieties that are required to be sterile in the pharmacopoeia are sterile. If the sample complies with the requirements of sterility test, it only indicates that no microbial contamination is found under the test condition. The sterility test should be conducted under the sterile condition, the test environment must meet the requirements of sterility test, and the whole testing process should strictly follow the sterile procedures to prevent microbial contamination, and the measures to prevent contamination should not affect the detection of microorganisms in the sample. Medium Thioglycollate fluid medium is mainly used for culturing anaerobic bacteria and it can also be used for culturing aerobic bacteria; trypsin liquid medium is used for culturing fungi and aerobic bacteria. Preparation and culturing conditions of medium The medium can be prepared according to the following prescription, and dehydrated medium or prepared medium which is produced according to the prescription and meets the specification can also be used. After preparation, a qualified sterilization procedure should be conducted for the sterilization. The finished medium should be stored in a dark place with the temperature of 2°C to 25°C, if stored in an unsealed container, generally it should be used within 3 weeks; and if stored in an airtight container, it can be used within a year.
Thioglycollate fluid medium Casein tryptone 15.0g Sodium chloride 2.5g Bacto-extract 5.0g Newly prepared 0.1% resazurin solution 1.0ml Anhydrous glucose 5.0g L-cystine 0.5g Agar 0.75g Sodium thiosulfate 0.5g Water 1000ml (Or thioglycollic acid) (0.3ml) Except for glucose and resazurin solution, mix the above ingredients, dissolve them at a low temperature, adjust pH to alkalescence, boiling and filtering. Add glucose and resazurin solution, shake them well and adjust pH to 7.1 土 0.2 at 25°C after sterilization. Then divide it into suitable containers, and the proportion of the quantity and the height of the container should be in accordance with that the oxide layer (pink) of the medium after culturing is not exceed 1/2 of its depth. Sterilizing. Before inoculation, the height of the oxide layer of the medium should not exceed 1/5 of the depth of the medium, otherwise it has to be heated in water bath at 100°C until the pink disappears (not more than 20 minutes), and cools rapidly, it should be heated only once and be protected from contamination. Unless otherwise specified, the thioglycollate fluid medium should be cultured between 30°C and 35°C.
Trypsin liquid medium Casein tryptone 17.0g Sodium chloride 5.0g Soybean papain hydrolysate 3.0g Dipotassium hydrogen phosphate 2.5g Glucose / Anhydrous glucose 2.5g / 2.3g Water 1000ml Except for glucose, mix the above ingredients, dissolve them at a low temperature, filtering, adjust pH to 7.3 土 0.2 at 25°C after sterilization, add glucose, subpackaging, sterilizing. Trypsin liquid medium should be cultured between 20°C and 25°C.
Neutralizer or inactivated medium According to the prescription and preparation method of the thioglycollate fluid medium or trypsin liquid medium, appropriate neutralizer, inactivator or surfactant can be added before the medium is sterilized or used, the dosage can refer to the method applicability test.
0.5% broth medium (used for sterility test of antibiotics like streptomycin sulfate) Peptone 10g Sodium chloride 5.0g Beef extract 3.0g Water 1000ml Glucose 5.0g Except for glucose, mix the above ingredients, dissolve them at a low temperature, adjust pH to alkalescence, boiling, add glucose and after the dissolution, shake them well, filtering, adjust pH to 7.2 土 0.2 at 25°C after sterilization, subpackaging, sterilizing.
Trypsin soy agar medium Casein tryptone 15.0g Agar 15.0g Soybean papain hydrolysate 5.0g Water 1000ml Glucose 5.0g Except for agar, mix the above ingredients, dissolve them at a low temperature, adjust pH to 7.3 土 0.2 at 25°C after sterilization, add agar and after heating and melting, shake them well, subpackaging, sterilizing.
Sabouraud glucose liquid medium Equivalent mixture of animal tissue pepsin hydrolysate and casein tryptone 10.0g Water 1000ml Glucose 20.0g Except for glucose, mix the above ingredients, dissolve them at a low temperature, adjust pH to 5.6 土 0.2 at 25°C after sterilization, add glucose, shake them well, subpackaging, sterilizing.
Sabouraud glucose agar medium Equivalent mixture of animal tissue pepsin hydrolysate and casein tryptone 10.0g Agar 15.0g Water 1000ml Glucose 40.0g Except for glucose and agar, mix the above ingredients, dissolve them at a low temperature, adjust pH to 5.6 土 0.2 at 25°C after sterilization, add agar and after heating and melting, add glucose, shake them well, subpackaging, sterilizing. Applicability test for the medium The thioglycollate fluid medium and trypsin liquid medium that are used for sterility test should conform to the requirements of sterility test and sensitivity test of the medium. Sterility test To randomly select no less than 5 medium (bottles) from each batch, and culture them at the specified temperature for 14 days with sterility
