71 STERILITY TESTS Change to read: Portions of this general chapter have been harmonized with the corresponding texts of the European Pharmacopeia and/or the Japanese Pharmacopeia. Those portions that are not harmonized are marked with symbols () to specify this fact. 6 These Pharmacopeial procedures are not by themselves designed to ensure that a batch of product is sterile or has been sterilized. This is accomplished primarily by validation of the sterilization process or of the aseptic processing procedures.The test is applied to substances, preparations, or articles which, according to the Pharmacopeia, are required to be sterile. However, a satisfactory result only indicates that no contaminating microorganism has been found in the sample examined under the conditions of the test.6 Add the following: PRECAUTIONS AGAINST MICROBIAL CONTAMINATION The test for sterility is carried out under aseptic conditions. In order to achieve such conditions, the test environment has to be adapted to the way in which the sterility test is performed. The precautions taken to avoid contamination are such that they do not affect any microorganisms that are to be revealed in the test. The working conditions in which the tests are performed are monitored regularly by appropriate sampling of the working area and by carrying out appropriate controls.6 Change to read: CULTURE MEDIA AND INCUBATION TEMPERATURES6 Media for the test may be prepared as described below or equivalent commercial media may be used provided that they comply with6 the requirements of the Growth Promotion Test of Aerobes, Anaerobes, and Fungi. 6 The following culture media have been found to be suitable for the test for sterility. Fluid Thioglycollate Medium is primarily intended for the culture of anaerobic bacteria. However, it will also detect aerobic bacteria. SoybeanCasein Digest Medium is suitable for the culture of both fungi and aerobic bacteria.Fluid Thioglycollate Medium l-Cystine 0.5 g Sodium Chloride 2.5 g Dextrose Monohydrate/Anhydrous6 5.5/5.0 g Agar 6 0.75 g Yeast Extract (water-soluble) 5.0 g Pancreatic Digest of Casein 15.0 g Sodium Thioglycollate 0.5 g or Thioglycolic Acid 0.3 mL Resazurin Sodium Solution (1 in 1000),freshly prepared 1.0 mL Purified Water 1000 mL pH after sterilization: 7.10.2.Mix the l-cystine, sodium chloride, dextrose, yeast extract, and pancreatic digest of casein with the purified water, and heat until solution is effected. Dissolve the sodium thioglycollate or thioglycolic acid in the solution and, if necessary, add 1 N sodium hydroxide so that, after sterilization, the solution will have a pH of 7.1 0.2. If filtration is necessary, heat the solution again without boiling, and filter while hot through moistened filter paper. Add the resazurin sodium solution, mix, and place the medium in suitable vessels that provide a ratio of surface to depth of medium such that not more than the upper half of the medium has undergone a color change indicative of oxygen uptake at the end of the incubation period. Sterilize using a validated process. If the medium is stored, store at a temperature between 2 and 25 in a sterile, airtight container. If more than the upper one-third of the medium has acquired a pink color, the medium may be restored once by heating the containers in a water-bath or in free-flowing steam until the pink color disappears and by cooling quickly, taking care to prevent the introduction of nonsterile air into the container. Do not use the medium for a longer storage period than has been validated.6 Fluid Thioglycollate Medium is to be incubated at 3035. For products containing a mercurial preservative that cannot be tested by the membrane filtration method, Fluid Thioglycollate Medium incubated at 2025 may be used instead of SoybeanCasein Digest Medium provided that it has been validated as described in Growth Promotion Test of Aerobes, Anaerobes, and Fungi. Where prescribed or justified and authorized, the following alternative thioglycollate medium might be used. Prepare a mixture having the same composition as that of the Fluid Thioglycollate Medium, but omitting the agar and the resazurin sodium solution. Sterilize as directed above. The pH after sterilization is 7.1 0.2. Heat in a water bath prior to use and incubate at 3035 under anaerobic conditions.6 SoybeanCasein Digest Medium Pancreatic Digest of Casein 17.0 g Papaic Digest of Soybean Meal 3.0 g Sodium Chloride 5.0 g Dibasic Potassium Phosphate 2.5 g Dextrose Monohydrate/Anhydrous6 2.5/2.3 g Purified Water 1000 mL pH after sterilization: 7.30.2.Dissolve the solids in the Purified Water, heating slightly to effect a solution. Cool the solution to room temperature, and adjust the pH with 1 N sodium hydroxide so that, after sterilization, it will have a pH of 7.3 0.2. Filter, if necessary to clarify, dispense into suitable containers, and sterilize us