发布时间 : 星期日 文章美国药典USP31 71 无菌检查法 双语版更新完毕开始阅读ad54fea69b89680202d8250b
并用经过验证的程序消毒。如果不立刻使用，则在2 到25 之间以无菌且密闭良好的容器保存。
Soybean–Casein Digest Medium is to be incubated at 22.5 ± 2.5 .
大豆-酪蛋白消化物培养基将在22.5 ± 2.5 条件下培养。
Media for Penicillins or Cephalosporins 用于青霉素和头孢菌素的培养基
Where sterility test media are to be used in the Direct Inoculation of the Culture Medium method under Test for Sterility of the Product to be Examined, modify the preparation of Fluid
Thioglycollate Medium and the Soybean–Casein Digest Medium as follows. To the containers of each medium, transfer aseptically a quantity of -lactamase sufficient to inactivate the amount of antibiotic in the specimen under test. Determine the quantity of -lactamase required to inactivate the antibiotic by using a -lactamase preparation that has been assayed previously for its penicillin- or cephalosporin-inactivating power. [NOTE—Supplemented -lactamase media can also be used in the membrane filtration test.]
当无菌检查培养基用于供试产品无菌检查项下的培养基直接接种法时，按如下内容变更巯基醋酸盐液体培养基和大豆-酪蛋白消化物培养基的 制备方法。向每一种培养基的容器中，以无菌操作转移足够灭活供试样品中所存在抗生素的 -内酰胺酶。使用此前已经对其青霉素或头孢菌素灭活能力进行了测定的 -内酰胺酶配制品，来测定灭活该抗生素所必需的 -内酰胺酶数量。[注意：补充的 -内酰胺酶培养基也可以用于膜过滤试验]
Alternatively (in an area completely separate from that used for sterility testing), confirm that an appropriate amount of -lactamase is incorporated into the medium, following either method under Validation Test, using less than 100 colony-forming units (cfu) of Staphylococcus aureus (see Table 1) as the challenge. Typical microbial growth of the inoculated culture must be observed as a confirmation that the -lactamase concentration is appropriate.
或者（在与无菌试验所用场所彻底隔离的区域中），按照验证试验项下的任意一种方法，使用少于100个菌落（cfu）的金黄色葡萄球菌（见表1）作为验证菌，来确认适当数量的 -内酰胺酶已经被整合到该培养基中。必须观测到接种后培养物中出现典型微生物生长，才能确认 -内酰胺酶浓度是适当的。
Table 1. Strains of the Test Microorganisms Suitable for Use in the Growth Promotion Test and
the
Validation Test
表1 适合用于生长促进试验和验证试验中的试验微生物的菌株
Aerobic bacteria好氧菌 Staphylococcus aureus 1 ATCC 6538, CIP 4.83, NCTC 10788, NCIMB 9518 金黄色葡萄球菌 Bacillus subtilis枯草芽孢杆菌 Pseudomonas aeruginosa 2 ATCC 6633, CIP 52.62, NCIMB 8054 ATCC 9027, NCIMB 8626, CIP 82.118 绿脓杆菌 Anaerobic bacterium厌氧菌 Clostridium sporogenes 3 ATCC 19404, CIP 79.3, NCTC 532 or ATCC 11437 产芽胞梭状芽胞杆菌 Fungi霉菌 Candida albicans白色念珠菌 Aspergillus niger黑曲霉 1 ATCC 10231, IP 48.72, NCPF 3179 ATCC 16404, IP 1431.83, IMI 149007 An alternative to Staphylococcus aureus is Bacillus subtilis (ATCC 6633).可替代金黄色葡萄球菌的是枯草杆菌(ATCC 6633) 2 An alternative microorganism is Micrococcus luteus (Kocuria rhizophila), ATCC 9341. 替代微生物是藤黄微球菌（Kocuria rhizophila），ATCC 9341。 3 An alternative to Clostridium sporogenes, when a nonspore-forming microorganism is desired, is Bacetroides vulgatus (ATCC 8482). 当需要不形成芽孢微生物时，产芽胞梭状芽胞杆菌的替代微生物是Bacetroides vulgatus [NOTE—Seed-lot culture maintenance techniques (seed-lot systems) are used so that the viable microorganisms used for inoculation are not more than five passages removed from the original master seed lot.] [注意：采用适宜的菌种保藏技术，以确保用于接种的微生物的传代次数不超过5代] Suitability Tests 适合性试验
The media used comply with the following tests, carried out before, or in parallel, with the test on the product to be examined.
所使用的培养基须符合下列试验，这些试验应在检验供试产品之前或者同时进行。
STERILITY 无菌状态
Confirm the sterility of each sterilized batch of medium by incubating a portion of the media at the specified incubation temperature for 14 days. No growth of microorganisms occurs.
通过在指定培养温度下将一部分培养基培养14天，来确认每一批已灭菌培养基的无菌状态。不得出现微生物生长。
GROWTH PROMOTION TEST OF AEROBES, ANAEROBES, and FUNGI
好氧菌、厌氧菌、霉菌的生长促进试验
Test each lot of ready-prepared medium and each batch of medium prepared either from
dehydrated medium or from ingredients 1 . Suitable strains of microorganisms are indicated in Table 1.
检查每一批已经配制好的培养基和每一批用脱水培养基或配料制备的培养基 1 。适当微生物菌株见表1。
Inoculate portions of Fluid Thioglycollate Medium with a small number (not more than 100 cfu) of the following microorganisms, using a separate portion of medium for each of the following species of microorganism: Clostridium sporogenes, Pseudomonas aeruginosa, and
Staphylococcus aureus. Inoculate portions of Alternative Fluid Thioglycollate Medium with a small number (not more than 100 cfu) of Clostridium sporogenes. Inoculate portions of
Soybean–Casein Digest Medium with a small number (not more than 100 cfu) of the following microorganisms, using a separate portion of medium for each of the following species of
microorganism: Aspergillus niger, Bacillus subtilis, and Candida albicans. Incubate for not more than 3 days in the case of bacteria and not more than 5 days in the case of fungi.
在部分巯基醋酸盐液体培养基上接种少量（不超过100cfu）下列微生物，每一种微生物均使用单独一部分培养基：产芽胞梭状芽胞杆菌、绿脓杆菌、金黄色葡萄球菌。 在部分替代巯基醋酸盐液体培养基上接种少量（不超过100cfu）产芽胞梭状芽胞杆菌。 在部分大豆-酪蛋白消化物培养基上接种少量（不超过100cfu）下列微生物，每一种微生物均使用单独一部分的培养基：黑曲霉、枯草芽孢杆菌、白色念珠菌。细菌培养时间不超过3天，霉菌培养时间不超过5天。
The media are suitable if a clearly visible growth of the microorganisms occurs. 如果出现清晰可见的微生物生长，则该培养基是适合的。
STORAGE 保存
If prepared media are stored in unsealed containers, they can be used for 1 month, provided that they are tested for growth promotion within 2 weeks of the time of use and that color indicator requirements are met. If stored in tight containers, the media can be used for 1 year, provided that they are tested for growth promotion within 3 months of the time of use and that the color indicator requirements are met.
如果配制好的培养基保存于未密闭的容器中，只要在使用时间的2周内对其进行了生长促进试验并且符合颜色指示剂的要求，它们就可以使用1个月。如果保存在密闭的容器中，只要在使用时间的3个月内对其进行了生长促进试验并且符合颜色指示剂的要求，则该培养基可以使用1年。
DILUTING AND RINSING FLUIDS FOR MEMBRANE FILTRATION
用于膜过滤的稀释和冲洗液
Fluid A 液体A PREPARATION 配制品
Dissolve 1 g of peptic digest of animal tissue in water to make 1 L, filter or centrifuge to clarify, if necessary, and adjust to a pH of 7.1 ± 0.2. Dispense into containers, and sterilize using a validated process.
将1g动物组织胃蛋白酶消化物溶于1L水中，如果需要则通过滤或离心使其澄清，再调节pH值至7.1 ± 0.2。分装入容器中，并用经过验证的工艺灭菌。
PREPARATION FOR PENICILLINS OR CEPHALOSPORINS
用于青霉素或头孢菌素的配制品
Aseptically add to the above Preparation, if necessary, a quantity of sterile -lactamase sufficient to inactivate any residual antibiotic activity on the membranes after the solution of the test specimen has been filtered (see Media for Penicillins or Cephalosporins).
在供试样品溶液已经过滤（见用于青霉素或头孢菌素的培养基）之后，如果需要，向上述配制品中，以无菌操作加入数量足够灭活滤膜上残余抗生素活性的 -内酰胺酶。
Fluid D 液体D