美国药典USP3171⽆菌检查法双语版
正数减负数怎么算
美国药典USP31-NF26⽆菌检查法《71》.doc
71 STERILITY TESTS ⽆菌检查法
Portions of this general chapter have been harmonized with the corresponding texts of the European Pharmacopeia and/or the Japane Pharmacopeia. Tho portions that are not harmonized are marked with symbols () to specify this fact.
此通则的各部分已经与欧洲药典和/或⽇本药典的对应部分做了协调。不⼀致的部分⽤符号（）来标明。
The following procedures are applicable for determining whether a Pharmacopeial article purporting to be sterile complies with the requirements t forth in the individual monograph with respect to the test for sterility. Pharmacopeial articles are to be tested by the Membrane Filtration method under Test for Sterility of the Product to be Examined where the nature of the product permits. If the membrane filtration technique is unsuitable, u the Direct Inoculation of the Culture Medium method under Test for Sterility of the Product to be Examined. All devices, with the exception of Devices with Pathways Labeled Sterile, are tested using the Direct Inoculation of the Culture Medium method. Provisions for retesting are included under Obrvation and Interpretation of Results.
下⾯这些步骤适⽤于测定是否某个⽤于⽆菌⽤途的药品是否符合其具体的各论中关于⽆菌检查的要求。只要其性质许可，这些药品将使⽤供试产品⽆菌检查法项下的膜过滤法来检测。如果膜过滤技术是不适合的，则使⽤在供试产品⽆菌检查法项下的培养基直接接种法。除了具有标记为⽆菌通道的设备之外，所有的设备均须使⽤培养基直接接种法进⾏检测。在结果的观测与理解项下包含了复验的规定。
Becau sterility testing is a very exacting procedure, where apsis of the procedure must be ensured for a correct interpretation of results, it is important that personnel be properly trained and qualified. The test for sterility is carried out under aptic 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.
由于⽆菌检查法是⼀个⾮常精确的程序，在此过程中程序的⽆菌状态必须得到确保以实现对结果的正确理解，因此⼈员经过适当的培训并取得资质是⾮常重要的。⽆菌检查在⽆菌条件下进⾏。为了实现这样的条件，试验环境必须调整到适合进⾏⽆菌检查的⽅式。为避免污染⽽采取的特定预防措施应不
会对任何试图在检查中发现的微⽣物产⽣影响。通过在⼯作区域作适当取样并进⾏适当控制，来定期监测进⾏此试验的⼯作条件。
The Pharmacopeial procedures are not by themlves 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 aptic processing procedures.
这些药典规定程序⾃⾝的设计不能确保⼀批产品⽆菌或已经灭菌。这主要是通过灭菌⼯艺或者⽆菌操作程序的验证来完成。
When evidence of microbial contamination in the article is obtained by the appropriate Pharmacopeial method, the result so obtained is conclusive evidence of failure of the article to meet the requirements of the test for sterility, even if a different result is obtained by an alternative procedure. For additional information on sterility testing, e Sterilization and Sterility Assurance of Compendial Articles 1211 .
当通过适当的药典⽅法获得了某物品中微⽣物污染的证据，这样获得的结果是该物品未能达到⽆菌检验要求的结论性证据，即便使⽤替代程序得到了不同的结果也⽆法否定此结果。如要获得关于⽆菌检验的其他信息，见药品的灭菌和⽆菌保证<1211>
MEDIA 培养基
Prepare media for the tests as described below, or dehydrated formulations may be ud provided that, when reconstituted as directed by the manufacturer or distributor, they meet the requirements of the Growth Promotion Test of Aerobes, Anaerobes, and Fungi. Media are sterilized using a validated process.
按照下⾯描述的⽅法配制实验⽤培养基；或者使⽤脱⽔培养基，只要根据其制造商或者分销商说明进⾏恢复之后，其能够符合好氧菌、厌氧菌、霉菌⽣长促进试验的要求即可。使⽤经过验证的⼯艺对培养基进⾏灭菌操作。
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. Soybean–Cain Digest Medium is suitable for the culture of both fungi and aerobic bacteria.
下⾯的培养基已经被证实适合进⾏⽆菌检查。巯基醋酸盐液体培养基主要⽤于厌氧菌的培养。但其也⽤于检测好氧菌。⼤⾖酪蛋⽩消化物培养基适合于培养霉菌和好氧菌。
Fluid Thioglycollate Medium 巯基醋酸盐液体培养基
Mix the L-cystine, sodium chloride, dextro, yeast extract, and pancreatic digest of cain 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 vesls 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.皮肤黑
将L-胱氨酸、氯化钠、葡萄糖、酵母提取物、酪蛋⽩胰酶消化物与纯净⽔混合，并加热⾄实现溶解。将巯基⼄酸钠或者巯基⼄酸溶解于该溶液，如果需要可再加⼊1N氢氧化钠，以便在灭菌后该溶液呈pH值7.1 ± 0.2。如需要则过滤，再次加热该溶液但不得煮沸，并趁热以湿润滤纸将该溶液过滤。加⼊刃天青钠溶液，混匀，并将该培养基置于适当容器中，该容器应为培养基提供特定的⾯积－深度⽐，以使在培养期末表明氧⽓摄⼊的变⾊部分不超过培养基的上半部分。使⽤经过验证的⼯艺进⾏灭菌。
如果需要储存该培养基，将其置于⽆菌、⽓密容器中，在2 ⾄25 之间储藏。如果超过上部三分之⼀的培养基已经呈粉⾊，可以⽤以下⽅法恢复该培养基⼀次：在⽔浴锅中或者⾃由流动蒸⽓中加热该容器，直⾄粉⾊消失，并迅速放凉，须⼩⼼防⽌⾮⽆菌空⽓进⼊到容器中。
Fluid Thioglycollate Medium is to be incubated at 32.5 ± 2.5 .
巯基醋酸盐液体培养基将在32.5 ± 2.5 条件下进⾏培养。
Alternative Thioglycollate Medium 替代巯基醋酸盐培养基
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, and allow to cool prior to u. The pH after sterilization is 7.1 ± 0.2. Incubate under anaerobic conditions for the duration of the incubation period.
配制与巯基醋酸盐液体培养基成分相同，但省略了琼脂和刃天青钠溶液的混合物，按上述⽅法灭菌，并在使⽤前静置⾄凉。灭菌后pH值为7.1 ± 0.2。在厌氧条件下培养，培养时间同培养期。
Alternative Fluid Thioglycollate Medium is to be incubated at 32.5 ± 2.5 .
替代性巯基醋酸盐培养基将在32.5 ± 2.5 条件下进⾏培养。
Soybean–Cain Digest Medium ⼤⾖-酪蛋⽩消化物培养基
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, dispen into suitable containers, and sterilize using a validated procedure. Store at a temperature between 2 and 25 in a sterile
well-clod container, unless it is intended for immediate u.
将固体物质溶解于纯净⽔，轻微加热以实现溶解。放凉溶液⾄室温，并⽤1N氢氧化钠调整pH值，以便在灭菌后其pH值呈7.3± 0.2。过滤，如需要则使之澄清，分装⼊适合的容器，
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–Cain 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 ud 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–Cain Digest Medium as follows. To the containers of each medium, transfer aptically a quantity of -lactama sufficient to inactivate the amount of antibiotic in the specimen under test. Determine the quantity of -lactama required to inactivate the antibiotic by using a -lactama preparation that has been assayed previously for its penicillin- or cephalosporin-inactivating power. [NOTE—Supplemented -lactama media can also be ud 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 parate from that ud for sterility testing), confirm that an appropriate amount of -lactama is incorporated into the medium, following either method under Validation Test, using less than 100 colony-forming units (cfu) of Staphylococcus aureus (e Table 1)
as the challenge. Typical microbial growth of the inoculated culture must be obrved as a confirmation that the -lactama 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 U 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
Suitability Tests 适合性试验
The media ud 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。
猕猴桃品种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尤尤我心
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 ingredients1 . 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 parate portion of medium for each of the following species of microorganism: Clostridium sporogenes, Pudomonas 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–Cain Digest Medium with a small number (not more than 100 cfu) of the following microorganisms, using a parate 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 ca of bacteria and not more than 5 days in the ca 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）产芽胞梭状芽胞杆怎么吹葫芦丝
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 unaled containers, they can be ud for 1 month, provided that they are tested for growth promotion within 2 weeks of the time of u and that color indicator requirements are met. If stored in tight containers, the media can be ud for 1 year, provided that they are tested for growth promotion within 3 months of the time of u 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
lol高清壁纸PREPARATION 配制品
跑步的呼吸方法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. Dispen into containers, and sterilize using a validated p
rocess.
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
Aptically add to the above Preparation, if necessary, a quantity of sterile -lactama sufficient to inactivate any residual antibiotic activity on the membranes after the solution of the test specimen has been filtered (e 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
To each L of Fluid A add 1 mL of polysorbate 80, adjust to a pH of 7.1 ± 0.2, dispen into containers, and sterilize using a validated process. U this fluid for articles containing lecithin or oil, or for devices labeled as ―sterile pathway.‖
向每升液体A中，加⼊1mL聚⼭梨酯80，调节pH值⾄7.1 ± 0.2，分装⼊容器中，并使⽤经过验证的⼯艺灭菌。此液体⽤于含有卵磷脂或油脂的物品，或⽤于标为―⽆菌通道‖的设备。美国的教育
Fluid K 液体K
Dissolve 5.0 g of peptic digest of animal tissue, 3.0 g of beef extract, and 10.0 g of polysorbate 80 in water to make 1 L. Adjust the pH to obtain, after sterilization, a pH of 6.9 ± 0.2. Dispen into containers, and sterilize using a validated process.将5.0g动物组织胃蛋⽩酶消化物、3.0g⽜⾁提取物、10.0g聚⼭梨酯80溶解于1L⽔中。调节pH值，以便使pH值在灭菌后呈6.9± 0.2。分装⼊容器中，并使⽤经过验证的⼯艺灭菌。
VALIDATION TEST验证试验
Carry out a test as described below under Test for Sterility of the Product to be Examined using exactly the same methods, except for the following modifications.
按照下⾯供试产品⽆菌检查项下的描述，使⽤除了下⾯变更之外完全相同的⽅法，进⾏试验。
Membrane Filtration 膜过滤
