2024年5月12日发(作者：贯晴霞)
USP<71> 无菌检查法Sterility Tests
无菌检查法系用于检查药典要求无菌的药物、制剂产品和其他物品是否无菌的一种方法。
若供试品符合无菌检查法的规定，仅表明供试品在该检验条件下未发现微生物污染。
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.
1 预防微生物污染
Precaotions Against Microbial Contamination
无菌检查应在无菌条件下进行，为了达到条件，检测环境应符合无菌检查的规定。防止
污染的措施不得影响供试品中微生物的检出。检测环境应定期抽样监测并进行适当控制。
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.
2 培养基和培养温度
Culture Media and Incubation
无菌检查需制备下表所述培养基，或者是能够符合需氧菌、厌氧菌、真菌促生长试验要
求的同等的商用培养基。
Media for the test may be prepared as described below or equivalent commercial media may
be used provided that they comply with the requirements of the Growth Promotion Test of
Aerobes, Anaerobes, and Fungi.
以下培养基已经被证实适合用于无菌检查。硫乙醇酸盐流体培养基主要用于厌氧菌的培
养，但其也可用于需氧菌培养。大豆酪蛋白培养基适合于培养真菌和需氧菌。
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–Casein Digest Medium is suitable for the culture of
both fungi and aerobic bacteria.
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化水混合，并加热
至溶解。将硫乙醇酸钠或硫乙醇酸溶解于该溶液，如果需要可再加入1mol/L氢氧化钠
溶液，以便在灭菌后该溶液呈pH值7.1±0.2。如需要过滤，再次加热该溶液但不得煮沸，
并趁热以润湿滤纸将该溶液过滤。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，以使在培养期结束后能明确显示氧气
1 / 20
摄入的变色部分不超过培养基的上半部分二分之一。使用经过验证的工艺进行灭菌。如
果需要贮存该培养基，应将其置于无菌、气密容器中，在2~25℃贮藏。如果超过上部三
分之一的培养基已经呈粉红色，可以用以下方法恢复该培养基功能，但每批培养基仅能
恢复一次：在水浴锅中或者自由流动蒸汽中加热该容器，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。灭菌后培养基存放时间超过验证期限时，不得使
用。
Mix the L-cystine, agar, 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.
硫乙醇酸盐流体培养基
Fluid Thioglycollate Medium
L-胱氨酸L-cystine
氯化钠Sodium Chloride
水合葡萄糖/无水葡萄糖Dextrose Monohydrate/Anhydrous
琼脂Agar
酵母提取物（水溶的）Yeast Extract (water-soluble)
胰酶消化酪蛋白胨Pancreatic Digest of Casein
硫乙醇酸钠Sodium Thioglycollate
或硫乙醇酸or Thioglycolic Acid
新配制的刃天青水溶液（1:1000）
Resazurin Sodium Solution (1 in 1000),freshly prepared
纯化水Purified Water
灭菌后pH：7.1±0.2
pH after sterilization: 7.1±0.2.
硫乙醇酸盐流体培养基应在30~35℃条件下进行培养。含有汞制剂防腐剂的产品不能使
用膜过滤方法检测。经需氧菌、厌氧菌、真菌促生长试验验证，在20~25℃培养时，大
豆酪蛋白消化培养基可以替代硫乙醇酸盐流体培养基。经合理授权，不含琼脂和刃天青
钠溶液配制的硫乙醇酸盐流体培养基可以替代硫乙醇酸盐流体培养基使用。按上述方法
灭菌，灭菌后pH值为7.1±0.2。使用前使用水浴加热，置于30~35℃厌氧条件下培养。
0.5g
2.5g
5.5/5.0g
0.75g
5.0g
15.0g
0.5g
0.3ml
1.0ml
1000ml
2 / 20
2024年5月12日发(作者：贯晴霞)
USP<71> 无菌检查法Sterility Tests
无菌检查法系用于检查药典要求无菌的药物、制剂产品和其他物品是否无菌的一种方法。
若供试品符合无菌检查法的规定，仅表明供试品在该检验条件下未发现微生物污染。
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.
1 预防微生物污染
Precaotions Against Microbial Contamination
无菌检查应在无菌条件下进行，为了达到条件，检测环境应符合无菌检查的规定。防止
污染的措施不得影响供试品中微生物的检出。检测环境应定期抽样监测并进行适当控制。
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.
2 培养基和培养温度
Culture Media and Incubation
无菌检查需制备下表所述培养基，或者是能够符合需氧菌、厌氧菌、真菌促生长试验要
求的同等的商用培养基。
Media for the test may be prepared as described below or equivalent commercial media may
be used provided that they comply with the requirements of the Growth Promotion Test of
Aerobes, Anaerobes, and Fungi.
以下培养基已经被证实适合用于无菌检查。硫乙醇酸盐流体培养基主要用于厌氧菌的培
养，但其也可用于需氧菌培养。大豆酪蛋白培养基适合于培养真菌和需氧菌。
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–Casein Digest Medium is suitable for the culture of
both fungi and aerobic bacteria.
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化水混合，并加热
至溶解。将硫乙醇酸钠或硫乙醇酸溶解于该溶液，如果需要可再加入1mol/L氢氧化钠
溶液，以便在灭菌后该溶液呈pH值7.1±0.2。如需要过滤，再次加热该溶液但不得煮沸，
并趁热以润湿滤纸将该溶液过滤。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，以使在培养期结束后能明确显示氧气
1 / 20
摄入的变色部分不超过培养基的上半部分二分之一。使用经过验证的工艺进行灭菌。如
果需要贮存该培养基，应将其置于无菌、气密容器中，在2~25℃贮藏。如果超过上部三
分之一的培养基已经呈粉红色，可以用以下方法恢复该培养基功能，但每批培养基仅能
恢复一次：在水浴锅中或者自由流动蒸汽中加热该容器，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。灭菌后培养基存放时间超过验证期限时，不得使
用。
Mix the L-cystine, agar, 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.
硫乙醇酸盐流体培养基
Fluid Thioglycollate Medium
L-胱氨酸L-cystine
氯化钠Sodium Chloride
水合葡萄糖/无水葡萄糖Dextrose Monohydrate/Anhydrous
琼脂Agar
酵母提取物（水溶的）Yeast Extract (water-soluble)
胰酶消化酪蛋白胨Pancreatic Digest of Casein
硫乙醇酸钠Sodium Thioglycollate
或硫乙醇酸or Thioglycolic Acid
新配制的刃天青水溶液（1:1000）
Resazurin Sodium Solution (1 in 1000),freshly prepared
纯化水Purified Water
灭菌后pH：7.1±0.2
pH after sterilization: 7.1±0.2.
硫乙醇酸盐流体培养基应在30~35℃条件下进行培养。含有汞制剂防腐剂的产品不能使
用膜过滤方法检测。经需氧菌、厌氧菌、真菌促生长试验验证，在20~25℃培养时，大
豆酪蛋白消化培养基可以替代硫乙醇酸盐流体培养基。经合理授权，不含琼脂和刃天青
钠溶液配制的硫乙醇酸盐流体培养基可以替代硫乙醇酸盐流体培养基使用。按上述方法
灭菌，灭菌后pH值为7.1±0.2。使用前使用水浴加热，置于30~35℃厌氧条件下培养。
0.5g
2.5g
5.5/5.0g
0.75g
5.0g
15.0g
0.5g
0.3ml
1.0ml
1000ml
2 / 20