USP 62 非无菌产品微生物限度检查(控制菌) USP38

62 MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: TESTS FOR SPECIFIED

MICROORGANISMS

非无菌产品微生物限度检查:控制菌（USP38）

1.INTRODUCTION导言

The tests described hereafter will allow determination of the absence of, or limited occurrence of, specified microorganisms that may be detected under the conditions described.

控制菌检查法系用于在规定的试验条件下，检查供试品中是否存在特定的微生物。

The tests are designed primarily to determine whether a substance or preparation complies with an established specification for microbiological quality. When used for such purposes, follow the instructions given below, including the number of samples to be taken, and interpret the results as stated below.

当本法用于检查非无菌制剂及其原辅料是否符合相应的微生物限度标准时,应按下列规定进行检验，,包括样品的取样量,结果的判断.

Alternative microbiological procedures, including automated methods, may be used, provided that their equivalence to the Pharmacopeial method has been demonstrated.

可以使用包括自动化法在内的方法,需确认与药典方法的等同性.

2.GENERAL PROCEDURES通用规程

The preparation of samples is carried out as described in Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests 61.

供试品制备,同USP<61>

If the product to be examined has antimicrobial activity, this is insofar as possible removed or neutralized as described in Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests 61.

若供试品有抗菌活性,应尽可能中和或去除,中和或去除的方法同USP<61>

If surface-active substances are used for sample preparation, their absence of toxicity for microorganisms and their compatibility with any inactivators used must be demonstrated as described in Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests 61.

若供试品制备过程中使用了表面活性剂,应确认其对微生物的无毒性以及与所用的中和剂/灭活剂的相容性，同USP<61>

3.GROWTH-PROMOTING AND INHIBITORY PROPERTIES OF THE MEDIA, SUITABILITY

OF THE TEST AND NEGATIVE CONTROLS 培养基适用性检查，控制菌检查方法的适用性

确认，阴性对照

The ability of the test to detect microorganisms in the presence of the product to be tested must be established. Suitability must be confirmed if a change in testing performance or a change in the product that may affect the outcome of the test is introduced.

在有供试品存在的情况下，所建立的方法应能检测微生物。若检测程序或产品发生变化可能影响检测结果时，控制菌检查方法应重新进行适用性试验。

3.1.Preparation of Test Strains试验菌液的制备

Use standardized stable suspensions of test strains as stated below. 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.

使用菌株标准化稳定的悬浮液。采用适宜的菌种保藏技术（种子批系统），确保接种的培养物自主种子批开始传代不超过5代。

3.1.1.AEROBIC MICROORGANISMS需氧菌

Grow each of the bacterial test strains separately in containers containing Soybean–Casein Digest Broth or on Soybean–Casein Digest Agar at 30° to 35° for 18 to 24 hours. Grow the test strain for Candida albicans separately on Sabouraud Dextrose Agar or in Sabouraud Dextrose Broth at 20° to 25° for 2 to 3 days.

将各试验菌株分别接种于TSB或TSA培养基，30~35℃培养18~24小时。将白色念珠菌接种于SDA或SDB培养基， 20~25℃培养2~3天。

Staphylococcus aureus金黄色葡萄球菌such as A TCC 6538, NCIMB 9518, CIP 4.83, or

NBRC 13276

Pseudomonas aeruginosa铜绿假单胞菌such as ATCC 9027, NCIMB 8626, CIP 82.118, or

NBRC 13275

Escherichia coli大肠埃希菌such as ATCC 8739, NCIMB 8545, CIP 53.126, or

NBRC 3972

Salmonella enterica subsp. enterica serovar

Typhimurium or, as an alternative,

肠道沙门氏菌肠道亚种鼠伤寒血清型（伤寒沙门氏

菌），或

such as ATCC 14028

Salmonella enterica subsp. enterica serovar Abony

肠道沙门氏菌肠道亚种阿邦尼血清型（阿邦尼沙门氏菌清型）such as NBRC 100797, NCTC 6017, or CIP 80.39

Candida albicans 白色念珠菌such as A TCC 10231, NCPF 3179, IP 48.72, or NBRC 1594

Use Buffered Sodium Chloride–Peptone Solution pH 7.0 or Phosphate Buffer Solution pH 7.2 to make test suspensions. Use the suspensions within 2 hours or within 24 hours if stored at 2° to 8°.

上述培养物用PH7.0的氯化钠-蛋白胨缓冲液或PH7.2磷酸盐缓冲液配制成适宜浓度的菌悬液。

菌液制备后若在室温下放置，应在2小时内使用，若保存在2~8℃，可在24小时内使用。

3.1.2.CLOSTRIDIA梭菌

Use Clostridium sporogenes such as ATCC 11437 (NBRC 14293, NCIMB 12343, CIP 100651) or ATCC 19404 (NCTC 532 or CIP 79.3). Grow the clostridial test strain under anaerobic conditions in Reinforced Medium for Clostridia at 30° to 35° for 24 to 48 hours. As an alternative to preparing and then diluting down a fresh suspension of vegetative cells of Cl. sporogenes, a stable spore suspension is used for test inoculation. The stable spore suspension may be maintained at 2° to 8°for a validated period.

梭菌，菌种ATCC 11437 (NBRC 14293, NCIMB 12343, CIP 100651) or ATCC 19404 (NCTC 532 or CIP 79.3)

将梭菌接种于梭菌增菌培养基中，置厌氧条件下30~35℃培养24~48小时。稳定的梭菌孢子混悬液可替代新鲜的菌悬液。孢子混悬液可保存在2~8℃，在验证过的期限内使用。

3.2.Negative Control阴性对照

To verify testing conditions, a negative control is performed using the chosen diluent in place of the test preparation. There must be no growth of microorganisms. A negative control is also performed when testing the products as described under Testing of Products. A failed negative control requires an investigation.

为确认试验条件是否符合要求，应用所选择的稀释剂替代供试品做阴性对照试验。阴性对照应无菌生长。若有菌生长，应进行偏差调查。