The tests described hereafter will allow quantitative enumeration of mesophilic bacteria and fungi that may grow under aerobic conditions.

微生物计数法系用于能在有氧条件下生长的嗜温细菌和霉菌的定量计数。

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、辅料等是否符合相应的微生物限度标准时，应按照下述规定进行检验，包括样品的取样量，结果的判断。

The methods are not applicable to products containing viable microorganisms as active ingredients.

本法不适用于活菌制剂的检查。

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

可使用包括自动化法在内的替代方法，需确认其与药典方法的等同性。

Carry out the determination under conditions designed to avoid extrinsic microbial contamination of the product to be examined. The precautions taken to avoid contamination must be such that they do not affect any microorganisms that are to be revealed in the test.

微生物计数环境应能防止外来微生物对供试品的污染。防止污染的措施不能影响供试品中微生物的检出。

If the product to be examined has antimicrobial activity, this is, insofar as possible, removed or neutralized. If inactivators are used for this purpose, their efficacy and their absence of toxicity for microorganisms must be demonstrated.

如果供试品有抗菌活性，应尽可能去除或中和。若使用了中和剂或灭活剂，需确认其有效性及对微生物无毒性。

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.

供试品制备过程中若使用了表面活性剂，需确认其对微生物无毒性以及与所使用的中和剂或灭活剂的相容性。

Use the Membrane Filtration method or one of the Plate-Count Methods, as directed. The Most-Probable-Number (MPN) Method is generally the least accurate method for microbial counts; however, for certain product groups withvery low bioburden, it may be the most appropriate method.

计数方法包括薄膜过滤法、平皿法和最可能数法（Most-Probable-Number 简称MPN)。MPN法用于微生物计数时精确度最差，但用于微生物污染量小的供试品，MPN法可能是最合适的方法。（MPN法不适用于霉菌的检测，仅在供试品总需氧菌数没有适应计数方法的情况下使用。）

The choice of a method is based on factors such as the nature of the product and the required limit of microorganisms. The method chosen must allow testing of a sufficient sample size to judge compliance with the specification. The suitability of the chosen method must be established.

供试品检查时，应根据供试品理化特性和微生物限度标准等因素选择计数方法。所选择的计数方法应能够通过检测足量的供试品，判断与质量标准的符合性。且该计数方法的适用性须经确认。

The ability of the test to detect microorganisms in the presence of 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，所选用的计数方法需能够检测微生物。若检测程序或产品发生变化可能影响检测结果时，计数方法需重新进行适用性确认。

Use standardized stable suspensions of test strains or prepare 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. Grow each of the bacterial and fungal test strains separately as described in Table 1.

试验菌液：使用试验菌株的标准化稳定悬浮液或按下述规定制备。按表1规定程序分别培养各试验菌液。

菌株：采用适宜的菌种保藏技术（种子批系统），试验用菌株传代次数自主种子批（第0代）算起不得超过5代。

Table 1. Preparation and Use of Test Microorganisms

Use Buffered Sodium Chloride–Peptone Solution pH 7.0 or Phosphate Buffer Solution pH 7.2 to make test suspensions; to suspend A. brasiliensis spores, 0.05% of polysorbate 80 may be added to the buffer. Use the suspensions within 2 hours, or within 24 hours if stored between 2°and 8°. As an alternative to preparing and then diluting a fresh suspension of vegetative cells of A. brasiliensis or B. subtilis, a stable spore suspension is prepared and then an appropriate volume of the spore suspension is used for test inoculation. The stable spore suspension may be maintained at 2 to 8 for a validated period of time.

菌液制备：取表1各试验菌株的新鲜培养物，用PH7.0的氯化钠-蛋白胨缓冲液或PH7.2的磷酸缓冲液制备混悬液。制备黑曲霉孢子悬液，需在缓冲液中加入0.05%的聚山梨酯80。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。

黑曲霉和枯草杆菌新鲜细胞的混悬液，可用其孢子混悬液替代用于试验。该孢子混悬液可保存在2~8℃，在验证过的储存期限内使用。

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，需用选择的稀释剂代替供试品进行阴性对照。阴性对照应无菌生长。如阴性对照有菌生长，应进行偏差调查。

供试品检测时也需要进行阴性对照。

Test each batch of ready-prepared medium and each batch of medium prepared either from dehydrated medium or from the ingredients described.

微生物计数用的制备培养基、脱水培养基或按处方配置的培养基，每批均需进行适用性检查

Inoculate portions/plates of Soybean–Casein Digest Broth and Soybean–Casein Digest Agar with a small number(not more than 100 cfu) of the microorganisms indicated in Table 1, using a separate portion/plate of medium foreach. Inoculate plates of Sabouraud Dextrose Agar with a small number (not more than 100 cfu) of the microorganisms indicated in Table 1, using a separate plate of medium for each. Incubate according to the conditions described in Table 1.

按表1规定，接种不大于100cfu的指定菌株至TSA或TSB或SDA培养基。在表1规定的条件下培养。对于TSA或TSB培养基，每一菌株使用单独的培养基或培养基上单独的区域。对于SDA培养基，每一菌株使用单独的培养基。

For solid media, growth obtained must not differ by a factor greater than 2 from the calculated value for a standardized inoculum. For a freshly prepared inoculum, growth of the microorganisms comparable to that previously obtained with a previously tested and approved batch of medium occurs. Liquid media are suitable if clearly visiblegrowth of the microorganisms comparable to that previously obtained with a previously tested and approved batch ofmedium occurs.

被检固体培养基上的菌落数与参比培养基（标准培养物）上的菌落相比，差异因子不能大于2（50%~200%的范围内）；被检新鲜制备的培养物与参比培养基（之前检测并批准批次的培养基）比较，应有菌生长；被检液体培养基管与参比培养基管（之前检测并批准批次的培养基）比较，试验菌应生长良好 。

The method for sample preparation depends on the physical characteristics of the product to be tested. If none of the procedures described below can be demonstrated to be satisfactory, a suitable alternative procedure must be developed.

根据供试品的理化性质，采用适宜的方法进行供试品制备。如以下制备方法均不适用，可采用其他合适的方法。

Water-Soluble Products— Dissolve or dilute (usually a 1 in 10 dilution is prepared) the product to be examined inBuffered Sodium Chloride–Peptone Solution pH 7.0, Phosphate Buffer Solution pH 7.2, or Soybean–Casein Digest Broth. If necessary, adjust to a pH of 6 to 8. Further dilutions, where necessary, are prepared with the same diluent.

水溶性供试品——取供试品，用PH7.0 氯化钠-蛋白胨缓冲液，或PH7.2磷酸盐缓冲液，或TSB溶解或稀释成1:10供试品溶液。若需要，调节供试品溶液PH值至6~8.必要时，用同一稀释液进一步稀释（10倍系列稀释）。

Nonfatty Products Insoluble in Water— Suspend the product to be examined (usually a 1 in 10 dilution is prepared)in Buffered Sodium Chloride–Peptone Solution pH 7.0, Phosphate Buffer Solution pH 7.2, or Soybean–Casein Digest Broth. A surface-active agent such as 1 g per L of polysorbate 80 may be added to assist the suspensionof poorly wettable substances. If necessary, adjust to a pH of 6 to 8. Further dilutions, where necessary, areprepared with the same diluent.

水不溶性非脂类供试品——取供试品，用PH7.0 氯化钠-蛋白胨缓冲液，或PH7.2磷酸盐缓冲液，或TSB配制成1:10供试品液。分散性差的供试品，可在稀释液中加入表面活性剂如1g/L的聚山梨酯80. 若需要，调节供试品溶液PH值至6~8.必要时，用同一稀释液进一步稀释（10倍系列稀释）。

Fatty Products— Dissolve in isopropyl myristate sterilized by filtration, or mix the product to be examined with the minimum necessary quantity of sterile polysorbate 80 or another noninhibitory sterile surface-active reagent heated, if necessary, to not more than 40 or, in exceptional cases, to not more than 45. Mix carefully and if necessary maintain the temperature in a water bath. Add a sufficient quantity of the prewarmed chosen diluent to make a 1 in 10 dilution of the original product. Mix carefully, while maintaining the temperature for the shortest timenecessary for the formation of an emulsion. Further serial 10-fold dilutions may be prepared using the chosen diluent containing a suitable concentration of sterile polysorbate 80 or another noninhibitory sterile surface-active reagent.

油脂类供试品——取供试品，加入经无菌过滤的十四酸异丙酯， 或与最少量（并能使供试品乳化）的无菌聚山梨酯80或其他无抑菌性的无菌表面活性剂混合，必要时加热，温度一般不得超过40℃（特殊情况，不得超过45℃）。小心混合，如若必要，可在水浴中进行，然后加入预热的稀释剂，将供试品进行1:10的稀释。保温，小心混合，并在最短时间内形成乳状液。必要时，用含上述无菌表面活性剂的稀释剂进一步稀释（10倍系列稀释）。

Fluids or Solids in Aerosol Form— Aseptically transfer the product into a membrane filter apparatus or a sterile container for further sampling. Use either the total contents or a defined number of metered doses from each of the containers tested.

固体或液体气溶胶——无菌操作，将供试品转移至膜过滤装置或无菌容器，进一步取样。从每一被测容器中取全部内容物或剂量的倍数进行检测。

Transdermal Patches— Remove the protective cover sheets (“release liners”) of the transdermal patches and place them, adhesive side upwards, on sterile glass or plastic trays. Cover the adhesive surface with a suitable sterile porous material (e.g., sterile gauze) to prevent the patches from sticking together, and transfer the patches to a suitable volume of the chosen diluent containing inactivators such as polysorbate 80 and/or lecithin. Shake the preparation vigorously for at least 30 minutes.

透皮贴剂——取供试品，去掉保护层（即释放衬板），将粘贴面朝上放置在无菌玻璃或塑料器皿上，在粘贴面上覆盖一层适宜的无菌多孔材料（如无菌纱布），防止贴膏剂粘在一起。将处理好的贴膏剂放入盛有适量体积并含灭活剂如聚山梨酯80和/或卵磷脂的容器中。剧烈振摇至少30分钟。

Add to the sample prepared as directed above and to a control (with no test material included) a sufficient volumeof the microbial suspension to obtain an inoculum of not more than than 100 cfu. The volume of the suspension of the inoculum should not exceed 1% of the volume of diluted product.

取上述制备好的供试品液，加入试验菌液，使供试品液中含菌量不超过100cfu。（下文称为试验组）

取对照液（不含供试品），同法操作。（下文称为菌液组）

所加菌液的体积，不得超过供试品液体积的1%。

To demonstrate acceptable microbial recovery from the product, the lowest possible dilution factor of the prepared sample must be used for the test. Where this is not possible due to antimicrobial activity or poorsolubility, further appropriate protocols must be developed. If inhibition of growth by the sample cannot otherwisebe avoided, the aliquot of the microbial suspension may be added after neutralization, dilution, or filtration.

为保证供试品中微生物的回收率，需对最低稀释级别的供试品液进行计数方法的适用性试验。若因供试品抗菌活性或溶解性差不能用最低稀释级别的供试品液进行适用性试验的，应采用适宜的方法对供试品液进行进一步的处理。若供试品的抗菌活性无法去除，先对供试品液进行中和，稀释，或膜过滤的处理，再加入菌液。

The number of microorganisms recovered from the prepared sample diluted as described in Inoculation and Dilution and incubated following the procedure described in Recovery of Microorganisms in the Presence of Product, is compared to the number of microorganisms recovered from the control preparation.

供试品经上述方法稀释接种后，按下列“供试品中微生物的回收”规定的方法进行培养，(试验组)回收的菌落数与菌液组进行比较。

If growth is inhibited (reduction by a factor greater than 2), then modify the procedure for the particular enumeration test to ensure the validity of the results. Modification of the procedure may include, for example,

1.  An increase in the volume of the diluent or culture medium;

2.  Incorporation of a specific or general neutralizing agents into the diluent;

3.  Membrane filtration; or

4.  A combination of the above measures.

如果微生物生长被抑制（以大于2的因子减少，即小于50%），可采用以下方法消除供试品的抑菌活性，以确保验证结果有效：

1.        增加稀释剂或培养基体积；

2.        稀释剂中加入适宜的中和剂（或灭活剂）；

3.        采用薄膜过滤法；

4.        以上几种方法的联合使用

Neutralizing Agents— Neutralizing agents may be used to neutralize the activity of antimicrobial agents (see Table 2). They may be added to the chosen diluent or the medium preferably before sterilization. If used, their efficacy and their absence of toxicity for microorganisms must be demonstrated by carrying out a blank with neutralizer and without product.

中和剂（或灭活剂）（见表2）可用于消除干扰物的抑菌活性，最好在稀释剂或培养基灭菌前加入。若使用中和剂（或灭活剂），试验中应设空白对照组，确认中和剂（或灭活剂）的有效性和对微生物的无毒性。

Table 2. Common Neutralizing Agents/Methods for Interfering Substances

If no suitable neutralizing method can be found, it can be assumed that the failure to isolate the inoculated organism is attributable to the microbicidal activity of the product. This information serves to indicate that the article is not likely to be contaminated with the given species of the microorganism. However, it is possible that the product inhibits only some of the microorganisms specified herein, but does not inhibit others not included among the test strains or those for which the latter are not representative. Then, perform the test with the highest dilution factor compatible with microbial growth and the specific acceptance criterion.

若没有适合消除供试品抑菌活性的方法，对特定试验菌的回收的失败，可假设是由于供试品对该试验菌具有较强的抗菌活性，同时也表明供试品不易被该类微生物污染。但是，供试品也可能仅对供试菌株中的某些具有抑制作用，而对供试菌株以外的微生物没有抑制作用或此类微生物对其不具代表性。因此，根据供试品需符合的微生物限度标准，应采用能使微生物生长的最高稀释级别的供试品溶液进行计数方法适用性试验。

For each of the microorganisms listed, separate tests are performed. Only microorganisms of the added test strain are counted.

表1所列的各试验菌应逐一进行微生物回收试验。只计数加入的试验菌液的菌落数。微生物的回收可用平皿法，薄膜过滤法或MPN法。

Membrane Filtration— Use membrane filters having a nominal pore size not greater than 0.45 μm. The type of filter material is chosen in such a way that the bacteria-retaining efficiency is not affected by the components of the sample to be investigated. For each of the microorganisms listed, one membrane filter is used.

薄膜过滤法——薄膜过滤法采用的滤膜孔径不得大于0.45μm，供试品应不影响滤膜材质对微生物的截留。表1所列的各试验菌，使用单独的滤膜。按如下所述操作：

Transfer a suitable quantity of the sample prepared as described under Preparation of the Sample, Inoculation and Dilution, and Neutralization/Removal of Antimicrobial Activity (preferably representing 1 g of the product, or less if large numbers of cfu are expected) to the membrane filter, filter immediately, and rinse the membrane filter with an appropriate volume of diluent.

取照上述“供试品的制备”“接种和稀释”“抗菌活性的去除或灭活”制备的供试品液适量（一般取相当于1g供试品的量，若供试品中所含的菌数较多，供试品液可酌情减量），转移至滤膜，立即过滤，用适量的稀释剂冲洗滤膜。

For the determination of total aerobic microbial count (TAMC), transfer the membrane filter to the surface of theSoybean–Casein Digest Agar. For the determination of total combined yeasts and molds count (TYMC), transfer the membrane to the surface of the Sabouraud Dextrose Agar. Incubate the plates as indicated in Table 1. Perform the counting.

若测定总需氧菌数，转移滤膜菌面朝上贴于TSA培养基；若测定总酵母菌和霉菌，转移滤膜菌面朝上贴于SDA培养基，按表1规定条件培养，计数 。

Plate-Count Methods— Perform plate-count methods at least in duplicate for each medium, and use the mean count of the result.

平皿法——平皿法包括倾注法和涂布法。每种培养基至少制备两个平皿，以平均值作为计数结果。

Pour-Plate Method— For Petri dishes 9 cm in diameter, add to the dish 1 mL of the sample prepared as described under Preparation of the Sample, Inoculation and Dilution, and Neutralization/Removal ofAntimicrobial Activity and 15 to 20 mL of Soybean–Casein Digest Agar or Sabouraud Dextrose Agar, both media maintained at not more than 45°.

倾注法——取照上述“供试品的制备”“接种和稀释”“抗菌活性的去除或灭活”制备的供试品液1 mL, 置9cm直径的无菌培养皿中，注入15~20mL 加热熔融不超过45℃的TSA或SDA培养基。

If larger Petri dishes are used, the amount of agar medium is increased accordingly. For each of the microorganisms listed in Table 1, at least two Petri dishes are used.

若使用直径较大的平皿，培养基的用量应相应增加。表1所列各实验菌株，每个菌株至少制备两个平皿。

Incubate the plates as indicated in Table 1. Take the arithmetic mean of the counts per medium, and calculate the number of cfu in the original inoculum.

按表1规定培养，取每培养基的算术平均值，并计算原培养物的菌落数。

Surface-Spread Method— For Petri dishes 9 cm in diameter, add 15 to 20 mL of Soybean–Casein Digest Agaror Sabouraud Dextrose Agar at about 45℃ to each Petri dish, and allow to solidify. If larger Petri dishes are used, the volume of the agar is increased accordingly. Dry the plates, for example, in a laminar-airflow cabinet or in an incubator. For each of the microorganisms listed in Table 1, at least two Petri dishes are used. Spread a measured volume of not less than 0.1 mL of the sample, prepared as directed under Preparation of the Sample, Inoculation and Dilution, and Neutralization/Removal of Antimicrobial Activity over the surface of the medium. Incubate and count as directed for Pour-Plate Method.

涂布法——取15~20mL 加热熔融不超过45℃的TSA或SDA培养基，注入直径9cm的无菌平皿，凝固，制成平板，采用适宜的方法使培养基凝固，如在通风厨或培养箱中干燥。若使用直径较大的平皿，培养基的用量也应相应增加。每一平板表面涂布上述“供试品的制备”“接种和稀释”“抗菌活性的去除或灭活”制备的供试品液至少0.1 mL，与倾注法同法培养，计数。表1所列各实验菌株，每个菌株至少制备两个平皿。

Most-Probable-Number (MPN) Method— The precision and accuracy of the MPN Method is less than that of theMembrane Filtration method or the Plate-Count Method. Unreliable results are obtained particularly for the enumeration of molds. For these reasons, the MPN Method is reserved for the enumeration of TAMC in situations where no other method is available. If the use of the method is justified, proceed as follows.

MPN法——MPN法的精密度和准确度不及薄膜过滤法和平皿计数法，仅在供试品总需氧菌数没有适宜计数方法的情况下使用。本法不适用于霉菌计数。若采用MPN法，按下列步骤进行:

Prepare a series of at least three serial 10-fold dilutions of the product as described for Preparation of the Sample, Inoculation and Dilution, and Neutralization/Removal of Antimicrobial Activity. From each level of dilution, three aliquots of 1 g or 1 mL are used to inoculate three tubes with 9 to 10 mL of Soybean–Casein Digest Broth. If necessary a surface-active agent such as polysorbate 80, or an inactivator of antimicrobial agents maybe added to the medium. Thus, if three levels of dilution are prepared, nine tubes are inoculated.

取照上述“供试品的制备”“接种和稀释”“抗菌活性的去除或灭活”制备的供试品液至少3个连续稀释级，每一稀释级别取3份1g或1mL供试品液分别接种至装有9~10mL TSB培养基的3个试管中。必要时，可在培养基中加入表面活性剂（如聚山梨酯80），中和剂或灭活剂。如上操作，制备三个稀释级别，将9根接种管按如下规定培养：

Incubate all tubes at 30° to 35° for not more than 3 days. If reading of the results is difficult or uncertain owing to the nature of the product to be examined, subculture in the same broth or in Soybean–Casein Digest Agar for 1 to 2 days at the same temperature, and use these results. From Table 3, determine the most probable number of microorganisms per g or mL of the product to be examined.

接种管置30~35℃至多培养3天，逐日观察各管微生物生长情况。如果由于供试品的原因使得结果难以判断，可将培养物接种至TSB或TSA培养基，在相同条件下培养1~2天，用再培养的结果计数。根据微生物生长情况，从表3查被测供试品每1g或每1mL中需氧菌总数的最可能数。

Table 3. Most-Probable-Number Values of Microorganisms

When verifying the suitability of the Membrane Filtration method or the Plate-Count Method, a mean count of any of the test organisms not differing by a factor greater than 2 from the value of the control defined in Inoculation and Dilution in the absence of product must be obtained. When verifying the suitability of the MPN Method, the calculated value from the inoculum must be within 95% confidence limits of the results obtained with the control.

计数方法适用性试验中，采用薄膜过滤法或平皿法时，试验组菌落的平均计数与“培养与稀释”项下制备的菌液组（不加入供试品）相比，差异因子不得大于2.（在50%~200%的范围内） 采用MPN法时，各菌种的试验组菌落数应在菌液组95%置信区间内。

If the above criteria cannot be met for one of more of the organisms tested with any of the described methods, the method and test conditions that come closest to the criteria are used to test the product.

若采用上述方法还有一株或多株试验菌的回收达不到要求，那么采用回收率最接近要求的方法和试验条件进行供试品的检查。

Unless otherwise directed, use 10 g or 10 mL of the product to be examined taken with the precautions referred to above. For fluids or solids in aerosol form, sample 10 containers. For transdermal patches, sample 10 patches.

除另有规定外，一般供试品的检测量为10g或10mL。液体或固定气溶胶为10个容器，透皮贴剂为10片。

The amount to be tested may be reduced for active substances that will be formulated in the following conditions: the amount per dosage unit (e.g., tablet, capsule, injection) is less than or equal to 1 mg, or the amount per g or mL (for preparations not presented in dose units) is less than 1 mg. In these cases, the amount of sample to betested is not less than the amount present in 10 dosage units or 10 g or 10 mL of the product.

原料药用于以下组方的制剂时，检测量可以酌减：单位剂量小于等于1mg（如片剂，胶囊剂，注射剂）；每1g或每1mL中（对于不按剂量单位使用的制剂）少于1mg。检测量不小于10个单位剂量或10g或10mL制剂中包含的量。

For materials used as active substances where the sample quantity is limited or batch size is extremely small (i.e., less than 1000 mL or 1000 g), the amount tested shall be 1% of the batch unless a lesser amount is prescribed or justified and authorized.

原料药样品量有限，或批量极小（如小于1000mL或1000g），检测量一般为批量的1%，更少的检测量需额外规定，合理性说明并获得批准。

For products where the total number of entities in a batch is less than 200 (e.g., samples used in clinical trials), the sample size may be reduced to two units, or one unit if the size is less than 100.

制剂批量小于200（例如临床试验的样品），检测量为2个单位，若批量小于100，则为1个单位。

Select the sample(s) at random from the bulk material or from the available containers of the preparation. To obtain the required quantity, mix the contents of a sufficient number of containers to provide the sample.

随机取样。混合，取规定量供试品进行检验。

Use a filtration apparatus designed to allow the transfer of the filter to the medium. Prepare the sample using a method that has been shown to be suitable as described in Growth Promotion Test and Suitability of theCounting Method, transfer the appropriate amount to each of two membrane filters, and filter immediately. Wash each filter following the procedure shown to be suitable.

采用的薄膜过滤装置应可以将滤液转移至培养基。照“促生长实验”和“计数方法适用性”确认的方法制备供试品溶液，并转移适量供试品液至两份滤膜，立即过滤。采用适宜的方法冲洗滤膜。

For the determination of TAMC, transfer one of the membrane filters to the surface of Soybean–Casein Digest Agar. For the determination of TYMC, transfer the other membrane to the surface of Sabouraud Dextrose Agar.Incubate the plate of Soybean–Casein Digest Agar at 30° to 35° for 3 to 5 days and the plate of Sabouraud Dextrose Agar at 20° to 25° for 5 to 7 days. Calculate the number of cfu per g or per mL of product.

取总需氧菌检测的滤膜，菌面朝上贴于TSA上培养，在30~35℃培养3~5天，取总酵母菌，霉菌检测的滤膜，菌面朝上贴于SDA培养基，在20~25℃培养5~7天。计数每g或每mL供试品中的菌落。

When examining transdermal patches, separately filter 10% of the volume of the preparation described forPreparation of the Sample through each of two sterile filter membranes. Transfer one membrane to Soybean–Casein Digest Agar for TAMC and the other membrane to Sabouraud Dextrose Agar for TYMC.

若检测透皮贴剂，各取10%“供试品制备”项下的供试品液至两份无菌滤膜，过滤。取出滤膜，菌面朝上贴于TSA和SDA培养基上培养，分别用于总需氧菌和总酵母菌霉菌的检测。

Pour-Plate Method— Prepare the sample using a method that has been shown to be suitable as described inGrowth Promotion Test and Suitability of the Counting Method. Prepare for each medium at least two Petridishes for each level of dilution. Incubate the plates of Soybean–Casein Digest Agar at 30 to 35 for 3 to 5 days and the plates of Sabouraud Dextrose Agar at 20 to 25 for 5 to 7 days. Select the plates corresponding to a given dilution and showing the highest number of colonies less than 250 for TAMC and 50 for TYMC. Take the arithmetic mean per culture medium of the counts, and calculate the number of cfu per g or per mL of product.

倾注法——取规定量样品，按”促生长实验”和”计数方法适用性试验”确认的方法进行供试品液的制备。每个稀释级别每种培养基至少制备2个平皿。TSA培养基在30~35℃培养3~5天。SDA培养基在20~25℃培养7天。总需氧菌数的测定应选取菌落数不超过250的稀释级用于计数，总酵母菌和霉菌的测定应选取菌落数不超过50的稀释级用于计数。取每培养基计数的算术平均值，并计算每克或每毫升供试品中的菌落数。

Surface-Spread Method— Prepare the sample using a method that has been shown to be suitable as describedin Growth Promotion Test and Suitability of the Counting Method. Prepare at least two Petri dishes for eachmedium and each level of dilution. For incubation and calculation of the number of cfu, proceed as directed for thePour-Plate Method.

涂布法——取规定量样品，按”促生长实验’和”计数方法适用性试验”确认的方法进行供试品液的制备。每个稀释级别每种培养基至少制备2个平皿。培养条件和计数方法，同倾注法。

Prepare and dilute the sample using a method that has been shown to be suitable as decribed in GrowthPromotion Test and Suitability of the Counting Method. Incubate all tubes for 3 to 5 days at 30° to 35°.Subculture if necessary, using the procedure shown to be suitable. Record for each level of dilution the number of tubes showing microbial growth. Determine the most probable number of microorganisms per g or mL of the product to be examined from Table 3.

取规定量样品，按”促生长实验”和”计数方法适用性试验”确认的方法进行供试品液的制备。所有接种管在30~35℃条件下培养3~5天。如必要，采用适宜的方法再培养。记录每个稀释级别微生物生长的管数。从表3查每克或每毫升供试品中最可能数的菌落数。

The total aerobic microbial count (TAMC) is considered to be equal to the number of cfu found usingSoybean–Casein Digest Agar; if colonies of fungi are detected on this medium, they are counted as part ofTAMC. The total combined yeasts and molds count (TYMC) is considered to be equal to the number of cfu found using Sabouraud Dextrose Agar; if colonies of bacteria are detected on this medium, they are counted as part of TYMC. When the TYMC is expected to exceed the acceptance criterion due to the bacterial growth, Sabouraud Dextrose Agar containing antibiotics may be used. If the count is carried out by the MPN Method, the calculated value is TAMC.

总需氧菌数是指TSA培养基上生长的总菌落数（包括霉菌菌落数）；总酵母菌和霉菌数指的是SDA培养基上生长的总菌落数（包括细菌菌落数）。若因SDA培养基上生长的细菌使总酵母菌和霉菌总数超出接收限度，可使用含抗生素的SDA培养基。若采用MPN法，测定结果为总需氧菌数。

When an acceptance criterion for microbiological quality is prescribed, it is interpreted as follows:

各品项下规定的微生物限度标准解释如下：

— 101 cfu: maximum acceptable count = 20;

101 cfu：可接受的最大菌数为20；

— 102 cfu: maximum acceptable count = 200;

102 cfu：可接受的最大菌数为200；

— 103 cfu: maximum acceptable count = 2000;

103 cfu：可接受的最大菌数为2000；

and so forth.

以此类推

The recommended solutions and media are described in Tests for Specified Microorganisms 62.

推荐的溶液和培养基详见USP<62> 控制菌的检测