美國(guó)藥典USP31-NF26無(wú)菌檢查法《71》.doc

71 STERILITY TESTS 無(wú)菌檢查法

Portions of this general chapter have been harmonized with the corresponding texts of the European Pharmacopeia and/or the Japanese Pharmacopeia. Those portions that are not harmonized are marked with symbols ( ) to specify this fact.

此通則的各部分已經(jīng)與歐洲藥典和/或日本藥典的對(duì)應(yīng)部分做了協(xié)調(diào)。不一致的部分用符號(hào)（ ）來(lái)標(biāo)明。

The following procedures are applicable for determining whether a Pharmacopeial article purporting to be sterile complies with the requirements set 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, use 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 Observation and Interpretation of Results.

下面這些步驟適用于測(cè)定是否某個(gè)用于無(wú)菌用途的藥品是否符合其具體的各論中關(guān)于無(wú)菌檢查的要求。只要其性質(zhì)許可，這些藥品將使用供試產(chǎn)品無(wú)菌檢查法項(xiàng)下的膜過(guò)濾法來(lái)檢測(cè)。如果膜過(guò)濾技術(shù)是不適合的，則使用在供試產(chǎn)品無(wú)菌檢查法項(xiàng)下的培養(yǎng)基直接接種法。除了具有標(biāo)記為無(wú)菌通道的設(shè)備之外，所有的設(shè)備均須使用培養(yǎng)基直接接種法進(jìn)行檢測(cè)。在結(jié)果的觀測(cè)與理解項(xiàng)下包含了復(fù)驗(yàn)的規(guī)定。

Because sterility testing is a very exacting procedure, where asepsis 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 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.

由于無(wú)菌檢查法是一個(gè)非常精確的程序，在此過(guò)程中程序的無(wú)菌狀態(tài)必須得到確保以實(shí)現(xiàn)對(duì)結(jié)果的正確理解，因此人員經(jīng)過(guò)適當(dāng)?shù)呐嘤?xùn)并取得資質(zhì)是非常重要的。無(wú)菌檢查在無(wú)菌條件下進(jìn)行。為了實(shí)現(xiàn)這樣的條件，試驗(yàn)環(huán)境必須調(diào)整到適合進(jìn)行無(wú)菌檢查的方式。為避免污染而采取的特定預(yù)防措施應(yīng)不會(huì)對(duì)任何試圖在檢查中發(fā)現(xiàn)的微生物產(chǎn)生影響。通過(guò)在工作區(qū)域作適當(dāng)取樣并進(jìn)行適當(dāng)控制，來(lái)定期監(jiān)測(cè)進(jìn)行此試驗(yàn)的工作條件。

These Pharmacopeial procedures are not by themselves 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 aseptic processing procedures.

這些藥典規(guī)定程序自身的設(shè)計(jì)不能確保一批產(chǎn)品無(wú)菌或已經(jīng)滅菌。這主要是通過(guò)滅菌工藝或者無(wú)菌操作程序的驗(yàn)證來(lái)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, see Sterilization and Sterility Assurance of Compendial Articles 1211 .

當(dāng)通過(guò)適當(dāng)?shù)乃幍浞椒ǐ@得了某物品中微生物污染的證據(jù)，這樣獲得的結(jié)果是該物品未能達(dá)到無(wú)菌檢驗(yàn)要求的結(jié)論性證據(jù)，即便使用替代程序得到了不同的結(jié)果也無(wú)法否定此結(jié)果。 如要獲得關(guān)于無(wú)菌檢驗(yàn)的其他信息，見藥品的滅菌和無(wú)菌保證<1211>

MEDIA 培養(yǎng)基

Prepare media for the tests as described below, or dehydrated formulations may be used 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實(shí)驗(yàn)用培養(yǎng)基；或者使用脫水培養(yǎng)基，只要根據(jù)其制造商或者分銷商說(shuō)明進(jìn)行恢復(fù)之后，其能夠符合好氧菌、厭氧菌、霉菌生長(zhǎng)促進(jìn)試驗(yàn)的要求即可。使用經(jīng)過(guò)驗(yàn)證的工藝對(duì)培養(yǎng)基進(jìn)行滅菌操作。

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–Casein Digest Medium is suitable for the culture of both fungi and aerobic bacteria.

下面的培養(yǎng)基已經(jīng)被證實(shí)適合進(jìn)行無(wú)菌檢查。巰基醋酸鹽液體培養(yǎng)基主要用于厭氧菌的培養(yǎng)。但其也用于檢測(cè)好氧菌。大豆酪蛋白消化物培養(yǎng)基適合于培養(yǎng)霉菌和好氧菌。

Fluid Thioglycollate Medium 巰基醋酸鹽液體培養(yǎng)基

Mix the L-cystine, 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.

將L-胱氨酸、氯化鈉、葡萄糖、酵母提取物、酪蛋白胰酶消化物與純凈水混合，并加熱至實(shí)現(xiàn)溶解。將巰基乙酸鈉或者巰基乙酸溶解于該溶液，如果需要可再加入1N氫氧化鈉，以便在滅菌后該溶液呈pH值7.1 ± 0.2。如需要?jiǎng)t過(guò)濾，再次加熱該溶液但不得煮沸，并趁熱以濕潤(rùn)濾紙將該溶液過(guò)濾。加入刃天青鈉溶液，混勻，并將該培養(yǎng)基置于適當(dāng)容器中，該容器應(yīng)為培養(yǎng)基提供特定的面積－深度比，以使在培養(yǎng)期末表明氧氣攝入的變色部分不超過(guò)培養(yǎng)基的上半部分。使用經(jīng)過(guò)驗(yàn)證的工藝進(jìn)行滅菌。如果需要儲(chǔ)存該培養(yǎng)基，將其置于無(wú)菌、氣密容器中，在2 至25 之間儲(chǔ)藏。如果超過(guò)上部三分之一的培養(yǎng)基已經(jīng)呈粉色，可以用以下方法恢復(fù)該培養(yǎng)基一次：在水浴鍋中或者自由流動(dòng)蒸氣中加熱該容器，直至粉色消失，并迅速放涼，須小心防止非無(wú)菌空氣進(jìn)入到容器中。

Fluid Thioglycollate Medium is to be incubated at 32.5 ± 2.5 .

巰基醋酸鹽液體培養(yǎng)基將在32.5 ± 2.5 條件下進(jìn)行培養(yǎng)。

Alternative Thioglycollate Medium 替代巰基醋酸鹽培養(yǎng)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 use. The pH after sterilization is 7.1 ± 0.2. Incubate under anaerobic conditions for the duration of the incubation period.

配制與巰基醋酸鹽液體培養(yǎng)基成分相同，但省略了瓊脂和刃天青鈉溶液的混合物，按上述方法滅菌，并在使用前靜置至涼。滅菌后pH值為7.1 ± 0.2。在厭氧條件下培養(yǎng)，培養(yǎng)時(shí)間同培養(yǎng)期。

Alternative Fluid Thioglycollate Medium is to be incubated at 32.5 ± 2.5 .

替代性巰基醋酸鹽培養(yǎng)基將在32.5 ± 2.5 條件下進(jìn)行培養(yǎng)。

Soybean–Casein Digest Medium 大豆-酪蛋白消化物培養(yǎng)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, dispense into suitable containers, and sterilize using a validated procedure. Store at a temperature between 2 and 25 in a sterile well-closed container, unless it is intended for immediate use.

將固體物質(zhì)溶解于純凈水，輕微加熱以實(shí)現(xiàn)溶解。放涼溶液至室溫，并用1N氫氧化鈉調(diào)整pH值，以便在滅菌后其pH值呈7.3 ± 0.2。過(guò)濾，如需要?jiǎng)t使之澄清，分裝入適合的容器，并用經(jīng)過(guò)驗(yàn)證的程序消毒。如果不立刻使用，則在2 到25 之間以無(wú)菌且密閉良好的容器保存。

Soybean–Casein Digest Medium is to be incubated at 22.5 ± 2.5 .

大豆-酪蛋白消化物培養(yǎng)基將在22.5 ± 2.5 條件下培養(yǎng)。

Media for Penicillins or Cephalosporins 用于青霉素和頭孢菌素的培養(yǎng)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.]

當(dāng)無(wú)菌檢查培養(yǎng)基用于供試產(chǎn)品無(wú)菌檢查項(xiàng)下的培養(yǎng)基直接接種法時(shí)，按如下內(nèi)容變更巰基醋酸鹽液體培養(yǎng)基和大豆-酪蛋白消化物培養(yǎng)基的制備方法。向每一種培養(yǎng)基的容器中，以無(wú)菌操作轉(zhuǎn)移足夠滅活供試樣品中所存在抗生素的 -內(nèi)酰胺酶。使用此前已經(jīng)對(duì)其青霉素或頭孢菌素滅活能力進(jìn)行了測(cè)定的 -內(nèi)酰胺酶配制品，來(lái)測(cè)定滅活該抗生素所必需的 -內(nèi)酰胺酶數(shù)量。[注意：補(bǔ)充的 -內(nèi)酰胺酶培養(yǎng)基也可以用于膜過(guò)濾試驗(yàn)]

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（在與無(wú)菌試驗(yàn)所用場(chǎng)所徹底隔離的區(qū)域中），按照驗(yàn)證試驗(yàn)項(xiàng)下的任意一種方法，使用少于100個(gè)菌落（cfu）的金黃色葡萄球菌（見表1）作為驗(yàn)證菌，來(lái)確認(rèn)適當(dāng)數(shù)量的 -內(nèi)酰胺酶已經(jīng)被整合到該培養(yǎng)基中。必須觀測(cè)到接種后培養(yǎng)物中出現(xiàn)典型微生物生長(zhǎng)，才能確認(rèn) -內(nèi)酰胺酶濃度是適當(dāng)?shù)摹?/P>

Table 1. Strains of the Test Microorganisms Suitable for Use in the Growth Promotion Test and the
Validation Test

表1 適合用于生長(zhǎng)促進(jìn)試驗(yàn)和驗(yàn)證試驗(yàn)中的試驗(yàn)微生物的菌株

Suitability Tests 適合性試驗(yàn)

The media used comply with the following tests, carried out before, or in parallel, with the test on the product to be examined.

所使用的培養(yǎng)基須符合下列試驗(yàn)，這些試驗(yàn)應(yīng)在檢驗(yàn)供試產(chǎn)品之前或者同時(shí)進(jìn)行。

STERILITY 無(wú)菌狀態(tài)

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.

通過(guò)在指定培養(yǎng)溫度下將一部分培養(yǎng)基培養(yǎng)14天，來(lái)確認(rèn)每一批已滅菌培養(yǎng)基的無(wú)菌狀態(tài)。不得出現(xiàn)微生物生長(zhǎng)。

GROWTH PROMOTION TEST OF AEROBES, ANAEROBES, and FUNGI

好氧菌、厭氧菌、霉菌的生長(zhǎng)促進(jìn)試驗(yàn)

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.

檢查每一批已經(jīng)配制好的培養(yǎng)基和每一批用脫水培養(yǎng)基或配料制備的培養(yǎng)基 ¹ 。適當(dāng)微生物菌株見表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.

在部分巰基醋酸鹽液體培養(yǎng)基上接種少量（不超過(guò)100cfu）下列微生物，每一種微生物均使用單獨(dú)一部分培養(yǎng)基：產(chǎn)芽胞梭狀芽胞桿菌、綠膿桿菌、金黃色葡萄球菌。 在部分替代巰基醋酸鹽液體培養(yǎng)基上接種少量（不超過(guò)100cfu）產(chǎn)芽胞梭狀芽胞桿菌。 在部分大豆-酪蛋白消化物培養(yǎng)基上接種少量（不超過(guò)100cfu）下列微生物，每一種微生物均使用單獨(dú)一部分的培養(yǎng)基：黑曲霉、枯草芽孢桿菌、白色念珠菌。細(xì)菌培養(yǎng)時(shí)間不超過(guò)3天，霉菌培養(yǎng)時(shí)間不超過(guò)5天。

The media are suitable if a clearly visible growth of the microorganisms occurs.

如果出現(xiàn)清晰可見的微生物生長(zhǎng)，則該培養(yǎng)基是適合的。

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.

如果配制好的培養(yǎng)基保存于未密閉的容器中，只要在使用時(shí)間的2周內(nèi)對(duì)其進(jìn)行了生長(zhǎng)促進(jìn)試驗(yàn)并且符合顏色指示劑的要求，它們就可以使用1個(gè)月。如果保存在密閉的容器中，只要在使用時(shí)間的3個(gè)月內(nèi)對(duì)其進(jìn)行了生長(zhǎng)促進(jìn)試驗(yàn)并且符合顏色指示劑的要求，則該培養(yǎng)基可以使用1年。

DILUTING AND RINSING FLUIDS FOR MEMBRANE FILTRATION

用于膜過(guò)濾的稀釋和沖洗液

Fluid A 液體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.

將1g動(dòng)物組織胃蛋白酶消化物溶于1L水中，如果需要?jiǎng)t通過(guò)濾或離心使其澄清，再調(diào)節(jié)pH值至7.1 ± 0.2。分裝入容器中，并用經(jīng)過(guò)驗(yàn)證的工藝滅菌。

PREPARATION FOR PENICILLINS OR CEPHALOSPORINS

用于青霉素或頭孢菌素的配制品

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).

在供試樣品溶液已經(jīng)過(guò)濾（見用于青霉素或頭孢菌素的培養(yǎng)基）之后，如果需要，向上述配制品中，以無(wú)菌操作加入數(shù)量足夠滅活濾膜上殘余抗生素活性的 -內(nèi)酰胺酶。

Fluid D 液體D

To each L of Fluid A add 1 mL of polysorbate 80, adjust to a pH of 7.1 ± 0.2, dispense into containers, and sterilize using a validated process. Use this fluid for articles containing lecithin or oil, or for devices labeled as “sterile pathway.”

向每升液體A中，加入1mL聚山梨酯80，調(diào)節(jié)pH值至7.1 ± 0.2，分裝入容器中，并使用經(jīng)過(guò)驗(yàn)證的工藝滅菌。此液體用于含有卵磷脂或油脂的物品，或用于標(biāo)為 “無(wú)菌通道”的設(shè)備。

Fluid K 液體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. Dispense into containers, and sterilize using a validated process.

將5.0g動(dòng)物組織胃蛋白酶消化物、3.0g牛肉提取物、10.0g聚山梨酯80溶解于1L水中。調(diào)節(jié)pH值，以便使pH值在滅菌后呈6.9 ± 0.2。分裝入容器中，并使用經(jīng)過(guò)驗(yàn)證的工藝滅菌。

VALIDATION TEST 驗(yàn)證試驗(yàn)

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.

按照下面供試產(chǎn)品無(wú)菌檢查項(xiàng)下的描述，使用除了下面變更之外完全相同的方法，進(jìn)行試驗(yàn)。

Membrane Filtration 膜過(guò)濾

After transferring the content of the container or containers to be tested to the membrane, add an inoculum of a small number of viable microorganisms (not more than 100 cfu) to the final portion of sterile diluent used to rinse the filter.

在將一個(gè)或多個(gè)供試容器中的內(nèi)容物轉(zhuǎn)移到濾膜之后，在最后一次的沖洗液中加入少量（不超過(guò)100cfu）試驗(yàn)菌.

Direct Inoculation 直接接種

After transferring the contents of the container or containers to be tested (for catgut and other surgical sutures for veterinary use: strands) to the culture medium, add an inoculum of a small number of viable microorganisms (not more than 100 cfu) to the medium.

在將一個(gè)或多個(gè)供試容器（對(duì)于獸醫(yī)的腸線和其他外科縫合用線：若干股線）中的內(nèi)容物轉(zhuǎn)移至培養(yǎng)基之后，將少量試驗(yàn)菌（不超過(guò)100cfu）加入至培養(yǎng)基中。

In both cases use the same microorganisms as those described above under Growth Promotion Test of Aerobes, Anaerobes, and Fungi. Perform a growth promotion test as a positive control. Incubate all the containers containing medium for not more than 5 days.

在這兩種情況中，均按照上述好氧菌、厭氧菌、霉菌生長(zhǎng)促進(jìn)試驗(yàn)項(xiàng)下的描述，使用同樣的微生物。進(jìn)行一個(gè)生長(zhǎng)促進(jìn)試驗(yàn)作為陽(yáng)性對(duì)照。培養(yǎng)所有含有培養(yǎng)基的容器，培養(yǎng)時(shí)間不超過(guò)5天。

If clearly visible growth of microorganisms is obtained after the incubation, visually comparable to that in the control vessel without product, either the product possesses no antimicrobial activity under the conditions of the test or such activity has been satisfactorily eliminated. The test for sterility may then be carried out without further modification.

如果在培養(yǎng)后得到清晰可見的微生物生長(zhǎng)，看起來(lái)與沒(méi)有產(chǎn)品的對(duì)照容器中的生長(zhǎng)類似，則該產(chǎn)品在此試驗(yàn)條件下沒(méi)有任何抗微生物活性，或者此活性已經(jīng)被令人滿意地消除了。然后，無(wú)菌試驗(yàn)可以進(jìn)行，而無(wú)需進(jìn)一步的變更。

If clearly visible growth is not obtained in the presence of the product to be tested, visually comparable to that in the control vessels without product, the product possesses antimicrobial activity that has not been satisfactorily eliminated under the conditions of the test. Modify the conditions in order to eliminate the antimicrobial activity, and repeat the validation test.

如果用肉眼與沒(méi)有產(chǎn)品的對(duì)照容器比較，無(wú)法在存在供試產(chǎn)品的情況下得到清晰可見的生長(zhǎng)，則該產(chǎn)品在試驗(yàn)條件下所具有的抗微生物活性尚未令人滿意地消除。變更條件以便消除抗微生物活性，并重復(fù)驗(yàn)證試驗(yàn)。

This validation is performed (a) when the test for sterility has to be carried out on a new product; and (b) whenever there is a change in the experimental conditions of the test. The validation may be performed simultaneously with the Test for Sterility of the Product to be Examined.

當(dāng)(a)一個(gè)新產(chǎn)品必須進(jìn)行無(wú)菌試驗(yàn)時(shí)，和(b)無(wú)論何時(shí)該試驗(yàn)的試驗(yàn)條件發(fā)生改變時(shí)，則需進(jìn)行此驗(yàn)證試驗(yàn)。該驗(yàn)證可以與供試產(chǎn)品的無(wú)菌試驗(yàn)同時(shí)進(jìn)行。

TEST FOR STERILITY OF THE PRODUCT TO BE EXAMINED

供試產(chǎn)品的無(wú)菌檢查

Number of Articles to Be Tested 供試物品數(shù)量

Unless otherwise specified elsewhere in this chapter or in the individual monograph, test the number of articles specified in Table 3. If the contents of each article are of sufficient quantity (see Table 2), they may be divided so that equal appropriate portions are added to each of the specified media. [NOTE—Perform sterility testing employing two or more of the specified media.] If each article does not contain sufficient quantities for each medium, use twice the number of articles indicated in Table 3.

除非在此章節(jié)的其他位置或在具體的各論中另有規(guī)定，供試物品的數(shù)量遵照表3中的規(guī)定。如果每個(gè)物品的內(nèi)容物有足夠數(shù)量（見表2），可以將其分成若干等份，將適當(dāng)?shù)牡确菁尤氲矫總�(gè)指定的培養(yǎng)基。[注意：使用兩個(gè)或更多指定培養(yǎng)基，來(lái)進(jìn)行無(wú)菌試驗(yàn)。]如果每個(gè)物品內(nèi)容物的數(shù)量不夠每個(gè)培養(yǎng)基的用量，使用表3中所規(guī)定物品數(shù)量的2倍。

Table 2. Minimum Quantity to be Used for Each Medium

表2：用于每個(gè)培養(yǎng)基的最小數(shù)量

Table 3. Minimum Number of Articles to be Tested in Relation to the Number of Articles in the Batch 表3：與物品批量相關(guān)的最小供試物品數(shù)量

The test may be carried out using the technique of Membrane Filtration or by Direct Inoculation of the Culture Medium with the product to be examined. Appropriate negative controls are included. The technique of membrane filtration is used whenever the nature of the product permits; that is, for filterable aqueous preparations, for alcoholic or oily preparations, and for preparations miscible with, or soluble in, aqueous or oily solvents, provided these solvents do not have an antimicrobial effect in the conditions of the test.

此試驗(yàn)可以使用膜過(guò)濾法或培養(yǎng)基直接接種法進(jìn)行。應(yīng)包括多個(gè)適當(dāng)?shù)年幮詫?duì)照。只要該產(chǎn)品的性質(zhì)許可，就應(yīng)使用膜過(guò)濾法；這些性質(zhì)是，可過(guò)濾的水溶性配制品、酒精或油性配制品、易混合或溶解于水或油性溶劑的配制品，只要這些溶劑在試驗(yàn)條件下沒(méi)有抗生素效果。

Membrane Filtration 膜過(guò)濾

Use membrane filters having a nominal pore size not greater than 0.45 µm whose effectiveness to retain microorganisms has been established. Cellulose nitrate filters, for example, are used for aqueous, oily, and weakly alcoholic solutions; and cellulose acetate filters, for example, are used for strongly alcoholic solutions. Specially adapted filters may be needed for certain products (e.g., for antibiotics).

使用標(biāo)稱孔徑不大于0.45 µm的膜過(guò)濾器，此孔徑已知能夠有效截留微生物。例如，硝酸纖維素過(guò)濾器可用于水、油、稀醇溶液；而醋酸纖維素可用于濃醇溶液。特定產(chǎn)品（例如，抗生素）可能需要特別改造過(guò)的過(guò)濾器。

The technique described below assumes that membranes about 50 mm in diameter will be used. If filters of a different diameter are used, the volumes of the dilutions and the washings should be adjusted accordingly. The filtration apparatus and membrane are sterilized by appropriate means. The apparatus is designed so that the solution to be examined can be introduced and filtered under aseptic conditions: it permits the aseptic removal of the membrane for transfer to the medium, or it is suitable for carrying out the incubation after adding the medium to the apparatus itself.

下面描述的方法所使用直徑約50mm的濾膜。如果使用不同直徑的過(guò)濾器，稀釋液和洗液的體積應(yīng)當(dāng)作相應(yīng)調(diào)節(jié)。以適當(dāng)方法將過(guò)濾設(shè)備和濾膜滅菌。該設(shè)備設(shè)計(jì)用于在無(wú)菌條件下加入和過(guò)濾供試溶液：其使得在無(wú)菌狀態(tài)下將濾膜摘掉轉(zhuǎn)移至培養(yǎng)基成為可能，或者其適合于將培養(yǎng)基加入該設(shè)備自身之中，并進(jìn)行培養(yǎng)。

AQUEOUS SOLUTIONS 水性溶液

If appropriate, transfer a small quantity of a suitable, sterile diluent such as Fluid A (see Diluting and Rinsing Fluids for Membrane Filtration) onto the membrane in the apparatus and filter. The diluent may contain suitable neutralizing substances and/or appropriate inactivating substances, for example, in the case of antibiotics.

如果適當(dāng)，將少量適當(dāng)?shù)臒o(wú)菌稀釋劑，例如液體A（見用于膜過(guò)濾的稀釋和沖洗液），轉(zhuǎn)移至設(shè)備中的濾膜上并過(guò)濾。該稀釋劑可以含有適當(dāng)?shù)闹泻臀镔|(zhì)和/或適當(dāng)?shù)臏缁钗镔|(zhì)，例如針對(duì)抗生素。

Transfer the contents of the container or containers to be tested to the membrane or membranes, if necessary, after diluting to the volume used in the Validation Test with the chosen sterile diluent, but using not less than the quantities of the product to be examined prescribed in Tables 2 and 3. Filter immediately. If the product has antimicrobial properties, wash the membrane not less than three times by filtering through it each time the volume of the chosen sterile diluent used in the Validation Test. Do not exceed a washing cycle of 5 times 200 mL, even if during validation it has been demonstrated that such a cycle does not fully eliminate the antimicrobial activity. Transfer the whole membrane to the culture medium or cut it aseptically into two equal parts, and transfer one half to each of two suitable media. Use the same volume of each medium as in the Validation Test. Alternatively, transfer the medium onto the membrane in the apparatus. Incubate the media for not less than 14 days.

將一個(gè)或多個(gè)供試容器的內(nèi)容物轉(zhuǎn)移到濾膜，如需要可先使用選定的無(wú)菌稀釋劑稀釋至驗(yàn)證試驗(yàn)中所用體積，但須使用不少于表2和3中規(guī)定的供試產(chǎn)品數(shù)量。立即過(guò)濾。如果該產(chǎn)品具有抗微生物特性，沖洗濾膜不少于3次，每次均將驗(yàn)證試驗(yàn)中所使用的無(wú)菌稀釋劑體積濾過(guò)該濾膜。即便驗(yàn)證中顯示5次200mL的沖洗循環(huán)沒(méi)有完全消除抗微生物活性，也不要超越這樣一個(gè)循環(huán)。轉(zhuǎn)移整個(gè)濾膜至培養(yǎng)基，或以無(wú)菌操作將其切開至相等的2部分，并將每一部分轉(zhuǎn)移至適當(dāng)?shù)呐囵B(yǎng)基中。每個(gè)培養(yǎng)基的體積與驗(yàn)證試驗(yàn)所用的一樣。或者，將培養(yǎng)基轉(zhuǎn)移至設(shè)備中的濾膜上。培養(yǎng)該培養(yǎng)基，不少于14天。

SOLUBLE SOLIDS (other than antibiotics) 可溶固體（非抗生素）

Use for each medium not less than the quantity prescribed in Tables 2 and 3 of the product dissolved in a suitable solvent, such as Fluid A (Diluting and Rinsing Fluids for Membrane Filtration), and proceed with the test as described above for Aqueous Solutions using a membrane appropriate to the chosen solvent.

在每個(gè)培養(yǎng)基中，使用不少于表2和3規(guī)定的產(chǎn)品數(shù)量溶于適當(dāng)溶劑，例如溶液A（用于膜過(guò)濾的稀釋和沖洗液），并按照上述關(guān)于水性溶液的描述，使用適合所選溶劑的濾膜，繼續(xù)進(jìn)行該試驗(yàn)。

OILS and OILY SOLUTIONS 油和油性溶液

Use for each medium not less than the quantity of the product prescribed in Tables 2 and 3. Oils and oily solutions of sufficiently low viscosity may be filtered without dilution through a dry membrane. Viscous oils may be diluted as necessary with a suitable sterile diluent such as isopropyl myristate shown not to have antimicrobial activity in the conditions of the test. Allow the oil to penetrate the membrane by its own weight, and then filter, applying the pressure or suction gradually. Wash the membrane at least three times by filtering through it each time about 100 mL of a suitable sterile solution such as Fluid A (see Diluting and Rinsing Fluids for Membrane Filtration) containing a suitable emulsifying agent at a concentration shown to be appropriate in the validation of the test, for example polysorbate 80 at a concentration of 10 g per L (Fluid K) . Transfer the membrane or membranes to the culture medium or media, or vice versa, as described above for Aqueous Solutions, and incubate at the same temperatures and for the same times.

在每個(gè)培養(yǎng)基中，使用不少于表2和3中描述的產(chǎn)品用量。粘性足夠低的油和油性溶液可能在不經(jīng)稀釋的情況下濾過(guò)干燥濾膜。如需要，粘稠油質(zhì)可以用適合的無(wú)菌稀釋劑進(jìn)行稀釋，例如已證實(shí)在該試驗(yàn)條件下不具有抗微生物活性的豆蔻酸異丙酯。使該油質(zhì)依靠其自身的重量穿過(guò)濾膜，然后逐漸應(yīng)用壓力或抽吸過(guò)濾。每次過(guò)濾約100mL適當(dāng)?shù)臒o(wú)菌溶液，例如液體A（見用于膜過(guò)濾的稀釋和沖洗液），并含有適當(dāng)乳化劑且其濃度已證實(shí)適用于該試驗(yàn)的驗(yàn)證，例如濃度為10克每升的聚山梨酯80（液體K）。將一個(gè)或多個(gè)濾膜轉(zhuǎn)移到一個(gè)或多個(gè)培養(yǎng)基，或反之，如上面關(guān)于水性溶液的描述，并在相同溫度下培養(yǎng)同樣的時(shí)間。

OINTMENTS and CREAMS 油膏和乳膏

Use for each medium not less than the quantities of the product prescribed in Tables 2 and 3. Ointments in a fatty base and emulsions of the water-in-oil type may be diluted to 1% in isopropyl myristate as described above, by heating, if necessary, to not more than 40 . In exceptional cases it may be necessary to heat to not more than 44 . Filter as rapidly as possible, and proceed as described above for Oils and Oily Solutions.

在每個(gè)培養(yǎng)基中，使用不少于表2和3中描述的產(chǎn)品用量。脂肪狀的油膏和水在油中形態(tài)乳化劑可以按照上面所述，在豆蔻酸異丙酯中稀釋至1%，如需要可加熱至不高于40 。在特別情況下，其可能必須加熱到不超過(guò)44 。盡可能迅速地過(guò)濾，并按照上面針對(duì)油和油性溶液所述內(nèi)容繼續(xù)操作。

PREFILLED SYRINGES 預(yù)裝填的注射器

For prefilled syringes without attached sterile needles, expel the contents of each syringe into one or two separate membrane filter funnels or into separate pooling vessels prior to transfer. If a separate sterile needle is attached, directly expel the syringe contents as indicated above, and proceed as directed for Aqueous Solutions. Test the sterility of the needle, using Direct Inoculation under Validation Test.

對(duì)于沒(méi)有附無(wú)菌針頭的預(yù)裝填注射器，在轉(zhuǎn)移之前，將每個(gè)注射器的內(nèi)容物排出至一個(gè)或兩個(gè)單獨(dú)的膜過(guò)濾器漏斗，或至若干單獨(dú)的合并容器。如果附了單獨(dú)的滅菌針頭，直接按照上面的規(guī)定將注射器內(nèi)容物直接排出，并按照關(guān)于水性溶液的規(guī)定繼續(xù)進(jìn)行。使用驗(yàn)證試驗(yàn)項(xiàng)下的直接接種，檢查針頭的無(wú)菌情況。

SOLIDS FOR INJECTION OTHER THAN ANTIBIOTICS 除了抗生素之外的注射用固體

Constitute the test articles as directed on the label, and proceed as directed for Aqueous Solutions or Oils and Oily Solutions, whichever applies. [NOTE—If necessary, excess diluent can be added to aid in the constitution and filtration of the constituted test article.]

按照其標(biāo)簽上的規(guī)定配制供試物品，并按照適用的關(guān)于水性溶液或油和油性溶液的規(guī)定繼續(xù)進(jìn)行。[注意：如需要，可以加入額外的稀釋劑以幫助對(duì)已配制的供試物品進(jìn)行再配制和過(guò)濾]

ANTIBIOTIC SOLIDS FOR INJECTION 用于注射的抗生素

Pharmacy Bulk Packages, < 5 g— From each of 20 containers, aseptically transfer about 300 mg of solids, into a sterile 500-mL conical flask, dissolve in about 200 mL of Fluid A (see Diluting and Rinsing Fluids for Membrane Filtration), and mix; or constitute, as directed in the labeling, each of 20 containers and transfer a quantity of liquid or suspension, equivalent to about 300 mg of solids, into a sterile 500-mL conical flask, dissolve in about 200 mL of Fluid A, and mix. Proceed as directed for Aqueous Solutions or Oils and Oily Solutions, whichever applies.

藥房散裝< 5 g：取20個(gè)容器，每個(gè)容器均以無(wú)菌操作將約300mg固體轉(zhuǎn)移至一個(gè)無(wú)菌的500mL錐形燒瓶，溶解于約200mL液體A中（見用于膜過(guò)濾的稀釋和沖洗液），并混勻；或取20個(gè)容器，每個(gè)容器均按照標(biāo)簽上的規(guī)定配制，并將相當(dāng)于大約300mg固體的液體或懸浮液轉(zhuǎn)移至一個(gè)無(wú)菌的500mL錐形燒瓶，溶解于約200mL液體A中，并混勻。按照適合的關(guān)于水性溶液或油和油性溶液的規(guī)定，繼續(xù)操作。

Pharmacy Bulk Packages, 5 g— From each of 6 containers, aseptically transfer about 1 g of solids into a sterile 500-mL conical flask, dissolve in about 200 mL of Fluid A, and mix; or constitute, as directed in the labeling, each of 6 containers and transfer a quantity of liquid, equivalent to about 1 g of solids, into a sterile 500-mL conical flask, dissolve in about 200 mL of Fluid A, and mix. Proceed as directed for Aqueous Solutions.

藥方散裝 5 g：取6個(gè)容器，每個(gè)均以無(wú)菌操作將大約1克的固體轉(zhuǎn)移至一個(gè)無(wú)菌的500mL錐形燒瓶，溶解于約200mL液體A中，并混勻；或取6個(gè)容器，每個(gè)均按照標(biāo)簽的規(guī)定配制，將相當(dāng)于1g固體的液體轉(zhuǎn)移至一個(gè)無(wú)菌的500mL錐形燒瓶，溶解于200mL液體A中，并混勻。按照關(guān)于水性溶液的規(guī)定，繼續(xù)操作。

ANTIBIOTIC SOLIDS, BULKS, and BLENDS 抗生素固體、散裝品、混合品

Aseptically remove a sufficient quantity of solids from the appropriate amount of containers (see Table 2), mix to obtain a composite, equivalent to about 6 g of solids, and transfer to a sterile 500-mL conical flask. Dissolve in about 200 mL of Fluid A, and mix. Proceed as directed for Aqueous Solutions.

以無(wú)菌操作從適當(dāng)數(shù)量的容器中（見表2）取出足夠數(shù)量的固體，混勻以獲得等同于6g固體的混合物，轉(zhuǎn)移至一個(gè)無(wú)菌的500mL錐形燒瓶。溶解于約200mL液體A中，并混勻。按照關(guān)于水性溶液的規(guī)定，繼續(xù)進(jìn)行。

STERILE AEROSOL PRODUCTS 無(wú)菌氣(噴)霧劑產(chǎn)品

For fluid products in pressurized aerosol form, freeze the containers in an alcohol-dry ice mixture at least at –20 for about 1 hour. If feasible, allow the propellant to escape before aseptically opening the container, and transfer the contents to a sterile pooling vessel. Add 100 mL of Fluid D to the pooling vessel, and mix gently. Proceed as directed for Aqueous Solutions or Oils and Oily Solutions, whichever applies.

對(duì)于以加壓氣(噴)霧劑形式存在的液體產(chǎn)品，在大約–20 的酒精－干冰混合物中冷凍容器約1小時(shí)。如果可行，在以無(wú)菌操作打開容器之前使推進(jìn)劑散發(fā)掉，并將內(nèi)容物轉(zhuǎn)移至一個(gè)無(wú)菌的合并容器。加入100mL液體D至該合并容器，并輕輕混勻。按照適合的關(guān)于水性溶液或油和油性溶液的規(guī)定，繼續(xù)進(jìn)行。

DEVICES WITH PATHWAYS LABELED STERILE 具有導(dǎo)管的醫(yī)療器具供試品

Aseptically pass not less than 10 pathway volumes of Fluid D through each device tested. Collect the fluids in an appropriate sterile vessel, and proceed as directed for Aqueous Solutions or Oils and Oily Solutions, whichever applies.

以無(wú)菌操作用10個(gè)通道體積的液體D通過(guò)供試設(shè)備。在適當(dāng)?shù)臒o(wú)菌容器中收集沖洗液，并按照適合的關(guān)于水性溶液或油和油性溶液的規(guī)定，繼續(xù)進(jìn)行。

In the case of sterile, empty syringes, draw sterile diluent into the barrel through the sterile needle, if attached, or th, rough a sterile needle attached for the purpose of the test, and express the contents into a sterile pooling vessel. Proceed as directed above.

對(duì)于無(wú)菌的空注射器，如果附有無(wú)菌針頭，通過(guò)其將無(wú)菌稀釋劑吸取至管中，或者使用一個(gè)專為此試驗(yàn)準(zhǔn)備的無(wú)菌針頭，并將內(nèi)容物壓出至合并容器中。按照上述內(nèi)容繼續(xù)進(jìn)行。

Direct Inoculation of the Culture Medium 培養(yǎng)基的直接接種

Transfer the quantity of the preparation to be examined prescribed in Tables 2 and 3 directly into the culture medium so that the volume of the product is not more than 10% of the volume of the medium, unless otherwise prescribed.

If the product to be examined has antimicrobial activity, carry out the test after neutralizing this with a suitable neutralizing substance or by dilution in a sufficient quantity of culture medium. When it is necessary to use a large volume of the product, it may be preferable to use a concentrated culture medium prepared in such a way that it takes into account the subsequent dilution. Where appropriate, the concentrated medium may be added directly to the product in its container.

按照表2和3規(guī)定的供試配制品的數(shù)量，將該配制品直接轉(zhuǎn)移到培養(yǎng)基中，除非另有規(guī)定，產(chǎn)品體積不得超過(guò)該培養(yǎng)基體積的10%。如果該供試產(chǎn)品具有抗微生物活性，通過(guò)使用適當(dāng)?shù)闹泻臀镔|(zhì)或在充足數(shù)量的培養(yǎng)基中稀釋以中和其抗菌活性之后，進(jìn)行該試驗(yàn)。當(dāng)必須使用大量產(chǎn)品時(shí)，最好使用在配制時(shí)考慮了后續(xù)的稀釋需要的濃縮培養(yǎng)基。在適當(dāng)時(shí)，該濃縮培養(yǎng)基可以直接加入到放在其容器中的產(chǎn)品。

OILY LIQUIDS 油性液體

Use media to which have been added a suitable emulsifying agent at a concentration shown to be appropriate in the validation of the test, for example polysorbate 80 at a concentration of 10 g per L.

使用已經(jīng)加入了適當(dāng)乳化劑的培養(yǎng)基，乳化劑濃度需已經(jīng)證實(shí)適于該試驗(yàn)的驗(yàn)證，例如濃度為10克每升的聚山梨酯80。

OINTMENTS and CREAMS 油膏和乳膏

Prepare by diluting to about 1 in 10 by emulsifying with the chosen emulsifying agent in a suitable sterile diluent such as Fluid A (see Diluting and Rinsing Fluids for Membrane Filtration). Transfer the diluted product to a medium not containing an emulsifying agent.

通過(guò)將選定的乳化劑在適當(dāng)?shù)臒o(wú)菌稀釋液中乳化，例如液體A（見用于膜過(guò)濾的稀釋和沖洗液），稀釋至約1比10，來(lái)配制該產(chǎn)品。轉(zhuǎn)移稀釋后的產(chǎn)品至不含乳化劑的一個(gè)培養(yǎng)基中。

Incubate the inoculated media for not less than 14 days. Observe the cultures several times during the incubation period. Shake cultures containing oily products gently each day. However, when thioglycollate medium or other similar medium is used for the detection of anaerobic microorganisms, keep shaking or mixing to a minimum in order to maintain anaerobic conditions.

將接種后的培養(yǎng)基培養(yǎng)不少于14天。在培養(yǎng)期中觀察培養(yǎng)基若干次。每天輕輕搖動(dòng)含有油性產(chǎn)品的培養(yǎng)基一次。但是，當(dāng)使用巰基醋酸鹽培養(yǎng)基或其他相似培養(yǎng)基檢測(cè)厭氧微生物時(shí)，將搖動(dòng)或混合保持到最少，以維持厭氧條件。

CATGUT and OTHER SURGICAL SUTURES FOR VETERINARIAN USE

獸醫(yī)用腸線和其他外科縫合線

Use for each medium not less than the quantities of the product prescribed in Tables 2 and 3. Open the sealed package using aseptic precautions, and remove three sections of the strand for each culture medium. Carry out the test on three sections, each 30-cm long, which have been cut off from the beginning, the center, and the end of the strand. Use whole strands from freshly opened cassette packs. Transfer each section of the strand to the selected medium. Use sufficient medium to cover adequately the material to be tested (20 mL to 150 mL).

在每個(gè)培養(yǎng)基中，使用不少于表2和3中所規(guī)定數(shù)量的產(chǎn)品。采取無(wú)菌預(yù)防措施，打開封閉的包裝，并取該股線的3個(gè)部分，分別至每個(gè)培養(yǎng)基。使用三節(jié)，每節(jié)30cm長(zhǎng)，并分別從該股線的前端、中間、末端截取的產(chǎn)品，進(jìn)行該試驗(yàn)。使用從剛剛打開的包裝盒中取出的整股線。將該股線的每個(gè)部分轉(zhuǎn)移至選定的培養(yǎng)基。使用充足的培養(yǎng)基，以充分覆蓋該供試物料（20mL至150mL）

SOLIDS 固體

Transfer a quantity of the product in the form of a dry solid (or prepare a suspension of the product by adding sterile diluent to the immediate container), corresponding to not less than the quantity indicated in Tables 2 and 3. Transfer the material so obtained to 200 mL of Fluid Thioglycollate Medium, and mix. Similarly, transfer the same quantity to 200 mL of Soybean–Casein Digest Medium, and mix. Proceed as directed above.

轉(zhuǎn)移干燥固體形態(tài)的產(chǎn)品（或通過(guò)加入無(wú)菌稀釋劑至中間容器中配制該產(chǎn)品的懸浮液），數(shù)量不少于表2和3中的規(guī)定。轉(zhuǎn)移如此獲得的物料至200mL巰基醋酸鹽液體培養(yǎng)基中，并混勻。同法轉(zhuǎn)移同樣數(shù)量的物料至200mL大豆－酪蛋白消化物培養(yǎng)基，并混勻。按照上述規(guī)定繼續(xù)進(jìn)行。

PURIFIED COTTON, GAUZE, SURGICAL DRESSINGS, and RELATED ARTICLES

脫脂棉花、紗布、外科敷料、相關(guān)物品

From each package of cotton, rolled gauze bandage, or large surgical dressings being tested, aseptically remove two or more portions of 100- to 500-mg each from the innermost part of the sample. From individually packaged, single-use materials, aseptically remove the entire article. Immerse the portions or article in each medium, and proceed as directed above.

對(duì)于待檢的每個(gè)包裝中的棉花、卷狀紗布繃帶、大塊外科敷料，以無(wú)菌操作從該樣品最核心的部位，取出2個(gè)或更多部分，每個(gè)部分100-500mg。從單個(gè)包裝、一次性使用的物料中，以無(wú)菌操作取出整個(gè)物品。將這些部分或單個(gè)物品浸沒(méi)在每個(gè)培養(yǎng)基中，并繼續(xù)按照上述內(nèi)容操作。

STERILE DEVICES 無(wú)菌設(shè)備

Articles can be immersed intact or disassembled. To ensure that device pathways are also in contact with the media, immerse the appropriate number of units per medium in a volume of medium sufficient to immerse the device completely, and proceed as directed above. For extremely large devices, immerse those portions of the device that are to come into contact with the patient in a volume of medium sufficient to achieve complete immersion of those portions.

若干物品可以完整地或在拆開后浸沒(méi)在培養(yǎng)基中。為確保設(shè)備通道與培養(yǎng)基接觸，使用體積足夠浸沒(méi)整個(gè)設(shè)備的培養(yǎng)基，在每個(gè)培養(yǎng)基中浸入適當(dāng)數(shù)量的部件，并按照上述內(nèi)容繼續(xù)操作。對(duì)于極大的設(shè)備，將該設(shè)備中將要與患者接觸的那些部分，浸入到體積足夠完全浸沒(méi)這些部分的培養(yǎng)基中。

For catheters where the inside lumen and outside are required to be sterile, either cut them into pieces such that the medium is in contact with the entire lumen or fill the lumen with medium, and then immerse the intact unit.

對(duì)于內(nèi)部的腔體和外部均要求無(wú)菌的導(dǎo)尿管，將它們切成片，這樣培養(yǎng)基就可以接觸整個(gè)腔體，或者用培養(yǎng)基填充腔體，然后浸沒(méi)整個(gè)設(shè)備。

OBSERVATION AND INTERPRETATION OF RESULTS 結(jié)果的觀測(cè)和理解

At intervals during the incubation period and at its conclusion, examine the media for macroscopic evidence of microbial growth. If the material being tested renders the medium turbid so that the presence or absence of microbial growth cannot be readily determined by visual examination, 14 days after the beginning of incubation transfer portions (each not less than 1 mL) of the medium to fresh vessels of the same medium, and then incubate the original and transfer vessels for not less than 4 days.

在培養(yǎng)期中間的觀測(cè)點(diǎn)和作結(jié)論時(shí)，檢查該培養(yǎng)基是否有肉眼可見的微生物生長(zhǎng)的證據(jù)。如果供試物料導(dǎo)致培養(yǎng)基混濁，從而使得無(wú)法通過(guò)肉眼觀察來(lái)確定是否存在微生物生長(zhǎng)，在開始培養(yǎng)14天之后，將該培養(yǎng)基的若干部分（每個(gè)部分不少于1mL）轉(zhuǎn)移至裝有相同培養(yǎng)基的新鮮容器中，然后將原有和轉(zhuǎn)移的容器培養(yǎng)不少于4天。

If no evidence of microbial growth is found, the product to be examined complies with the test for sterility. If evidence of microbial growth is found, the product to be examined does not comply with the test for sterility, unless it can be clearly demonstrated that the test was invalid for causes unrelated to the product to be examined. The test may be considered invalid only if one or more of the following conditions are fulfilled:

如果沒(méi)有找到微生物生長(zhǎng)的證據(jù)，則該供試產(chǎn)品符合無(wú)菌檢查。如果找到了微生物生長(zhǎng)的證據(jù)，則該供試產(chǎn)品不符合無(wú)菌檢查，除非能夠清楚地證實(shí)此次試驗(yàn)無(wú)效且無(wú)效原因與該供試產(chǎn)品無(wú)關(guān)。該試驗(yàn)僅可能在下面一個(gè)或多個(gè)條件被滿足的情況下，才有可能認(rèn)為無(wú)效：

a. The data of the microbiological monitoring of the sterility testing facility show a fault.該無(wú)菌試驗(yàn)設(shè)施的微生物監(jiān)控?cái)?shù)據(jù)顯示有缺陷。

b. A review of the testing procedure used during the test in question reveals a fault.對(duì)該試驗(yàn)過(guò)程中存有疑問(wèn)的試驗(yàn)步驟進(jìn)行審核后揭示出缺陷。

c. Microbial growth is found in the negative controls.在陰性對(duì)照中發(fā)現(xiàn)微生物生長(zhǎng)。

d. After determination of the identity of the microorganisms isolated from the test, the growth of this species (or these species) may be ascribed unequivocally to faults with respect to the material and or the technique used in conducting the sterility test procedure.在從試驗(yàn)中分離出的微生物確定之后，此種（或這些種）微生物的生長(zhǎng)可以毫不含糊地歸咎于與物料和/或者進(jìn)行無(wú)菌試驗(yàn)過(guò)程中所使用的方法。

If the test is declared to be invalid, it is repeated with the same number of units as in the original test. If no evidence of microbial growth is found in the repeat test, the product examined complies with the test for sterility. If microbial growth is found in the repeat test, the product examined does not comply with the test for sterility.

如果該試驗(yàn)證明無(wú)效，應(yīng)用與原試驗(yàn)同樣數(shù)量的產(chǎn)品進(jìn)行復(fù)檢。如果在復(fù)檢中未發(fā)現(xiàn)微生物生長(zhǎng)的證據(jù)，則該供試產(chǎn)品符合無(wú)菌試驗(yàn)的要求。如果在復(fù)檢中發(fā)現(xiàn)了微生物生長(zhǎng)，則該產(chǎn)品不符合無(wú)菌試驗(yàn)。

APPLICATION OF THE TEST TO PARENTERAL PREPARATIONS, OPHTHALMIC, AND OTHER NONINJECTABLE PREPARATIONS REQUIRED TO COMPLY WITH THE TEST FOR STERILITY

此試驗(yàn)在注射藥品、眼科和其他必須符合無(wú)菌試驗(yàn)的非注射藥品中的應(yīng)用

When using the technique of membrane filtration, use, whenever possible, the whole contents of the container, but not less than the quantities indicated in Tables 2 and 3, diluting where necessary to about 100 mL with a suitable sterile solution, such as Fluid A (see Diluting and Rinsing Fluids for Membrane Filtration).

當(dāng)使用膜過(guò)濾法時(shí)，只要可能，就要使用該容器的全部?jī)?nèi)容物，但不少于表2和3中規(guī)定的數(shù)量，必需時(shí)以適當(dāng)?shù)臒o(wú)菌溶液將其稀釋至約100mL，例如液體A（見用于膜過(guò)濾的稀釋和沖洗液）。

When using the technique of direct inoculation of media, use the quantities shown in Tables 2 and 3, unless otherwise justified and authorized. The tests for bacterial and fungal sterility are carried out on the same sample of the product to be examined. When the volume or the quantity in a single container is insufficient to carry out the tests, the contents of two or more containers are used to inoculate the different media.

當(dāng)使用培養(yǎng)基直接接種法時(shí)，除非另有證據(jù)或授權(quán)，使用表2和3中顯示的數(shù)量。用于檢驗(yàn)細(xì)菌和霉菌的試驗(yàn)使用供試產(chǎn)品的同一個(gè)樣品。當(dāng)單一容器中的產(chǎn)品體積或數(shù)量不足以進(jìn)行該試驗(yàn)時(shí)，適用2個(gè)或更多容器的內(nèi)容物來(lái)接種不同的培養(yǎng)基。

¹  In appropriate cases, periodic testing of the different batches prepared from the same lot of dehydrated medium is acceptable.

在適當(dāng)?shù)那闆r下，用同一個(gè)批次的脫水培養(yǎng)基配制的不同批次的定期試驗(yàn)是可以接受的。

Auxiliary Information— Staff Liaison : Radhakrishna S Tirumalai, Scientist

Expert Committee : (MSA05) Microbiology and Sterility Assurance

USP30–NF25 Page 97

Phone Number : 1-301-816-8339