培养基欧洲药典ep8.0 2.6.1无菌检验 sterility中英文翻译

2.6.1.STERILITY

2.6. 1 无菌检查法

The test is appl ied to substances, preparations or articles which,according to the Pharmacopoeia, are requi red to be ster i le. However,a satisfactory result only indicates that no contaminating micro-organism has been found in the sample examined in the conditions ofthe test.

本检查方法适用于按照药典要求应当无菌的原料、制剂或其他物质。但是如果按照本无菌检查法的结果符合要求仅表明在该检查条件下未发现微生物污染。

PRECAUT IONS AGAINST MICROBIAL CONTAMINAT ION

微生物污染防范

The test for ster i l ity is carr ied out under aseptic conditions.In order to achieve such conditions, the test envi ronment has to beadapted to the way in which the ster i l ity test is performed. Theprecautions taken to avoid contamination are such that they do notaffect any micro-organisms which are to be revealed in the test. Theworking conditions in which the tests are performed are monitoredregular ly by appropr iate sampl ing of the working area and by carryingout appropr iate controls.

无菌检测试验应在无菌的条件下进行。为了达到这样的条件检测环境应当与无菌检测的操作要求相适应。避免污染的防范措施应当不对本检查方法进行检测的微生物造成影响应并不影响用本检查法检测的微生物 。通过对工作区域的适当取样以及进行适当的控制来对无菌检查的工作环境进行例行监测。

CULTURE MEDIA AND INCUBATION TEMPERATURES

培养基和培养温度

Media for the test may be prepared as descr ibed below, orequivalent commercial media may be used provided that they complywith the growth promotion test.

应按下面描述的方法制备无菌检查的培养介质如果满足生长促进试验要求 与本处所述培养基相当的商业化培养基也可以采用也可采用与本处…… 。

The fol lowing culture media have been found to be suitable forthe test for ster i l ity. Fluid thioglycol late medium is pr imar i lyintended for the culture of anaerobic bacter ia; however, it wi l l alsodetect aerobic bacter ia. Soya -bean casein digest medium is suitablefor the culture of both fungi and aerobic bacter ia.

下述的培养基已被证明经证明适用于无菌检查。硫乙醇酸盐流体培养基主要用于厌氧菌培养但是也适用于需氧菌检测。大豆酪蛋白消化物培养基适用于真菌和需氧菌培养。

Mix the L-cystine, agar, sodium chlor ide, glucose, water-solubleyeast extract and pancreatic digest of casein with the water R andheat unti l solution is effected. Dissolve the sodium thioglycol lateor thioglycol l ic acid in the solution and, if necessary, add 1 Msodium hydroxide so that, after ster i l isation, the solution wi l l have

a pH of 7. 1±0.2. If fi ltration is necessary, heat the solution againwithout boi l ing and f i lter whi le hot through moistened f i lter paper.Add the resazur in sodium solution, mix and place the medium insuitable vessels which provide a ratio of surface to depth of mediumsuch that not more than the upper half of the medium has undergon e a

colour change indicative of oxygen uptake at the end of theincubation per iod. Ster i l ise using a val idated process. If the mediumis stored, store at a temperature between 2 ° C and 25 ° C in aster i le, ai rtight container. If more than the upper one-thi rd of themedium has acqui red a pink colour, the medium may be restored once byheating the containers in a water-bath or in free-f lowing steam unti lthe pink colour disappears and cool ing quickly, taking care toprevent the introduction of non -ster i le ai r into the container.

Do not use the medium for a longer storage per iod than has beenval idated.

将L-胱氨酸、琼脂、氯化钠、葡萄糖、水溶性酵母提取物以及酪蛋白胰酶消化物与水R混合加热至溶解。将硫乙醇酸钠或硫乙醇酸用上述溶液溶解必要时用1M氢氧化钠调节pH值使灭菌后培养基溶液的pH值为7. 1±0.2。如需要过滤处理将溶液在此加热加热此溶液 但不得煮到沸腾乘热采用经润湿的滤纸进行过滤。加入刃天青钠溶液混合均匀将制备的培养基装入合适的容器中。在该容器中培养基的表面和高度应具有恰当的比例 以便在灭菌结束后指示氧气摄入的颜色变化不超过培养基的上半部分。采用经验证的工艺灭菌。如果需要保存将培养基装入无菌、气密容器并在2-25° C 之间储存。如果培养基的上面超过1/3的部分已经出现粉红色将装有培养基的容器采用水浴或自由流动蒸气加热直到粉红颜色消失之后快速冷却注意预防非无菌的气体被引入装培养基的容器以此进行培养基再生处理。如果培养基保存的时间超过经验证的保存期限不得使用。 禁止使用超过验证储存期限的培养基

Fluid thioglycol late medium is to be incubated at 30-35 ° C. Forproducts containing a mercur ial preservative that cannot be tested bythe membrane-f i ltration method, f luid thioglycol late medium incubatedat 20

-25 ° C may be used instead of soya-bean casein digest mediumprovided that it has been val idated as descr ibed in growth promotiontest.

硫乙醇酸盐流体培养基用于应在30-35° C条件下培养。对于含有汞类防腐剂无法采用薄膜过滤法进行检查的产品如果已按照生长促进试验所述方法验证硫乙醇酸盐流体培养基可代替替代重复大豆酪蛋白消化物培养基在20-25° C条件下进行培养。

Where prescr ibed or justif ied and author ised, the fol lowingalternative thioglycol late medium may be used. Prepare a mixturehaving the same composition as that of the f luid thioglycol latemedium, but omitting the agar and the resazur in sodium solution,ster i l ise as di rected above. The pH after ster i l isation is 7. 1 ± 0.2.Heat in a water-bath pr ior to use and incubate at 30-35 ° C underanaerobic conditions.

按照规定或者证明合理并获得主管机构许可时如果有经批准的规定或正当理由 也可以使用以下替代硫乙醇酸盐流体培养基。配制与硫乙醇酸盐流体培养基成分相同的混合物但是不包括琼脂和刃天青钠溶液按照上面说明的方法进行灭菌。灭菌后培养基的pH值为7. 1 ± 0.2使用前采用水浴加热在厌氧及30-35° C条件下培养。

Soya-bean casein digest medium

大豆-酪蛋白消化物培养基

Pancreatic digest of casein 17.0 g

酪蛋白胰酶消化物17.0g

Papaic digest of soya-bean meal 3.0 g

大豆粉木瓜蛋白酶消化物3.0g

Sodium chlor ide 5.0 g

氯化钠5.0g

Dipotassium hydrogen phosphate 2.5 g

磷酸氢二钾2.5g

Glucose monohydrate/anhydrous 2.5 g/2.3 g

葡萄糖一水合物/无水葡萄糖2.5 g/2.3 g

Water R 1000 mL

水 R 1000mlpH after ster i l isation 7.3 ± 0.2

灭菌后pH 7.3 ± 0.2

Dissolve the sol ids in water R, warming sl ightly to effectsolution. Cool the solution to room temperature. Add 1M sodiumhydroxide, if necessary, so that after ster i l isation the solutionwi l l have a pH of 7.3 ± 0.2. Fi lter, if necessary, to clar ify,distr ibute into suitable vessels and ster i l ise using a val idatedprocess. Store at a temperature between 2 ° C and 25 ° C in a ster i lewel l-closed container, unless it is intended for immediate use. Donot use the medium for a longer storage per iod than has beenval idated.

将上述固体用水R溶解微微加热直到溶解再放至室温。必要时用1M氢氧化钠调节pH值使灭菌后培养基溶液的pH值为7.3±0.2。如需要过滤使培养基溶液澄清再将其装入合适的容器中并采用经验证的工艺灭菌。除非立即使用应将培养基装入无菌、气密容器并在2-25° C 之间储存。如果培养基保存的时间超过经验证的保存期限不得使用。 禁止使用超过验证储存期限的培养基

Soya-bean casein digest medium is to be incubated at 20-25 ° C.The media used comply with the fol lowing tests, carr ied out before orin paral lel with the test on the product to be examined.

大豆-酪蛋白消化物培养基用于在20-25° C条件下培养。使用的培养基应满足以下试验要求相关检查可以在使用前或者和待测样品同时进行。

Ster i l ity. Incubate portions of the media for 14 days. No growthof micro-organisms occurs.

无菌 取部分培养基培养14天不得出现微生物生长。

Growth promotion test of aerobes, anaerobes and fungi .

需氧菌、厌氧菌和真菌的生长促进试验

Test each batch of ready-prepared medium and each batch of mediumprepared either from dehydrated medium or from ingredients. Suitablestrains of micro-organisms are indicated in Table 2.6. 1 .-1 .

对每一批配好待用的培养基以及每一批采用脱水培养基或成分配制的培养基进行检测。适合的微生物菌株见表2.6. 1 .-1 .

Inoculate portions of f luid thioglycol late medium with a smal lnumber (not more than 100 CFU) of the fol lowing micro -organisms,using a separate portion of medium for each of the fol lowing speciesof micro-organism: Clostr idium sporogenes, Pseudomonasaeruginosa,Staphylococcus aureus. Inoculate portions of soya-beancasein digest medium with a smal l number (not more than 100 CFU) ofthe fol lowing micro-organisms, using a separate portion of medium foreach of the fol lowing species of micro-organism: Aspergi l lusbrasi l iensis, Baci l lus subti l is, Candida albicans. Incubate fornotmorethan3days in the case of bacter ia and not more than 5 days inthe case of fungi .

取部分硫乙醇酸盐流体培养基接种少量不超过100CFU下述试验菌生孢梭菌、铜绿假单胞菌以及金黄色葡萄球菌每种试验菌均使用单独一份培养基。取部分大豆-酪蛋白消化物培养基接种少量不超过100CFU下述试验菌黑曲霉、枯草芽孢杆菌以及白色念珠菌每种试验菌均使用单独一份培养基。细菌培养不超过3天真菌培养不超过5天。

Seed lot culture maintenance techniques (seed-lot systems) areused so that the viable micro-organisms used for inoculation are notmore than 5 passages removed from the or iginal master seed-lot.

采用菌种保藏技术种子-批系统 以确保用于接种的活试验菌从原始主种子批的传代数不超过5。

The media are suitable if a clear ly visible growth of the micro-organisms occurs.

如果出现清晰可见的微生物生长则该培养基是适合的。

METHOD SUITABILITY TEST

方法适用性检测

Carry out a test as descr ibed below under Test for ster i l ity ofthe product to be examined using exactly the same methods except forthe fol lowing modif ications.