歐洲藥典EP80261無菌檢驗(yàn)sterility中英文翻譯

1、2.6.1. STERILITY 2.6.1 無菌檢查法The test is applied to substances, preparations or articles which, according to the Pharmacopoeia, are required to be sterile. However, a satisfactory result only indicates that no contaminating micro-organism has been found in the sample examined in the conditions of the

2、 test. 本檢查方法適用于按照藥典要求應(yīng)當(dāng)無菌的原料、制劑或其他物質(zhì)。但是，如果按照本無菌檢查法的結(jié)果符合要求，僅表明在該檢查條件下未發(fā)現(xiàn)微生物污染。PRECAUTIONS AGAINST MICROBIAL CONTAMINATION 微生物污染防范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 sterilit

3、y test is performed. The precautions taken to avoid contamination are such that they do not affect any micro-organisms which 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

4、 appropriate controls. 無菌檢測試驗(yàn)應(yīng)在無菌的條件下進(jìn)行。為了達(dá)到這樣的條件，檢測環(huán)境應(yīng)當(dāng)與無菌檢測的操作要求相適應(yīng)。避免污染的防范措施應(yīng)當(dāng)不對(duì)本檢查方法進(jìn)行檢測的微生物造成影響（應(yīng)并不影響用本檢查法檢測的微生物）。通過對(duì)工作區(qū)域的適當(dāng)取樣以及進(jìn)行適當(dāng)?shù)目刂苼韺?duì)無菌檢查的工作環(huán)境進(jìn)行例行監(jiān)測。CULTURE MEDIA AND INCUBATION TEMPERATURES 培養(yǎng)基和培養(yǎng)溫度Media for the test may be prepared as described below, or equivalent commercial media may be

5、 used provided that they comply with the growth promotion test. 應(yīng)按下面描述的方法制備無菌檢查的培養(yǎng)介質(zhì)，如果滿足生長促進(jìn)試驗(yàn)要求，與本處所述培養(yǎng)基相當(dāng)?shù)纳虡I(yè)化培養(yǎng)基也可以采用（也可采用與本處）。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 bact

6、eria; however, it will also detect aerobic bacteria. Soya -bean casein digest medium is suitable for the culture of both fungi and aerobic bacteria. 下述的培養(yǎng)基已被證明（經(jīng)證明）適用于無菌檢查。硫乙醇酸鹽流體培養(yǎng)基主要用于厭氧菌培養(yǎng)，但是，也適用于需氧菌檢測。大豆酪蛋白消化物培養(yǎng)基適用于真菌和需氧菌培養(yǎng)。Fluid thioglycollate medium硫乙醇酸鹽流體培養(yǎng)基L-Cystine 0.5 gL-胱氨酸 0.5g Agar 0.75

7、 g瓊脂 0.75gSodium chloride 2.5 g氯化鈉 2.5gGlucose monohydrate/anhydrous 5.5 g/5.0 g葡萄糖一水合物/無水葡萄糖5.5 g/5.0 gYeast extract (water-soluble) 5.0 g酵母提取物（水溶性） 5.0gPancreatic digest of casein 15.0 g酪蛋白胰酶消化物 15.0gSodium thioglycollate or 0.5 g 硫乙醇酸鈉 0.5gThioglycollic acid 0.3 mL硫乙醇酸 0.3mlResazurin sodium solut

8、ion (1 g/L of resazurin1.0 mL sodium), freshly prepared 刃天青鈉溶液（刃天青鈉1 g/L），新鮮配制Water R 1000 mL水 R 1000mlpH after sterilisation 7.1 0.2 滅菌后的pH 7.1 0.2Mix the L-cystine, agar, sodium chloride, glucose, water-soluble yeast extract and pancreatic digest of casein with the water R and heat until solution

9、is effected. Dissolve the sodium thioglycollate or thioglycollic acid in the solution and, if necessary, add 1 M sodium hydroxide so that, after sterilisation, the solution will have a pH of 7.10.2. If filtration is necessary, heat the solution again without boiling and filter while hot through mois

10、tened filter paper. Add the resazurin sodium solution, mix and place the medium in suitable vessels which provide a ratio of surface to depth of medium such that not more than the upper half of the medium has undergon e a colour change indicative of oxygen uptake at the end of the incubation period.

11、 Sterilise using a validated process. If the medium is stored, store at a temperature between 2 C and 25 C in a sterile, airtight container. If more than the upper one-third of the medium has acquired a pink colour, the medium may be restored once by heating the containers in a water-bath or in free

12、-flowing steam until the pink colour disappears and cooling quickly, taking care to prevent the introduction of non -sterile air into the container. Do not use the medium for a longer storage period than has been validated. 將L-胱氨酸、瓊脂、氯化鈉、葡萄糖、水溶性酵母提取物以及酪蛋白胰酶消化物與水R混合，加熱至溶解。將硫乙醇酸鈉或硫乙醇酸用上述溶液溶解，必要時(shí)用1M氫氧化

13、鈉調(diào)節(jié)pH值，使滅菌后培養(yǎng)基溶液的pH值為7.10.2。如需要過濾處理，將溶液在此加熱（加熱此溶液），但不得煮到沸騰，乘熱采用經(jīng)潤濕的濾紙進(jìn)行過濾。加入刃天青鈉溶液，混合均勻，將制備的培養(yǎng)基裝入合適的容器中。在該容器中，培養(yǎng)基的表面和高度應(yīng)具有恰當(dāng)?shù)谋壤?，以便在滅菌結(jié)束后指示氧氣攝入的顏色變化不超過培養(yǎng)基的上半部分。采用經(jīng)驗(yàn)證的工藝滅菌。如果需要保存，將培養(yǎng)基裝入無菌、氣密容器并在2-25C 之間儲(chǔ)存。如果培養(yǎng)基的上面超過1/3的部分已經(jīng)出現(xiàn)粉紅色，將裝有培養(yǎng)基的容器采用水浴或自由流動(dòng)蒸氣加熱，直到粉紅顏色消失，之后快速冷卻，注意預(yù)防非無菌的氣體被引入裝培養(yǎng)基的容器，以此進(jìn)行培養(yǎng)基再生處理。如

14、果培養(yǎng)基保存的時(shí)間超過經(jīng)驗(yàn)證的保存期限，不得使用。（禁止使用超過驗(yàn)證儲(chǔ)存期限的培養(yǎng)基）Fluid thioglycollate medium is to be incubated at 30-35 C. For products containing a mercurial preservative that cannot be tested by the membrane-filtration method, fluid thioglycollate medium incubated at 20-25 C may be used instead of soya-bean casein dig

15、est medium provided that it has been validated as described in growth promotion test. 硫乙醇酸鹽流體培養(yǎng)基用于（應(yīng)）在30-35C條件下培養(yǎng)。對(duì)于含有汞類防腐劑無法采用薄膜過濾法進(jìn)行檢查的產(chǎn)品，如果已按照生長促進(jìn)試驗(yàn)所述方法驗(yàn)證，硫乙醇酸鹽流體培養(yǎng)基可代替替代重復(fù)大豆酪蛋白消化物培養(yǎng)基在20-25C條件下進(jìn)行培養(yǎng)。Where prescribed or justified and authorised, the following alternative thioglycollate medium may b

16、e used. Prepare a mixture having the same composition as that of the fluid thioglycollate medium, but omitting the agar and the resazurin sodium solution, sterilise as directed above. The pH after sterilisation is 7.1 0.2. Heat in a water-bath prior to use and incubate at 30-35 C under anaerobic con

17、ditions. 按照規(guī)定或者證明合理并獲得主管機(jī)構(gòu)許可時(shí)（如果有經(jīng)批準(zhǔn)的規(guī)定或正當(dāng)理由），也可以使用以下替代硫乙醇酸鹽流體培養(yǎng)基。配制與硫乙醇酸鹽流體培養(yǎng)基成分相同的混合物，但是不包括瓊脂和刃天青鈉溶液，按照上面說明的方法進(jìn)行滅菌。滅菌后培養(yǎng)基的pH值為7.1 0.2，使用前采用水浴加熱，在厭氧及30-35C條件下培養(yǎng)。Soya-bean casein digest medium大豆-酪蛋白消化物培養(yǎng)基Pancreatic digest of casein 17.0 g 酪蛋白胰酶消化物 17.0gPapaic digest of soya-bean meal 3.0 g 大豆粉木瓜蛋白酶消

18、化物 3.0gSodium chloride 5.0 g 氯化鈉 5.0gDipotassium hydrogen phosphate 2.5 g 磷酸氫二鉀 2.5gGlucose monohydrate/anhydrous 2.5 g/2.3 g 葡萄糖一水合物/無水葡萄糖2.5 g/2.3 gWater R 1000 mL 水 R 1000mlpH after sterilisation 7.3 0.2 滅菌后pH 7.3 0.2Dissolve the solids in water R, warming slightly to effect solution. Cool the so

19、lution to room temperature. Add 1M sodium hydroxide, if necessary, so that after sterilisation the solution will have a pH of 7.3 0.2. Filter, if necessary, to clarify, distribute into suitable vessels and sterilise using a validated process. Store at a temperature between 2 C and 25 C in a sterile

20、well-closed container, unless it is intended for immediate use. Do not use the medium for a longer storage period than has been validated. 將上述固體用水R溶解，微微加熱直到溶解，再放至室溫。必要時(shí)用1M氫氧化鈉調(diào)節(jié)pH值，使滅菌后培養(yǎng)基溶液的pH值為7.30.2。如需要，過濾使培養(yǎng)基溶液澄清，再將其裝入合適的容器中并采用經(jīng)驗(yàn)證的工藝滅菌。除非立即使用，應(yīng)將培養(yǎng)基裝入無菌、氣密容器并在2-25C 之間儲(chǔ)存。如果培養(yǎng)基保存的時(shí)間超過經(jīng)驗(yàn)證的保存期限，不得使用。

21、（禁止使用超過驗(yàn)證儲(chǔ)存期限的培養(yǎng)基） Soya-bean casein digest medium is to be incubated at 20-25 C. The media used comply with the following tests, carried out before or in parallel with the test on the product to be examined. 大豆-酪蛋白消化物培養(yǎng)基用于在20-25C條件下培養(yǎng)。使用的培養(yǎng)基應(yīng)滿足以下試驗(yàn)要求，相關(guān)檢查可以在使用前或者和待測樣品同時(shí)進(jìn)行。Sterility. Incubate portion

22、s of the media for 14 days. No growth of micro-organisms occurs.無菌 取部分培養(yǎng)基培養(yǎng)14天，不得出現(xiàn)微生物生長。 Growth promotion test of aerobes, anaerobes and fungi. 需氧菌、厭氧菌和真菌的生長促進(jìn)試驗(yàn)Test each batch of ready-prepared medium and each batch of medium prepared either from dehydrated medium or from ingredients. Suitable str

23、ains of micro-organisms are indicated in Table 2.6.1.-1. 對(duì)每一批配好待用的培養(yǎng)基以及每一批采用脫水培養(yǎng)基或成分配制的培養(yǎng)基進(jìn)行檢測。適合的微生物菌株見表2.6.1.-1.Table 2.6.1.-1 Strains of the test micro-organisms suitable for use in the growth promotion test and the method validation表2.6.1.-1.生長促進(jìn)試驗(yàn)及方法驗(yàn)證中使用的試驗(yàn)微生物菌株Aerobic bacteria需氧菌Staphylococcu

24、s aureus 金黃色葡萄球菌 ATCC 6538, CIP 4.83, NCTC 10788, NCIMB 9518, NBRC 13276Bacillus subtilis枯草芽孢桿菌ATCC 6633, CIP 52.62, NCIMB 8054, NBRC 3134Pseudomonas aeruginosa 銅綠假單胞菌ATCC 9027, NCIMB 8626, CIP 82.118, NBRC 13275Anaerobic bacterium 厭氧菌Clostridium sporogenes 生孢梭菌ATCC 19404, CIP 79.3, NCTC 532, ATCC 1

25、1437, NBRC 14293Fungi 真菌Candida albicans白色念珠菌ATCC 10231, IP 48.72, NCPF 3179, NBRC 1594Aspergillus brasiliensis 黑曲霉菌ATCC 16404, IP 1431.83, IMI 149007, NBRC 9455Inoculate portions of fluid thioglycollate medium with a small number (not more than 100 CFU) of the following micro -organisms, using a se

26、parate portion of medium for each of the following species of micro-organism: Clostridium sporogenes, Pseudomonas aeruginosa,Staphylococcus aureus. Inoculate portions of soya-bean casein digest medium with a small number (not more than 100 CFU) of the following micro-organisms, using a separate port

27、ion of medium for each of the following species of micro-organism: Aspergillus brasiliensis, Bacillus subtilis, Candida albicans. Incubate for notmorethan3days in the case of bacteria and not more than 5 days in the case of fungi. 取部分硫乙醇酸鹽流體培養(yǎng)基，接種少量（不超過100CFU）下述試驗(yàn)菌：生孢梭菌、銅綠假單胞菌以及金黃色葡萄球菌，每種試驗(yàn)菌均使用單獨(dú)一份培

28、養(yǎng)基。取部分大豆-酪蛋白消化物培養(yǎng)基，接種少量（不超過100CFU）下述試驗(yàn)菌：黑曲霉、枯草芽孢桿菌以及白色念珠菌，每種試驗(yàn)菌均使用單獨(dú)一份培養(yǎng)基。細(xì)菌培養(yǎng)不超過3天，真菌培養(yǎng)不超過5天。Seed lot culture maintenance techniques (seed-lot systems) are used so that the viable micro-organisms used for inoculation are not more than 5 passages removed from the original master seed-lot. 采用菌種保藏技術(shù)（種

29、子-批系統(tǒng)），以確保用于接種的活試驗(yàn)菌從原始主種子批的傳代數(shù)不超過5。The media are suitable if a clearly visible growth of the micro-organisms occurs. 如果出現(xiàn)清晰可見的微生物生長，則該培養(yǎng)基是適合的。METHOD SUITABILITY TEST 方法適用性檢測Carry out a test as described below under Test for sterility of the product to be examined using exactly the same methods excep

30、t for the following modifications. 除了以下一些變動(dòng)，其余完全按照供試品的無菌檢測項(xiàng)下描述的方法進(jìn)行檢測。（完全按照，以下變動(dòng)除外：）Membrane filtration. After transferring the contents of the container or containers to be tested to the membrane add an inoculum of a small number of viable micro-organisms (not more than 100 CFU) to the final portio

31、n of sterile diluent used to rinse the filter. 薄膜過濾法 : 將待測容器的內(nèi)容物轉(zhuǎn)至薄膜后，采用沖洗濾器的最后一部分無菌稀釋劑接種少量（不超過100CFU）活試驗(yàn)菌。Direct inoculation. After transferring the content of the container or containers to be tested (for catgut and other surgical sutures for veterinary use: strands) to the culture medium add an i

32、noculum of a small number of viable micro -organisms (not more than 100 CFU) to the medium. 直接接種法: 將待測容器的內(nèi)容物（對(duì)于腸線或其它獸用手術(shù)縫合線：若干股線）轉(zhuǎn)至培養(yǎng)基，再向培養(yǎng)基中接種少量（不超過100CFU）活試驗(yàn)菌。In both cases use the same micro-organisms as those described above under Growth promotion test of aerobes, anaerobes and fungi. Perform a

33、growth promotion test as a positive control. Incubate all the containers containing medium for not more than 5 days. 在這兩種方法，采用的都是 需氧菌、厭氧菌和真菌的生長促進(jìn)試驗(yàn) 項(xiàng)下提到的試驗(yàn)菌里的同一種菌株。進(jìn)行生長促進(jìn)試驗(yàn)，作為陽性對(duì)照。所有含培養(yǎng)基容器的培養(yǎng)時(shí)間不超過5天。If clearly visible growth of micro-organisms is obtained after the incubation, visually comparable to

34、 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. 如果供試品在接種后可觀察到清晰可見的微生物生長，目測與不含供試品的對(duì)照容器

35、相當(dāng)。說明供試品在測定試驗(yàn)條件下無抗微生物活性，或者該活性被有效消除了。不需要對(duì)方法進(jìn)行改變就可以用于無菌檢查不需要對(duì)方法進(jìn)行改進(jìn)就可以用于無菌檢查。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 satisfa

36、ctorily eliminated under the conditions of the test. Modify the conditions in order to eliminate the antimicrobial activity and repeat the method suitability test. 如果含有供試品的容器中未觀察到清晰可見的微生物生長，目測與不含供試品的對(duì)照容器相當(dāng)。則供試品具有抗微生物活性，在測定試驗(yàn)條件下未能有效去除。需要變更測定條件以消除其抗微生物活性，并重復(fù)方法適用性檢測。This method suitability test is perf

37、ormed: 方法適用性檢測在以下情況下應(yīng)進(jìn)行：a) when the test for sterility has to be carried out on a new product; 當(dāng)無菌檢查法用于新的產(chǎn)品檢測時(shí)；b) whenever there is a change in the experimental conditions of the test. 每當(dāng)檢測試驗(yàn)條件發(fā)生改變的時(shí)候。The method suitability test may be performed simultaneously with the test for sterility of the prod

38、uct to be examined. 方法適用性檢測可與供試品無菌檢查同時(shí)進(jìn)行。TEST FOR STERILITY OF THE PRODUCT TO BE EXAMINED 供品的無菌檢查The test may be carried out using the technique of membrane filtration or by direct inoculation of the culture media with the product to be examined. Appropriate negative controls are included. The techn

39、ique 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 th

40、e conditions of the test. 供試品的無菌檢查可以采用薄膜過濾法或培養(yǎng)基直接接種法進(jìn)行測定，應(yīng)當(dāng)包含適當(dāng)?shù)年幮詫?duì)照。如果供試品中的溶劑在測定條件下無抗微生物活性，當(dāng)只要產(chǎn)品性質(zhì)允許時(shí)的話，薄膜過濾法可廣泛應(yīng)用于可過濾的水性制劑、醇溶性或油溶性制劑以及可與水性或油性溶劑混合或溶解的制劑。Membrane filtration. Use membrane filters having a nominal pore size not greater than 0.45 m whose effectiveness to retain micro-organisms has bee

41、n established. Cellulose nitrate filters, for example, are used for aqueous, oily and weakly alcoholic solutions and cellulose acetate filters, for example, for strongly alcoholic solutions. Specially adapted filters may be needed for certain products, e.g. for antibiotics. 薄膜過濾法 使用標(biāo)示孔徑不大于0.45m的、已被證

42、實(shí)可有效截留微生物的膜過濾器經(jīng)確定可有效截留微生物的膜過濾器。比如，硝酸纖維素過濾器可用于水性、油性以及弱醇性溶液，而醋酸纖維素可用于強(qiáng)醇性溶液。對(duì)于某些產(chǎn)品，比如抗生素，可能需要特定的過濾器。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 acc

43、ordingly. The filtration apparatus and membrane are sterilised 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

44、carrying out the incubation after adding the medium to the apparatus itself. 以下描述的方法中使用的是50mm的薄膜，如果使用不同尺寸的薄膜，稀釋和沖洗的體積應(yīng)當(dāng)相應(yīng)進(jìn)行調(diào)節(jié)稀釋和沖洗的體積應(yīng)當(dāng)進(jìn)行相應(yīng)調(diào)節(jié)。過濾的設(shè)備和薄膜應(yīng)當(dāng)采用適當(dāng)?shù)姆绞竭M(jìn)行滅菌。應(yīng)對(duì)設(shè)備進(jìn)行設(shè)計(jì)，以便供試品溶液可以加入并在無菌條件下進(jìn)行過濾；設(shè)備應(yīng)支持無菌狀態(tài)下去除薄膜來進(jìn)行培養(yǎng)基轉(zhuǎn)運(yùn)，或者適合將培養(yǎng)基加入設(shè)備中后進(jìn)行培養(yǎng)。Aqueous solutions. If appropriate, transfer a small quantity o

45、f a suitable, sterile diluent such as a 1 g/L neutral solution of meat or casein peptone pH 7.1 0.2 onto the membrane in the apparatus and filter. The diluent may contain suitable neutralising substances and/or appropriate inactivating substances for example in the case of antibiotics. 水性溶液 如合適，將少量適

46、當(dāng)?shù)臒o菌稀釋劑（比如1 g/L中性肉或酪蛋白胨，pH 7.1 0.2）轉(zhuǎn)至設(shè)備的濾膜上并進(jìn)行過濾。在產(chǎn)品為抗生素等情形中，稀釋劑可以包含適當(dāng)具有中和作用和/或具有滅活作用的物質(zhì)。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 method suitability test with the chosen sterile diluent but i

47、n any case using not less than the quantities of the product to be examined prescribed in Table 2.6.1.-2. Filter immediately. If the product has antimicrobial properties, wash the membrane not less than 3 times by filtering through it each time the volume of the chosen sterile diluent used in the me

48、thod suitability test. Do not exceed a washing cycle of 5 times 100 mL per filter, even if during the method suitability test 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 2

49、 equal parts and transfer one half to each of 2 suitable media. Use the same volume of each medium as in the method suitability test. Alternatively, transfer the medium onto the membrane in the apparatus. Incubate the media for not less than 14 days. 將供試品容器內(nèi)容物轉(zhuǎn)至濾膜表面，必要時(shí)可采用選定的無菌稀釋劑稀釋至方法適用性檢測中使用的體積，但任

50、何時(shí)候所檢測的檢供試品的數(shù)量均不得低于表2.6.1.-2規(guī)定的量。立即過濾。如果供試品具有抗微生物活性，采用選用的稀釋劑過濾沖洗濾膜至少3次，每次沖洗的體積按照方法法適用性檢測中使用的體積。每個(gè)沖洗循環(huán)不要超過5次及100mL/次，即使方法適用性檢測已證明這樣的循環(huán)并不能完全消除供試品的抗微生物活性。轉(zhuǎn)移整個(gè)濾膜到培養(yǎng)基中或者將其在無菌條件下剪切成大小相同的量塊然后將每一部分分別轉(zhuǎn)移到2種適合的培養(yǎng)基中。每種培養(yǎng)基使用的體積和方法適用性檢測中的一致。也可以將培養(yǎng)基加入設(shè)備中的濾膜上。所得培養(yǎng)基培養(yǎng)時(shí)間不得少于14天。Soluble solids. Use for each medium not

51、 less than the quantity prescribed in Table 2.6.1.-2 of the product dissolved in a suitable solvent such as the solvent provided with the preparation, water for injections, saline or a 1 g/L neutral solution of meat or casein peptone and proceed with the test as described above for aqueous solutions

52、 using a membrane appropriate to the chosen solvent. 水溶性固體 對(duì)于每個(gè)培養(yǎng)基，需要將不少于表2.6.1.-2中規(guī)定數(shù)量的供試品采用合適的溶劑（比如，產(chǎn)品伴有的溶劑、注射用水、生理鹽水以及1 g/L中性肉或酪蛋白胨）溶解，采用適用于選用溶劑的薄膜按照上述水性溶液部分描述的方法進(jìn)行測定。Oils and oily solutions. Use for each medium not less than the quantity of the product prescribed in Table 2.6.1.-2. Oils and oily

53、 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 th

54、e membrane by its own weight then filter, applying the pressure or suction gradually. Wash the membrane at least 3 times by filtering through it each time about 100 mL of a suitable sterile solution such as 1 g/L neutral meat or casein peptone containing a suitable emulsifying agent at a concentrati

55、on shown to be appropriate in the method suitability test, for example polysorbate 80 at a concentration of 10 g/L. 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 temperature and for the same times.油及

56、油性溶液 對(duì)于每個(gè)培養(yǎng)基，需要選用不少于表2.6.1.-2中規(guī)定數(shù)量的供試品進(jìn)行檢測。粘度十分低的油和油性溶液可不經(jīng)稀釋直接采用干燥薄膜過濾。粘性較大的油需要選用十四烷酸異丙酯等合適的、在試驗(yàn)測定條件無抗微生物活性的合適無菌稀釋劑進(jìn)行稀釋。讓油依靠自身重力作用滲透金薄膜，然后逐漸通過壓力或吸力進(jìn)行過濾。用100mL含有適當(dāng)乳化劑的1 g/L中性肉或酪蛋白胨等合適無菌溶液過濾沖洗濾膜至少3次，該溶劑中包含適量的乳化劑，其濃度（適用于方法適用性檢測）在方法驗(yàn)證試驗(yàn)中表面適用，例如聚山梨醇酯80的濃度選用10 g/L,按照以上水溶液項(xiàng)下的方法同樣操作，轉(zhuǎn)移薄膜至培養(yǎng)基中，并在相同的溫度溫孵培養(yǎng)相同的

57、時(shí)間。Table 2.6.1.-2 Minimum quantity to be used for each medium表2.6.1.-2用于每個(gè)培養(yǎng)基的最小數(shù)量Quantity per container每個(gè)容器中的數(shù)量Minimum quantity to be used for each medium unless otherwise justified and authorized最小使用數(shù)量(除非另有依據(jù)并獲得許可)Liquids液體 less than 1 mL 小于1 mL 1-40 mL 1-40 mL greater than 40 mL and not greater t

58、han 100 mL 大于40mL，不大于100mLgreater than 100 mL 大于100mLAntibiotic liquids 抗生素溶液The whole contents of each container 每個(gè)容器的總內(nèi)容物Half the contents of each container but not less than 1 mL 每個(gè)容器中內(nèi)容物的一半，但不得少于1mL20 mL 10 per cent of the contents of the container but not less than 20 mL 該容器內(nèi)容物的10%，但不得少于20mL1mL Insoluble preparations, creams and ointments to be suspended or emulsified 待懸浮或乳化的不溶性配制品、乳膏、油膏The whole co