微生物限度培训课件.doc

1 61 MICROBIAL LIMIT TESTS 微生物限度检查 This chapter provides tests for the estimation of the number of viable aerobic microorganisms present and for freedom from designated microbial species in pharmaceutical articles of all kinds from raw materials to the finished forms An automated method may be substituted for the tests presented here provided it has been properly validated as giving equivalent or better results In preparing for and in applying the tests observe aseptic precautions in handling the specimens Unless otherwise directed where the procedure specifies simply incubate hold the container in air that is thermostatically controlled at a temperature between 30 and 35 for a period of 24 to 48 hours The term growth is used in a special sense herein i e to designate the presence and presumed proliferation of viable microorganisms 本章为估计具有繁殖能力的有氧微生物数量和保证从原料到最终剂型都不含有各种药物条 款所指定的微生物种群提供检查方法 若某种自动化方法已经证明能提供等效或更好的检 查效果 那么可以取代此处所说的检查方法 在准备和开始检查时 观察样品处理过程的 无菌预防措施 除非有另外的说明过程中何处只需 培养 的指导 否则将容器置于温度 恒控制在 30 35 之间的空气中 24 48 小时 生长 在此为专用的标明存在和可能存在 的有繁殖能力的微生物的增殖的术语 PREPARATORY TESTING 预备检验 The validity of the results of the tests set forth in this chapter rests largely upon the adequacy of a demonstration that the test specimens to which they are applied do not of themselves inhibit the multiplication under the test conditions of microorganisms that may be present Therefore preparatory to conducting the tests on a regular basis and as circumstances require subsequently inoculate diluted specimens of the material to be tested with separate viable cultures of Staphylococcus aureus Escherichia coli Pseudomonas aeruginosa and Salmonella This can be done by adding 1 mL 2 of not less than 10 3 dilution of a 24 hour broth culture of the microorganism to the first dilution in pH 7 2 Phosphate Buffer Fluid Soybean Casein Digest Medium or Fluid Lactose Medium of the test material and following the test procedure Failure of the organism s to grow in the relevant medium invalidates that portion of the examination and necessitates a modification of the procedure by 1 an increase in the volume of diluent the quantity of test material remaining the same or by 2 the incorporation of a sufficient quantity of suitable inactivating agent s in the diluents or by 3 an appropriate combination of modifications 1 and 2 so as to permit growth of the inocula The following are examples of ingredients and their concentrations that may be added to the culture medium to neutralize inhibitory substances present in the sample soy lecithin 0 5 and polysorbate 20 4 0 Alternatively repeat the test as described in the preceding paragraph using Fluid Casein Digest Soy Lecithin Polysorbate 20 Medium to demonstrate neutralization of preservatives or other antimicrobial agents in the test material Where inhibitory substances are contained in the product and the latter is soluble a suitable validated adaptation of a procedure set forth in the section Membrane Filtration under Test for Sterility of the Product to be Examined under Sterility Tests 71 may be used 本章所述的检查结果的有效性很大程度上取决于实证的适当性 即实证中所用的检验样品 的本身在检验条件下不会抑制可能存在的微生物增殖 因此 正在筹备的检验应该依据基 本规则及所要求的条件下进行 分别用单独的葡萄球菌 大肠埃希氏菌 假单胞菌和沙门 氏菌属的培养物接种待检验物质的稀释样品 通过下面的方法可以得到 向待检验的物质 的第一份稀释液中加入 毫升的稀释度不低于 10 3的含微生物的 小时肉汤培养基后继 续后面的检验程序 生物体在相应的培养基中增殖的失败会使部分检查无效 同时也使修 改检验方法成为必要的 修改检验方法可通过以下方式 1 增加稀释液的体积 待检验 物质的质量不变 2 向稀释液中加入足够量的适当的破坏活性制剂 3 方法 1 和 2 的适当结合使接种物增殖 下面是一些可能加入到培养基中中和样品中存在的抑制性物质 的成分和浓聚物的例子 大豆卵磷脂 0 5 聚山梨醇酯 20 4 0 另有一种方法可供选 择 重复前述段落所述的检验 用液态酪蛋白水解液 大豆卵磷脂 聚山梨醇酯 20 培养 3 基验证检验物质中的防腐剂或其他抗菌剂 若产物中含有可抑制性物质且可溶 那么无菌 检验 的待检产品的无菌检验的膜过滤章节中所述的方法可用 If in spite of the incorporation of suitable inactivating agents and a substantial increase in the volume of diluent it is still not possible to recover the viable cultures described above and where the article is not suitable for employment of membrane filtration it can be assumed that the failure to isolate the inoculated organism is attributable to the bactericidal activity of the product This information serves to indicate that the article is not likely to be contaminated with the given species of microorganism Monitoring should be continued in order to establish the spectrum of inhibition and bactericidal activity of the article 如果适当的破坏活性剂的结合及稀释液体积的增加仍然无法重新得到上面所描述的具繁殖 能力的培养物及药物中不适合使用薄膜过滤法的地方 可以假设隔离接种生物体的失败是 由于产物的灭菌活性 该信息表明该药物不可能受到所给的微生物种群的污染 继续监测 以建立药物的抑制和灭菌活性谱 BUFFER SOLUTION AND MEDIA 缓冲液和培养基 Culture media may be prepared as follows or dehydrated culture media may be used provided that when reconstituted as directed by the manufacturer or distributor they have similar ingredients and or yield media comparable to those obtained from the formulas given herein 依据下列方法准备培养基 或在厂商或销售商的指示下恢复的脱水培养基与公式中得到的 具有类似的成分与收率时 那么也可以使用脱水培养基 In preparing media by the formulas set forth herein dissolve the soluble solids in the water using heat if necessary to effect complete solution and add solutions of hydrochloric acid or sodium hydroxide in quantities sufficient to yield the desired pH in the medium when it is ready for use Determine the pH at 25 2 依照此处所述公式准备培养基时 将可溶固体溶解在水中 若需要可以加热促进完全溶解 使用时加入足够量的盐酸或氢氧化钠以得到所要求的 pH 值 确定 pH 值为 25 2 4 Where agar is called for in a formula use agar that has a moisture content of not more than 15 Where water is called for in a formula use Purified Water PH 7 2 Phosphate Buffer Stock Solution Dissolve 34 g of monobasic potassium phosphate in about 500 mL of water contained in a 1000 mL volumetric flask Adjust to pH 7 2 0 1 by the addition of sodium hydroxide TS about 175 mL add water to volume and mix Dispense and sterilize Store under refrigeration For use dilute the Stock Solution with water in the ratio of 1 to 800 and sterilize 配方中需要琼脂的地方 使用水分不超过 15 的琼脂 需要水的地方使用蒸馏水 磷酸缓冲液的 PH 值为 7 2 原液 将 34g 磷酸二氢钾溶解在装有 500 mL 水的 1000 mL 烧瓶中 加入氢氧化钠调节 pH 值到 7 2 0 1 再加入水定容并搅拌 分配并灭菌 冷冻储藏 使用时 按照 1 800 的比率加水稀释原液后灭菌 Media 培养基 Unless otherwise indicated the media should be sterilized by heating in an autoclave see Steam Sterilization under Sterilization 1211 the exposure time depending on the volume to be sterilized 除非有其他的指导 否则培养基应该置于高压锅中加热灭菌 暴露的时间由需要灭菌的体 积决定 I Fluid Casein Digest Soy Lecithin Polysorbate 20 Medium Pancreatic Digest of Casein 20 g Soy Lecithin5 g Polysorbate 2040 mL Water960 mL Dissolve the pancreatic digest of casein and soy lecithin in 960 mL of water heating in a water bath at 48 to 50 for about 30 minutes to effect solution Add 40 mL of polysorbate 20 Mix and dispense as desired I 液态酪蛋白消化液 大豆卵磷脂 聚山梨醇酯 20 培养基 酪蛋白胰消化液 大豆卵磷脂 聚山梨醇酯 20 5 水 将酪蛋白胰消化液和大豆卵磷脂溶解在 的水中 在 48 50 的水浴中加热 分钟促进溶解 加入 的聚山梨醇酯 20 混合 按照所要求的分配 II Soybean Casein Digest Agar Medium Pancreatic Digest of Casein15 0 g Papaic Digest of Soybean Meal 5 0 g Sodium Chloride5 0 g Agar15 0 g Water1000 mL pH after sterilization 7 3 0 2 大豆 酪蛋白消化液琼脂培养基 酪蛋白胰消化液15 0 g 大豆粉的 Papaic 消化液5 0 g 氯化钠5 0 g 琼脂5 0 g 水1000 mL 灭菌后的 pH 值 7 3 0 2 III Fluid Soybean Casein Digest Medium Prepare as directed for Soybean Casein Digest Medium under Sterility Tests 71 液态大豆 酪蛋白消化液培养基 依照无菌检验 的大豆 酪蛋白消化液培养基制备 IV Mannitol Salt Agar Medium Pancreatic Digest of Casein5 0 g Peptic Digest of Animal Tissue 5 0 g Beef Extract1 0 g D Mannitol10 0 g Sodium Chloride75 0 g Agar15 0 g Phenol Red0 025 g Water1000 mL Mix then heat with frequent agitation and boil for 1 minute to effect solution pH after sterilization 7 4 0 2 甘露醇 盐琼脂培养基 酪蛋白胰消化液5 0 g 动物组织胃蛋白酶消化液5 0 g 牛肉提取物1 0 g D 甘露醇10 0 g 6 氯化钠75 0 g 琼脂15 0 g 酚红0 025 g 水1000 mL 混合 加热并频繁搅拌 加热到沸腾一分钟以促进溶解 灭菌后的 PH 值为 7 4 0 2 V Baird Parker Agar Medium Pancreatic Digest of Casein 10 0 g Beef Extract5 0 g Yeast Extract1 0 g Lithium Chloride5 0 g Agar20 0 g Glycine12 0 g Sodium Pyruvate10 0 g Water950 mL Heat with frequent agitation and boil for 1 minute Sterilize cool to between 45 and 50 and add 10 mL of sterile potassium tellurite solution 1 in 100 and 50 mL of egg yolk emulsion Mix intimately but gently and pour into plates Prepare the egg yolk emulsion by disinfecting the surface of whole shell eggs aseptically cracking the eggs and separating out intact yolks into a sterile graduated cylinder Add sterile saline TS to obtain a 3 to 7 ratio of egg yolk to saline Add to a sterile blender cup and mix at high speed for 5 seconds pH after sterilization 6 8 0 2 Baird Parker 琼脂培养基 酪蛋白胰消化液10 0 g 牛肉提取物5 0 g 酵母菌提取物1 0 g 氯化锂5 0 g 琼脂20 0 g 甘氨酸12 0 g 丙酮酸钠10 0 g 水950 mL 加热并频繁搅拌 沸腾 1 分钟 灭菌 冷却至 45 50 后加入 10 mL 的无菌亚碲酸钠溶液 1 和 50 mL 的蛋黄乳浊液 逐渐充分混合后倒入皿中 准备蛋黄乳浊液时 给整 颗蛋的外壳消毒 在无菌条件下敲破蛋壳并将其内的蛋黄倒入一个无菌量筒中 加入无菌 的盐试液使蛋黄和盐的比例为 3 7 将该溶液倒入一个无菌的搅拌器并在高速下搅拌 5 秒 种 灭菌后的 PH 值为 6 8 0 2 7 VI Vogel Johnson Agar Medium Pancreatic Digest of Casein10 0 g Yeast Extract5 0 g Mannitol10 0 g Dibasic Potassium Phosphate 5 0 g Lithium Chloride5 0 g Glycine10 0 g Agar16 0 g Phenol Red25 0 mg Water1000 mL Boil the solution of solids for 1 minute Sterilize cool to between 45 and 50 and add 20 mL of sterile potassium tellurite solution 1 in 100 pH after sterilization 7 2 0 2 VI Vogel Johnson 琼脂培养基 酪蛋白胰消化液10 0 g 牛肉提取物5 0 g 甘露醇10 0 g 磷酸二氢钾5 0 g 氯化锂5 0 g 甘氨酸10 0 g 琼脂16 0 g 酚红25 0 mg 水1000 mL 煮沸含固体的溶液 1 分钟 灭菌后冷却至 45 50 加入 20ml 的无菌亚碲酸钠溶液 1 灭菌后的 PH 值为 7 2 0 2 VII Cetrimide Agar Medium Pancreatic Digest of Gelatin20 0 g Magnesium Chloride1 4 g Potassium Sulfate10 0 g Agar13 6 g Cetyl Trimethylammonium Bromide Cetrimide 0 3 g Glycerin10 0 mL Water1000 mL Dissolve all solid components in the water and add the glycerin Heat with frequent agitation and boil for 1 minute to effect solution pH after sterilization 7 2 0 2 VII 溴化十六烷基三甲铵琼脂培养基 明胶胰消化液20 0 g 8 氯化镁1 4 g 硫酸钾10 0 g 琼脂13 6 g 溴化十六烷基三甲铵0 3 g 甘氨酸10 0 mL 水1000 mL 将所有的固体成分溶解在水中 加入甘油 加热 频繁搅拌 沸腾 1 分钟促进溶解 灭菌 后的 PH 值为 7 2 0 2 VIII Pseudomonas Agar Medium for Detection of Fluorescin Pancreatic Digest of Casein10 0 g Peptic Digest of Animal Tissue10 0 g Anhydrous Dibasic Potassium Phosphate 1 5 g Magnesium Sulfate MgSO4 7H2O 1 5 g Glycerin10 0 mL Agar15 0 g Water1000 mL Dissolve the solid components in the water before adding the glycerin Heat with frequent agitation and boil for 1 minute to effect solution pH after sterilization 7 2 0 2 VIII 探测二氢荧光素的假单胞菌琼脂培养基 酪蛋白胰消化液10 0 g 动物组织胃蛋白酶消化液10 0 g 无水磷酸二氢钾1 5 g 硫酸镁 MgSO4 7H2O 1 5 g 甘氨酸10 0 mL 琼脂15 0 g 水1000 mL 在加入甘油前将固体成分溶解在水中 加热并频繁搅拌 沸腾 1 分钟以促进溶解 灭菌后 的 PH 值为 7 2 0 2 IX Pseudomonas Agar Medium for Detection of Pyocyanin Pancreatic Digest of Gelatin20 0 g Anhydrous Magnesium Chloride 1 4 g Anhydrous Potassium Sulfate10 0 g Agar15 0 g Glycerin10 0 mL Water1000 mL Dissolve the solid components in the water before adding the glycerin Heat with frequent agitation and boil for 1 minute to effect solution pH after sterilization 7 2 0 2 9 IX 探测绿菌素的假单胞菌琼脂培养基 明胶胰消化液20 0 g 无水氯化镁1 4 g 无水硫酸钾10 0 g 琼脂15 0 g 甘氨酸10 0 mL 水1000 mL 在加入甘油前将固体成分溶解在水中 加热并频繁搅拌 沸腾 1 分钟以促进溶解 灭菌后 的 PH 值为 7 2 0 2 X Fluid Lactose Medium Beef Extract3 0 g Pancreatic Digest of Gelatin 5 0 g Lactose5 0 g Water1000 mL Cool as quickly as possible after sterilization pH after sterilization 6 9 0 2 X 液态乳糖培养基 牛肉提取物3 0 g 明胶胰消化液5 0 g 乳糖5 0 g 水1000 mL 灭菌后尽可能迅速的冷却 灭菌后的pH 为 6 9 0 2 XI Fluid Selenite Cystine Medium Pancreatic Digest of Casein 5 0 g Lactose4 0 g Sodium Phosphate10 0 g Sodium Acid Selenite4 0 g L Cystine10 0 mg Water1000 mL Final pH 7 0 0 2 Mix and heat to effect solution Heat in flowing steam for 15 minutes Do not sterilize XI 液态亚硒酸盐 胱氨酸培养基 酪蛋白胰消化液5 0 g 乳糖4 0 g 磷酸钠10 0 g 10 亚硒酸钠4 0 g L 胱氨酸10 0 mg 水1000 mL 最终 pH 值 7 0 0 2 搅拌并加热以促进溶解 在流动蒸汽中加热 15 分钟 不需灭菌 XII Fluid Tetrathionate Medium Pancreatic Digest of Casein2 5 g Peptic Digest of Animal Tissue 2 5 g Bile Salts1 0 g Calcium Carbonate10 0 g Sodium Thiosulfate30 0 g Water1000 mL Heat the solution of solids to boiling On the day of use add a solution prepared by dissolving 5 g of potassium iodide and 6 g of iodine in 20 mL of water Then add 10 mL of a solution of brilliant green 1 in 1000 and mix Do not heat the medium after adding the brilliant green solution XII 液态连四硫酸盐培养基 酪蛋白胰消化液2 5 g 动物组织胃蛋白消化液2 5 g 胆汁盐1 0 g 碳酸钙10 0 g 硫代硫酸钠30 0 g 水1000 mL 加热含固体的溶液至沸腾 到使用的时候加入用 5g 碘化钠和 6g 碘溶解在 20ml 的水中制 成的溶液 然后加入 10ml 的亮绿溶液 1 并搅拌 加入亮绿溶液后不能加热培养基 XIII Brilliant Green Agar Medium Yeast Extract3 0 g Peptic Digest of Animal Tissue 5 0 g Pancreatic Digest of Casein5 0 g Lactose10 0 g Sodium Chloride5 0 g Sucrose10 0 g Phenol Red80 mg Agar20 0 g Brilliant Green12 5 mg Water1000 mL Boil the solution of solids for 1 minute Sterilize just prior to use melt the medium pour into petri dishes and allow to cool 11 pH after sterilization 6 9 0 2 XIII 亮绿琼脂培养基 酵母菌提取物3 0 g 动物组织胃蛋白消化液5 0 g 酪蛋白胰消化液5 0 g 乳糖10 0 g 氯化钠5 0 g 蔗糖10 0 g 酚红80 mg 琼脂20 0 g 亮绿12 5 mg 水1000 mL 煮沸含固体的溶液 1 分钟 使用前先灭菌 溶化培养基后倒入培养皿中冷却 灭菌后的 PH 值为 6 9 0 2 XIV Xylose Lysine Desoxycholate Agar Medium Xylose3 5 g L Lysine5 0 g Lactose7 5 g Sucrose7 5 g Sodium Chloride5 0 g Yeast Extract3 0 g Phenol Red80 mg Agar13 5 g Sodium Desoxycholate2 5 g Sodium Thiosulfate6 8 g Ferric Ammonium Citrate 800 mg Water1000 mL Final pH 7 4 0 2 Heat the mixture of solids and water with swirling just to the boiling point Do not overheat or sterilize Transfer at once to a water bath maintained at about 50 and pour into plates as soon as the medium has cooled XIV 木糖 赖氨酸 去氧胆酸盐琼脂培养基 木糖3 5 g L 赖氨酸5 0 g 乳糖7 5 g 蔗糖7 5 g 氯化钠5 0 g 酵母菌提取物3 0 g 酚红80 mg 12 琼脂13 5 g 去氧胆酸钠2 5 g 硫代硫酸钠6 8 g 柠檬酸铁胺800 mg 水1000 mL 最终 pH 7 4 0 2 加热固体与水的混合物 搅拌直到刚刚达到沸点 不要过度加热或灭菌 立即转入 50 的 水浴中 当培养基冷却时立即倒入培养皿中 XV Bismuth Sulfite Agar Medium Beef Extract5 0 g Pancreatic Digest of Casein5 0 g Peptic Digest of Animal Tissue 5 0 g Dextrose5 0 g Sodium Phosphate4 0 g Ferrous Sulfate300 mg Bismuth Sulfite Indicator8 0 g Agar20 0 g Brilliant Green25 mg Water1000 mL Final pH 7 6 0 2 Heat the mixture of solids and water with swirling just to the boiling point Do not overheat or sterilize Transfer at once to a water bath maintained at about 50 and pour into plates as soon as the medium has cooled XV 亚硫酸铋琼脂培养基 牛肉提取物5 0 g 酪蛋白胰消化液5 0 g 动物组织胃蛋白消化液5 0 g 葡萄糖5 0 g 磷酸钠4 0 g 硫酸亚铁300 mg 亚硫酸铋指示剂8 0 g 琼脂20 0 g 亮绿25 mg 水1000 mL 最终 pH 7 6 0 2 加热固体与水的混合物 搅拌直到刚刚达到沸点 不要过度加热或灭菌 立即转入 50 的 水浴中 当培养基冷却时立即倒入培养皿中 XVI Triple Sugar Iron Agar Medium 13 Pancreatic Digest of Casein10 0 g Pancreatic Digest of Animal Tissue 10 0 g Lactose10 0 g Sucrose10 0 g Dextrose1 0 g Ferrous Ammonium Sulfate200 mg Sodium Chloride5 0 g Sodium Thiosulfate200 mg Agar13 0 g Phenol Red25 mg Water1000 mL pH after sterilization 7 3 0 2 XVI 三糖 铁 琼脂培养基 酪蛋白胰消化液10 0 g 动物组织胃蛋白消化液10 0 g 乳糖10 0 g 蔗糖10 0 g 葡萄糖1 0 g 硫酸亚铁胺200 mg 氯化钠5 0 g 硫代硫酸钠200 mg 琼脂13 0 g 酚红25 mg 水1000 mL 灭菌后的 pH 值为 7 3 0 2 XVII MacConkey Agar Medium Pancreatic Digest of Gelatin17 0 g Pancreatic Digest of Casein1 5 g Peptic Digest of Animal Tissue 1 5 g Lactose10 0 g Bile Salts Mixture1 5 g Sodium Chloride5 0 g Agar13 5 g Neutral Red30 mg Crystal Violet1 0 mg Water1000 mL Boil the mixture of solids and water for 1 minute to effect solution pH after sterilization 7 1 0 2 XVII MacConkey 琼脂培养基 明胶胰消化液17 0 g 酪蛋白胰消化液1 5 g 14 动物组织胃蛋白消化液1 5 g 乳糖10 0 g 胆汁盐混合物1 5 g 氯化钠5 0 g 琼脂13 5 g 中性红30 mg 水晶紫1 0 mg 水1000 mL 煮沸固体与水的混合物 1 分钟 以促进溶解 灭菌后的 pH 值为 7 1 0 2 XVIII Levine Eosin Methylene Blue Agar Medium Pancreatic Digest of Gelatin10 0 g Dibasic Potassium Phosphate 2 0 g Agar15 0 g Lactose10 0 g Eosin Y400 mg Methylene Blue65 mg Water1000 mL Dissolve the pancreatic digest of gelatin the dibasic potassium phosphate and the agar in the water with warming and allow to cool Just prior to use liquefy the gelled agar solution add the remaining ingredients as solutions in the following amounts and mix for each 100 mL of the liquefied agar solution 5 mL of lactose solution 1 in 5 2 mL of the eosin Y solution 1 in 50 and 2 mL of methylene blue solution 1 in 300 The finished medium may not be clear pH after sterilization 7 1 0 2 XVIII 列文伊红 亚甲基蓝琼脂培养基 明胶胰消化液 10 0 g 磷酸二氢钾2 0 g 琼脂15 0 g 乳糖10 0 g Y 伊红400 mg 亚甲基蓝65 mg 水1000 mL 将明胶胰消化液 磷酸二氢钾和琼脂溶解在水中 加热促进溶解后再冷却 使用前 使凝 固的琼脂溶液液化 加入下列成分 每 100 mL 液化的琼脂溶液 5 mL 的乳糖溶液 体积 比 1 5 2 mL 的伊红 Y 溶液 体积比 1 50 2 mL 的亚甲基蓝溶液 体积比 1 300 最终的培养基可能不清澈 15 灭菌后的 pH 值为 7 1 0 2 XIX Sabouraud Dextrose Agar Medium Dextrose40 g Mixture of equal parts of Peptic Digest of Animal Tissue and Pancreatic Digest of Casein10 g Agar15 g Water1000 mL Mix and boil to effect solution pH after sterilization 5 6 0 2 XIX 萨布罗右旋糖琼脂培养基 右旋糖40 g 等量的动物组织胃蛋白酶消化液和酪蛋白胰消化液的混合物 10 g 琼脂15 g 水1000 mL 搅拌并使溶液沸腾促进溶解 灭菌后的 PH 值为 5 6 0 2 XX Potato Dextrose Agar Medium Cook 300 g of peeled and diced potatoes in 500 mL of water prepared by distillation filter through cheesecloth add water prepared by distillation to make 1000 mL and add the following Agar15 g Glucose20 g Dissolve by heating and sterilize pH after sterilization 5 6 0 2 For use just prior to pouring the plates adjust the melted and cooled to 45 medium with sterile tartaric acid solution 1 in 10 to a pH of 3 5 0 1 Do not reheat the pH 3 5 medium XX 马铃薯葡萄糖琼脂培养基 将 300g 已削皮并切成块状的马铃薯放在 500ml 的蒸馏水中煮 然后用纱布过滤 并用 蒸馏水稀释到 1000ml 再加入下列物质 琼脂15 g 右旋糖20 g 加热溶解并灭菌 灭菌后的 pH 为 5 6 0 2 使用时 在倒入培养皿前先溶解并冷却到 45 向培养基中加入无菌酒石酸溶液使其 pH 值为 3 5 0 1 pH 为 3 5 的培养基无需再加热 16 SAMPLING 样品 Provide separate 10 mL or 10 g specimens for each of the tests called for in the individual monograph 为在单独专论中的每个测试提供 10 mL 或 10 g 的样品 PROCEDURE 方法 Prepare the specimen to be tested by treatment that is appropriate to its physical characteristics and that does not alter the number and kind of microorganisms originally present in order to obtain a solution or suspension of all or part of it in a form suitable for the test procedure s to be carried out For a solid that dissolves to an appreciable extent but not completely reduce the substance to a moderately fine powder suspend it in the vehicle specified and proceed as directed under Total Aerobic Microbial Count and under Test for Staphylococcus aureus and Pseudomonas aeruginosa and Test for Salmonella species and Escherichia coli 准备待测试的样品时要采取适合其自然特点并且不改变其原来所有的微生物数量和种类的 方法 目的是得到与要实行的实验过程相配合的全部或部分样品的溶液或悬浮液 对于溶 解到可估计但不完全的程度的固体 将该物质磨成一定精细的粉末后悬浮在指定的溶剂中 再依照总有氧微生物计数 葡萄球菌和假单胞菌的检验及沙门氏菌属和大肠杆菌的检验的 指导操作 For a fluid specimen that consists of a true solution or a suspension in water or a hydroalcoholic vehicle containing less than 30 percent of alcohol and for a solid that dissolve