USP36 1117 优良微生物检测规范中英文版
USP36 1117 优良微生物检测规范中英文版
USP36 1117 优良微生物检测规范（中英文1/ 2）2013-08-09 15:30:46| 分类：USP|举报|字号订阅1117 MICROBIOLOGICAL BEST LABORATORY PRACTICES 优良微生物检测规范INTRODUCTION 介绍Good laboratory practices in a microbiology laboratory consist of activities that depend on several principles: aseptic technique, control of media, control of test strains, operation and control of equipment, diligent recording and evaluation of data, and training of the laboratory staff. Because of the inherent risk of variability in microbiology data, reliability and reproducibility are dependent on the use of accepted methods and adherence to good laboratory practices.优良微生物检测规范由一些活动组成，这些活动依赖于几个基本要素：无菌技术、培养基控制、检测用菌株控制、设备操作和控制、完善的记录和数据评估、化验室员工的培训。
由于微生物数据具有天生的不确定性，数据的可靠性和重复性取决于是否使用被接受的方法，以及是否严格遵守化验室规范。
MEDIA PREPARATION AND QUALITY CONTROL 培养基制备和质量控制Media Preparation 培养基制备Culture media are the basis for most microbiological tests. Safeguarding the quality of the media is therefore critical to the success of the microbiology laboratory. Media preparation, proper storage, and quality control testing can ensure a consistent supply of high-quality media.培养基是大多数微生物测试的基础。
USP-35-1113 微生物鉴定、鉴别及菌株分型指导原则翻译稿
通过表型进行微生物鉴定
表型方法是通过基因表达产物区分不同的微生物。通常这需要相当数量的单菌落纯培养 物。微生物活化、繁殖和鉴定受到培养条件的限制，事实上许多环境中的微生物目前还不能 通过常规培养条件分离。此外，由原代微生物传代的分离的得到的新鲜子代培养物，其表型 性质可能不会充分表达。但是，很多鉴定系统都是基于碳源利用、生理生化反应而设计的，
这些函数的数学推导见表 4。
表 4 两行两列联表采用培养方法和替代 PCR 方法（ISO 5725-1 和 5725-2 2004）*
PCR 方法
培养方法
阳性
阴性
合计
阳性
a 阳性
b 假阴性
a+b
阴性
c 假阳性
d 阴性
c+d
合计
a +c
b+d
* ISO 5725-1:1994 准确度（真实性和精密）的测量方法和结果-第 1 部分：总则和定义， ISO 5725-2:1994 准确度（真实性和精密）的测量方法和结果-第 2 部分：标准测量方法的重复 性和再现性的基本方法。
分类 菌落 形态 生理特性 生化特性 抑制 血清 化学分类 生态学
表 1 表型特征用于微生物分类学 特征特性
菌落形态, 菌落颜色, 形状和大小, 色素产生 菌体形态, 菌体大小, 菌体形状, 鞭毛类型, 储存物质, 革兰氏染色, 芽孢染色, 产孢情况
耐氧性, pH 范围, 最适温度和范围, 抗盐性 碳源利用, 碳水化合物氧化和发酵, 酶反应
美国药典USP31-71-无菌检查法-双语版
美国药典USP31-71-⽆菌检查法-双语版美国药典USP31-NF26⽆菌检查法《71》.doc71 STERILITY TESTS ⽆菌检查法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.此通则的各部分已经与欧洲药典和/或⽇本药典的对应部分做了协调。
不⼀致的部分⽤符号（）来标明。
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.下⾯这些步骤适⽤于测定是否某个⽤于⽆菌⽤途的药品是否符合其具体的各论中关于⽆菌检查的要求。
USP37-1111非无菌药品微生物检查：药用制剂和药用物质可接受标准(中英文)
USP37 <1111>非无菌药品微生物检查：药用制剂和药用物质可接受标准（中英文）(2014-06-06 14:19:33)转载▼分类：食品药品<1111> MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS:ACCEPTANCE CRITERIA FOR PHARMACEUTICAL PREPARATIONS ANDSUBSTANCES FOR PHARMACEUTICAL USE非无菌药品微生物检查：药用制剂和药用物质可接受标准The presence of certain microorganisms in nonsterile preparations may have the potential to reduce or even inactivate the therapeutic activity of the product and has a potential to adversely affect the health of the patient. Manufacturers have therefore to ensure a low bioburden of finished dosage forms by implementing current guidelines on Good Manufacturing Practice during the manufacture, storage, and distribution of pharmaceutical preparations.在非无菌制剂中如果出现特定的微生物，可能会降低甚至失去治疗活性，对患者健康产生潜在负面影响。
因此，生产商需要通过在生产、存贮制剂的过程中实施GMP现行指南，保证制剂的生物负载在一个较低的水平。
Microbial examination of nonsterile products is performed according to the methods given in the texts on Microbial Enumeration Tests 61 and Tests for Specified Microorganisms 62 . Acceptance criteria for nonsterile pharmaceutical products based upon the total aerobic microbial count (TAMC) and the total combined yeasts and molds count (TYMC) are givenin Tables 1 and 2. Acceptance criteria are based on individual results or on the average of replicate counts when replicate counts are performed (e.g., direct plating methods).非无菌药品的微生物检查应根据<61 微生物限度检查>和<62 控制菌测试>进行。
(完整word版)USP38通用章节目录中文
USP38-通用章节指导目录(附录)Guide to General Chapters 通用章节指导General Requirements for Test and Assays检查与含量分析的一般要求＜1＞INJECTIONS AND IMPLANTED DRUG PRODUCTS (PARENTERALS)—PRODUCT QUALITY TESTS 注射和植入药物产品(注射用) —产品质量测试＜1＞INJECTIONS注射剂＜2＞ORAL DRUG PRODUCTS—PRODUCT QUALITY TESTS 口服药物产品质量测试＜3＞TOPICAL AND TRANSDERMAL DRUG PRODUCTS—PRODUCT QUALITY TESTS 局部和透皮药物产品—产品质量测试＜4＞MUCOSAL DRUG PRODUCTS—PRODUCT QUALITY TESTS 粘膜药物产品质量测试＜5＞INHALATION AND NASAL DRUG PRODUCTS—GENERAL INFORMATION AND PRODUCT QUALITY TESTS 吸入剂产品—产品质量测试＜7＞LABELING 标签＜11＞USP REFERENCE STANDARDS USP标准品Apparatus for Test and Assays用于检查与含量分析的器具＜17＞PRESCRIPTION CONTAINER LABELING处方容器标签＜21＞THERMOMETERS温度计＜31＞VOLUMETRIC APPARATUS容量器具＜41＞BALANCES天平Microbiological Tests 微生物检查法＜51＞ANTIMICROBIAL EFFECTIVENESS TESTING抗菌剂有效性检查法＜55＞BIOLOGICAL INDICATORS—RESISTANCE PERFORMANCE TESTS生物指示剂－耐药性实验＜61＞MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: MICROBIAL ENUMERATION TESTS非无菌产品的微生物限度检查：微生物列举检查法＜62＞MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: TESTS FOR SPECIFIED MICROORGANISMS 非无菌产品的微生物限度检查：特定微生物检查法＜63＞MYCOPLASMA TESTS 支原体检查法＜71＞STERILITY TESTS无菌检查法Biological tests and assays生物检查法与测定法＜81＞ANTIBIOTICS—MICROBIAL ASSAYS抗生素－微生物测定＜85＞BACTERIAL ENDOTOXINS TEST细菌内毒素检查法＜87＞BIOLOGICAL REACTIVITY TESTS, IN VITRO体外的生物反应性检查法＜88＞BIOLOGICAL REACTIVITY TESTS, IN VIVO 体内的生物反应性检查法＜89＞ENZYMES USED AS ANCILLARY MATERIALS IN PHARMACEUTICAL MANUFACTURING 药品生产中酶作为辅料所使用＜90＞FETAL BOVINE SERUM—QUALITY ATTRIBUTES AND FUNCTIONALITY TESTS 牛胎儿血清-质量品质和功能检查法＜91＞CALCIUM PANTOTHENATE ASSAY泛酸钙测定法＜92＞GROWTH FACTORS AND CYTOKINES USED IN CELL THERAPY MANUFACTURING 在细胞疗法中使用生长因子和细胞因子＜111＞DESIGN AND ANALYSIS OF BIOLOGICAL ASSAYS 生物测定法的设计与分析＜115＞DEXPANTHENOL ASSAY右泛醇（拟胆碱药）测定法＜121＞INSULIN ASSAYS胰岛素测定法＜121.1＞PHYSICOCHEMICAL ANALYTICAL PROCEDURES FOR INSULINS胰岛素的物理化学分析程序＜123＞GLUCAGON BIOIDENTITY TESTS 高血糖素的生物鉴别检查法＜124＞ERYTHROPOIETIN BIOASSAYS 红细胞生成素的微生物测定＜126＞SOMATROPIN BIOIDENTITY TESTS 生长激素的生物鉴别检查法＜130＞PROTEIN A QUALITY ATTRIBUTES 蛋白质A的质量特征＜151＞PYROGEN TEST热原检查法＜161＞TRANSFUSION AND INFUSION ASSEMBLIES AND SIMILAR MEDICAL DEVICES 输血输液用具以及相类似的医疗器械＜171＞VITAMIN B12 ACTIVITY ASSAY……2548维生素B12活性测定法Chemical Tests and assays化学实验检查与测定法鉴别检查＜181＞IDENTIFICATION—ORGANIC NITROGENOUS BASES鉴别－有机氮碱化合物＜191＞IDENTIFICATION TESTS—GENERAL鉴别实验－通用＜193＞IDENTIFICATION—TETRACYCLINES鉴别－四环素类＜197＞SPECTROPHOTOMETRIC IDENTIFICATION TESTS分光光度计鉴别实验＜201＞THIN-LAYER CHROMATOGRAPHIC IDENTIFICATION TEST薄层色谱鉴别实验Limit Tests 限度检查法＜206＞ALUMINUM铝＜207＞TEST FOR 1,6-ANHYDRO DERIV ATIVE FOR ENOXAPARIN SODIUM依诺肝素钠的酐类衍生物实验＜208＞ANTI-FACTOR Xa AND ANTI-FACTOR IIa ASSAYS FOR UNFRACTIONATED AND LOW MOLECULAR WEIGHT HEPARINS普通肝素和低分子肝素产品中抗体Xa和抗体IIa测定＜209＞LOW MOLECULAR WEIGHT HEPARIN MOLECULAR WEIGHT DETERMINATIONS 低分子肝素钠分子量测定＜211＞ARSENIC砷＜221＞CHLORIDE AND SULFATE氯和硫＜223＞DIMETHYLANILINE二甲基苯胺＜226＞4-EPIANHYDRO-TETRACYCLINE4－？－四环素＜227＞4-AMINOPHENOL IN ACETAMINOPHEN-CONTAINING DRUG PRODUCTS 对乙酰氨酚药物产品中氨基酚＜228＞ETHYLENE OXIDE AND DIOXANE 环氧乙烷和二氧六环＜231＞HEA VY METALS重金属（删除）＜232＞ELEMENTAL IMPURITIES—LIMITS 元素杂质-限度＜233＞ELEMENTAL IMPURITIES—PROCEDURES 元素杂质-规程＜241＞IRON铁＜251＞LEAD铅＜261＞MERCURY汞＜267＞POROSIMETRY BY MERCURY INTRUSION 水银孔隙仪＜268＞POROSITY BY NITROGEN ADSORPTION–DESORPTION 氮吸附-解吸测定孔隙率＜271＞READILY CARBONIZABLE SUBSTANCES TEST易碳化物检查法＜281＞RESIDUE ON IGNITION炽灼残渣＜291＞SELENIUM硒Other Tests and Assays 其它检查法与测定法＜301＞ACID-NEUTRALIZING CAPACITY酸中和容量＜311＞ALGINATES ASSAY藻酸盐测定法＜341＞ANTIMICROBIAL AGENTS—CONTENT 抗菌剂－含量＜345＞Assay for Citric Acid/Citrate and Phosphate 柠檬酸/柠檬酸盐和磷酸盐的测定＜351＞ASSAY FOR STEROIDS类固醇（甾类化合物）测定法＜361> BARBITURATE ASSAY 巴比妥类药物测定法＜371＞COBALAMIN RADIOTRACER ASSAY钴铵素放射性跟踪剂测定法＜381＞ELASTOMERIC CLOSURES FOR INJECTIONS 注射剂的弹性密封件＜391＞EPINEPHRINE ASSAY肾上腺素测定法＜401＞FATS AND FIXED OILS脂肪与混合油＜411＞FOLIC ACID ASSAY叶酸测定法＜413＞IMPURITIES TESTING IN MEDICAL GASES 医用气体杂质检查＜415＞MEDICAL GASES ASSAY 医用气体含量检查＜425＞IODOMETRIC ASSAY—ANTIBIOTICS碘量检查法－抗生素＜429＞LIGHT DIFFRACTION MEASUREMENT OF PARTICLE SIZE粒径的光衍射测量法＜431＞METHOXY DETERMINATION甲氧基测定法＜441＞NIACIN OR NIACINAMIDE ASSAY 烟酰或烟酰胺测定法＜451＞NITRITE TITRATION亚硝酸盐滴定＜461＞NITROGEN DETERMINATION氮测定法＜466＞ORDINARY IMPURITIES一般杂质＜467＞RESIDUAL SOLVENTS残留溶剂＜469＞ETHYLENE GLYCOL, DIETHYLENE GLYCOL, AND TRIETHYLENE GLYCOL IN ETHOXYLATED SUBSTANCES乙氧基物质中乙二醇、二甘醇、三甘醇测定＜471＞OXYGEN FLASK COMBUSTION氧瓶燃烧法＜481＞RIBOFLAVIN ASSAY核黄素（维生素B2）测定法＜501＞SALTS OF ORGANIC NITROGENOUS BASES有机氮盐＜503＞ACETIC ACID IN PEPTIDES 多肽类中乙酸测定＜511＞SINGLE-STEROID ASSAY单一的类固醇测定法＜525＞SULFUR DIOXIDE 二氧化硫＜531＞THIAMINE ASSAY硫胺素测定法＜541＞TITRIMETRY滴定法＜551＞VITAMIN E ASSAY维生素E测定法＜561＞ARTICLES OF BOTANICAL ORIGIN植物起源的药品＜563＞IDENTIFICATION OF ARTICLES OF BOTANICAL ORIGIN植物药品的鉴别＜565＞BOTANICAL EXTRACTS植物提取＜571＞VITAMIN A ASSAY维生素A测定法＜581＞VITAMIN D ASSAY维生素D测定法＜591＞ZINC DETERMINATION锌的测定法Physical Test and Determinations物理检查与测定法＜601＞INHALATION AND NASAL DRUG PRODUCTS: AEROSOLS, SPRAYS, AND POWDERS—PERFORMANCE QUALITY TESTS吸入剂、鼻雾剂：气溶胶，喷雾，干粉-质量通则＜602＞PROPELLANTS 推进剂＜603＞TOPICAL AEROSOLS 局部喷雾剂＜604＞LEAK RATE 渗漏率＜610＞ALTERNATIVE MICROBIOLOGICAL SAMPLING METHODS FOR NONSTERILE INHALED AND NASAL PRODUCTS非无菌吸入和鼻雾剂可供选择的微生物取样方法＜611＞ALCOHOL DETERMINATION乙醇测定法＜616＞BULK DENSITY AND TAPPED DENSITY堆密度与振实密度＜621＞CHROMATOGRAPHY色谱法＜631＞COLOR AND ACHROMICITY呈色与消色＜641＞COMPLETENESS OF SOLUTION溶解度＜643＞TOTAL ORGANIC CARBON总有机碳＜645＞W ATER CONDUCTIVITY水电导率＜651＞CONGEALING TEMPERATURE凝点温度＜659＞PACKAGING AND STORAGE REQUIREMENTS 包装和储藏要求＜660＞CONTAINERS—GLASS 容器-玻璃＜661＞CONTAINERS—PLASTICS容器-塑料＜670＞AUXILIARY PACKAGING COMPONENTS 辅助包装部件＜671＞CONTAINERS—PERFORMANCE TESTING容器－性能测试＜691＞COTTON棉花＜695＞CRYSTALLINITY结晶度＜696＞CHARACTERIZATION OF CRYSTALLINE SOLIDS BY MICROCALORIMETRY AND SOLUTION CALORIMETRY 通过溶液量热学测定结晶性＜697＞CONTAINER CONTENT FOR INJECTIONS 注射剂容器容积＜698＞DELIVERABLE VOLUME抽取体积＜699＞DENSITY OF SOLIDS固体密度＜701＞DISINTEGRATION崩解时限＜705＞QUALITY ATTRIBUTES OF TABLETS LABELED AS HA VING A FUNCTIONAL SCORE ？＜711＞DISSOLUTION 溶出度＜721＞DISTILLING RANGE馏程＜724＞DRUG RELEASE药物释放度＜729＞GLOBULE SIZE DISTRIBUTION IN LIPID INJECTABLE EMULSIONS脂类可注射的乳剂的粒径分布＜730＞Plasma Spectrochemistry 血浆光谱化学？＜731＞LOSS ON DRYING4干燥失重＜733＞LOSS ON IGNITION灼烧失重＜735＞X-RAY FLUORESCENCE SPECTROMETRY X射线光谱＜736＞MASS SPECTROMETRY 质谱＜741＞MELTING RANGE OR TEMPERATURE熔距或熔点＜751＞METAL PARTICLES IN OPHTHALMIC OINTMENTS眼用软膏中的金属粒子＜755＞MINIMUM FILL最低装量＜761＞NUCLEAR MAGNETIC RESONANCE核磁共振＜771＞OPHTHALMIC OINTMENTS眼用软膏＜776＞OPTICAL MICROSCOPY光学显微镜＜781＞OPTICAL ROTATION旋光度＜785＞OSMOLALITY AND OSMOLARITY渗透压＜786＞PARTICLE SIZE DISTRIBUTION ESTIMATION BY ANALYTICAL SIEVING 筛分法估算粒径分布＜787＞SUBVISIBLE PARTICULATE MATTER IN THERAPEUTIC PROTEIN INJECTIONS显微计数法在治疗性蛋白注射剂中应用＜788＞PARTICULATE MATTER IN INJECTIONS注射剂中的不溶性微粒＜789＞PARTICULATE MATTER IN OPHTHALMIC SOLUTIONS眼用溶液中的不溶性微粒＜790＞VISIBLE PARTICULATES IN INJECTIONS 注射剂中可见异物＜791＞pH＜795＞PHARMACEUTICAL COMPOUNDING—NONSTERILE PREPARATIONS药物混合－非无菌制剂＜797＞PHARMACEUTICAL COMPOUNDING—STERILE PREPARATIONS药物混合－无菌制剂＜801＞POLAROGRAPHY极谱法＜811＞POWDER FINENESS粉剂细度＜821＞RADIOACTIVITY放射性＜823＞POSITRON EMISSION TOMOGRAPHY DRUGS FOR COMPOUNDING, INVESTIGATIONAL, AND RESEARCH USES用于正电子发射断层造影术的放射性药物＜831＞REFRACTIVE INDEX折光率＜841＞SPECIFIC GRAVITY比重＜846＞SPECIFIC SURFACE AREA 比表面积＜851＞SPECTROPHOTOMETRY AND LIGHT-SCATTERING分光光度计与光散射＜852＞ATOMIC ABSORPTION SPECTROSCOPY 原子吸收光谱＜853＞FLUORESCENCE SPECTROSCOPY 荧光光谱＜854＞MID-INFRARED SPECTROSCOPY 中红外光谱＜857＞ULTRAVIOLET-VISIBLE SPECTROSCOPY 紫外可见光谱＜861＞SUTURES—DIAMETER缝线－直径？＜871＞SUTURES—NEEDLE ATTACHMENT缝线－穿孔实验＜881＞TENSILE STRENGTH张力＜891＞THERMAL ANALYSIS热分析＜905＞UNIFORMITY OF DOSAGE UNITS制剂单位的含量均匀度＜911＞VISCOSITY—CAPILLARY METHODS黏度-毛细管法＜912＞VISCOSITY—ROTATIONAL METHODS 黏度-旋转法＜913＞VISCOSITY—ROLLING BALL METHOD 黏度-球法＜921＞W ATER DETERMINATION水分测定＜941＞CHARACTERIZATION OF CRYSTALLINE AND PARTIALLY CRYSTALLINE SOLIDS BY X-RAY POWDER DIFFRACTION (XRPD)X光衍射General Information通用信息＜1005＞ACOUSTIC EMISSION 声频发射＜1010＞ANALYTICAL DATA—INTERPRETATION AND TREATMENT分析数据－解释与处理＜1015＞AUTOMATED RADIOCHEMICAL SYNTHESIS APPARATUS放射性自动合成装置＜1024＞BOVINE SERUM 牛血清＜1027＞FLOW CYTOMETRY 流式细胞仪＜1030＞BIOLOGICAL ASSAY CHAPTERS—OVERVIEW AND GLOSSARY生物测定章节-综述和术语＜1031＞THE BIOCOMPATIBILITY OF MATERIALS USED IN DRUG CONTAINERS, MEDICAL DEVICES, AND IMPLANTS用于药物容器、医疗设施和植入剂的材料的生物相容性＜1034＞ANALYSIS OF BIOLOGICAL ASSAYS 生物测定分析＜1035＞BIOLOGICAL INDICATORS FOR STERILIZATION灭菌用生物指示剂＜1041＞BIOLOGICS生物制剂＜1043＞Ancillary Material for Cell, Gene, and Tissue-Engineered Products细胞，基因与组织设计产品的辅助材料＜1044＞CRYOPRESERV ATION OF CELLS 细胞低温保存＜1045＞BIOTECHNOLOGY-DERIVED ARTICLES生物技术提取产品＜1046＞CELLULAR AND TISSUE-BASED PRODUCTS细胞与组织产品＜1047＞GENE THERAPY PRODUCTS 基因治疗产品＜1048＞QUALITY OF BIOTECHNOLOGICAL PRODUCTS: ANALYSIS OF THE EXPRESSION CONSTRUCT IN CELLS USED FOR PRODUCTION OF r-DNA DERIVED PROTEIN PRODUCTS 生物技术产品的质量：从蛋白质产品中提取的r-DNA产品在细胞中表达结构的分析＜1049＞QUALITY OF BIOTECHNOLOGICAL PRODUCTS: STABILITY TESTING OF BIOTECHNOLOGICAL/BIOLOGICAL PRODUCTS生物技术产品的质量：生物技术/生物产品的稳定性实验＜1050＞VIRAL SAFETY EV ALUATION OF BIOTECHNOLOGY PRODUCTS DERIVED FROM CELL LINES OF HUMAN OR ANIMAL ORIGIN从人或动物细胞中提取的生物技术产品的病毒安全性评估＜1051＞CLEANING GLASS APPARATUS玻璃容器的清洗＜1052＞BIOTECHNOLOGY-DERIVED ARTICLES—AMINO ACID ANALYSIS生物技术提取法-氨基酸测定＜1053＞CAPILLARY ELECTROPHORESIS 毛细管电泳法＜1054＞BIOTECHNOLOGY-DERIVED ARTICLES—ISOELECTRIC FOCUSING生物技术提取法-等电点聚集＜1055＞BIOTECHNOLOGY-DERIVED ARTICLES—PEPTIDE MAPPING生物技术提取法-肽谱＜1056＞BIOTECHNOLOGY-DERIVED ARTICLES—POLYACRYLAMIDE GEL ELECTROPHORESIS 生物技术提取法-凝胶电泳＜1057＞BIOTECHNOLOGY-DERIVED ARTICLES—TOTAL PROTEIN ASSAY生物技术提取法-总蛋白测定＜1058＞ANALYTICAL INSTRUMENT QUALIFICATION 分析仪器要求＜1059＞EXCIPIENT PERFORMANCE 赋形剂＜1061＞COLOR—INSTRUMENTAL MEASUREMENT显色－仪器测量＜1065＞Ion Chromatography 离子色谱法＜1066＞PHYSICAL ENVIRONMENTS THAT PROMOTE SAFE MEDICATION USE物理环境促使安全使用药物＜1072＞DISINFECTANTS AND ANTISEPTICS 消毒剂和防腐剂＜1074＞EXCIPIENT BIOLOGICAL SAFETY EV ALUATION GUIDELINES赋形剂（辅料）生物安全性评估指导＜1078＞GOOD MANUFACTURING PRACTICES FOR BULK PHARMACEUTICAL EXCIPIENTS 批药品赋形剂的生产管理规范＜1079＞Good Storage and Shipping Practices 良好的贮存与运输规范＜1080＞BULK PHARMACEUTICAL EXCIPIENTS—CERTIFICATE OF ANALYSIS批药品赋形剂-COA＜1084＞GLYCOPROTEIN AND GLYCAN ANALYSIS—GENERAL CONSIDERATIONS 糖蛋白和多糖分析-一般通则＜1086＞IMPURITIES IN DRUG SUBSTANCES AND DRUG PRODUCTS药物和药物产品中的杂质＜1087＞APPARENT INTRINSIC DISSOLUTION—DISSOLUTION TESTING PROCEDURES FOR ROTATING DISK AND STATIONARY DISK内部的溶出度-旋转和静止溶出检测程序？＜1088＞IN VITRO AND IN VIVO EV ALUATION OF DOSAGE FORMS体内与体外的剂型的评估＜1090＞ASSESSMENT OF DRUG PRODUCT PERFORMANCE-BIOAV AILABILITY, BIOEQUIV ALENCE, AND DISSOLUTION药物产品性能评估：生物利用度、生物等效性和溶出＜1091＞LABELING OF INACTIVE INGREDIENTS非活性成分的标示＜1092＞THE DISSOLUTION PROCEDURE: DEVELOPMENT AND V ALIDATION溶出程序：开发与验证＜1094＞CAPSULES—DISSOLUTION TESTING AND RELATED QUALITY ATTRIBUTES 胶囊-关于产品质量的溶出测定＜1097＞BULK POWDER SAMPLING PROCEDURES：粉末样品取样程序＜1102＞IMMUNOLOGICAL TEST METHODS—GENERAL CONSIDERATIONS免疫测试方法-总则＜1103＞IMMUNOLOGICAL TEST METHODS—ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) 免疫学测试方法-酶联免疫吸附测定＜1104＞IMMUNOLOGICAL TEST METHODS—IMMUNOBLOT ANALYSIS免疫测试方法-免疫印迹法＜1105＞IMMUNOLOGICAL TEST METHODS—SURFACE PLASMON RESONANCE 免疫测试方法-表面等离子体共振＜1106＞IMMUNOGENICITY ASSAYS—DESIGN AND VALIDATION OF IMMUNOASSAYS TO DETECT ANTI-DRUG ANTIBODIES？＜1111＞MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: ACCEPTANCE CRITERIA FOR PHARMACEUTICAL PREPARATIONS AND SUBSTANCES FOR PHARMACEUTICAL USE非无菌产品的微生物学检查：药用制剂和制药过程使用的物质接受标准＜1112＞MICROBIAL CHARACTERIZATION, IDENTIFICATION, AND STRAIN TYPING 非无菌药物产品水活性测定应用＜1113＞MICROBIOLOGICAL ATTRIBUTES OF NONSTERILE PHARMACEUTICAL PRODUCTS 非无菌药品中的微生物分布＜1115＞BIOBURDEN CONTROL OF NONSTERILE DRUG SUBSTANCES AND PRODUCTS 非无菌药物和产品的生物负载控制＜1116＞MICROBIOLOGICAL CONTROL AND MONITORING OF ASEPTIC PROCESSING ENVIRONMENTS洁净的房间与其它可控环境的微生物评估＜1117＞MICROBIOLOGICAL BEST LABORATORY PRACTICES 微生物最优实验室规范＜1118＞MONITORING DEVICES—TIME, TEMPERATURE, AND HUMIDITY监控装置－时间、温度与湿度＜1119＞NEAR-INFRARED SPECTROPHOTOMETRY近红外分光光度测定法＜1120＞Raman Spectrophotometry 拉曼分光光度测定法＜1121＞NOMENCLATURE命名＜1125＞NUCLEIC ACID-BASED TECHNIQUES—GENERAL 核酸技术-通则＜1126＞NUCLEIC ACID-BASED TECHNIQUES—EXTRACTION, DETECTION, AND SEQUENCING 核酸技术-提取、检测、测序＜1127＞NUCLEIC ACID-BASED TECHNIQUES—AMPLIFICATION 核酸技术-扩增＜1128＞NUCLEIC ACID-BASED TECHNIQUES—MICROARRAY 核酸技术-微阵列＜1129＞NUCLEIC ACID-BASED TECHNIQUES—GENOTYPING 核酸技术-基因分型＜1130＞NUCLEIC ACID-BASED TECHNIQUES—APPROACHES FOR DETECTING TRACE NUCLEIC ACIDS (RESIDUAL DNA TESTING)核酸技术-探测微量核酸的应用（残留DNA测试）＜1136＞PACKAGING AND REPACKAGING—SINGLE-UNIT CONTAINERS包装和再包装－单一容器＜1151＞PHARMACEUTICAL DOSAGE FORMS药物剂型＜1152＞ANIMAL DRUGS FOR USE IN ANIMAL FEEDS兽药在动物饲料中的使用＜1160＞PHARMACEUTICAL CALCULATIONS IN PRESCRIPTION COMPOUNDING 按处方混合的药物的计算＜1163＞QUALITY ASSURANCE IN PHARMACEUTICAL COMPOUNDING按处方混合的药物的质量保证＜1171＞PHASE-SOLUBILITY ANALYSIS相溶解分析＜1174＞Powder Flow 粉末流动性＜1176＞PRESCRIPTION BALANCES AND VOLUMETRIC APPARATUS 处方天平与容量器具＜1177＞Good Packaging Practices 良好的包装操作＜1178＞Good Repackaging Practices 良好的再包装操作＜1180＞HUMAN PLASMA 人血浆＜1181＞SCANNING ELECTRON MICROSCOPY扫描电子显微镜＜1184＞SENSITIZATION TESTING 致敏测试＜1191＞STABILITY CONSIDERATIONS IN DISPENSING PRACTICE分装操作中稳定性考察＜1195＞SIGNIFICANT CHANGE GUIDE FOR BULK PHARMACEUTICAL EXCIPIENTS 散装药用辅料更换指导原则＜1197＞GOOD DISTRIBUTION PRACTICES FOR BULK PHARMACEUTICAL EXCIPIENTS 散装药用辅料良好的分装操作＜1207＞STERILE PRODUCT PACKAGING—INTEGRITY EV ALUATION无菌产品包装－完整性评估＜1208＞STERILITY TESTING—V ALIDATION OF ISOLATOR SYSTEMS无菌实验－隔离系统的验证＜1209＞STERILIZATION—CHEMICAL AND PHYSICOCHEMICAL INDICATORS AND INTEGRATORS灭菌－化学与物理化学的指示剂以及二者的综合＜1211＞STERILIZATION AND STERILITY ASSURANCE OF COMPENDIAL ARTICLES 药典物品中的灭菌与灭菌保证＜1216＞TABLET FRIABILITY片剂的脆碎度＜1217＞TABLET BREAKING FORCE 片剂断裂力＜1222＞TERMINALLY STERILIZED PHARMACEUTICAL PRODUCTS—PARAMETRIC RELEASE 药品终端灭菌－放行参数＜1223＞V ALIDATION OF ALTERNATIVE MICROBIOLOGICAL METHODS可供选择的微生物学方法的验证＜1224＞TRANSFER OF ANALYTICAL PROCEDURES 分析方法转移＜1225＞V ALIDATION OF COMPENDIAL PROCEDURES药典方法的验证＜1226＞VERIFICATION OF COMPENDIAL PROCEDURES 药典方法的确认＜1227＞V ALIDATION OF MICROBIAL RECOVERY FROM PHARMACOPEIAL ARTICLES从药物中回收微生物的验证＜1229＞STERILIZATION OF COMPENDIAL ARTICLES 药典灭菌过程＜1229.1＞STEAM STERILIZATION BY DIRECT CONTACT 直接蒸汽灭菌＜1229.2＞MOIST HEAT STERILIZATION OF AQUEOUS LIQUIDS 水溶液的湿热灭菌＜1229.3＞MONITORING OF BIOBURDEN 生物负载监控＜1229.4＞STERILIZING FILTRATION OF LIQUIDS 溶液的无菌过滤器＜1229.6＞LIQUID-PHASE STERILIZATION 液态灭菌＜1229.7＞GASEOUS STERILIZATION 气态灭菌＜1229.8＞DRY HEAT STERILIZATION 干热灭菌＜1229.10＞RADIATION STERILIZATION 辐射灭菌＜1230＞W ATER FOR HEMODIALYSIS APPLICATIONS 血液透析过程用水＜1231＞W ATER FOR PHARMACEUTICAL PURPOSES制药用水＜1234＞VACCINES FOR HUMAN USE—POLYSACCHARIDE AND GLYCOCONJUGATE VACCINES 人用疫苗-多糖和糖复合物疫苗＜1235＞V ACCINES FOR HUMAN USE—GENERAL CONSIDERATIONS 人用疫苗-通则＜1237＞VIROLOGY TEST METHODS 病毒测试方法＜1238＞V ACCINES FOR HUMAN USE—BACTERIAL V ACCINES 人用疫苗-细菌疫苗＜1240＞VIRUS TESTING OF HUMAN PLASMA FOR FURTHER MANUFACTURE下一步使用人血浆的病毒测试＜1241＞W ATER–SOLID INTERACTIONS IN PHARMACEUTICAL SYSTEMS在药物系统中水与固体的相互作用＜1251＞WEIGHING ON AN ANALYTICAL BALANCE关于分析天平的称重＜1265＞Written Prescription Drug Information－Guidelines 书面的处方药信息－指南＜1285＞PREPARATION OF BIOLOGICAL SPECIMENS FOR HISTOLOGIC AND IMMUNOHISTOCHEMICAL ANALYSIS为了组织和免疫组织分析的生物标本制备＜1285.1＞HEMATOXYLIN AND EOSIN STAINING OF SECTIONED TISSUE FOR MICROSCOPIC EXAMINATION显微镜观察用苏木精和伊红染色的切片＜1601＞PRODUCTS FOR NEBULIZATION—CHARACTERIZATION TESTS 产品雾化状态-性状描述＜1644＞THEORY AND PRACTICE OF ELECTRICAL CONDUCTIVITY MEASUREMENTS OF SOLUTIONS溶液电导值测量方法的理论与实践＜1660＞EV ALUATION OF THE INNER SURFACE DURABILITY OF GLASS CONTAINERS 玻璃容器内表面耐久性评估＜1724＞SEMISOLID DRUG PRODUCTS—PERFORMANCE TESTS 半固态药物产品-性能测试＜1736＞APPLICATIONS OF MASS SPECTROMETRY 质谱应用＜1761＞APPLICATIONS OF NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY核磁共振光谱应用＜1787＞MEASUREMENT OF SUBVISIBLE PARTICULATE MATTER IN THERAPEUTIC PROTEIN INJECTIONS用显微镜测量方法测量治疗性蛋白注射剂的不溶性微粒＜1788＞METHODS FOR THE DETERMINATION OF PARTICULATE MATTER IN INJECTIONS AND OPHTHALMIC SOLUTIONS注射剂和眼用溶液的不溶性微粒测定的方法选择＜1852＞ATOMIC ABSORPTION SPECTROSCOPY—THEORY AND PRACTICE原子吸收光谱-理论与实践＜1853＞FLUORESCENCE SPECTROSCOPY—THEORY AND PRACTICE荧光光谱-理论与实践＜1854＞MID-INFRARED SPECTROSCOPY—THEORY AND PRACTICE中红外光谱-理论与实践＜1857＞ULTRA VIOLET-VISIBLE SPECTROSCOPY—THEORY AND PRACTICE紫外可见光谱-理论与实践＜1911＞RHEOMETRY 流变测定Dietary Supplements营养补充剂General Tests and Assays 一般检查法与测定法＜2021＞MICROBIAL ENUMERATION TESTS—NUTRITIONAL AND DIETARY SUPPLEMENTS…3080微生物数量实验－营养与食品添加剂＜2022＞MICROBIOLOGICAL PROCEDURES FOR ABSENCE OF SPECIFIED MICROORGANISMS—NUTRITIONAL AND DIETARY SUPPLEMENTS (3083)不得检出特定微生物的程序－营养与营养补充剂＜2023>MICROBIOLOGICAL ATTRIBUTES OF NONSTERILE NUTRITIONAL AND DIETARY SUPPLEMENTS……3087非无菌的营养与食品添加剂中的微生物分布<2040>DISINTEGRATION AND DISSOLUTION OF DIETARY SUPPLEMENTS (3089)食品添加剂的崩解与溶出<2091>WEIGHT V ARIATION OF DIETARY SUPPLEMENTS……3092食品添加剂的重量差异<2750>MANUFACTURING PRACTICES FOR DIETARY SUPPLEMENTS (3093)食品添加剂的生产操作。
微生物化验室的检查指南翻译稿-7页文档资料
微生物化验室的检查指南质检化验室的检查指南重点强调了有关化学分析方面的内容，对于微生物检查的内容涉及比较少，本文将作为化验室检查中对微生物检查方面的指南。
对于任何一次质检室的检查，我们都建议需要有熟悉这方面知识的分析学家或者微生物学家参与检查。
由于种种理由，我们发现一些外用制剂、滴鼻剂和吸入剂会产生微生物污染的问题。
但USP中有关微生物检测的章节<1111>对此方面只提到了“对于非无菌药品的微生物特性，应根据产品用途、产品特性以及对使用者产生的潜在危害进行评价”。
USP建议对某些种类的药品应进行微生物污染总数及微生物鉴定方面的检测。
例如，对植物药、动物药和某些矿物药应进行沙门氏菌的检测，对口服液应进行大肠杆菌的检测，对外用制剂应进行绿脓杆菌和金黄色葡萄球菌的检测，对供直肠、尿道或阴道用的制剂应进行酵母菌和霉菌的检测。
在药典的一些各论中也包括了特定的微生物限度要求。
作为药品微生物污染可接受水平和类型的总指南，FDA药物局的Dunnigan博士对微生物所产生的健康危害作了评论。
在1970年，他指出使用被革兰氏阴性菌污染的外用制剂会产生中度到重度的健康为害。
通过文献的报道及我们的调查发现，多种感染都与外用制剂的革兰氏阴性菌污染有关。
最典型的例子就是几年前在麻萨诸塞州的一家医院发生的洋葱假单胞菌污染聚维酮碘事件。
因此，每家公司都应该为他们的非无菌药品制定微生物限度标准。
USP微生物限度章节<61>也提供了检测一些微生物的方法，但并不包括所有的致病菌。
例如，在外用制剂或者滴鼻剂中如存在大量的洋葱假单胞菌会对人体产生危害，但是，在USP中却未能提供鉴别该微生物的方法。
有关这个问题的相应例子还有对间羟异丙肾上腺素吸入剂的召回事件。
USPXXII专论中规定对这种产品不需要进行微生物检测。
但FDA将被唐菖蒲假单胞菌污染的间羟异丙肾上腺素吸入剂作为I类召回。
不良反应评估指出这些污染菌对肺部感染的风险非常大，并且对患有慢性阻塞性呼吸道疾病、囊肿性纤维化和免疫功能低下的病人具有潜在的生命威胁。
微生物实验室良好规范 USP_1117
公用系统取样：如，水，无菌取样方式
微生物实验室布局与操作
环境监测取样
仪器设备的摆放与移除对无菌操作要求较低 设备仪器应同时与微生物培养基同时摆放
各类无菌检验：应在控制环境中进行
无菌制剂 原料药 发酵菌种 生物制品菌种
隔离器 <1208>
样品操作
样品操作
操作与储存条件：关键 微生物样品：水、环境监测样、生物负荷样 关键参数： 样品成分 包材成份 储存时间 储存温度 取样操作至检验开始的时间 严格控制 期间的储存条件 样品转移到较远处检测 运输条件确认：时间、温度等 USP <1231> 医药用水
检验时间大于72小时，按天表达
30-35℃, 3 – 5天 上午或下午开始培养，也应同样时间结束
人员培训
人员培训
微生物学教育背景 员工 主管 经理 职责明确 操作规程的培训 检验方法描述 文件编号便于检索化验员接受的培训内容 建立个人培训档案 确认后方能独立操作 培训记录时效性：现行版本与SOP
化验室记录的维护
化验室记录应包括的要素
日期 样品名称 化验员姓名 规程号 记录检验结果 偏差 参数记录
设备、微生物菌种、培养基批号
主管/复核人签名
化验室记录的维护
记录 关键设备，按期校验、有维保记录、使用日志 设备温度（水浴、培养箱、灭菌柜），可追溯 SOP SOP名称与版本号 方法描述应具体详细 书写修改： 单线划掉，签字、日期 原数据不得擦掉、或涂抹 检验结果 原始平板计数：便于复核 图表：装订应安全、数据可读；骑缝签字
首次结果无效
做出结论 进行复验（按复验SOP） 必要时重新取样
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人员培训
微生物检测美国药典
微生物检测非无菌供试品的微生物检测：微生物计数检测修改：生长促进实验，计数方法的适应性以及阴性对照概论在供试品存在的情况下发现微生物检验能力必须被确定。
如果检验过程中发生变更或者供试品变更，且这些变更可能影响检验结果，适应性必须被确认。
检验菌株的准备使用稳定的标准菌悬液或者按照下面所述备制。
使用菌种保存技术（种子批系统），以便用于接种的可萌发微生物从最初的主种子批开始不超过5代。
每种细菌和霉菌菌株的生长分别按照表一中的描述进行。
表一测试微生物的备制和使用使用pH7.0的缓冲氯化钠蛋白胨溶液或者pH7.2的磷酸盐缓冲溶液备制测试菌悬液；备制黑曲霉孢子悬浮液时，0.05%的聚山梨脂80可以被添加到缓冲液中。
测试菌悬液应在两小时内使用，或者在2-8℃的条件下24小时内使用。
也可以通过制备并稀释枯草芽胞杆菌营养细胞的新鲜悬液进行替代，制备稳定的胞子悬液，在接种测试中使用适当体积的胞子菌悬液，稳定的胞子悬液在2-8℃保存，保存期是经过验证的。
阴性对照为了确定检测条件，用选择好的稀释液代替测试备制来进行阴性对照。
必须没有微生物的生长。
当按照供试品检测中的描述进行检验时，也需要进行阴性对照。
如果阴性对照不合格需要进行调查。
培养基的生长促进检测每个批次的已经备制好的培养基以及通过脱水培养基或者描述的配料备制的每个批次的培养基。
在部分/盘大豆酪蛋白消化肉汤培养基以及大豆酪蛋白消化琼脂上接种少量（不超过100CFU）微生物，按表一中所示，每一种微生物应使用单独一部分/一盘培养基。
在沙氏葡萄糖琼脂上接种少量（不超过100CFU）微生物，按表一中所示，每一种微生物应使用单独的一盘培养基。
按照表一中所述的条件进行接种。
固体培养基的生长通过一个不大于2的系数的调节必须不能与标准接种体计算得到的数值有区别。
新鲜备制的接种体微生物的生长应与上一批通过检测的培养基的生长情形相同。
如果肉眼能清晰看到的微生物的生长与上一批通过检测的培养基的生长情形相同的话，液体培养基是适合的。
美国药典36版目录
固体的疏松密度和叩击密 度测定
色谱法 呈色与消色 溶解的完全性检查 总有机炭测定 水导电性测定 冻凝温度的测定 包转和贮藏要求 玻璃容器 塑料容器 附加包装构成 容器性能检测
用于无菌固体和液体溶剂 形式的单独单位容器和单 位剂量容器的重新分配
<691> 棉花
<695> <696> <698>
<55>
生物指示剂：耐受性能检 测
<61>
非无菌制品的微生物检 查：计数试验
<62>
非无菌制品的特定菌检查
<63>
支原体检测
<71>
无菌检测
生物学试验和检定
<81>
抗生素微生物检定
<85>
细菌内毒素检测
如大肠 杆菌、 金葡菌 、沙门 氏菌等
<87>
体外生物反应性试验
<88>
体内生物反应性试验
<90> <91> <92> <111> <115> <121> <123> <130> <151>
美国药典目录
检测和含量分析的一般要求
<1>
注射剂
<3>
局部和经皮的药物产品-产 品质量检测
<11>
USP对照标准品
检测和含量分析的仪器
<16>
自动分析方法
<17>
处方容器具标签
<21>
测温仪
USP351113中文版
自己翻译，非专业，有错误改正，勿拍砖〈1113〉细菌鉴定35（1）版167页，该章包括标题正在修订。
本章阐述微生物鉴定方法，方法的选用及验证。
〈1113〉微生物特性确定、鉴定及菌株分型前言在检测药品原辅料，生产用水，生产环境，中间体以及终端产品市的微生物时，需要对其特性进行确定。
这些包括适当的鉴定及菌株分型（标注，一些项目在章节末端提及）。
日常的微生物特性确定包括菌落形态、细胞形态（杆菌、球菌、细胞集合、产胞方式），革兰氏染色或者其他染色方法，和特征生化反应（氧化酶，过氧化氢酶，凝固酶反应）。
在非无菌药品生产过程和一些无菌制剂的生产环境中，微生物鉴定到这个水平足以达到风险评估的目的。
在某些情况下，可以用一些更确定的方法鉴定微生物种属。
除此之外，有些方法进行菌株鉴定对于确定微生物来源的研究很有用。
当微生物在以异常高频率或者数量超趋势出现在特殊种类的产品中时，微生物鉴定试验尤为频繁。
另外，微生物鉴定可用于无菌生产，当无菌实验中出现的阳性和从失败的无菌工艺模拟收回的污染样品如培养基灌装试验，都需要进行鉴定。
微生物鉴定系统是基于不同的分析方法，其限制可能源于固定的方法或者/和数据库的限制。
微生物鉴定是通过一个既定的标准菌株根据匹配特性（基因型和/或表型）确定其种属。
如果某种微生物在数据库中不存在，无法鉴定，制造商应当回顾所建立鉴定系统的数据库的宽度及适用性。
使用者应当考虑哪种系统是最符合实际使用要求的。
考虑到这些限制和鉴定所要达到的等级（属，种，株），使用者还要选择合适的方法用于日常药品的微生物鉴定。
在USP通用测试章特别提到的非无菌产品的微生物检验：指定微生物检查62，这一章表示用于鉴定实验确认的微生物需要通过选择培养基或者特征培养基培养并具有确定的形态特征。
同样，在USP通用测试章无菌试验71中规定：在从试验中分离出的微生物确定之后，此种（或这些种）微生物的生长可以毫不含糊地归咎于与物料和/或者进行无菌试验过程中所使用的方法，该实验无效。
USP微生物限度检查中文
USP微生物限度检查中文USP微生物限度检查中文61）微生物限度检测（MICROBIAL LIMIT TESTS）此章提供方法来检测可能存在的好氧微生物其他制药过程中可能出现的微生物的数量，包括原材料和成品中的。
如果经过验证确认可以得到相同或更好的检测结论，也允许采用自动化的检测方法。
在样品检测过程中须进行无菌操作。
若无特别说明，则“培养（incubate）”一词指在30—35℃的培养箱内培养24至48小时；“生长（growth）”一词用于专门的判定，说明“存在和可能存在活的微生物”。
准备实验 (Preparatory Testing)本章涉及实验结果的有效性取决于：提供的被检测样品本身在实验条件下，被充分证明不会抑制可能存在的微生物的生长。
因此，在准备样品时，需要正规的实验操作和符合要求的实验条件，接种稀释样品到含有以下（微生物）培养物的培养基：金黄色（奥里斯）葡萄球菌（Staphylococcus aureus）,大肠埃希氏菌（Escherichia coli）, 铜绿假单胞菌(Pseudomonas aeruginosa), 和沙门氏菌（Salmonella）。
方法如下：将用肉汤培养基培养24小时后的（微生物）不小于10-3稀释的微生物培养物，加1 ml（微生物）培养液到磷酸(盐)缓冲液(pH 7.2)，液体大豆酪蛋白消化物培养基（Fluid Soybean-Casein Digest Medium），或者液体乳糖培养基（Fluid Lactose Medium）。
相应培养基培养失败则需要采取以下方法更改检测程序：（1）增加稀释液体积，检测样品加入量仍维持不变；或者（2）中和一定数量的干扰因子；或者（3）结合（1）、（2）得出适当条件，使接种物得以生长。
以下是一些物质的成分和浓度，该物质及浓度可用于加入培养基、阻止物质发挥抑菌作用：大豆卵磷脂（soy lecithin, 0.5%）或者聚山梨醇酯20（polysorbate 20, 4.0%）。
USP40版1117 良好微生物实验室规范
Ljungqvist B, Reinmüller B. Interaction between air movements and the dispersion of contaminants: clean zones with unidir-ectional air flow. J Parenter Sci Technol. 1993; 47(2):60–69.Ljungqvist B, Reinmüller B. Airborne viable particles and total number of airborne particles: comparative studies of active air sampling. PDA J Sci Technol. 2000; 54:112–116.Maruyama M, Matsuoka T, Deguchi M, Akers J. The application of robotics to aseptic surface monitoring. Pharm Technol. 2007; 32(7):40–44.Process simulation testing for sterile bulk pharmaceutical chemicals. PDA Technical Report No. 28. J Parenter Sci Technol. 1998; 52 S3.Reinmüller B. Dispersion and risk assessment of airborne contaminants in pharmaceutical cleanrooms. Building Serv Eng Bull (Sweden). 2001; Bulletin No. 56.Stewart SL, Grinshpun SA, Willeke K, Terzieva S, Ulevicius V, Donnelly J. Effect of impact stress on microbial recovery on an agar surface. Appl Environ Micro. 1995; 61:1232–1239.Whyte W. Reduction of microbial dispersion by clothing. J Parenter Sci Technol. 1985; 39(1):51–60.á1117ñ MICROBIOLOGICAL BEST LABORATORY PRACTICESINTRODUCTIONGood laboratory practices in a microbiology laboratory consist of activities that depend on several principles: aseptic techni-que, control of media, control of test strains, operation and control of equipment, diligent recording and evaluation of data, and training of the laboratory staff. Because of the inherent risk of variability in microbiology data, reliability and reproducibili-ty are dependent on the use of accepted methods and adherence to good laboratory practices.MEDIA PREPARATION AND QUALITY CONTROLMedia PreparationCulture media are the basis for most microbiological tests. Safeguarding the quality of the media is therefore critical to the success of the microbiology laboratory. Media preparation, proper storage, and quality control testing can ensure a consistent supply of high-quality media.It is important to choose the correct media or components in making media based on the use of accepted sources or refer-ences for formulas. The manufacturer's formula and instructions for preparation routinely accompany dehydrated media and ready-made media. Because different media types may have different preparation requirements (e.g., heating, additives, and pH adjustment), it is important to follow these instructions to ensure preparation of acceptable media quality. A certificate of analysis describing expiration dating and recommended storage conditions accompanies ready-made media, as well as the quality control organisms used in growth-promotion and selectivity testing of that media.Water is the universal diluent for microbiological media. Purified Water is most often used for media preparation, but in cer-tain cases the use of deionized or distilled water may be appropriate. Water of lesser quality should not be used for microbio-logical media preparation. The volume of the water used should be recorded.Consistent preparation of media requires accurate weighing of dehydrated media or media constituents. A calibrated bal-ance with the appropriate weight range for the ingredients should be used (See Weighing on an Analytical Balance á1251ñ). Clean weighing containers and tools (such as spatulas) should be used to prevent foreign substances from entering the formu-lation. The weight of the components should be recorded.Dehydrated media should be thoroughly dissolved in water before dispensing and sterilization. If heating is necessary to help dissolve the media, care should be taken not to overheat media, because all culture media, to a greater or lesser extent, are heat-sensitive. Equipment used in the preparation of media should be appropriate to allow for controlled heating, constant agitation, and mixing of the media. Darkening of media (Maillard-type reaction or nonenzymatic browning) is a general indi-cation of overheating. When adding required supplements to media, adequate mixing of the medium after adding the supple-ment should be performed.Preparation of media in poorly cleaned glassware can allow inhibitory substances to enter the media. Inhibitory substances can come from detergent residue after cleaning glassware or from prior materials used in the glassware. Be sure that the clean-ing process removes debris and foreign matter, and that the detergent is thoroughly rinsed out with Purified Water. See Clean-ing Glass Apparatus á1051ñ for additional guidance.Sterilization of media should be performed within the parameters provided by the manufacturer or validated by the user. Commercially prepared media should provide documentation of the sterilization method used. Autoclaving by moist heat is the preferred sterilization technique, except in instances when boiling is required in order to avoid deterioration of heat-labilecomponents of the media. Sterilization by filtration may also be appropriate for some formulations.USP 40General Information / á1117ñ Microbiological Best Laboratory Practices1443The effects of the sterilization method and conditions on the media should be validated by sterility and growth-promotion testing of the media. In addition, if sterilized by moist heat, the autoclave cycle should be validated to ensure proper heat dis-tribution for selected loads and volumes. Typically, manufacturers recommend using an autoclave cycle of 121° for 15 minutes using a validated autoclave. These conditions apply to time at temperature of the media. As container size and the load config-uration of the autoclave will influence the rate of heating, longer cycles may be required for larger loads. However, the sterili-zation time will be dependent on the media volume and autoclave load. Sterilization cycles in which the autoclave is slow to come up to temperature may result in overheating of the media. Therefore, care must be taken to validate a sterilization cycle,balancing the need for sterile media against the tendency of the media to degrade under excessive heating. Storage of the media in the autoclave after the liquid cycle is completed is not recommended after cooling, as it may damage the media.Improper heating or sterilizing conditions—for commercially prepared or internally prepared media—may result in a difference in color change, loss of clarity, altered gel strength, or pH drift from the manufacturer's recommended range, as well as re-duced growth-promotion activity and/or selectivity.The pH of each batch of medium should be confirmed after it has cooled to room temperature (20°–25°) by aseptically withdrawing a sample for testing. Refrigerated purchased media should be allowed to warm up to ambient room temperature if it is to be checked for pH confirmation. A flat pH probe is recommended for agar surfaces, and an immersion probe is rec-ommended for liquids. See pH á791ñ for guidance with pH measurement and instrument calibration. The pH of media should be in a range of ±0.2 of the value indicated by the manufacturer, unless a wider range is acceptable by the validated method.Prepared media should be checked by appropriate inspection of plates and tubes for the following:•Cracked containers or lids•Unequal filling of containers•Dehydration resulting in cracks or dimpled surfaces on solid medium•Hemolysis•Excessive darkening or color change•Crystal formation from possible freezing•Excessive number of bubbles•Microbial contamination•Status of redox indicators (if appropriate)•Lot number and expiration date checked and recorded•Sterility of the media•Cleanliness of plates (lid should not stick to dish)Media StorageIt is prudent to consider how the manufacturer or supplier transports and stores media before distribution to the end user.Manufacturers of media should use transport and storage conditions that minimize the loss of moisture, control the tempera-ture, prevent microbial contamination, and provide mechanical protection to the prepared media.Media should be labeled properly with batch or lot numbers, preparation and expiration dates, and media identification.Media should be stored according to the manufacturer's instructions. Media prepared in house should be stored under valida-ted conditions. Do not store agar at or below 0°, as freezing could damage the gel structure. Protect stored media from expo-sure to light and excessive temperature. Before prolonged storage, agar plates should be placed into a sealed package or con-tainer to retard moisture loss.Remelting of an original container of solid media should be performed only once to avoid media whose quality is compro-mised by overheating or potential contamination. It is recommended that remelting be performed in a heated water bath or by using free-flowing steam. The use of microwave ovens and heating plates is common, but care should be taken to avoid damaging media by overheating and to avoid the potential injury to laboratory personnel from glass breakage and burns. The molten agar medium should be held in a monitored water bath at a temperature of 45° to 50° for not more than 8 hours.Caution should be taken when pouring the media from a container immersed in a water bath to prevent water from the bath commingling with the poured sterile media. Wiping the exterior of the container dry before pouring may be advisable.Disposal of used cultured media (as well as expired media) should follow local biological hazard safety procedures.Quality Control TestingAlthough growth media can be prepared in a laboratory from individual components, many laboratories, for ease of use, use dehydrated media or purchase commercially prepared media in plastic plates or glass containers. Manufacturers of media at-tempt to standardize raw materials from biological sources, but must constantly deal with unavoidable differences in raw ma-terials obtained from natural sources, and therefore, lot-to-lot variability of media must be considered. In addition, the per-formance of media prepared in a laboratory or by a manufacturer is highly dependent on preparation and storage conditions.Improper media preparation can cause unsatisfactory conditions for microbial growth or recovery and unreliable results.Therefore, quality control tests should be performed on all prepared media, including media associated with swabs or media in strips and other nontraditional formats. Tests routinely performed on in-house prepared media should include pH, growthpromotion, inhibition, and indicative properties (as appropriate), and periodic stability checks to confirm the expiration dating.1444 á1117ñ Microbiological Best Laboratory Practices / General Information USP 40When in-house prepared microbiological media are properly prepared and sterilized using a validated method, the growth-promotion testing may be limited to each incoming lot of dehydrated media, unless otherwise instructed by the relevant com-pendial method. If the media preparation procedure was not validated, then every batch of media should be subjected to growth-promotion testing. Test organisms may be selected from the appropriate compendial test chapter. In addition, micro-organisms used in growth-promotion testing may be based on the manufacturer's recommendation for a particular medium, or may include representative environmental isolates (but these latter are not to be construed as compendial requirements). Expiration dates on media should have supporting growth-promotion testing to indicate that the performance of the media still meets acceptance criteria up to and including the expiration date. The length of shelf life of a batch of media will depend on the stability of the ingredients and formulation under specified conditions, as well as the type of container and closure. When a batch of media does not meet the requirements of growth-promotion testing, an investigation should be initiated to identify the cause. This investigation should include a corrective action plan to prevent the recurrence of the problem. Any batch of media that fails growth-promotion testing is unsuitable for use. [NOTE—Failed growth-promotion test results may not be used to negate positive test results.]Some reagents are used for diagnostic purposes to help support identification of microbial organisms, e.g., Gram stain and oxidase test reagents. These may have attributes that can be quality control tested similar to microbiological media. Select the correct quality control standard microorganisms, following the manufacturer's instructions, and perform the testing before un-known sample diagnostic testing. All relevant diagnostic reagents should be subjected to incoming quality confirmation before use.Special care should be taken with media that is used in sterility tests (see Sterility Tests á71ñ for requirements) and in environ-mental monitoring studies. Media used for environmental monitoring of critical areas should preferably be double-wrapped and terminally sterilized. If terminal sterilization is not performed, media should be subjected to 100% pre-incubation and in-spection before use within a critical area. [N OTE—Growth-promotion testing for this media must be performed after the pre-incubation stage.] This will prevent extraneous contamination from being carried into controlled environments and will pre-vent false-positive results. A raised agar level for surface contact plates should be verified.MAINTENANCE OF MICROBIOLOGICAL CULTURESBiological specimens can be the most delicate standards to handle because their viability and characteristics are dependent on adequate handling and storage. Standardizing the handling and storage of cultures by the user laboratory should be done in a way that will minimize the opportunity for contamination or alteration of growth characteristics. The careful and consis-tent treatment of stock cultures is critically important to the consistency of microbiological test results. Cultures for use in com-pendial tests should be acquired from a national culture collection or a qualified secondary supplier. They can be acquired fro-zen, freeze-dried, on slants, or in ready-to-use forms. Confirmation of the purity of the culture and the identity of the culture should be performed before its use in quality control testing. Ready-to-use cultures should be subjected to incoming testing for purity and identity before use. The confirmation of identity for commonly used laboratory strains should ideally be done at the level of genus and species.Preparation and resuscitation of cultures should follow the instructions of the supplier or a validated, established method. The “Seed-Lot” technique is recommended for storage of stock cultures.The original sample from the national culture collection or a qualified secondary supplier is resuscitated and grown in an appropriate medium. Aliquots of this stock culture (the first transfer or passage) are suspended in a cryoprotective medium, transferred to vials, and frozen at –30° or below, until use. If stored at –70°, or in lyophilized form, strains may be kept indefi-nitely. These frozen stocks can then be used to inoculate monthly or weekly working cultures. Once opened, do not refreeze unused cell suspensions after culturing a working suspension. The unused portion should be discarded to minimize the risk of loss of viability and contamination of the stock.The number of transfers of working control cultures should be tracked to prevent excessive subculturing that increases the risk of phenotypic alteration or mutation. The number of transfers allowable for specific compendial tests may be specified in that test. One passage is defined as the transfer of organisms from a viable culture to a fresh medium with growth of the mi-croorganisms. Any form of subculturing is considered to be a transfer/passage.LABORATORY EQUIPMENTMost equipment (incubators, water baths, and autoclaves) is subject to standard validation practices of incoming qualifica-tion, operational qualification, and performance qualification. Additionally, periodic calibration (generally annually) is com-monly required. New equipment, critical to the operation of the laboratory, should be qualified according to a protocol ap-proved by the quality assurance unit (QAU). In addition, regular cleaning and sanitization of equipment such as incubators, refrigerators, and water baths should be performed to minimize the potential for contamination in the laboratory. Door seals of incubators and refrigerators should be cleaned and checked for state of repair.Instruments (pH meters and spectrophotometers) used in a microbiology laboratory should be calibrated on a regular schedule and tested to verify performance on a routine basis. The frequency of calibration and performance verification willvary based on the type of instrument and the importance of that equipment to the generation of data in the laboratory. USP 40General Information / á1117ñ Microbiological Best Laboratory Practices1445Equipment that is difficult to sanitize (such as refrigerators and incubators) should be dedicated to aseptic operations (such as storage of media for testing and incubation of sterility test samples) and live culture operations to minimize the potential for inadvertent contamination of the tests.Autoclaves are central to the operation of the laboratory and must have proper validation in place to demonstrate adequate sterilization for a variety of operations. Autoclave resources must be available (and validated) to sterilize waste media (if per-formed in that laboratory) as well as the media prepared in that laboratory. The choice of one or several autoclaves is not driv-en by a need to separate aseptic and live operations (everything in the properly maintained autoclave is sterile after the cycle)but rather driven by resource considerations (see below).LABORATORY LAYOUT AND OPERATIONSLaboratory layout and design should carefully consider the requirements of good microbiological practices and laboratory safety. It is essential that cross-contamination of microbial cultures be minimized to the greatest extent possible, and it is also important that microbiological samples be handled in an environment that makes contamination highly unlikely.In general, a laboratory should be divided into clean or aseptic areas and live culture areas. Areas in which environmental or sterile product samples are handled and incubated should be maintained completely free of live cultures, if possible. If com-plete separation of live and clean culture zones cannot be accomplished, then other barriers and aseptic practices should be employed to reduce the likelihood of accidental contamination. These barriers include protective clothing, sanitization and dis-infection procedures, and biological safety cabinets designated for clean or aseptic operations only. Procedures for handling spills or mishaps with live cultures should be in place, and all relevant technical personnel should be trained regarding thesemethods.Some samples will demonstrate microbial growth and require further laboratory analysis to identify the contaminants. Whengrowth is detected, the sample should be taken from the clean section of the laboratory to the live culture section without undue delay. Subculturing, staining, microbial identification, or other investigational operations should be undertaken in thelive culture section of the laboratory. If possible, any sample found to contain growing colonies should not be opened in theclean zone of the laboratory. Careful segregation of contaminated samples and materials will reduce false-positive results.Staff engaged in sampling activities should not enter or work in the live culture handling section of a laboratory unless spe-cial precautions are taken, including wearing protective clothing and gloves and careful sanitizing of hands upon exiting. Ideal-ly, staff assigned to sampling activities, particularly those in support of aseptic processing, should not work in the vicinity of live culture laboratory operations.It is important to consider that microbial contamination of samples, which leads to false-positive results, is always possible unless careful aseptic precautions are taken. Facilities should be designed so that raw material and excipient sampling can be done under controlled conditions, including proper gowning and sterilized sampling equipment. It may not always be possible to sample utility systems, such as water systems, under full aseptic conditions; however, it should be noted that when samples are not taken aseptically, their reliability is inevitably compromised.Environmental sampling methods should require minimal aseptic handling in loading and unloading sampling instruments.Whenever possible, sampling equipment should be loaded with its microbiological recovery media in the environment that is to be sampled.All testing in laboratories used for critical testing procedures, such as sterility testing of final dosage forms, bulk product,seed cultures for biological production, or cell cultures used in biological production, should be performed under controlled conditions. Isolator technology is also appropriate for critical, sterile microbiological testing. Isolators have been shown to have lower levels of environmental contamination than manned clean rooms, and therefore, are generally less likely to produce false-positive results. Proper validation of isolators is critical both to ensure environmental integrity and to prevent the possibili-ty of false-negative results as a result of chemical disinfection of materials brought into or used within isolators (see Sterility Testing—Validation of Isolator Systems á1208ñ).SAMPLE HANDLINGViable microorganisms in most microbiology samples, particularly water, environmental monitoring and bioburden samples,are sensitive to handling and storage conditions. Critical parameters in these conditions include product (or sample) composi-tion, container composition, time of storage, and temperature of storage. Therefore, it is important to minimize the amount of time between the sampling event and the initiation of testing and to control, as much as possible, the conditions of storage. If the sample is to be transported to a distant location for testing, then the conditions of transport (time, temperature, etc.)should be qualified as suitable for that test and sample. Guidance for water testing in this regard can be found in Water for Pharmaceutical Purposes á1231ñ. Product mixing before sampling may need to be evaluated and applied in order to ensure mi-crobial dispersement and representation in the sample aliquot.All microbiological samples should be taken using aseptic techniques, including those taken in support of nonsterile prod-ucts. If possible, all microbiological samples should be taken under full aseptic conditions in specialized sampling areas. The areas should be as close to the point of use as possible to minimize contamination during transit.Samples submitted to the microbiology laboratory should be accompanied by documentation detailing source of the sam-ple, date the sample was taken, date of sample submission, person or department responsible for the submission, and any1446 á1117ñ Microbiological Best Laboratory Practices / General Information USP 40potentially hazardous materials associated with the sample. The testing department should acknowledge receipt of the sample and reconcile the identity and number of samples as part of this sample documentation.MICROBIOLOGICAL MEDIA INCUBATION TIMESIncubation times for microbiological tests of less than 3 days' duration should be expressed in hours: e.g., “Incubate at 30°to 35° for 18 to 72 hours”. Tests longer than 72 hours' duration should be expressed in days: e.g., “Incubate at 30° to 35° for 3 to 5 days”. For incubation times expressed in hours, incubate for the minimum specified time, and exercise good microbio-logical judgment when exceeding the incubation time. For incubation times expressed in days, incubations started in the morning or afternoon should generally be concluded at that same time of day.TRAINING OF PERSONNELEach person engaged in each phase of pharmaceutical manufacture should have the education, training, and experience to do his or her job. The demands of microbiological testing require that the core educational background of the staff, supervi-sors, and managers be in microbiology or a closely related biological science. They should be assigned responsibilities in keep-ing with their level of skill and experience.A coherent system of standard operating procedures (SOPs) is necessary to run the microbiology laboratory. These proce-dures serve two purposes in a training program. Firstly, these SOPs describe the methodology that the microbiologist will fol-low to obtain accurate and reproducible results, and so serve as the basis for training. Secondly, by tracking the procedures in which a particular microbiologist has demonstrated proficiency, the procedure number or title also serves to identify what training the microbiologist has received specific to his or her job function.Training curricula should be established for each laboratory staff member specific to his or her job function. He or she should not independently conduct a microbial test until qualified to run the test. Training records should be current, documenting the microbiologist's training in the current revision to the particular SOP.Periodic performance assessment is a wise investment in data quality. This performance testing should provide evidence of competency in core activities of the microbiology laboratory such as hygiene, plating, aseptic technique, documentation, and others as suggested by the microbiologist's job function.Microbiologists with supervisory or managerial responsibilities should have appropriate education and in-house training in supervisory skills, laboratory safety, scheduling, budgeting, investigational skills, technical report writing, relevant SOPs, and other critical aspects of the company's processes as suggested in their role of directing a laboratory function.Competency may be demonstrated by specific course work, relevant experience, and routinely engaging in relevant con-tinuing education. Achieving certification through an accredited body is also a desirable credential. Further, it is expected that laboratory supervisors and managers have a demonstrated level of competence in microbiology at least as high as those they supervise. Expertise in microbiology can be achieved by a variety of routes in addition to academic course work and accredita-tion. Each company is expected to evaluate the credentials of those responsible for designing, implementing, and operating the microbiology program. Companies can thus ensure that those responsible for the program understand the basic principles of microbiology, can interpret guidelines and regulations based on good science, and have access to individuals with theoreti-cal and practical knowledge in microbiology to provide assistance in areas in which the persons responsible for the program may not have adequate knowledge and understanding. It should be noted that microbiology is a scientifically based discipline that deals with biological principles substantially different from those of analytical chemistry and engineering disciplines. Many times it is difficult for individuals without specific microbiological training to make the transition.LABORATORY RESOURCESThe laboratory management is responsible for ensuring that the laboratory has sufficient resources to meet the existing test-ing requirements. This requires some proficiency in budget management and in determining appropriate measures of labora-tory performance. A measure of laboratory performance is the number of investigations performed on tests conducted by the laboratory, but this measure alone is not sufficient. In addition to tracking investigations, the period of time between sample submission and initiation of testing should be tracked, as well as the period of time between end of test and report release (or test closure). Significant delays in these measures are also indications of an under-resourced laboratory staff.The laboratory management should have sufficient budget to meet testing requirements. Particular measures of budgetary requirements will be specific to the given laboratory, but budgetary considerations related directly to the need of the laborato-ry for sufficient resources must be addressed to ensure reliable testing results.DOCUMENTATIONDocumentation should be sufficient to demonstrate that the testing was performed in a laboratory and by methods that were under control. This includes, but is not limited to, documentation of the following:•Microbiologist training and verification of proficiencyUSP 40General Information / á1117ñ Microbiological Best Laboratory Practices1447。
（最新）美国药典微生物检测精要
美国药典微生物检测精要培训小结一微生物实验室规范重要的控制点：无菌技术、培养基控制、菌种控制、设备操作与控制、数据记录与实验室布局和运作、员工培训等。
1无菌技术：防止和保持无菌物品及无菌区域不被污染的操作方法和管理方法。
可通过无菌环境、灭菌、消毒、卫生要求、无菌操作技术等手段来实现。
2培养基：是微生物实验工作的核心。
包括培养基制备、培养基储存、培养基质量控制。
2.1培养基制备2.1.1根据既定的目的选用正确的培养基2.1.2参考生产商的ＣＯＡ和说明书：关于配制、储存和菌种选用等。
2.1.3所用的水应为去离子水或蒸馏水，应记录用量。
2.1.4成分的称量应用校准过的天平，根据称量量的不同选择合适的天平。
使用清洁的容器和工具（<1051>玻璃器皿清洁）以防外来污染和抑制剂。
应记录称量量。
2.1.5加热帮助溶解时应防止过度加热，通常培养基颜色变深说明加热过度了。
2.1.6灭菌条件：●灭菌参数应验证，验证应同时考虑无菌和培养基生长能力●采用高压蒸汽灭菌或过滤灭菌。
湿热灭菌应考虑装载分布，加热速度过慢会使培养基过度加热，通常为121℃15min，此时间是指培养基温度达到121℃后15min。
●保证最低SAL，在无菌和过度加热之间平衡。
●与污染物分开灭菌●消毒结束后，不应在灭菌柜中储存培养基，过度加热会影响培养基颜色、澄清、pH和凝固能力。
2.2培养基储存2.2.1制备培养基的储存●琼脂培养基易受冷冻影响，低于0℃会破坏凝胶结构。
●应避光避热，密封防止水分蒸发（建议使用螺旋盖的瓶子密封，保存时间长）。
2.2.2培养基融化●不能超过一次，以防过度加热或潜在污染。
●融化可采用水浴、流通蒸汽或微波炉加热。
微波炉加热宜采用少量多次，以防过度。
●培养基融化后在40-50℃水浴中储存不应超过8小时，浇制平皿前应擦干以防污染。
2.2.3培养基标示●培养基名称●批号、制备批号●制备日期、有效期2.2.4有效期确定●根据成分、配方、容器、储存条件等确定●有生长能力试验数据支持2.2.5应在验证过的条件下储存2.2.6重要区域环控用培养基必须●双层包装●最终灭菌，否则进行全数培养及用前检查2.2.7根据当地生物危险品安全程序处置过期培养基2.3培养基质控2.3.1灭菌后培养基检查2.3.1.1生长能力●每批制备的培养基均需进行●测试菌种根据药典，供应商说明书及环境中分离得到●测试不合格应进行调查，如无原因，或无有效纠正措施，不得使用该批号2.3.1.2无菌2.3.1.3pH2.3.1.4容器/平皿的完整性2.3.1.5抑制或指示能力2.3.2定期稳定性检查以确认储存有效期3菌种保藏3.1建立标准程序处理、保存菌种，防止污染和特性变化3.2使用前确认菌种身份和纯度3.3根据生产商说明书复苏菌种，使用接种技术3.4-70℃以下或冷冻干燥保存储备菌种，可延长保存周期3.5开启后不要重新冷冻菌种，剩余部分应弃置3.6传代次数（每接种一次即为一代）不超过5次3.7保藏时间根据具体保藏条件确定4设备维护4.1定期校准、保养4.2定期性能检查4.3定期清洁、消毒5实验室布局和运作5.1防止交叉污染5.2活动分区：无菌、环控样品的处理和培养应在无菌环境中进行5.3当发现微生物生长，后续的工作应转移至“阳性”区5.4环控设备应放置在待取样区域环境中并小心操作5.5应在受控条件下小心取样6样品处理6.1大部分样品、水或环控样品中的微生物对操作、储存环境敏感6.2减少样品，取样与测试间的时间间隔6.3长时间的转移需确认转移条件对测试及样品的影响6.4在无菌条件下，使用无菌技术取样，即使是非无菌样品6.5取样记录：样品信息、取样日期、送样日期、人员/部门、实验室样品接受及确认7培养基培养时间7.1短于3天，用小时表示7.2长于3天，用天表示（上午、下午，相同时间段）8人员培训8.1每个岗位应有相应培训要求，进行上岗前资质确认9实验室文件9.1微生物人员培训和能力确认（上岗资质）9.2设备验证、校准和维护9.3测试中的设备性能（如温度记录）9.4培养基制备、无菌检查、生长能力测试、选择性测试9.5培养基清单与控制9.6根据程序（ＳＯＰ）进行测试的重要数据9.7数据和计算的确认9.8经ＱＡ人员或相应领导审核过的报告9.9超标结果调查10实验室结果维护10.1检测报告至少应包括：●日期●测试样品●微生物检验人员名字●程序编号●测试结果●偏差（如有）●测试参数（设备、菌种、培养基）●管理人员审核签名10.2数据纠错：错误的地方划一横线，签名和日期，必要时说明原因10.3测试结果●应包含平皿计数结果（如有）●数据分析方法应罗列在相关ＳＯＰ中●对粘贴的图表、数据骑缝签名10.4存档：记录应存档并防止丢失，应有记录留存计划11测试结果解释11.1微生物数据有时不容易解释●与人类相关的微生物群落被广泛的用于许多测试●人员污染是始终被关心的问题●样品或环境中的微生物并非均匀分布●微生物检测可变范围大：可能在+/-0.5log10单位左右11.2不符合的结果可能是由2个原因造成的：实验室错误或产品不符合。
美国药典34版71无菌检测中文
71 STERILITY TESTS 无菌检测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 notharmonized are marked with symbols to specify this fact.此通则的各部分已经与欧洲药典和/或日本药典的对应部分做了协调。
不一致的部分用符号来标明。
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.这些药典规定程序自身的设计不能确保一批产品无菌或已经灭菌。
这主要是通过灭菌工艺或者无菌操作程序的验证来完成。
The test is applied to substance, preparations, or articles which, according to the Pharmacopeia, are required to be sterile. However, a satisfactory result only indicates that no contaminating microorganism has been found in the sample examined under the conditions of the test.检测应用于根据药典要求无菌的如物质、药剂或商品。
USP微生物检测精要
口服固体 口服液体 直肠制剂
1000 100 1000
100 10 100
大肠 大肠 —
局部及鼻腔制剂 100
阴道制剂 吸入剂
10 10
10
金黄 铜绿
金黄 铜绿 白念 金黄 铜绿 耐胆盐G-
—产品用途（眼，鼻，等） —产品特点：支持生长吗？或有防腐性 —应用的方法 —药物接受者：对婴儿、幼儿及衰弱患者的风险是不 同的 —存在疾病、伤口、器官损伤
相应因素的风险评估由经授权的有资质 的人员进行。
药物制剂和药用物质的接受标准
适当的书面程序必须建立并遵循以防止 非无菌药品中的有害微生物。
不仅仅是不存在USP规定的微生物
环境的微生物控制 生产过程的微生物控制 成品的微生物控制 人员的微生物控制 所有微生物控制的整合
USP概述
原料和组分的微生物控制的USP观点及 文件
<61>非无菌产品的微生物检查：微生物计数测 试 <62>非无菌产品的微生物检查：控制菌检查 <1111>非无菌药品的微生物评估
USP概述
环境微生物控制的USP观点及文件
微生物实验室规范
样品处理
—大部分样品，水或环控样品中的微生物对操作、储 存环境敏感。 产品处方、容器材质、储存时间/温度 —减少样品，取样与测试间的间隔时间，并控制储存 条件。 —远距离的转移需确认转移条件对测试及样品的影响。 —可能的话，在无菌条件下，使用无菌技术取样，即 使是非无菌样品。 —取样记录 样品信息、取样日期、送样日期、人员/部门，实 验室样品，接受及确认
药物制剂和药用物质的接受标准
微生物计数检查法USP61中英对照版
非无菌产品微生物学检查：微生物计数检查法USP61中英对照版<61> MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: MICROBIAL ENUMENRATION TESTS非无菌产品微生物学检查：微生物计数检查法INTRODUCTION 导言The tests described hereafter will allow quantitative enumeration of mesophilic bacteria and fungi that may grow under aerobic conditions.以下所描述的这些检测将使得对在有氧的条件下生长的嗜温性细菌和真菌进行定量计数成为可能。
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.这些检测主要设计用于测定一种物质或制备品是否符合已确立的微生物质量标准。
当用于此类目的时，需遵照以下所给的说明，包括待取样品的数量，并且按照下面所述解释结果。
The methods are not applicable to products containing viable microorganisms as active ingredients.这些方法不适用于以活菌作为活性成分的产品。
USP 35 1113中文版
自己翻译，非专业，有错误改正，勿拍砖〈1113〉细菌鉴定35（1）版167页，该章包括标题正在修订。
本章阐述微生物鉴定方法，方法的选用及验证。
〈1113〉微生物特性确定、鉴定及菌株分型前言在检测药品原辅料，生产用水，生产环境，中间体以及终端产品市的微生物时，需要对其特性进行确定。
这些包括适当的鉴定及菌株分型（标注，一些项目在章节末端提及）。
日常的微生物特性确定包括菌落形态、细胞形态（杆菌、球菌、细胞集合、产胞方式），革兰氏染色或者其他染色方法，和特征生化反应（氧化酶，过氧化氢酶，凝固酶反应）。
在非无菌药品生产过程和一些无菌制剂的生产环境中，微生物鉴定到这个水平足以达到风险评估的目的。
在某些情况下，可以用一些更确定的方法鉴定微生物种属。
除此之外，有些方法进行菌株鉴定对于确定微生物来源的研究很有用。
当微生物在以异常高频率或者数量超趋势出现在特殊种类的产品中时，微生物鉴定试验尤为频繁。
另外，微生物鉴定可用于无菌生产，当无菌实验中出现的阳性和从失败的无菌工艺模拟收回的污染样品如培养基灌装试验，都需要进行鉴定。
微生物鉴定系统是基于不同的分析方法，其限制可能源于固定的方法或者/和数据库的限制。
微生物鉴定是通过一个既定的标准菌株根据匹配特性（基因型和/或表型）确定其种属。
如果某种微生物在数据库中不存在，无法鉴定，制造商应当回顾所建立鉴定系统的数据库的宽度及适用性。
使用者应当考虑哪种系统是最符合实际使用要求的。
考虑到这些限制和鉴定所要达到的等级（属，种，株），使用者还要选择合适的方法用于日常药品的微生物鉴定。
在USP通用测试章特别提到的非无菌产品的微生物检验：指定微生物检查62，这一章表示用于鉴定实验确认的微生物需要通过选择培养基或者特征培养基培养并具有确定的形态特征。
同样，在USP通用测试章无菌试验71中规定：在从试验中分离出的微生物确定之后，此种（或这些种）微生物的生长可以毫不含糊地归咎于与物料和/或者进行无菌试验过程中所使用的方法，该实验无效。
USP35-1116中文
《1116》无菌工艺环境的微生物控制和监测在药品行业，微生物控制环境有多种用途。
本章节提供了关于通过无菌工艺控制微生物污染的环境的信息和建议。
在这些环境中生产的产品包括无菌制剂、无菌原料药、无菌中间体、辅料和部分药用器械。
从患者风险的角度来说，无菌工艺环境比其他生产操作的控制环境要重要的多，例如设备和备件的准备、非无菌产品的生物负荷限度控制和最终灭菌产品工艺。
本章节中的无菌工艺环境有人和无人操作应区别对待。
一个高级无菌工艺是不需要和禁止穿常规洁净服的操作人员直接接触敞口容器或暴露的产品接触表面的。
（注意：本章所用的术语在章节最后的目录里有解释。
）本章节给出了指导意见和洁净室、限制进入隔离系统（RABS）和隔离器等无菌工艺的微生物评价的监控参数，其他不要求在无菌条件下生产的无菌产品的生产环境的污染控制要求可以根据ISO环境分类执行。
非无菌产品的生产环境要有不同的微生物控制等级。
大部分标有无菌的产品是在无菌工艺条件下生产的而不是最终灭菌。
因为无菌工艺不仅生产过程中将微生物从生产用蒸汽中排除，并在空的容器进入时也阻止微生物的进入，还将生产环境的生物负荷作为一个控制微生物污染的重要参数。
无菌和灭菌并不是一个同义词。
灭菌指的是将全部的微生物或组织进行杀灭，有复活的可能性。
从纯粹的微生物意义上说，一个无菌工艺可以通过排除微生物的方式防止污染。
在现代无菌药品制造业，无菌指的是在一个设计好的受控环境中处理已灭菌的物料使微生物污染处于风险最小的已知水平。
在有人进行操作的任何环境中，微生物污染在一定程度上是不可避免的。
如果有人员存在的话，即使最谨慎的洁净室环境设计和操作也不能避免微生物的掉落。
因此在所有无菌生产工艺操作过程的所有场所达到零污染在技术上是不可能的并且也是不现实的。
去证明一个无菌生产工艺的环境和在这个环境中产品接触的表面是已消毒的是没有意义的。
应当根据风险评估确定监测点。
虽然生产者会经常对环境监测结果进行回顾以确保生产操作在受控的验证状态，但是监测结果不能证明无菌或非无菌。
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USP36 1117 优良微生物检测规范（中英文1/ 2）2013-08-09 15:30:46| 分类：USP|举报|字号订阅1117 MICROBIOLOGICAL BEST LABORATORY PRACTICES 优良微生物检测规范INTRODUCTION 介绍Good laboratory practices in a microbiology laboratory consist of activities that depend on several principles: aseptic technique, control of media, control of test strains, operation and control of equipment, diligent recording and evaluation of data, and training of the laboratory staff. Because of the inherent risk of variability in microbiology data, reliability and reproducibility are dependent on the use of accepted methods and adherence to good laboratory practices.优良微生物检测规范由一些活动组成，这些活动依赖于几个基本要素：无菌技术、培养基控制、检测用菌株控制、设备操作和控制、完善的记录和数据评估、化验室员工的培训。
由于微生物数据具有天生的不确定性，数据的可靠性和重复性取决于是否使用被接受的方法，以及是否严格遵守化验室规范。
MEDIA PREPARATION AND QUALITY CONTROL 培养基制备和质量控制Media Preparation 培养基制备Culture media are the basis for most microbiological tests. Safeguarding the quality of the media is therefore critical to the success of the microbiology laboratory. Media preparation, proper storage, and quality control testing can ensure a consistent supply of high-quality media.培养基是大多数微生物测试的基础。
保证培养基的质量因而成为微生物实验室成功的关键。
培养基的制备、合适的存贮和质量控制检测可以保证持续高质量培养基供应。
It is important to choose the correct media or components in making media based on the use of accepted sources or references for formulas. The manufacturer's formula and instructions for preparation routinely accompany dehydrated media andready-made media. Because different media types may have different preparation requirements (e.g., heating, additives, and pH adjustment), it is important to follow these instructions to ensure preparation of acceptable media quality. A certificate of analysis describing expiration dating and recommended storage conditions accompanies ready-made media, as well as the quality control organisms used in growth-promotion and selectivity testing of that media.在制备培养基过程中使用已接受的来源或配方标准，选择正确的培养基或成份是非常重要的。
一般生产商在供应培养基干粉和配制好的培养基时，其配方和配制指示都会随货发送。
由于不同的培养基类型可能有不同的配制要求（例如，加热、添加剂，和pH值调节），重要的一点是需要遵守其提供的配制指示以保证配制出的培养基质量。
如果是已制备好的培养基碟/瓶，随货会收到指明有效期和推荐的存贮条件的分析报告，以及促生产试验用微生物种类，和该培养基的选择性试验用微生物种类。
Water is the universal diluent for microbiological media. Purified Water is most often used for media preparation, but in certain cases the use of deionized or distilled water may be appropriate. Water of lesser quality should not be used for microbiological media preparation. The volume of the water used should be recorded.水是通用的微生物培养基稀释剂。
纯化水常用于培养基制备，但在某些情况下，也可以使用去离子水或蒸馏水。
更低品质的水不应用于微生物培养基制备。
水的用量应记录。
Consistent preparation of media requires accurate weighing of dehydrated media or media constituents. A calibrated balance with the appropriate weight range for the ingredients should be used (See Weighing on an Analytical Balance1251). Clean weighing containers and tools (such as spatulas) should be used to prevent foreign substances from entering the formulation. The weight of the components should be recorded.要保持培养基配制的一致性，需要对培养基干粉或培养基组份进行准确称量。
应使用在所需称量范围内经过校正的天平（见1251分析天平称量）。
应对称重容器和工具（例如料勺）进行清洁以保证无异物进入所配物料。
各成份的重量应进行记录。
Dehydrated media should be thoroughly dissolved in water before dispensing and sterilization. If heating is necessary to help dissolve the media, care should be taken not to overheat media, because all culture media, to a greater or lesser extent, areheat-sensitive. Equipment used in the preparation of media should be appropriate to allow for controlled heating, constant agitation, and mixing of the media.培养基干粉应在水中完全溶解，然后进行配制和灭菌。
如果需要加热助溶，要注意不能过热，因为所有的培养基，或多或少，都是对热敏感的。
用于培养基配制的设备应适当，以便控制加热、持续搅拌和培养基混合。
Darkening of media (Maillard-type reaction or nonenzymatic browning) is a general indication of overheating. When adding required supplements to media, adequate mixing of the medium after adding the supplement should be performed.培养基变黑（美拉德类型反应或非酶褐变）一般说明过热。
在向培养基中加入所需要的补充成分时，在加入后需要进行充分搅拌混合。
Preparation of media in poorly cleaned glassware can allow inhibitory substances to enter the media.在清洁不彻底的玻璃器皿中配制培养基会使得抑制性物质带入培养基。
Inhibitory substances can come from detergent residue after cleaning glassware or from prior materials used in the glassware. Be sure that the cleaning process removes debris and foreign matter, and that the detergent is thoroughly rinsed out with Purified Water. See Cleaning Glass Apparatus1051for additional guidance.抑制性物质可能来自于玻璃器皿中的清洁剂残留，或来自于玻璃器皿中上次所盛装的物料。
要保证清洗程序可以去除残渣和外来物质，并且清洁剂可以被纯化水彻底冲洗掉。
参见1051玻璃容器的清洁。
Sterilization of media should be performed within the parameters provided by the manufacturer or validated by the user. Commercially prepared media should provide documentation of the sterilization method used.培养基灭菌应在生产商提供的参数范围，或用户验证的参数范围内实施。