美国FDA药品质量控制微生物实验室检查指南1993年

美国FDA高纯水系统检查指南注释：这份文件是检查员和其他FDA人员的参考资料。

这份文件不约束FDA,不授予任何人任何权力、特权、利益或豁免权。

这份指南主要从微生物方面讨论对用于药物制剂和原料药生产的高纯水系统的检查及评价。

它也涉及了对不同类型系统的设计及一些与这些系统相关问题的检查。

如同其它指南一样，它并不是包含一切的，但是提供了对高纯水系统的检查和评价的背景及指导。

微生物实验室的药物质量控制检查指南（1993，5月）提供了另外的指导。

1、系统设计系统设计的一个基本考虑因素是所生产产品的种类。

对于注射剂关注的是热原，这就需要使用注射用水。

注射用水适用于产品配制，以及成分和生产设备的最终清洗。

蒸馏和反渗透膜过滤是USP中列出的生产注射用水的唯一可接受方法。

但是，在原料药和生物技术及一些国外公司中，超滤由于可以使内毒素量减到最少而用于那些针用原料药中。

对于一些眼科药物如眼用冲洗液和一些吸入产品如吸入的无菌水是有热原规定的，我们希望它们的配制使用注射用水。

但是对于大多数吸入剂和眼科用药，它们的配制中使用纯化水。

纯化水也适用于外用药、化妆品和口服制剂。

设计的另一个考虑因素是系统温度。

我们发现热（65-80℃）系统能自身消毒。

虽然公司采用其它系统的花费可能较低，但维护、检测和潜在问题的花费比能源节省的花费要大得多。

系统是否循环或单向也是设计中的一个重要考虑因素。

明显地，持续流动的水受高水平污染的可能性较低。

一个单向水系统基本上是一个死角。

最后，可能最重要的考虑因素是风险评估或要求的质量水平。

同时也应该认识到不同的产品要求不同质量的水。

注射剂要求无内毒素的高纯水。

外用和口服制剂要求较低纯度的水，并无内毒素要求。

即使对于外用和口服制剂也有许多因素决定了不同质量的水。

比如，抗酸剂中的保护剂并不是总是有效的，因此必须制定严格的微生物限度。

质量管理部门应该评估使用他们系统的水生产的每一个产品，并确定在微生物最敏感的产品基础上建立的微生物纠偏限度。

GUIDE TO INSPECTIONS OF MICROBIOLOGICAL PHARMACEUTICAL QUALITY CONTROL LABORATORIESNote: This document is reference material for investigators and other FDApersonnel. The document does not bind FDA, and does no confer any rights,privileges, benefits, or immunities for or on any person(s).I. INTRODUCTIONThe Guide to the Inspection of Pharmaceutical Quality Control Laboratoriesprovided very limited guidance on the matter of inspection ofmicrobiological laboratories. While that guide addresses many of the issuesassociated with the chemical aspect of laboratory analysis ofpharmaceuticals, this document will serve as a guide to the inspection ofthe microbiology analytical process. As with any laboratory inspection, itis recommended that an analyst (microbiologist) who is familiar with thetests being inspected participate in these inspections.II. MICROBIOLOGICAL TESTING OF NON-STERILE PRODUCTSFor a variety of reasons, we have seen a number of problems associated withthe microbiological contamination of topical drug products, nasal solutionsand inhalation products. The USP Microbiological Attributes Chapter <1111>provides little specific guidance other than "The significance ofmicroorganisms in non-sterile pharmaceutical products should be evaluated interms of the use of the product, the nature of the product, and thepotential hazard to the user." The USP recommends that certain categories beroutinely tested for total counts and specified indicator microbialcontaminants. For example natural plant, animal and some mineral productsfor Salmonella, oral liquids for E. Coli, topicals for P. aeruginosa and S.Aureus, and articles intended for rectal, urethral, or vaginaladministration for yeasts and molds. A number of specific monographs alsoinclude definitive microbial limits.As a general guide for acceptable levels and types of microbiologicalcontamination in products, Dr. Dunnigan of the Bureau of Medicine of the FDAcommented on the health hazard. In 1970, he said that topical preparationscontaminated with gram negative organisms are a probable moderate to serioushealth hazard. Through the literature and through our investigations, it hasbeen shown that a variety of infections have been traced to the gramnegative contamination of topical products. The classical example being thePseudomonas cepacia contamination of Povidone Iodine products reported by ahospital in Massachusetts several years ago.Therefore, each company is expected to develop microbial specifications fortheir non-sterile products. Likewise, the USP Microbial Limits Chapter <61>provides methodology for selected indicator organisms, but not allobjectionable organisms. For example, it is widely recognized thatPseudomonas cepacia is objectionable if found in a topical product or nasal solution in high numbers; yet, there are no test methods provided in the USP that will enable the identification of the presence of this microorganism.A relevant example of this problem is the recall of Metaproterenol Sulfate Inhalation Solution. The USP XXII monograph requires no microbial testingfor this product. The agency classified this as a Class I recall because the product was contaminated with Pseudomonas gladioli/cepacia. The healthhazard evaluation commented that the risk of pulmonary infection isespecially serious and potentially life-threatening to patients with chronic obstructive airway disease, cystic fibrosis, and immuno-compromisedpatients. Additionally, these organisms would not have been identified bytesting procedures delineated in the general Microbial Limits section of the Compendia.The USP currently provides for retests in the Microbial Limits section <61> however there is a current proposal to remove the retest provision. As with any other test, the results of initial test should be reviewed andinvestigated. Microbiological contamination is not evenly dispersedthroughout a lot or sample of product and finding a contaminant in onesample and not in another does not discount the findings of the initialsample results. Retest results should be reviewed and evaluated, andparticular emphasis should be placed on the logic and rationale forconducting the retest.In order to isolate specific microbial contaminants, FDA laboratories, aswell as many in the industry, employ some type of enrichment mediacontaining inactivators, such as Tween or lecithin. This is essential toinactivate preservatives usually present in these types of product andprovides a better medium for damaged or slow growing cells. Other growthparameters include a lower temperature and longer incubation time (at least 5 days) that provide a better survival condition for damaged or slow-growing cells.For example, FDA laboratories use the test procedures for cosmetics in the Bacteriological Analytical Manual (BAM), 6th Edition, to identifycontamination in non-sterile drug products. This testing includes anenrichment of a sample in modified letheen broth. After incubation, further identification is carried out on Blood Agar Plates and MacConkey AgarPlates. Isolated colonies are then identified. This procedure allows FDAmicrobiologists to optimize the recovery of all potential pathogens and to quantitate and speciate all recovered organisms. Another important aspect of procedures used by FDA analysts is to determine growth promotioncharacteristics for all of the media used.The selection of the appropriate neutralizing agents are largely dependent upon the preservative and formulation of the product under evaluation. Ifthere is growth in the enrichment broth, transfer to more selective agarmedia or suitable enrichment agar may be necessary for subsequentidentification.Microbiological testing may include an identification of colonies foundduring the Total Aerobic Plate Count test. Again, the identification should not merely be limited to the USP indicator organisms.The importance of identifying all isolates from either or both Total Plate Count testing and enrichment testing will depend upon the product and itsintended use. Obviously, if an oral solid dosage form such as a tablet istested, it may be acceptable to identify isolates when testing shows highlevels. However, for other products such as topicals, inhalants or nasalsolutions where there is a major concern for microbiological contamination, isolates from plate counts, as well as enrichment testing, should beidentified.III. FACILITIES, EQUIPMENT, ANDMEDIABegin the inspection with a review of analyses being conducted and inspect the plates and tubes of media being incubated (caution should be exercised not to inadvertently contaminate plates or tubes of media on test). Beparticularly alert for retests that have not been documented and "specialprojects" in which investigations of contamination problems have beenidentified. This can be evaluated by reviewing the ongoing analyses (product or environmental) for positive test results. Request to review the previous day's plates and media, if available and compare your observations to therecorded entries in the logs. Inspect the autoclaves used for thesterilization of media. Autoclaves may lack the ability to displace steamwith sterile filtered air. For sealed bottles of media, this would notpresent a problem. However, for non-sealed bottles or flasks of media,non-sterile air has led to the contamination of media. In addition,autoclaving less than the required time will also allow media associatedcontaminants to grow and cause a false positive result. These problems may be more prevalent in laboratories with a heavy workload.Check the temperature of the autoclave since overheating can denature andeven char necessary nutrients. This allows for a less than optimal recovery of already stressed microorganisms. The obvious problem with potential false positives is the inability to differentiate between inadvertent mediumcontamination and true contamination directly associated with the sampletested.IV. STERILITY TESTINGOn 10/11/91, the Agency published a proposed rule regarding the manufacture of drug products by aseptic processing and terminal sterilization. A list of contaminated or potentially contaminated drug products made by asepticprocessing and later recalled was also made available. Many of theinvestigations/inspections of the recalled products started with a list of initial sterility test failures. FDA review of the manufacturer'sproduction, controls, investigations and their inadequacies, coupled withthe evidence of product failure (initial sterility test failure) ultimately led to the action.The USP points out that the facilities used to conduct sterility testsshould be similar to those used for manufacturing product. The USP states,"The facility for sterility testing should be such as to offer no greater a microbial challenge to the articles being tested than that of an asepticprocessing production facility". Proper design would, therefore, include a gowning area and pass-through airlock. Environmental monitoring and gowning should be equivalent to that used for manufacturing product.Since a number of product and media manipulations are involved in conducting a sterility test, it is recommended that the inspection include actualobservation of the sterility test even though some companies have tried to discourage inspection on the grounds that it may make the firm's analystnervous. The inspection team is expected to be sensitive to this concern and make the observations in a manner that will create the least amount ofdisruption in the normal operating environment. Nevertheless, such concerns are not sufficient cause for you to suspend this portion of the inspection.One of the most important aspects of the inspection of a sterilityanalytical program is to review records of initial positive sterility test results. Request lists of test failures to facilitate review of production and control records and investigation reports. Particularly, for the highrisk aseptically filled product, initial positive sterility test results and investigations should be reviewed. It is difficult for the manufacturer to justify the release of a product filled aseptically that fails an initialsterility test without identifying specific problems associated with thecontrols used for the sterility test.Examine the use of negative controls. They are particularly important to a high quality sterility test. Good practice for such testing includes the use of known terminally sterilized or irradiated samples as a system control.Alternatively, vials or ampules filled during media fills have also beenused.Be especially concerned about the case where a manufacturer of aseptically filled products has never found an initial positive sterility test. Whilesuch situations may occur, they are rare. In one case, a manufacturer'srecords showed that they had never found a positive result; their recordshad been falsified. Also, the absence of initial positives may indicate that the test has not been validated to demonstrate that there is no carryover of inhibition from the product or preservative.Inspect robotic systems or isolation technology, such as La Calhene unitsused for sterility testing. These units allow product withdrawal in theabsence of people. If an initial test failure is noted in a sample tested in such a system, it could be very difficult to justify release based on aretest, particularly if test controls are negative.Evaluate the time period used for sterility test sample incubation. Thisissue has been recently clarified. The USP states that samples are to beincubated for at least 7 days, and a proposal has been made to change theUSP to require a period of 14 days incubation. You are expected to evaluate the specific analytical procedure and the product for the proper incubation period. Seven days may be insufficient, particularly when slow growingorganisms have been identified. Media fill, environmental, sterility testresults and other data should be reviewed to assure the absence of slowgrowing organisms. Also, you should compare the methods being used forincubation to determine if they conform to those listed in approved orpending applications.V. METHODOLOGY ANDDetermine the source of test procedures. Manufacturers derive testprocedures from several sources, including the USP, BAM and othermicrobiological references. It would be virtually impossible to completely validate test procedures for every organism that may be objectionable.However, it is a good practice to assure that inhibitory substances insamples are neutralized.During inspections, including pre-approval inspections, evaluate themethodology for microbiological testing. For example, we expect test methods to identify the presence of organisms such as Pseudomonas cepacia or other Pseudomonas species that may be objectional or present a hazard to the user. Where pre-approval inspections are being conducted, compare the method being used against the one submitted in the application. Also verify that thelaboratory has the equipment necessary to perform the tests and that theequipment was available and in good operating condition on the dates ofcritical testing.The USP states that an alternate method may be substituted for compendialtests, provided it has been properly validated as giving equivalent orbetter results.You may find that dehydrated media are being used for the preparation of<<< Continued to next message >>><<< This message is part 2 of a previous message >>>media. Good practice includes the periodic challenge of prepared media with low levels of organisms. This includes USP indicator organisms as well asnormal flora. The capability of the media to promote the growth of organisms may be affected by the media preparation process, sterilization(overheating) and storage. These represent important considerations in any inspection and in the good management of a microbiology laboratory.VI. DATA STORAGEEvaluate the test results that have been entered in either logbooks or onloose analytical sheets. While some manufacturers may be reluctant toprovide tabulations, summaries, or printouts of microbiological testresults, this data should be reviewed for the identification of potentialmicrobial problems in processing. When summaries of this data are notavailable the inspection team is expected to review enough data to construct their own summary of the laboratory test results and quality controlprogram.Some laboratories utilize preprinted forms only for recording test data.Some laboratories have also pointed out that the only way microbiologicaltest data could be reviewed during inspections would be to review individual batch records. However, in most cases, preprinted forms are in multiplecopies with a second or third copy in a central file. Some companies uselog-books for recording data. These logbooks should also be reviewed.Additionally, many manufacturers are equipped with an automated microbialsystem, such as a Vitek, for the identification of microorganisms. Logs of such testing, along with the identification of the source of the sample, are also of value in the identification of potential microbial problems inprocessing.The utilization of automated systems for the identification ofmicroorganisms is relatively common in the parenteral manufacturer whereisolates from the environment, water systems, validation and people areroutinely identified.Microbiologists in our Baltimore District are expert on the use of automated microbic analytical systems. They were the first FDA laboratory to use such equipment and have considerable experience in validating methods for these pieces of equipment. Contact the Baltimore District laboratory forinformation or questions about these systems. Plants with heavy utilization of these pieces of equipment should be inspected by individuals from theBaltimore District laboratory.VII. MANAGEMENT REVIEWMicrobiological test results represent one of the more difficult areas for the evaluation and interpretation of data. These evaluations requireextensive training and experience in microbiology. Understanding themethodology, and more importantly, understanding the limitations of the test present the more difficult issues. For example, a manufacturer found highcounts of Enterobacter cloacae in their oral dosage form product derivedfrom a natural substance. Since they did not isolate E. coli, they released the product. FDA analysis found E. cloacae in most samples from the batchand even E. coli in one sample. In this case management failed to recognize that microbiological contamination might not be uniform, that otherorganisms may mask the presence of certain organisms when identificationprocedures are performed, and that microbiological testing is far fromabsolute. The inspection must consider the relationship between theorganisms found in the samples and the potential for the existence of other objectionable conditions. For example, it is logical to assume that if the process would allow E. cloacae to be present, it could also allow thepresence of the objectionable indicator organism. The microbiologist should evaluate this potential by considering such factors as methodology, and the growth conditions of the sample as well as other fundamental factorsassociated with microbiological analysis.Evaluate management's program to audit the quality of the laboratory workperformed by outside contractors.VIII. CONTRACT TESTINGLABORATORIESMany manufacturers contract with private or independent testing laboratories to analyze their products. Since, these laboratories will conduct only thetests that the manufacturer requests, determine the specific instructionsgiven to the contractor. Evaluate these instructions to assure thatnecessary testing will be completed. For example, in a recent inspection of a topical manufacturer, total plate count and testing for the USP indicator organisms were requested. The control laboratory performed this testing only and did not look for other organisms that would be objectionable based onthe product's intended use.Analytical results, particularly for those articles in which additional or retesting is conducted, should be reviewed. Test reports should be provided to the manufacturer for tests conducted. It is not unusual to see contract laboratories fail to provide complete results, with both failing as well as passing results.Bacteriostasis/fungiostasis testing must be performed either by the contract lab or the manufacturer. These test results must be negative otherwise any sterility test results obtained by the contractor on the product may not be valid.There are no references from this document.。

高纯水系统检查指南1GUIDE TO INSPECTIONS OF HIGH PURITY WATER SYSTEMSNote: This document is reference material for investigators and other FDA personnel. The document does not bind FDA, and does no confer any rights, privileges, benefits, or immunitiesfor or on any person(s).注释：该文件只是检查官和FDA职员的参考资料。

本指南不具法律约束力，也未授于任何人任何形式的特权，利益或豁免权。

This guide discusses, primarily from a microbiological aspect, the review and evaluation of high purity water systems that are used for the manufacture of drug products and drug substances. It also includes a review of the design of the various types of systems and some of the problems that have been associated with these systems. As with other guides, it is not all-inclusive, but provides background and guidance for the review and evaluation of high purity water systems. The Guide To Inspections of Microbiological Pharmaceutical Quality Control Laboratories (May, 1993) provides additional guidance.本指南主要是从微生物方面，讨论和评估了原料药和制剂生产中用到的高纯水系统。

Evaluation, V alidation and Implementation of New Microbiological Testing Methods新微生物测试方法的评估、验证和执行Technical Report No. 33 第33号技术报告PDA目录第一部分: 新微生物方法的选择1.0 简介. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11.1 文件范围. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2 文件目的. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.3 文件结构概览. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22.0 技术概要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22.1微生物方法类型的一般说明. . . . . .. . . . . . . . . . .. . . . . . . . .. . 22.2 技术审核. . .. . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . .. . . . 32.3 基于生长的技术. . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. . . .. .32.3.1 ATP生物发光. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ..32.3.2 CO2生产的色度检测. . . . . . . . . . . . . . . . . . . . . . . .. . . .42.3.3 顶部空间压力变更的测量. . . .. . . . . . . . . . . . . .. .. . . .. .42.3.4阻抗. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .42.3.5 生物化学分析. . .. . . . . .. . . . . . .. . . . . . . . . . . . . . . .42.4 基于活性的技术. . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . 62.4.1 固相血细胞计数. . . . . .. . . . . . . . . . . . . . . . . . . . . . . . 62.4.2 流式荧光细胞计. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .62.5 人工技术. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.1 脂肪酸概要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.2 质谱学. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.3 ELISA酶联免疫吸附测定. . . . . . . . . . . . . . . . . .. . . . . . 72.5.4 荧光探针检测. . . .. . . . . . . . . . . . . . . . . . . .. . . . . . . . 72.5.5 细菌内毒素-鲎变形细胞溶解物测试. . . . . . . . . . . . . ..72.6 基于核酸的技术. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. (8)2.6.1 探针. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .82.6.2 核糖型/分子型. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . 82.6.3 聚合酶链反应. . .. . . . . .. . . . . . . . . . . . . . . . . . . . . .93.0监管审查. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103.1微生物测试总分类. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.1 加工过程中测试. . .. . . . . . . . . . . . . . . . . . . . . . (10)3.1.2 产品测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.3 定性测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.4 定量测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.2 药典微生物测试方法参考文件. . .. . . . . . . . . . . . . . . . . . . . 103.2.1 制药用水. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..103.2.2 抗菌效力测试. . . . .. . . . . . . . . . . . . . . . . . . . . . . (10)3.2.3 微生物限度测试. . . . . .. . . . . . . . . . . . . . . . . . (11)3.2.4 无菌测试. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . (11)3.2.5 环境监测. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (11)3.2.6 微生物鉴定. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .113.3 改变微生物测试方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.3.1 新微生物测试方法介绍的监管观点 . . . . . . . . . . . . . ..113.3.2 药典观点. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (12)3.4 药典变更. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (13)3.5 新微生物方法的管理者评估. . . . . . . . . . . . . . . . . . . . .13 第二部分: 如何验证并执行新微生物测试方法4.0 验证流程. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .154.1 设备确认模型. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . .164.1.1 供应商/说明要求. . . . . . . . . . . . . . . . . . . . . . . . . . 164.1.1.1 测试方法选择. . . . . . . . . . . . . . . . . . . . .174.1.1.2 供应商选择. . . . . . . . . . . . . . . . . . . . . . .174.1.2 验证方法设计. . . . . . . . . . . . . . . . . . . . . . . . . . (18)4.1.3 安装确认. . . . . . . . . . . . . . . . . . . . . . . . . . . . (18)4.1.4 运行确认. . . . . . . . . . . . . . . . . . . . . . . . . . . . (20)4.1.5 性能确认. . . . . . . . . . . . . . . . . . . . . . . . . . ....... (20)4.2验证标准. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (21)4.2.1测试样品制备. . .. . . . . . . . . . . . . . . . . ....... . . . (23)4.2.2 微生物方法的活性：特别注意. . . . . . . . . . . . . . . . 254.2.2.1 样品分布错误. . .. . . . . . . . . . . . . . . (25)4.2.2.2 细胞形态. . . . . . . . . . . . . . . . . . . . . . . . 254.2.2.3 代谢活动. . .. . . . . . . . . . . . . . . . . . . . . 254.2.3使用推荐验证标准的方案设计. .. . . . . . . . . . . . . . .264.3微生物方法验证的特殊考虑. . . . . . . . . . . . . . . . . . . . . . . 294.3.1在实验室和公司内使用多件相同设备. . . . . . . . . . . . 294.3.2微生物设备的独特测试要求. .. . . . . . . . . . . . . . . . . 305.0 术语. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32第一部分新微生物测量方法的选择1.0简介1.1文件范围本文件的目的是对制药、生物技术和医药设备工业为确保产品质量进行成功评估、验证和实施所需新微生物方法提供指南。

行业指南：药品检验结果超标调查美国健康与人服务部食品药品监督管理局(FDA) 药物评价与研究中心(CDER) 2006.10目录1. 引言2. 背景3. 超标检测结果的鉴定和评估—第一阶段:实验室调查A. 分析员的责任B. 实验室监督员的责任4. 超标检测结果的调查—第二阶段:全面超标调查A. 生产工艺回顾B. 附加的实验室检验C. 检验结果报告5. 调查结论A. 调查结果的解释B. 警戒/或译保证?C. 分布警示报告本指南代表了FDA当前关于这个话题的想法.它不会创造或赠与任何人任何权力，也不会约束FDA或公众。

如果其他可选择的相接近的指南能满足适用的法令和法规的要求，也可以使用。

如果你想讨论一种替代方法,请与负责执行这指南的FDA工作人员联系,如果你不能找到专门人员,请致电到这指南扉页上的号码.1. 前言本行业指南反应了FDA目前关于如何评估超标检验结果的看法, 从此文件的目的出发，术语OOS 结果包含了所有的超出新药申请、药物主文件DMF,官方手册或者生产企业制定的规格或可接受标准的检验结果。

此术语也适用于所有超出既定标准的进行中的实验室测试.该指南适用于由CDER规定的基于化学的实验室药物检测.它是针对传统药物检测和释放(release)方法.这些检测是对适用于CGMP规范(21 CFR parts210 and 211)和联邦食品药物和化妆品法(section 501(a)(2)(B))的活性药物成份、辅料和其他成分，加工材料以及成品药进行的.这个指南的原则也适用于内部测试外购药物成分.本指南意见也可被用于合同公司从事生产和/或化验的责任.本指南具体讨论了如何调查OOS检验结果，包括了实验室人员的责任、实验室阶段调查、必须的额外试验，何时进行实验室范围外的调查和对所有检验结果的最终评估。

该机构按照其在2002年8月“为了21世纪的药物CGMP”的倡议,鼓励采用现代方法制造,监测和控制以提高过程预见性和效率.工艺过程分析技术(PAT),采取了一种不同的办法以质量保证通过使用工艺过程控制和过程的数据作为释放规格,而不是依靠单一的实验室检测做出批可接受性决定. 本指南不打算陈述PAT方法,因为在日常的进行过程中使用这些方法可能包括其他因素. 对于信息关于实时性在过程测试,见CGMP指南题为PAT--一个为创新药物研究、生产和质量保证的框架.FDA的指导性文件,包括这个指导,没有建立法律约束力的责任.反而, 指导性文件叙述的是目前该机构的在一主题的想法,并且应只作为建议,除非是特有管理或法定要求的引用.在FDA机构should这个词的使用意味着一些建议或推荐,但不是必要的.2. 背景CGMP规范(§§ 211.160 and 211.165)所要求的实验室检测活动是确认药物成分、容器与密封材料、中间产物及最终成品是否符合标准要求包括稳定性要求的必要手段。

美国FDA非肠道用冻干制剂检查指南1993年引言冻干是指药品冷冻后在真空状态下不经液态，直接从固态升华至气态，由此去除水分的作业过程。

该过程包括三个彼此独立又相互依赖的步骤：冷冻、一级干燥(升华)以及二级干燥 (解吸附)。

冻干工艺的优点在于：液体加工方便，简化了无菌作业过程；提高了干粉的稳定性；无需过热处理就能去除产品中的水分；增强了冻干产品的稳定性及复水(溶解)性。

冻干工艺的缺点在于：加工处理过程所需时间长；复水时需用无菌稀释液；设备复杂且生产成本高。

冻干工艺一般包括以下步骤：(1)将药品和赋形剂溶解于适当的溶剂中，通常使用注射用水。

(2)将药液通过一个0．22μm的微孔除菌过滤器，以此灭菌。

(3)灌装到各个已灭菌的容器中，并在无菌条件下半压塞。

(4)在无菌条件下将半压塞的容器转移至冻干腔室内。

(5)冷冻该溶液：把半压塞的容器置于冻干腔室的冷冻搁板上，或在另一腔室内作预冷冻。

(6)将腔室抽真空并将冷冻搁板升温，以便在冷冻状态下通过蒸发除去水分。

(7)全压塞密封：通常由安装在冻干机内的液压式或螺杆式压塞装置来完成。

现在有许多新的非肠道药品，如抗感染药物、生物制剂、体外诊断用药等等都采用冻干工艺来加工。

调查及研究业已发现与冻干工艺及其控制相关的冻干剂在疗效、无菌性和稳定性方面存在问题。

为此，本检查指南对冻干产品有关的生产工艺和缺陷进行了讨论，以便为调研人员提供这方面的信息和指导。

人们已认识到冻干药品的生产和控制是一项很复杂的工艺技术。

冻干作业的重要内容包括药液的制备；西林瓶灌装及其验证；冻干机的灭菌和设计；冻干工艺的放大生产(批量放大)及其验证；最终产品的检验等等。

冻干剂成品生产和控制相关的问题是本文的重点内容。

药品种类和配方药品需经冻干是因为其溶液状态下的不稳定性。

有许多抗生素，如某些半合成的青霉素、头孢菌素和红霉素、强力霉素、氯霉素的盐类都是由冻干工艺制造的。

可以预计在生产过程中这类产品的带菌量很低，因为它们是抗生素。

GUIDE TO INSPECTIONS OF MICROBIOLOGICAL PHARMACEUTICALQUALITY CONTROL LABORATORIES制药行业微生物实验室现场检查指南Note: This document is reference material for investigators and other FDA personnel. The document does not bind FDA, and does not confer any rights, privileges, benefits, or immunities for or on anyperson(s).注：该指南是检察官和其他FDA人员的参考材料。

该指南不约束FDA，也不授予任何权利，特权，利益或者任何人的豁免权。

I. INTRODUCTION介绍The Guide to the Inspection of Pharmaceutical Quality Control Laboratories provided very limited guidance on the matter of inspection of microbiological laboratories. While that guide addresses many of the issues associated with the chemical aspect of laboratory analysis of pharmaceuticals, this document will serve as a guide to the inspection of the microbiology analytical process. As with any laboratory inspection, it is recommended that an analyst (microbiologist) who is familiar with the tests being inspected participate in these inspections.“制药行业分析实验室现场检查指南”对如何进行微生物实验室的检查仅提供了非常有限的指导，那份指南文件主要涉及医药行业分析实验室化学分析方面的很多内容，而本指南主要讨论的是微生物分析过程的现场检查指导。

纯化水系统检查指南提示：该文献参考研究者及其它FDA人员的资料，它不属于FDA且未授予个人任何专利。

该指南主要从微生物的影响来评价用于药品和药物生产的高纯化水系统，并评论了不同型号系统的设计及这些产品的一些问题，和其它指南相同，该指南只对高纯化水系统的评估起指导作用但并不包括全部指标。

另外可参考药物质量控制实验室的微生物检查指南（May，1993）。

1．系统设计在设计一个系统时最初首先要考虑的是生产的产品类型。

对注射用药物来说，由于涉及到热源质，所以得用注射用水。

产品制剂、最后的组分洗涤及生产中所用仪器的洗涤都要用到注射用水。

USP中规定的生产注射用水方法只有蒸馏法和反渗透（逆向渗透）法。

但是，在大量制药工业，生物技术工业及一些国外公司中，都用超滤法将用于注射给药的药物中的内毒素减到最少。

在一些眼药产品（如眼药水）及一些吸入产品（如吸入用的消毒水）中，由于有热原质规格，所以在其制剂中要用注射用水。

但是，大部分吸入及眼科产品的制剂都使用纯化水，纯化水还用于局部用药、化妆品及口服产品。

设计一个系统其次要考虑的是系统温度。

温度在65 - 80o C的系统被认为可自我消毒。

虽然对一个公司来说其它系统更便宜一些，但维护费，试验和潜在问题会比省下来的能量价值更高。

系统是循环还是单向也是设计系统时所要考虑的重点之一。

显然，让水持续流动是污染很少的方法，一个单向水系统基本上是“死角”("dead-leg")。

最后的也是最需要考虑的一点是风险评估或所期望的质量水平。

应该认识到不同产品需要不同质量的水。

注射用药需要无内毒素的高纯水，局部用药和口服用药所需水没有内毒素的要求，纯度要求稍低。

即使局部用药和口服用药也由于各种因素影响要用不同质量的水。

比如，在抗酸剂中防腐剂起一定作用，所以得更严格规定其微生物指标。

质量控制部门应该用系统中的水评估每步的产品生产并根据对微生物最敏感的产品确定微生物限值。

对于敏感的药品，在系统中水作用限制严的情况下，厂商可在生产过程中加一步除微生物操作。

清洁验证中的微生物问题------设备表面微生物限度的制定及其检验陈雯秋½-ËÕÎÞÎý 214092¶ÔÖÆÒ©É豸½øÐÐÇåÏ´ÊÇ·ÀÖ¹Ò©Æ·ÎÛȾºÍ½»²æÎÛȾµÄ±ØÒªÊֿɽ«É豸±íÃæµÄ²ÐÁôÎï¼õÉÙµ½²»»áÓ°ÏìÏÂÅú²úÆ·µÄÁÆЧÇå½àÑéÖ¤¾ÍÊÇͨ¹ý¿ÆѧµÄ·½·¨²É¼¯×ã¹»µÄÊý¾ÝÄÜʼÖÕÈçÒ»µØ´ïµ½Ô¤¶¨µÄÇå½à±ê×¼[1]¿É·ÖΪ»¯Ñ§ÎïÖʺÍ΢ÉúÎïÁ½´óÀà化学物质一般有活性药物成分制药企业一般能够根据最低日治疗剂量成品的杂质限量等参数制定出适宜的限度对于产品质量虽然多数法规也强调了其重要性清洁验证的文件指南设备清洁的微生物问题都必须加以考虑特别要采取措施防止在设备存放期间的微生物繁殖[2]È´ÉÙÓÐÎÄÏ×Ìá³öºÏÀíµÄÖƶ¨·½·¨ÆäÏ޶ȵÄÖƶ¨µÄÈ·ÓÐÒ»¶¨ÄѶȶøÔÚʵ¼ÊµÄÇå½àÑéÖ¤Öл¹ÐèÒª½áºÏ²úÆ·ÒªÇóºÍÉú²úÇé¿ö×ÃÇéÖƶ¨»¯Ñ§ÎïÖʾßÓÐÊýÁ¿ÉϵĶø¶ÔÓÚ΢ÉúÎïÀ´Ëµ±íÃæ²ÐÁôµÄ΢ÉúÎï»áÓÐÊýÁ¿Éϵı仯´ÓÉ豸±íÃæ»ãÈëÏÂÅú²úÆ·µÄ΢ÉúÎïÀýÈç»òÕßÏÂÅú²úÆ·Ô-ÁÏÖк¬ÒÖ¾ú¼Á¶øÈç¹ûÉ豸´æ·ÅÆÚ¼äÓÐÊÊÒ˵ÄÉú³¤Ìõ¼þÔò»áʹ֮¼±ËÙÔöÖ³¼´±ãÔÚ¸ÉÔïµÄÉ豸±íÃæÒàÄÜ´æ»î»òÕßÏÂÅú²úÆ·ÊǸÉÆ·1. 微生物限度的制定方法1-1 微生物限度的理论推导[3]由于微生物的特殊性本文分三种情况来讨论微生物限度的制定方法下批产品的原料及其生产过程既不引起微生物的增殖第二种情况而产品将接受最终灭菌下批产品生产前设备需灭菌因下批产品的原料和生产过程不引起微生物数量的变化首先确定下批产品的成品微生物限度CFUÖ®ºó¼ÆË㵥λÃæ»ýµÄ΢ÉúÎïÏÞ¶ÈÓÐÁ½¸öÏà¹ØÐÅÏ¢Ô´一般会有微生物限度的要求非无菌药品的微生物要求例如然后要考虑该限度中究竟有多少该于设备表面的微生物残留内包装材料等也是成品中微生物的来源举例来说算得由原料造成的最高的微生物污染为27CFU/g那么允许由设备表面造成的污染限度为70CFU/g¼ÙÉèÈ¡5也可以采用更高的安全因子当确定了下批产品中由设备表面造成的微生物污染限度之后CFU/cm2这就和化学残留物的计算方法完全一样了LSPÀ´×ÔÉ豸±íÃæµÄ²ÐÁô΢ÉúÎïÏÞ¶ÈMBS Minimum batch size产品接触的设备表面积　假设某产品的批量为200kg由刚才推导出的LSP 计算所得的限度值为11CFU/cm2ÏÂÅú²úÆ·Éú²úÇ°É豸²»Ãð¾úÕâÖÖÇé¿öϲúÆ·ÎÞ¾ú±£Ö¤ÖµµÄ´óСÓëÃð¾ú¹¤ÒÕµÄF0值以及灭菌前微生物的含量有关即N0知晓这个限度后确定该限度中于清洁后设备表面的比例然后以最小批量和接触面积算出最终的限度值设备清洁非常重要以保证灭菌或消毒能够达到一定的无菌保证值[2]第三种情况因此设备本身将接受灭菌因为在做该设备的灭菌验证时已经制定了表面微生物的限度只须以该限度值除以某安全因子1-2 结合实际情况制定微生物限度通常理论推导出的微生物限度往往大大高于实际可达到的清洁水平良好的清洁规程应能使接触碟每碟小于25CFUÒ»°ãÿµú»á´óÓÚ100CFU¼ÆËãËùµÃÏÞ¶ÈֵΪ11CFU/cm2ÔòÏ൱ÓÚÿµú275CFU显然太宽因为一般每碟最多只能数清250CFUÈçºÎÖƶ¨Ï޶ȲÅËãºÏÀíÄØ×ÛºÏȨ衡举例来说则应予以考虑无菌药品的生产对不同洁净区的动态微生物水平提出了建议值如静态关键表面和一般表面分别制定所谓关键表面显然设备表面是属于关键表面而该企业对十万级区的关键表面要求动态不大于每碟10CFU二者取其低另外例如如管路而这些地方正是最不易清洁的此时淋洗水样品的限度值就未必适用于擦拭样品宜将设备分为不同的表面分别制定合理的限度值未必从一开始就能制定出合理的限度限度也是需要验证的过宽的限度会给产品质量带来风险从验证过程中积累的数据会发现原定限度的不合理性但是无论限度的制定还是修改2. 设备表面残留微生物的取样及检验方法传统的测量表面微生物的取样及检验方法都可使用通常用膜过滤棉签擦拭然后用倾注平皿法以及接触碟等一般来说比较关注需氧菌三种取样方法应视情况结合使用但难以反应真实的污染情况能弥补淋洗水取样的不足但是如每碟微生物计数超过250CFU3. 结束语微生物的残留问题在设备清洁验证中不容忽视除使用清洁剂和充分淋洗外以及保证设备存放期间的环境微生物水平等应自设备刚刚清洁完毕起在整个存放期间评价微生物随时间的增殖情况确保设备表面微生物在存放期间不进入快速增殖期应该满足生产和质量控制的要求并且是检验方法能够检验的制药企业应该根据具体的产品和生产情况在验证方案中阐述合理的微生物限度参考文献。