美国FDA药品质量控制微生物实验室检查指南
FDA-QC微生物实验室检查指南
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.。
美国FDA药品质量控制实验室检查指南年
(1)不得进行两次复检和凭据三次化验的平均值对产物进行发放；
(2)不能用。outlinertest做化学查验；
(3)不能用重复取样的步伐假定取样或制备历程误差；
(4)当确认可以复检时(见另外的标准)，可以取同一样品中的差别药片做复检。
C．正式视察
超出厂实验室范畴的正式视察，必须依照一个提纲进行，并要特别注意整改步伐。
不切合规格标准结果可以分成三类：
一——实验室误差；
——一非生产工艺性误差大概称为操纵者误差；
———和生产工艺有关的误差大概称为生产工艺误差；
A．实验室误差
实验室误差产生于下列情况：化验员未能正确地按阐发要领操纵；使用不正确的标准和(或)简单地算错了数据。实验室误差必须通过一项视察来确定，以便判定不切合规格标准的原因。—旦不切合规格标准结果的性质被确定了，就可以把它归入上述三类中的一类。由于视察的目的不一样，查询可能很不相同。
我们希望实验室查验数据能直接记在记录本上，制止使用纸片或活页纸。这些知识性要领可以增强数据的准确性和完整性。
审查和评价实验室用于进行产物不合格视察的标准操纵步伐，对单一和多个不切合规格标准结果的视察应遵循差别的步伐。对单一不切合规格标准结果，视察应包罗下列步调，并且这些视察应当在该样品被复检之前进行：
法庭认为对不切合规格标准结果进行的复检只有在此种情况下才是适合的，即对不合格的视察正在进行且该视察部分地决定厂复检是否符合。当视察发明不切合规格标准结果确由化验员误差所致或对化验员事情的复查是“无结论性的(inconclusive)”情况下，复检是符合的。而对付众所周知，没有争议的与生产历程有关或无关的错误，复检是不符合的。
药物申请的阐发部分通常只包罗化验结果和用来得到这些结果的要领，并不要求卖力人提交所有的化验数据，因为这样做会使提交的资料体积太大，并可导致提供过多不须要的资料。卖力人可能有意无意地选择并陈诉那些能显示药物宁静有效并能得到批准的数据，而不陈诉那些证明该产物不能满足预先制定的规格标准的数据。查抄组必须确定这样做是否存在有效的、科学的解释。
FDAQC实验室检查指南
GUIDE TO INSPECTIONS OF 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).1. INTRODUCTIONThe pharmaceutical quality control laboratory serves one of the mostimportant functions in pharmaceutical production and control. A significantportion of the CGMP regulations (21 CFR 211) pertain to the quality controllaboratory and product testing. Similar concepts apply to bulk drugs.This inspection guide supplements other inspectional information containedin other agency inspectional guidance documents. For example, ComplianceProgram 7346.832 requiring pre-approval NDA/ANDA inspections containsgeneral instructions to conduct product specific NDA/ANDA inspection auditsto measure compliance with the applications and CGMP requirements. Thisincludes pharmaceutical laboratories used for in-process and finishedproduct testing.2. OBJECTIVEThe specific objective will be spelled out prior to the inspection. Thelaboratory inspection may be limited to specific issues, or the inspectionmay encompass a comprehensive evaluation of the laboratory's compliance withCGMP's. As a minimum, each pharmaceutical quality control laboratory shouldreceive a comprehensive GMP evaluation each two years as part of thestatutory inspection obligation.In general these inspections may include-- the specific methodology which will be used to test a new product-- a complete assessment of laboratory's conformance with GMP's-- a specific aspect of laboratory operations3. INSPECTION PREPARATIONFDA Inspection Guides are based on the team inspection approach and ourinspection of a laboratory is consistent with this concept. As part of oureffort to achieve uniformity and consistency in laboratory inspections, weexpect that complex, highly technical and specialized testing equipment,procedures and data manipulations, as well as scientific laboratoryoperations will be evaluated by an experienced laboratory analyst withspecialized knowledge in such matters.District management makes the final decision regarding the assignment ofpersonnel to inspections. Nevertheless, we expect investigators, analystsand others to work as teams and to advise management when additionalexpertise is required to complete a meaningful inspection.Team members participating in a pre-approval inspection must read and befamiliar with Compliance Program 7346.832, Pre-ApprovalInspections/Investigations. Relevant sections of the NDA or ANDA should bereviewed prior to the inspection; but if the application is not availablefrom any other source, this review will have to be conducted using thecompany's copy of the application.Team members should meet, if possible, prior to the inspection to discussthe approach to the inspection, to define the roles of the team members, andto establish goals for completion of the assignment. Responsibilities fordevelopment of all reports should also be established prior to theinspection. This includes the preparation of the FDA 483.The Center for Drug Evaluation and Research (CDER) may have issueddeficiency letters listing problems that the sponsor must correct prior tothe approval of NDA/ANDA's and supplements. The inspection team is expected to review such letters on file at the district office, and they are expectedto ask the plant for access to such letters. The team should evaluate thereplies to these letters to assure that the data are accurate and authentic.Complete the inspection even though there has been no response to theseletters or when the response is judged inadequate.4. INSPECTION APPROACHA. GeneralIn addition to the general approach utilized in a drug CGMP inspection, theinspection of a laboratory requires the use of observations of thelaboratory in operation and of the raw laboratory data to evaluatecompliance with CGMP's and to specifically carry out the commitments in anapplication or DMF. When conducting a comprehensive inspection of alaboratory, all aspects of the laboratory operations will be evaluated.Laboratory records and logs represent a vital source of information thatallows a complete overview of the technical ability of the staff and ofoverall quality control procedures. SOPs should be complete and adequate and the operations of the laboratories should conform to the written procedures.Specifications and analytical procedures should be suitable and, asapplicable, in conformance with application commitments and compendialrequirements.Evaluate raw laboratory data, laboratory procedures and methods, laboratoryequipment,including maintenance and calibration, and methods validation data to determine the overall quality of the laboratory operation and the abilityto comply with CGMP regulations.Examine chromatograms and spectra for evidence of impurities, poortechnique, or lack of instrument calibration.s use systems that provide for the investigation oflaboratory test failures. These are generally recorded in some type of log.Ask to see results of analyses for lots of product that have failed to meetspecifications and review the analysis of lots that have been retested,rejected, or reworked. Evaluate the decision to release lots of product whenthe laboratory results indicate that the lot failed to meet specificationsand determine who released them.B. Pre-ApprovalDocuments relating to the formulation of the product, synthesis of the bulkdrug substance, product specifications, analysis of the product, and othersare examined during the review process in headquarters. However, thesereviews and evaluations depend on accurate and authentic data that trulyrepresents the product.Pre-approval inspections are designed to determine if the data submitted inan application are authentic and accurate and if the procedures listed inthe application were actually used to produce the data contained in theapplication. Additionally, they are designed to confirm that plants(including the quality control laboratory) are in compliance with CGMPregulations.The analytical sections of drug applications usually contain only testresults and the methods used to obtain them. Sponsors are not required tofile all the test data because such action would require voluminoussubmissions and would often result in filing redundant information. Sponsors may deliberately or unintentionally select and report data showing that adrug is safe and effective and deserves to be approved. The inspection team must decide if there is valid and scientific justification for the failureto report data which demonstrates the product failed to meet itspredetermined specifications.Coordination between headquarters and the field is essential for a completereview of the application and the plant. Experienced investigators andanalysts may contact the review chemist (with appropriate supervisoryconcurrence) when questions concerning specifications and standards arise.Inspections should compare the results of analyses submitted with results of analysis of other batches that may have been produced. Evaluate the methods and note any exceptions to the procedures or equipment actually used fromthose listed in the application and confirm that it is the same methodlisted in the application. The analyst is expected to evaluate rawlaboratory data for tests performed on the test batches (biobatches andclinical batches) and to compare this raw data to the data filed in theapplication.5. FAILURE (OUT-OF-SPECIFICATION) LABORATORY RESULTSEvaluate the company's system to investigate laboratory test failures. These investigations represent a key issue in deciding whether a product may bereleased or rejected and form the basis for retesting, and resampling.In a recent court decision the judge used the term "out-of-specification"(OOS) laboratory result rather than the term "product failure" which is morecommon to FDA investigators and analysts. He ruled that an OOS resultidentified as a laboratory error by a failure investigation or an outliertest. The court provided explicit limitations on the use of outlier testsand these are discussed in a later segment of this document., or overcome byretesting. The court ruled on the use of retesting which is covered in alater segment of this document. is not a product failure. OOS results fallinto three categories:-- laboratory error-- non-process related or operator error-- process related or manufacturing process errorA. LABORATORY ERRORSLaboratory errors occur when analysts make mistakes in following the methodof analysis, use incorrect standards, and/or simply miscalculate the data.Laboratory errors must be determined through a failure investigation toidentify the cause of the OOS. Once the nature of the OOS result has beenidentified it can be classified into one of the three categories above. Theinquiry may vary with the object under investigation.B. LABORATORY INVESTIGATIONSThe exact cause of analyst error or mistake can be difficult to determinespecifically and it is unrealistic to expect that analyst error will alwaysbe determined and documented. Nevertheless, a laboratory investigationconsists of more than a retest. The inability to identify an error's causewith confidence affects retesting procedures, not the investigation inquiryrequired for the initial OOS result.The firm's analyst should follow a written procedure, checking off each stepas it is completed during the analytical procedure. We expect laboratorytest data to be recorded directly in notebooks; use of scrap paper and loosepaper must be avoided. These common sense measures enhance the accuracy and integrity of data.Review and evaluate the laboratory SOP for product failure investigations.Specific procedures must be followed when single and multiple OOS resultsare investigated. For the single OOS result the investigation should includethe following steps and these inquiries must be conducted before there is aretest of the sample:o the analyst conducting the test should report the OOS result to thesupervisoro the analyst and the supervisor should conduct an informal laboratoryinvestigation which addresses the following areas:1. discuss the testing procedure2. discuss the calculation3. examine the instruments4. review the notebooks containing the OOS resultAn alternative means to invalidate an initial OOS result, provided thefailure investigation proves inconclusive, is the "outlier" test. However,specific restrictions must be placed on the use of this test.1. Firms cannot frequently reject results on this basis.2. The USP standards govern its use in specific cases only.3. The test cannot be used for chemical testing results. An initial contentuniformity test was OOS followed by a passing retest. The initial OOS result was claimed the result of analyst error based on a statistical evaluation ofthe data. The court ruled that the use of an outlier test is inappropriatein this case..4. It is never appropriate to utilize outlier tests for a statisticallybased test, i.e., content uniformity and dissolution.Determine if the firm uses an outlier test and evaluate the SOP.Determine that a full scale inquiry has been made for multiple OOS results. This inquiry involves quality control and quality assurance personnel inaddition to laboratory workers to identify exact process or non processrelated errors.When the laboratory investigation is inconclusive (reason for the error isnot identified) the firm:1. Cannot conduct 2 retests and base release on average of three tests2. Cannot use outlier test in chemical tests3. Cannot use a re-sample to assume a sampling or preparation error4. Can conduct a retest of different tablets from the same sample when aretest is considered appropriate (see criteria elsewhere)C. FORMAL INVESTIGATIONSFormal investigations extending beyond the laboratory must follow an outline with particular attention to corrective action. The company must:1. State the reason for the investigation2. Provide summation of the process sequences that may have caused the problem3. Outline corrective actions necessary to save the batch and preventsimilar recurrence4. List other batches and products possibly affected, the results ofinvestigation of these batches and products, and any corrective action.Specifically:o examine other batches of product made by the errant employee or machine o examine other products produced by the errant process or operation5. Preserve the comments and signatures of all production and qualitycontrol personnel who conducted the investigation and approved anyreprocessed material after additional testingD. INVESTIGATION DOCUMENTATIONAnalyst's mistakes, such as undetected calculation errors, should bespecified with particularity and supported by evidence. Investigations alongwith conclusions reached must be preserved with written documentation that enumerates each step of the investigation. The evaluation, conclusion andcorrective action, if any, should be preserved in an investigation orfailure report and placed into a central file.E. INVESTIGATION TIME FRAMESAll failure investigations should be performed within 20 business days ofthe problem's occurrence and recorded and written into a failure orinvestigation report.6. PRODUCT FAILURESAn OOS laboratory result can be overcome (invalidated) when laboratory error has been documented. However, non-process and process related errorsresulting from operators making mistakes, equipment (other than laboratoryequipment) malfunctions, or a manufacturing process that is fundamentallydeficient, such as an improper mixing time, represent product failures.Examine the results of investigations using the guidance in section 5 aboveand evaluate the decision to release, retest, or rework products.7. RETESTINGEvaluate the company's retesting SOP for compliance with scientificallysound and appropriate procedures. A very important ruling in one recentcourt decision sets forth a procedure to govern the retesting program. Thisdistrict court ruling provides an excellent guide to use in evaluating someaspects of a pharmaceutical laboratory, but should not be considered as law, regulation or binding legal precedent. The court ruled that a firm shouldhave a predetermined testing procedure and it should consider a point atwhich testing ends and the product is evaluated. If results are notsatisfactory, the product is rejected.Additionally, the company should consider all retest results in the contextof the overall record of the product. This includes the history of theproduct. The court ordered a recall of one batch of product on the basis ofan initial content uniformity failure and no basis to invalidate the testresult and on a history of content uniformity problems with the product.,type of test performed, and in-process test results. Failing assay resultscannot be disregarded simply on the basis of acceptable content uniformity results.The number of retests performed before a firm concludes that an unexplained OOS result is invalid or that a product is unacceptable is a matter ofscientific judgment. The goal of retesting is to isolate OOS results butretesting cannot continue ad infinitum.In the case of nonprocess and process-related errors, retesting is suspect.Because the initial tests are genuine, in these circumstances, additionaltesting alone cannot contribute to product quality. The court acknowledgedthat some retesting may precede a finding of nonprocess or process-based errors. Once this determination is made, however, additional retesting forpurposes of testing a product into compliance is not acceptable.For example, in the case of content uniformity testing designed to detectvariability in the blend or tablets, failing and non-failing results are notinherently inconsistent and passing results on limited retesting do not ruleout the possibility that the batch is not uniform. As part of theinvestigation firms should consider the record of previous batches, sincesimilar or related failures on different batches would be a cause ofconcern.Retesting following an OOS result is ruled appropriate only after thefailure investigation is underway and the failure investigation determinesin part whether retesting is appropriate. It is appropriate when analysterror is documented or the review of analyst's work is "inconclusive" , butit is not appropriate for known and undisputed non-process or processrelated errors.The court ruled that retesting:o must be done on the same, not a different sampleo may be done on a second aliquot from the same portion of the sample that was the source of the first aliquoto may be done on a portion of the same larger sample previously collectedfor laboratory purposes8. RESAMPLINGFirms cannot rely on resampling. The court ordered the recall of one batchof product after having concluded that a successful resample result alonecannot invalidate an initial OOS result. to release a product that hasfailed testing and retesting unless the failure investigation disclosesevidence that the original sample is not representative or was improperlyprepared.Evaluate each resampling activity for compliance with this guidance.9. AVERAGING RESULTS OF ANALYSISAveraging can be a rational and valid approach when the object underconsideration is total product assay, but as a general rule this practiceshould be avoided. The court ruled that the firm must recall a batch thatwas released for content uniformity on the basis of averaged test results.because averages hide the variability among individual test results. Thisphenomenon is particularly troubling if testing generates both OOS andpassing individual results which when averaged are within specification.Here, relying on the average figure without examining and explaining theindividual OOS results is highly misleading and unacceptable.Content uniformity and dissolution results never should be averaged toobtain a passing value.In the case of microbiological turbidimetric and plate assays an average ispreferred by the USP. In this case, it is good practice to include OOSresults in the average unless an outlier test (microbiological assays)suggests the OOS is an anomaly.10. BLEND SAMPLING AND TESTINGThe laboratory serves a vital function in blend testing which is necessaryto increase the likelihood of detecting inferior batches. Blend uniformitytesting cannot be waived in favor of total reliance on finished producttesting because finished product testing is limited.One court has ruled that sample size influences ultimate blend test resultsand that the sample size should resemble the dosage size. Any other practice would blur differences in portions of the blend and defeat the object of thetest. If a sample larger than the unit must be taken initially, aliquotswhich resemble the dosage size should be carefully removed for the test,retests, and reserve samples. Obviously, the initial larger sample shouldnot be subjected to any additional mixing or manipulation prior to removingtest aliquots as this may obscure non-homogeneity.Multiple individual blend uniformity samples taken from different areascannot be composited. However when variation testing is not the object ofassay testing, compositing is permitted.If firms sample product from sites other than the blender, they mustdemonstrate through validation that their sampling technique isrepresentative of all portions and concentrations of the blend. This meansthat the samples must be representative of those sites that might beproblems; e.g. weak or hot spots in the blend.11. MICROBIOLOGICALThe review of microbiological data on applicable dosage forms is bestperformed by the microbiologist (analyst). Data that should be reviewedinclude preservative effectiveness testing, bioburden data, and productspecific microbiological testing and methods.Review bioburden (before filtration and/or sterilization) from both anendotoxin and sterility perspective. For drug substance labs evaluatemethods validation and raw data for sterility, endotoxin testing,environmental monitoring, and filter and filtration validation. Also,evaluate the methods used to test and establish bioburdens.Refer to the Microbiological Inspection Guide for additional informationconcerning the inspection of microbiological laboratories.12. SAMPLINGSamples will be collected on pre-approval inspections. Follow the samplingguidelines in CP 7346.832, Part III, pages 5 and 6.13. LABORATORY RECORDS AND DOCUMENTATIONReview personal analytical notebooks kept by the analysts in the laboratoryand compare them with the worksheets and general lab notebooks and records. Be prepared to examine all records and worksheets for accuracy andauthenticity and to verify that raw data are retained to support theconclusions found in laboratory results.Review laboratory logs for the sequence of analysis versus the sequence ofmanufacturing dates. Test dates should correspond to the dates when thesample should have been in the laboratory. If there is a computer data base,determine the protocols for making changes to the data. There should be anaudit trail for changes to data.We expect raw laboratory data to be maintained in bound, (not loose or scrapsheets of paper), books or on analytical sheets for which there isaccountability, such as prenumbered sheets. For most of those manufacturerswhich had duplicate sets of records or "raw data", non-numbered loose sheetsof paper were employed. Some companies use discs or tapes as raw data andfor the storage of data. Such systems have also been accepted provided theyhave been defined (with raw data identified) and validated.Carefully examine and evaluate laboratory logs, worksheets and other recordscontaining the raw data such as weighings, dilutions, the condition ofinstruments, and calculations. Note whether raw data are missing, if recordshave been rewritten, or if correction fluid has been used to conceal errors.Results should not be changed without explanation. Cross reference the datathat has been corrected to authenticate it. Products cannot be "tested intocompliance" by arbitrarily labeling out-of-specification lab results as"laboratory errors" without an investigation resulting in scientificallyvalid criteria.Test results should not have been transcribed without retention of theoriginal records, nor should test results be recorded selectively. Forexample, investigations have uncovered the use of loose sheets of paper withsubsequent selective transcriptions of good data to analyst worksheetsand/or workbooks. Absorbance values and calculations have even been found on desk calendars.Cut charts with injections missing, deletion of files in direct data entrysystems, indirect data entry without verification, and changes tocomputerized programs to override program features should be carefullyexamined. These practices raise questions about the overall quality of data.The firm should have a written explanation when injections, particularlyfrom a series are missing from the official work-sheets or from files andare included among the raw data. Multiple injections recorded should be inconsecutive files with consecutive injection times recorded. Expect to seewritten justification for the deletion of all files.Determine the adequacy of the firm's procedures to ensure that all validlaboratory data are considered by the firm in their determination ofacceptability of components, in-process, finished product, and retainedstability samples. Laboratory logs and documents when cross referenced may show that data has been discarded by company officials who decided torelease the product without a satisfactory explanation of the resultsshowing the product fails to meet the specifications. Evaluate thejustification for disregarding test results that show the product failed tomeet specifications.14. LABORATORY STANDARD SOLUTIONSAscertain that suitable standards are being used (i.e. in-date, storedproperly). Check for the reuse of stock solutions without assuring theirstability. Stock solutions are frequently stored in the laboratoryrefrigerator. Examine the laboratory refrigerators for these solutions andwhen found check for appropriate identification. Review records of standardsolution preparation to assure complete and accurate documentation. It ishighly unlikely that a firm can "accurately and consistently weigh" to thesame microgram. Therefore data showing this level of standardization orpattern is suspect and should be carefully investigated.15. METHODS VALIDATIONInformation regarding the validation of methods should be carefullyevaluated for completeness, accuracy and reliability. In particular, if acompendial method exists, but the firm chooses to use an alternate methodinstead, they must compare the two and demonstrate that the in-house method is equivalent or superior to the official procedure. For compendial methodsfirms must demonstrate that the method works under the actual conditions of use.Methods can be validated in a number of ways. Methods appearing in the USP are considered validated and they are considered validated if part of anapproved ANDA. Also a company can conduct a validation study on theirmethod. System suitability data alone is insufficient for and does notconstitute method validation.In the review of method validation data, it is expected that data forrepetitive testing be consistent and that the varying concentrations of testsolutions provide linear results. Many assay and impurity tests are nowHPLC, and it is expected that the precision of these assays be equal or lessthan the RSD's for system suitability testing. The analytical performanceparameters listed in the USP XXII, <1225>, under the heading of Validationof Compendial Methods, can be used as a guide for determining the analytical parameters (e.g., accuracy, precision, linearity, ruggedness, etc.) neededto validate the method.16. EQUIPMENTLaboratory equipment usage, maintenance, calibration logs, repair records,and maintenance SOPs also should be examined. The existence of the equipment specified in the analytical methods should be confirmed and its conditionnoted. Verify that the equipment was present and in good working order atthe time the batches were analyzed. Determine whether equipment is beingused properly.In addition, verify that the equipment in any application was in goodworking order when it was listed as used to produce clinical or biobatches.One would have to suspect the data that are generated from a piece ofequipment that is known to be defective. Therefore, continuing to use andrelease product on the basis of such equipment represents a seriousviolation of CGMP's.17. RAW MATERIAL TESTINGSome inspections include the coverage of the manufacturer of the drugsubstance. The safety and efficacy of the finished dosage form is largelydependent on the purity and quality of the bulk active drug substance.Examine the raw data reflecting the analysis of the drug substance includingpurity tests, charts, etc.Check the impurity profiles of the BPC used in the biobatch and clinicalproduction batches to determine if it is the same as that being used tomanufacture full scale production batches. Determine if the manufacturer hasa program to audit the certificate of analysis of the BPC, and, if so, checkthe results of these tests. Report findings where there is substantialdifference in impurity profiles and other test results.Some older compendial methods may not be capable of detecting impurities asnecessary to enable the control of the manufacturing process, and newermethods have been developed to test these products. Such methods must bevalidated to ensure that they are adequate for analytical purposes in thecontrol and validation of the BPC manufacturing process. The drug substancemanufacturer must have complete knowledge of the manufacturing process and the potential impurities that may appear in the drug substance. Theseimpurities cannot be evaluated without a suitable method and one that hasbeen validated.Physical tests such as particle size for raw materials, adhesion tests forpatches, and extrusion tests for syringes are essential tests to assureconsistent operation of the production and control system and to assurequality and efficacy. Some of these tests are filed in applications andothers may be established by the protocols used to manufacture the product.The validation of methods for such tests are as important as the test forchemical attributes.Physical properties tests often require the use of unique equipment andprotocols. These tests may not be reproducible in other laboratories,therefore, on site evaluation is essential.。
制药行业微生物实验室检查指南-翻译并解读
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。
美国FDA药品质量控制微生物实验室检查指南
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。
美国FDA药品质量控制微生物实验室检查指南
后， 方可转种。 ３ 用适宜的培养基将冻干菌种稀释、 ． 转种、 培 养、 保存。 该菌种经鉴定无误后可作为对照菌用。 如 发现菌株特性不符， 不得使用。 ４ 实验室菌种保存， ． 可用半固体、 营养琼脂斜 面接种， 戴橡胶塞， 置冰箱保存 （＇） ４ ，或接种于 Ｃ ４ ％甘油、 ０ 其他适宜的培养基于低温（ ０ －２ ＇ Ｃ以上） 保存。如半固体接种， 戴橡胶塞， 冰箱保存，－６ ５ 个 月再转种。 如营养琼脂斜面接种， 戴橡胶塞， 冰箱保 存，－４ ３ 个月再转种。 ５ 对照用菌种在使用过程中， ． 亦应检查其生物 学特性。 如菌落形态、 革兰染色、 镜检菌体形态、 主要 生化特性等有无变异或污染杂菌。如发现染菌或变 异、 衰退等现象时， 应及时分离、 纯化、 复壮。 ６ 保存的各菌种， ． 应控制传代次数， ５ 使用 －６
８ 中凶药吕标准 ２０ 年第 ‘ ０３ 拐第 ３ 期
（ １６ 总 ３）
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药品检验所微生物实验室质量管理规范问题的浅见
苏 德模。 胡昌勒 曹晓云 。 ． Ｏ ｉｉ ｓ Ｍｉ ｏｉ Ｌ ｂｒｔｒ Ｐ ａｔ ｅ Ｓｍ Ｉｓ ｔｔ ｆｒ ｕ Ｃ ｎｒｌ ｐｎｏ ｏ ｃ ｂ ｌ ｏａｏｙ ｃ ｃ ｉ ｏ ｅ ｔ ｕｅ ｏ Ｄ ｇ ｔ ｎ ｆ ｒ ａ ａ ｒ ｉ ｎ ｎ ｉ ｓ ｒ ｏ ｏ
系的组成部分。 也应包括组织机构、 责任、 程序、 过程 和资源。每个专业技术责任人的能力考核、 履行职 责， 按程序接受并检测样品， 审核原始记录及相关数 据， 出具检验报告书。一切设施和环境、 设备和标准
美国微生物检查要求及微生物监控体系
USDA FSIS：《Microbiology Laboratory Guidebook,MLG》 (3RD Edition,1998)
FDA：《Bacteriological Analysis Manual online, MAB》 (2001)
美国官方机构和青岛局食品实 验室对禽肉及产品微生物检测依据
对非商业无菌的即食产品中同时抽样检测沙门氏菌，美国对 即食产品中沙门氏菌的检测样品培养量与众不同，规定为325 克。官方也规定了对与食品接触的台面强制性实施李斯特氏菌 官方检查。要求加工厂实验室可使用国际认可的方法对食品接 触的表面的环境表面进行单核增生李斯特氏、李斯特氏属或类 似李斯特氏其中一种进行检查。但官方确认和调查检验中采用 的方法应为农业部实验室的方法检测单核增生李斯特氏。在从 事调查过程中食品安全局还会使用脉冲凝胶电泳技术，并使用 CDC（疾病控制中心）的数据库进行鉴定。
USDA FSIS对出口国肉类微生物检测要求
对非商业无菌的即食产品的检测频率是具有高风险可能的 产品单核增生李斯特氏菌每月必须抽取一个样品实施检测；如 果同一个工厂有其他产品出口美国还应每月在随即抽取另外一 个样品检测。在即食食品（不包括罐装肉制品）中检测单核增 生李斯特氏菌，检测样品25克，标准为不得检出。
USDA FSIS对实验室质量控制的要求——检测方法
沙门氏菌:
沙门氏菌是USDA FSIS官方实验室的重点检 测项目，同时也要求企业实验室实施该项目的 检测。对于生肉连续抽取51个样品检测，阳性 样品，不得检出。罐装食品不做沙 门氏菌检测。
法(参考采用FDA BAM Chapter4)、沙门氏菌的检测（参考采用FDA BAM Chapter 5）、 霍乱弧菌等致病弧菌的检测（参考采用FDA BAM Chapter 9 ）、单核增生李斯特氏菌 的检验和计数（参考采用FDA BAM Chapter 10）、设备校正、维护及试用验证（等同 采用USDA FSIS MLG） 、实验室培养基制备质量保证通则（参考采用ISO/TS111331:2000(E) & NCCLS M22-A2）、取样和样品保存（参考采用FAO Food and Nutrition Paper-1991）、3M Petrifilm Plate测试卡计数食品中的细菌总数、大肠杆菌、大肠菌群、 肠杆菌科、金黄葡萄球菌、霉菌和酵母菌（参考采用AOAC 963.33、AOAC989.10、 AOAC990.12、AOAC991.14、AOAC997.02、AOAC2000.05 、AOAC998.08）、MINIVIDAS筛选食品中的沙门氏菌、李斯特氏菌、O157、空肠弯曲菌（参考采用AOAC 2001.07、AOAC2001.08、AOAC2001.09、AOAC999.06、AOAC996.08 ）、培养基和 试剂（ 参考采用FDA BAM & USDA FSIS MLG ）、致泻性大肠杆菌的检测（参考采 用FDA BAM Chapter4 A）、菌种保藏、微生物安全手册。
美国FDA制剂厂检查指南CGMP
FDA 制剂厂检查指南对制剂企业的检查-CGMP备注：这个文件是对检查者和其他的FDA 官员的参照。
这份文件并不限制FDA，并不猎取任何利益，义务，权利，或豁免某人1简介这个文件介绍到一个大约的药厂检查指导他们是否符合药物的CGMP。
这个指导应当被用来作IOM 的介绍，其他的药物检查指导，和复核工程。
这个检查的指导相对于IOM 的第十章，一些指导是：A 原料药视察指导B 高纯水系统视察的指导C QC 试验室视察的指导D 微生物QC 试验室的指导E 冻干注射剂视察的指导F清洁验证视察的指导G 制药过程中的计算机系统的视察的指导H 一般工艺过程验证的指导2CGMP处方和非处方全部的药物的生产过程要对应于CGMP 否则就被认为在FDC 实施中掺假。
501(a)(2)(B)依据处方药物的记录肯定要比照于704(a)(1)(B)章。
假设药物是OTC 药物被NDA 或ANDA 掩盖，FDA 可以回忆，复制，验证在505(k)(2)章下的FDC。
然而，假设产品是在FDA 没注册过的OTC 药物。
并没有法定的严格要求在视察过程中给视察者看的的记录要按704 章FDC 的要求。
而且，全部的处方药和OTC 药的生产必需符合CGMP 的要求，包括那些记录。
视察者应当回忆这些记录作为打算是否符合CGMP 要求的一局部。
在很少的场合，可以拒绝回忆OTC 的记录由于并没有法定要求这样做。
当工厂在没有法定义务去供给回忆这些记录时，没有减轻需要符合在501(a)(2)(B)CGMP的要求，包括对主体文件的要求。
假设一个工厂拒绝回忆OTC 的记录，视察者应当被其他的视察方法打算特别符合CGMP 的要求。
视察者观看和查找对于处方药和非处方药并不依据FDA-483 确定的检查表中做的行为。
组织和人员工厂肯定要有QC 把握文件描述CFR 的责任和权威。
QC 文件肯定要在说明它的相对于生产文件的独立性，在书面说明它的责任权属。
说明操作者的官员的名字，头衔和内在的责任，其他的主要的职员在IOM 中列出。
美国药典微生物检测精要
美国药典微生物检测精要引言微生物检测在药品生产中起着至关重要的作用。
药品的微生物污染可能对患者的健康造成严重威胁。
为了确保药品的安全性、可靠性和有效性，美国药典（United States Pharmacopeia, USP）制定了一系列用于微生物检测的指南和标准。
本文将概述美国药典微生物检测的精要内容。
检测方法目视法目视法是最简单、最常用的微生物检测方法之一。
该方法通过对样品进行肉眼观察，检验人员可以检测到样品中是否存在明显的微生物污染。
目视法的优点是操作简单，不需要特殊的设备和试剂，适用于多种药品和原料的微生物检测。
然而，目视法存在主观性和局限性，无法检测到微生物的低水平污染。
营养物质法营养物质法是一种常用的培养基法。
该方法以适宜的培养基为基础，通过对样品进行培养，利用微生物的生长特性来检测样品中是否存在微生物污染。
营养物质法可以检测到微生物的低水平污染，并且可以进一步鉴定和计数微生物。
然而，营养物质法需要一定的培养时间，且存在一定的培养基选择性和适应性的局限性。
生物化学法生物化学法是一种通过检测微生物代谢产物来判断样品中是否存在微生物污染的方法。
该方法利用微生物的代谢反应，检测样品中的特定代谢产物的存在与否。
生物化学法可以快速、准确地检测微生物污染，并且不受培养基选择性和适应性的限制。
然而，生物化学法需要特定的试剂和设备，并且对检测人员的技术要求较高。
检测标准美国药典对药品的微生物检测制定了一系列的标准和要求。
这些标准主要包括微生物限度试验、总微生物数试验、鉴别试验和特定微生物试验等。
微生物限度试验微生物限度试验旨在确定样品中存在的微生物数量的限度。
该试验要求将样品接种在适当的培养基中，经过一定的培养时间后，观察并计数生长的微生物数量。
美国药典针对不同类型的药品和原料制定了不同的微生物限度指标，以确保药品的微生物安全性。
总微生物数试验总微生物数试验用于确定样品中所有可培养的微生物的数量。
该试验要求将样品接种在适当的培养基中，经过一定的培养时间后，观察并计数生长的微生物数量。
FDA 微生物学手册 6 非灭菌产品的微生物检查
FDA微生物学手册6非灭菌产品的微生物检查第二章：非灭菌产品的微生物检查本节包含了对活性微生物的定量计数和成品药品和原料中缺少指定微生物的确定的补充信息，以前称为微生物限度检测（MLT）o这些测试的详细程序不在本PMM章节中说明，因为它们在以下USP中查阅：USPv60＞非无菌产品的微生物检验:洋葱伯克霍尔德氏菌检验；USPv 61＞非无菌产品微生物检验:微生物计数检验；USP＜62＞非无菌产品的微生物检验:对指定微生物的检验。
＜60＞是一种检测制剂或水系辅用制剂中是否存在洋葱伯克霍尔德氏菌的试验。
吸入使用产品或口服、口腔粘膜、皮肤或鼻腔的水溶液辅用制剂。
＜61〉描述了从药品中计数微生物的方法，包括膜过滤、常规平板计数（包括倾注平板法、表面铺开法）和最可能数法（MPN）o ＜62＞描述了特定的富集程序，取决于产品专论要求的必须缺少的目标指定微生物（不得检出的控制菌）。
不溶于水或不分散于水的产品必须经过适当处理，以获得适合于试验程序的悬浮液。
＜1111＞被推荐用于各种药物剂型微生物的鉴定，这对解决问题是有益的。
注意这一点很重要：即使USP根据各论的要求描述了特定微生物的回收和鉴定方法，通常需要确定产品中是否也存在其他有害微生物，并在工作表上报告这些微生物。
在许多情况下，这些可能是机会性的或新出现的病原体，而不是USP＜62＞针对性回收的目标。
鉴别方法，如VITEK,应用于鉴别在USP＜62＞实验中回收的微生物。
替代方法，先进的分子方法（即PCR,测序等）或使用附加的通用富集琼脂平板或无选择性的肉汤，可能更适合检测样品的筛选。
这些附加琼脂或方法的应用可能需要根据分析中药物或产品的目标人群来考虑, 并可能需要与实验室主管进行沟通以获得附加说明。
A.产品的储存和处理样品应在包装标签或说明书规定的存储条件下保存。
1.在产品测试前，单元容器的外部应进行消毒，然后转移到工作站或HEPA过滤层流罩。
如果产品容器不是密封的，不要将产品容器浸泡在消毒溶液中，这样可能会使杀菌溶液进入产品。
FDA QC实验室检查指南
GUIDE TO INSPECTIONS OF 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).1. INTRODUCTIONThe pharmaceutical quality control laboratory serves one of the mostimportant functions in pharmaceutical production and control. A significantportion of the CGMP regulations (21 CFR 211) pertain to the quality controllaboratory and product testing. Similar concepts apply to bulk drugs.This inspection guide supplements other inspectional information containedin other agency inspectional guidance documents. For example, ComplianceProgram 7346.832 requiring pre-approval NDA/ANDA inspections containsgeneral instructions to conduct product specific NDA/ANDA inspection auditsto measure compliance with the applications and CGMP requirements. Thisincludes pharmaceutical laboratories used for in-process and finishedproduct testing.2. OBJECTIVEThe specific objective will be spelled out prior to the inspection. Thelaboratory inspection may be limited to specific issues, or the inspectionmay encompass a comprehensive evaluation of the laboratory's compliance withCGMP's. As a minimum, each pharmaceutical quality control laboratory shouldreceive a comprehensive GMP evaluation each two years as part of thestatutory inspection obligation.In general these inspections may include-- the specific methodology which will be used to test a new product-- a complete assessment of laboratory's conformance with GMP's-- a specific aspect of laboratory operations3. INSPECTION PREPARATIONFDA Inspection Guides are based on the team inspection approach and ourinspection of a laboratory is consistent with this concept. As part of oureffort to achieve uniformity and consistency in laboratory inspections, weexpect that complex, highly technical and specialized testing equipment,procedures and data manipulations, as well as scientific laboratoryoperations will be evaluated by an experienced laboratory analyst withspecialized knowledge in such matters.District management makes the final decision regarding the assignment ofpersonnel to inspections. Nevertheless, we expect investigators, analystsand others to work as teams and to advise management when additionalexpertise is required to complete a meaningful inspection.Team members participating in a pre-approval inspection must read and befamiliar with Compliance Program 7346.832, Pre-ApprovalInspections/Investigations. Relevant sections of the NDA or ANDA should be reviewed prior to the inspection; but if the application is not availablefrom any other source, this review will have to be conducted using thecompany's copy of the application.Team members should meet, if possible, prior to the inspection to discussthe approach to the inspection, to define the roles of the team members, and to establish goals for completion of the assignment. Responsibilities fordevelopment of all reports should also be established prior to theinspection. This includes the preparation of the FDA 483.The Center for Drug Evaluation and Research (CDER) may have issueddeficiency letters listing problems that the sponsor must correct prior to the approval of NDA/ANDA's and supplements. The inspection team is expected to review such letters on file at the district office, and they are expected to ask the plant for access to such letters. The team should evaluate thereplies to these letters to assure that the data are accurate and authentic. Complete the inspection even though there has been no response to theseletters or when the response is judged inadequate.4. INSPECTION APPROACHA. GeneralIn addition to the general approach utilized in a drug CGMP inspection, the inspection of a laboratory requires the use of observations of thelaboratory in operation and of the raw laboratory data to evaluatecompliance with CGMP's and to specifically carry out the commitments in an application or DMF. When conducting a comprehensive inspection of alaboratory, all aspects of the laboratory operations will be evaluated.Laboratory records and logs represent a vital source of information thatallows a complete overview of the technical ability of the staff and ofoverall quality control procedures. SOPs should be complete and adequate and the operations of the laboratories should conform to the written procedures. Specifications and analytical procedures should be suitable and, asapplicable, in conformance with application commitments and compendialrequirements.Evaluate raw laboratory data, laboratory procedures and methods, laboratory equipment,including maintenance and calibration, and methods validation data to determine the overall quality of the laboratory operation and the ability to comply with CGMP regulations.Examine chromatograms and spectra for evidence of impurities, poortechnique, or lack of instrument calibration.s use systems that provide for the investigation oflaboratory test failures. These are generally recorded in some type of log. Ask to see results of analyses for lots of product that have failed to meet specifications and review the analysis of lots that have been retested,rejected, or reworked. Evaluate the decision to release lots of product when the laboratory results indicate that the lot failed to meet specifications and determine who released them.B. Pre-ApprovalDocuments relating to the formulation of the product, synthesis of the bulk drug substance, product specifications, analysis of the product, and others are examined during the review process in headquarters. However, thesereviews and evaluations depend on accurate and authentic data that trulyrepresents the product.Pre-approval inspections are designed to determine if the data submitted in an application are authentic and accurate and if the procedures listed inthe application were actually used to produce the data contained in theapplication. Additionally, they are designed to confirm that plants(including the quality control laboratory) are in compliance with CGMPregulations.The analytical sections of drug applications usually contain only testresults and the methods used to obtain them. Sponsors are not required tofile all the test data because such action would require voluminoussubmissions and would often result in filing redundant information. Sponsors may deliberately or unintentionally select and report data showing that adrug is safe and effective and deserves to be approved. The inspection team must decide if there is valid and scientific justification for the failure to report data which demonstrates the product failed to meet itspredetermined specifications.Coordination between headquarters and the field is essential for a complete review of the application and the plant. Experienced investigators andanalysts may contact the review chemist (with appropriate supervisoryconcurrence) when questions concerning specifications and standards arise.Inspections should compare the results of analyses submitted with results of analysis of other batches that may have been produced. Evaluate the methods and note any exceptions to the procedures or equipment actually used fromthose listed in the application and confirm that it is the same methodlisted in the application. The analyst is expected to evaluate rawlaboratory data for tests performed on the test batches (biobatches andclinical batches) and to compare this raw data to the data filed in theapplication.5. FAILURE (OUT-OF-SPECIFICATION) LABORATORY RESULTSEvaluate the company's system to investigate laboratory test failures. These investigations represent a key issue in deciding whether a product may bereleased or rejected and form the basis for retesting, and resampling.In a recent court decision the judge used the term "out-of-specification"(OOS) laboratory result rather than the term "product failure" which is morecommon to FDA investigators and analysts. He ruled that an OOS resultidentified as a laboratory error by a failure investigation or an outliertest. The court provided explicit limitations on the use of outlier testsand these are discussed in a later segment of this document., or overcome by retesting. The court ruled on the use of retesting which is covered in alater segment of this document. is not a product failure. OOS results fall into three categories:-- laboratory error-- non-process related or operator error-- process related or manufacturing process errorA. LABORATORY ERRORSLaboratory errors occur when analysts make mistakes in following the method of analysis, use incorrect standards, and/or simply miscalculate the data. Laboratory errors must be determined through a failure investigation toidentify the cause of the OOS. Once the nature of the OOS result has beenidentified it can be classified into one of the three categories above. The inquiry may vary with the object under investigation.B. LABORATORY INVESTIGATIONSThe exact cause of analyst error or mistake can be difficult to determinespecifically and it is unrealistic to expect that analyst error will always be determined and documented. Nevertheless, a laboratory investigationconsists of more than a retest. The inability to identify an error's cause with confidence affects retesting procedures, not the investigation inquiry required for the initial OOS result.The firm's analyst should follow a written procedure, checking off each step as it is completed during the analytical procedure. We expect laboratorytest data to be recorded directly in notebooks; use of scrap paper and loose paper must be avoided. These common sense measures enhance the accuracy and integrity of data.Review and evaluate the laboratory SOP for product failure investigations. Specific procedures must be followed when single and multiple OOS resultsare investigated. For the single OOS result the investigation should include the following steps and these inquiries must be conducted before there is a retest of the sample:o the analyst conducting the test should report the OOS result to thesupervisoro the analyst and the supervisor should conduct an informal laboratoryinvestigation which addresses the following areas:1. discuss the testing procedure2. discuss the calculation3. examine the instruments4. review the notebooks containing the OOS resultAn alternative means to invalidate an initial OOS result, provided thefailure investigation proves inconclusive, is the "outlier" test. However, specific restrictions must be placed on the use of this test.1. Firms cannot frequently reject results on this basis.2. The USP standards govern its use in specific cases only.3. The test cannot be used for chemical testing results. An initial content uniformity test was OOS followed by a passing retest. The initial OOS result was claimed the result of analyst error based on a statistical evaluation of the data. The court ruled that the use of an outlier test is inappropriate in this case..4. It is never appropriate to utilize outlier tests for a statisticallybased test, i.e., content uniformity and dissolution.Determine if the firm uses an outlier test and evaluate the SOP.Determine that a full scale inquiry has been made for multiple OOS results. This inquiry involves quality control and quality assurance personnel inaddition to laboratory workers to identify exact process or non processrelated errors.When the laboratory investigation is inconclusive (reason for the error is not identified) the firm:1. Cannot conduct 2 retests and base release on average of three tests2. Cannot use outlier test in chemical tests3. Cannot use a re-sample to assume a sampling or preparation error4. Can conduct a retest of different tablets from the same sample when aretest is considered appropriate (see criteria elsewhere)C. FORMAL INVESTIGATIONSFormal investigations extending beyond the laboratory must follow an outline with particular attention to corrective action. The company must:1. State the reason for the investigation2. Provide summation of the process sequences that may have caused theproblem3. Outline corrective actions necessary to save the batch and preventsimilar recurrence4. List other batches and products possibly affected, the results ofinvestigation of these batches and products, and any corrective action.Specifically:o examine other batches of product made by the errant employee or machineo examine other products produced by the errant process or operation5. Preserve the comments and signatures of all production and qualitycontrol personnel who conducted the investigation and approved anyreprocessed material after additional testingD. INVESTIGATION DOCUMENTATIONAnalyst's mistakes, such as undetected calculation errors, should bespecified with particularity and supported by evidence. Investigations along with conclusions reached must be preserved with written documentation that enumerates each step of the investigation. The evaluation, conclusion andcorrective action, if any, should be preserved in an investigation orfailure report and placed into a central file.E. INVESTIGATION TIME FRAMESAll failure investigations should be performed within 20 business days ofthe problem's occurrence and recorded and written into a failure orinvestigation report.6. PRODUCT FAILURESAn OOS laboratory result can be overcome (invalidated) when laboratory error has been documented. However, non-process and process related errorsresulting from operators making mistakes, equipment (other than laboratory equipment) malfunctions, or a manufacturing process that is fundamentallydeficient, such as an improper mixing time, represent product failures.Examine the results of investigations using the guidance in section 5 above and evaluate the decision to release, retest, or rework products.7. RETESTINGEvaluate the company's retesting SOP for compliance with scientificallysound and appropriate procedures. A very important ruling in one recentcourt decision sets forth a procedure to govern the retesting program. This district court ruling provides an excellent guide to use in evaluating some aspects of a pharmaceutical laboratory, but should not be considered as law, regulation or binding legal precedent. The court ruled that a firm shouldhave a predetermined testing procedure and it should consider a point atwhich testing ends and the product is evaluated. If results are notsatisfactory, the product is rejected.Additionally, the company should consider all retest results in the context of the overall record of the product. This includes the history of theproduct. The court ordered a recall of one batch of product on the basis of an initial content uniformity failure and no basis to invalidate the testresult and on a history of content uniformity problems with the product.,type of test performed, and in-process test results. Failing assay results cannot be disregarded simply on the basis of acceptable content uniformity results.The number of retests performed before a firm concludes that an unexplained OOS result is invalid or that a product is unacceptable is a matter ofscientific judgment. The goal of retesting is to isolate OOS results butretesting cannot continue ad infinitum.In the case of nonprocess and process-related errors, retesting is suspect. Because the initial tests are genuine, in these circumstances, additionaltesting alone cannot contribute to product quality. The court acknowledged that some retesting may precede a finding of nonprocess or process-basederrors. Once this determination is made, however, additional retesting for purposes of testing a product into compliance is not acceptable.For example, in the case of content uniformity testing designed to detectvariability in the blend or tablets, failing and non-failing results are not inherently inconsistent and passing results on limited retesting do not rule out the possibility that the batch is not uniform. As part of theinvestigation firms should consider the record of previous batches, sincesimilar or related failures on different batches would be a cause ofconcern.Retesting following an OOS result is ruled appropriate only after thefailure investigation is underway and the failure investigation determines in part whether retesting is appropriate. It is appropriate when analysterror is documented or the review of analyst's work is "inconclusive" , but it is not appropriate for known and undisputed non-process or processrelated errors.The court ruled that retesting:o must be done on the same, not a different sampleo may be done on a second aliquot from the same portion of the sample that was the source of the first aliquoto may be done on a portion of the same larger sample previously collectedfor laboratory purposes8. RESAMPLINGFirms cannot rely on resampling. The court ordered the recall of one batch of product after having concluded that a successful resample result alonecannot invalidate an initial OOS result. to release a product that hasfailed testing and retesting unless the failure investigation disclosesevidence that the original sample is not representative or was improperlyprepared.Evaluate each resampling activity for compliance with this guidance.9. AVERAGING RESULTS OF ANALYSISAveraging can be a rational and valid approach when the object underconsideration is total product assay, but as a general rule this practiceshould be avoided. The court ruled that the firm must recall a batch thatwas released for content uniformity on the basis of averaged test results. because averages hide the variability among individual test results. Thisphenomenon is particularly troubling if testing generates both OOS andpassing individual results which when averaged are within specification.Here, relying on the average figure without examining and explaining theindividual OOS results is highly misleading and unacceptable.Content uniformity and dissolution results never should be averaged toobtain a passing value.In the case of microbiological turbidimetric and plate assays an average is preferred by the USP. In this case, it is good practice to include OOSresults in the average unless an outlier test (microbiological assays)suggests the OOS is an anomaly.10. BLEND SAMPLING AND TESTINGThe laboratory serves a vital function in blend testing which is necessary to increase the likelihood of detecting inferior batches. Blend uniformity testing cannot be waived in favor of total reliance on finished producttesting because finished product testing is limited.One court has ruled that sample size influences ultimate blend test results and that the sample size should resemble the dosage size. Any other practice would blur differences in portions of the blend and defeat the object of the test. If a sample larger than the unit must be taken initially, aliquotswhich resemble the dosage size should be carefully removed for the test,retests, and reserve samples. Obviously, the initial larger sample shouldnot be subjected to any additional mixing or manipulation prior to removing test aliquots as this may obscure non-homogeneity.Multiple individual blend uniformity samples taken from different areascannot be composited. However when variation testing is not the object ofassay testing, compositing is permitted.If firms sample product from sites other than the blender, they mustdemonstrate through validation that their sampling technique isrepresentative of all portions and concentrations of the blend. This means that the samples must be representative of those sites that might beproblems; e.g. weak or hot spots in the blend.11. MICROBIOLOGICALThe review of microbiological data on applicable dosage forms is bestperformed by the microbiologist (analyst). Data that should be reviewedinclude preservative effectiveness testing, bioburden data, and productspecific microbiological testing and methods.Review bioburden (before filtration and/or sterilization) from both anendotoxin and sterility perspective. For drug substance labs evaluatemethods validation and raw data for sterility, endotoxin testing,environmental monitoring, and filter and filtration validation. Also,evaluate the methods used to test and establish bioburdens.Refer to the Microbiological Inspection Guide for additional informationconcerning the inspection of microbiological laboratories.12. SAMPLINGSamples will be collected on pre-approval inspections. Follow the sampling guidelines in CP 7346.832, Part III, pages 5 and 6.13. LABORATORY RECORDS AND DOCUMENTATIONReview personal analytical notebooks kept by the analysts in the laboratory and compare them with the worksheets and general lab notebooks and records. Be prepared to examine all records and worksheets for accuracy andauthenticity and to verify that raw data are retained to support theconclusions found in laboratory results.Review laboratory logs for the sequence of analysis versus the sequence of manufacturing dates. Test dates should correspond to the dates when thesample should have been in the laboratory. If there is a computer data base, determine the protocols for making changes to the data. There should be an audit trail for changes to data.We expect raw laboratory data to be maintained in bound, (not loose or scrap sheets of paper), books or on analytical sheets for which there isaccountability, such as prenumbered sheets. For most of those manufacturers which had duplicate sets of records or "raw data", non-numbered loose sheets of paper were employed. Some companies use discs or tapes as raw data andfor the storage of data. Such systems have also been accepted provided they have been defined (with raw data identified) and validated.Carefully examine and evaluate laboratory logs, worksheets and other records containing the raw data such as weighings, dilutions, the condition ofinstruments, and calculations. Note whether raw data are missing, if records have been rewritten, or if correction fluid has been used to conceal errors. Results should not be changed without explanation. Cross reference the data that has been corrected to authenticate it. Products cannot be "tested into compliance" by arbitrarily labeling out-of-specification lab results as"laboratory errors" without an investigation resulting in scientificallyvalid criteria.Test results should not have been transcribed without retention of theoriginal records, nor should test results be recorded selectively. Forexample, investigations have uncovered the use of loose sheets of paper with subsequent selective transcriptions of good data to analyst worksheetsand/or workbooks. Absorbance values and calculations have even been found on desk calendars.Cut charts with injections missing, deletion of files in direct data entry systems, indirect data entry without verification, and changes tocomputerized programs to override program features should be carefullyexamined. These practices raise questions about the overall quality of data.The firm should have a written explanation when injections, particularlyfrom a series are missing from the official work-sheets or from files andare included among the raw data. Multiple injections recorded should be in consecutive files with consecutive injection times recorded. Expect to see written justification for the deletion of all files.Determine the adequacy of the firm's procedures to ensure that all validlaboratory data are considered by the firm in their determination ofacceptability of components, in-process, finished product, and retainedstability samples. Laboratory logs and documents when cross referenced may show that data has been discarded by company officials who decided torelease the product without a satisfactory explanation of the resultsshowing the product fails to meet the specifications. Evaluate thejustification for disregarding test results that show the product failed to meet specifications.14. LABORATORY STANDARD SOLUTIONSAscertain that suitable standards are being used (i.e. in-date, storedproperly). Check for the reuse of stock solutions without assuring theirstability. Stock solutions are frequently stored in the laboratoryrefrigerator. Examine the laboratory refrigerators for these solutions and when found check for appropriate identification. Review records of standard solution preparation to assure complete and accurate documentation. It ishighly unlikely that a firm can "accurately and consistently weigh" to the same microgram. Therefore data showing this level of standardization orpattern is suspect and should be carefully investigated.15. METHODS VALIDATIONInformation regarding the validation of methods should be carefullyevaluated for completeness, accuracy and reliability. In particular, if acompendial method exists, but the firm chooses to use an alternate methodinstead, they must compare the two and demonstrate that the in-house method is equivalent or superior to the official procedure. For compendial methods firms must demonstrate that the method works under the actual conditions of use.Methods can be validated in a number of ways. Methods appearing in the USP are considered validated and they are considered validated if part of anapproved ANDA. Also a company can conduct a validation study on theirmethod. System suitability data alone is insufficient for and does notconstitute method validation.In the review of method validation data, it is expected that data forrepetitive testing be consistent and that the varying concentrations of test solutions provide linear results. Many assay and impurity tests are nowHPLC, and it is expected that the precision of these assays be equal or less than the RSD's for system suitability testing. The analytical performanceparameters listed in the USP XXII, <1225>, under the heading of Validation of Compendial Methods, can be used as a guide for determining the analytical parameters (e.g., accuracy, precision, linearity, ruggedness, etc.) needed to validate the method.16. EQUIPMENTLaboratory equipment usage, maintenance, calibration logs, repair records, and maintenance SOPs also should be examined. The existence of the equipment specified in the analytical methods should be confirmed and its conditionnoted. Verify that the equipment was present and in good working order atthe time the batches were analyzed. Determine whether equipment is beingused properly.In addition, verify that the equipment in any application was in goodworking order when it was listed as used to produce clinical or biobatches. One would have to suspect the data that are generated from a piece ofequipment that is known to be defective. Therefore, continuing to use andrelease product on the basis of such equipment represents a seriousviolation of CGMP's.17. RAW MATERIAL TESTINGSome inspections include the coverage of the manufacturer of the drugsubstance. The safety and efficacy of the finished dosage form is largelydependent on the purity and quality of the bulk active drug substance.Examine the raw data reflecting the analysis of the drug substance including purity tests, charts, etc.Check the impurity profiles of the BPC used in the biobatch and clinicalproduction batches to determine if it is the same as that being used tomanufacture full scale production batches. Determine if the manufacturer has a program to audit the certificate of analysis of the BPC, and, if so, check the results of these tests. Report findings where there is substantialdifference in impurity profiles and other test results.Some older compendial methods may not be capable of detecting impurities as necessary to enable the control of the manufacturing process, and newermethods have been developed to test these products. Such methods must bevalidated to ensure that they are adequate for analytical purposes in thecontrol and validation of the BPC manufacturing process. The drug substance manufacturer must have complete knowledge of the manufacturing process and the potential impurities that may appear in the drug substance. Theseimpurities cannot be evaluated without a suitable method and one that hasbeen validated.Physical tests such as particle size for raw materials, adhesion tests for patches, and extrusion tests for syringes are essential tests to assureconsistent operation of the production and control system and to assurequality and efficacy. Some of these tests are filed in applications andothers may be established by the protocols used to manufacture the product. The validation of methods for such tests are as important as the test forchemical attributes.Physical properties tests often require the use of unique equipment andprotocols. These tests may not be reproducible in other laboratories,therefore, on site evaluation is essential.。
FDA微生物实验室检查指南
GUIDE TO INSPECTIONS OF MICROBIOLOGICAL PHARMACEUTICAL QUALITY CONTROL LABORATORIES微生物实验室检查指南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 any person(s).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 aguide 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.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。
美国FDA药品质量控制微生物实验室检查指南
美国FDA药品质量控制微生物实验室检查指南1993年I．导言《药品质量控制实验室检查指南》主要涉及许多有关药品实验室分析的化学方面的问题，对微生物实验室的检查仅提供了有限的指导，而本指南则是微生物分析检查过程的指导。
本指南建议，如同对任何实验室检查—样，在检查微生物实验室时，应有—名熟悉检验的分析学家(微生物学家)参与。
II．非无菌药品的微生物检验由于种种原因，局部药品、滴鼻剂和吸入剂存在许多微生物污染方面的问题.美国药典‘‘微生物属性”章(1111)指出“应该从药品用法、药品性质及时患者的潜在危害等方面评价微生物在非无菌药品中的重要性”，除此之外，没有提供具体的指导。
美国药典还建议应对某些种类的非无菌药品做常规的总菌数检验及某些特定的污染指示微生物的检验：例如：植物、动物和某些矿物质中的沙门氏菌属；口服液体中的大肠杆菌；局部用药品小的金黄色倘萄球菌和绿脓杆菌污染；以及：自肠、尿道、阴道用药中的酵母菌和霉菌：大量的专题文章还沦及厂微生物的限度。
作为非无菌药品受微生物污染的可接受程度和类型的—般性指导，美国食品和药品管理局药品分局的邓尼根博士曾强调其对健康的危害问题。
1970年他提出被革兰氏阴性细菌污染的局部制剂可能引起中度至重度的健康危害：文献和调查表明，许多感染都源于这种局部药品的革兰氏阴性细菌污染。
几年前马萨诸塞州的—家医院就报道过—宗络合碘(Povidonelodine)被洋葱假恤孢菌(Pseudomonas cepacia)污染的典型病例。
因此，每家公司都希望为自己的非无菌药品制订出一种关于微生物限度的标准，美国药典中“微生物限度”(USP61)提供了检验几种指示微生物的方法，但并末涉及所有有害微生物。
例如医药界普遍认为，洋葱假中孢菌在局部药品或滴鼻剂斗，大量存在是有害的，但美国药典没有提供证明这种微生物存在的检验方法。
间羟异丙肾上腺素硫酸盐吸入剂溶液的收回就是这方面的—个例子。
美国药典第XⅫ版各论部分没有要求对这种药品进行微生物检验。
美国FDA药品质量控制微生物实验室检查指南
美国FDA药品质量控制微生物实验室检查指南
钱维清;潘有友
【期刊名称】《中国药品标准》
【年(卷),期】2001(002)002
【摘要】@@ 译者按:rn值此2000年版药典出版发行、执行之际,为了更好地理解药典中无菌和微生物限度检查法的增、修订内容;了解药品微生物质量控制在制药行业及质量检验中的重要性;以及今后发展的方向,特将美国FDA<药品质量控制微生物实验室检查指南>一文翻译推荐给大家.通过该文,可以了解和借鉴FDA的一些先进的观点.如:对化学、生化药品是否需要进行微生物限度检查?如何评价微生物污染在非无菌药品中的重要性.制订非无菌药品微生物限度的原则.无菌和微生物限度检查的培养时间、初试阳性结果后,是否复试及如何复试的规定.微生物实验中建立阳性、阴性对照的重要性.参加检查或验收微生物实验室时应重视和注意的问题等.
【总页数】4页(P60-62,64)
【作者】钱维清;潘有友
【作者单位】上海市药品检验所,上海,200233;无锡华瑞制药有限公司,无
锡,214000
【正文语种】中文
【中图分类】R9
【相关文献】
1.对美国FDA《植物药品企业指南》(草案)几点剖视 [J], 祝国光
2.美国FDA公布药品安全信息沟通指南 [J], 郑晓红（摘）
3.美国FDA关于《由生物工程操作的植物来源的人与动物用药品、生物制品和医疗器械工业生产指南(草案)》(讨论稿)介绍 [J], 张雪梅
4.美国FDA公布了最新版《医疗器械、药品以及其他产品分类指南》 [J],
5.美国FDA发布3个药品风险管理指南 [J],
因版权原因，仅展示原文概要，查看原文内容请购买。
FDA检查员指导手册
原料药生产检查（药物质量保证）目录现场检查汇报规定 (55)第I部分背景 (56)第II部分实行 (57)第III部分检查 (58)第IV部分分析 (63)第V部分法规/行政方略 (65)第VI部分参照资料，附件和联络接触方式 (68)第VII部分中心旳职责 (69)附件A (69)附件B (72)现场检查汇报规定工艺专论汇报在API检查时，要使用下列旳分类进行汇报所检查旳工艺状况1．Non Sterile API by Chemical Synthesis CSN化学合成非无菌原料CSN2．Sterile API by Chemical Synthesis CSS化学合成无菌原料药CSS3．Non Sterile API by Fermentation CFN发酵生产旳非无菌原料CFN4．Sterile API by Fermentation CFS发酵生产旳无菌原料CFS5．Plant/Animal Extraction API CEX植物/动物提取原料药CEX6．Biotechnology API CBI生物技术生产旳原料药CBI第I部分――背景至八十年代后期以来，美国食品与药物管理局以强化了其对原料药(API)生产企业旳检查内容。
从部分方面来说，这归咎于对原料药质量在制剂旳质量、效力、和安全面所起旳重要作用认识旳提高。
例如，在配制成固体口服制剂，混悬剂和局部用药时原料药旳化学特性会对制剂旳溶出度/生物运用度产生不利影响。
此外，原料中旳少许没有鉴别出旳杂质或其特性未知旳杂质会给病人导致旳严重不良反应。
FDA长期以来一直认为，收载在制剂药物生产质量管理规范规定(21 CFR 210 and 211)中旳CGMP概念对原料药生产工艺同样有效。
这些概念包括，与其他一起，产品质量是生产出来旳，雇佣可以胜任和通过培训旳员工，建立合适旳书面程序和管理规定，建立一套在线测试和产品测试系统，工艺验证，和保证原料药在预期旳有效期内质量稳定。
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美国FDA药品质量控制微生物实验室检查指南1993年I．导言《药品质量控制实验室检查指南》主要涉及许多有关药品实验室分析的化学方面的问题，对微生物实验室的检查仅提供了有限的指导，而本指南则是微生物分析检查过程的指导。
本指南建议，如同对任何实验室检查—样，在检查微生物实验室时，应有—名熟悉检验的分析学家(微生物学家)参与。
II．非无菌药品的微生物检验由于种种原因，局部药品、滴鼻剂和吸入剂存在许多微生物污染方面的问题.美国药典‘‘微生物属性”章(1111)指出“应该从药品用法、药品性质及时患者的潜在危害等方面评价微生物在非无菌药品中的重要性”，除此之外，没有提供具体的指导。
美国药典还建议应对某些种类的非无菌药品做常规的总菌数检验及某些特定的污染指示微生物的检验：例如：植物、动物和某些矿物质中的沙门氏菌属；口服液体中的大肠杆菌；局部用药品小的金黄色倘萄球菌和绿脓杆菌污染；以及：自肠、尿道、阴道用药中的酵母菌和霉菌：大量的专题文章还沦及厂微生物的限度。
作为非无菌药品受微生物污染的可接受程度和类型的—般性指导，美国食品和药品管理局药品分局的邓尼根博士曾强调其对健康的危害问题。
1970年他提出被革兰氏阴性细菌污染的局部制剂可能引起中度至重度的健康危害：文献和调查表明，许多感染都源于这种局部药品的革兰氏阴性细菌污染。
几年前马萨诸塞州的—家医院就报道过—宗络合碘(Povidonelodine)被洋葱假恤孢菌(Pseudomonas cepacia)污染的典型病例。
因此，每家公司都希望为自己的非无菌药品制订出一种关于微生物限度的标准，美国药典中“微生物限度”(USP61)提供了检验几种指示微生物的方法，但并末涉及所有有害微生物。
例如医药界普遍认为，洋葱假中孢菌在局部药品或滴鼻剂斗，大量存在是有害的，但美国药典没有提供证明这种微生物存在的检验方法。
间羟异丙肾上腺素硫酸盐吸入剂溶液的收回就是这方面的—个例子。
美国药典第XⅫ版各论部分没有要求对这种药品进行微生物检验。
药品管理局将其列为一级收回，因为此药受到了洋葱假单孢菌污染。
健康危害评估表明，这种微生物对肺部感染的风险很大，特别是对某些患有慢性气管阻塞、囊性纤维变性和免疫缺陷等病的患者具有潜在的生命危险。
此外，药典的微生物限度部分所描述的检验程度不能鉴别这些微生物。
现行美国药典在“微生物限度”部分(61)做了复检的规定，但是许多建议要求取消这种复检条款。
类似于其他检验，初次检验结果应予以审查与研究。
微生物污染并不是均匀地分布在一批药品或样品中的。
如果在一个样品中发现污染而在另—次取样样品中没有发现，则不应忽视初次发现的结果。
复检的结果应予以审查与评估，并且应特别注意进行复检的逻辑性和合理性。
为了分离出特定的微生物污染物，FDA实验室和制药工业的许多实验室，使用某些自钝化剂如吐温或卵磷脂的营养培养基，这对于钝化药品中常有的防腐剂的作用是非常必要的，还能为受损的或生长缓慢的微生物提供更好的基质；其他生长参数包括降低培养温度以及延长培养时间(至少5天)，因为它们可以为这类微生物提供良好的生存条件。
例如：在《细菌学分析手册》(BAM)第Ⅵ版中，FDA实验室采用化妆品的检验方法鉴别非无菌药品中的污染物。
这项检验包括将样品置于改良Letheen肉汤中培养。
培养结束后，再用血琼脂平皿和麦球凯琼脂平皿进一步鉴定，然后再鉴别分离出的菌落。
FDA微生物学家用这种方法使所有潜在的病原体的复活达到最佳，并且测定复活的微生物数量及其类别。
FDA分析学家使用这种方法的另—个重要方面就是要确定所有用过的培养暴促进微生物生长的性能。
选择适当的中和剂很大程度上取决于防腐剂的性质和被评价药品所智的配方：如果在营养肉汤出现微生物生长，那么下一步的鉴别就可将样品转到更有选择性的琼脂培养基或适当的增菌琼脂上。
微生物检验可包括对在需氧菌总数检验小发现的菌落的鉴定。
另外上述鉴定不应仅限于美国药典中的指示微生物。
鉴别从微生物总数检验或(和)富集培养检验中分离的各种菌落的重要性，将取决于药品的种类及共其途。
很明显，如果检验—个口服固体制剂如片剂，当检验显示微生物污染量很大时，就可以考虑对分离出的菌落进行鉴别，然而对其他被微生物污染后易引起危害的药品，如局部用药品、吸人剂、滴鼻剂，应鉴别从前落计数及富集检验中分离的菌落。
III．设备、器材和培养基首先审查正在进行的分析，并检查培养中的培养皿．和试管(操作过程中不要疏忽大意而污染检验中的培养皿和试管)。
特别注意那些没有文字记录的复检和经凋查已经鉴别出有污染问题的“特殊项目”。
这种评价可以通过审查正在进行的分析是否有阳性结果来实现。
如有可能要审查前一天的培养皿和培养基，还要把观察结果与实验记录本中的记录进行比较。
检查用于培养基灭菌的高压蒸汽灭菌器。
高压蒸汽灭菌器可能缺少以无菌过滤的空气来代替蒸汽的能力。
对于密封的培养瓶，不会出现这种问题；然而对于不密封的培养瓶，非无菌的空气会导致培养基的污染。
此外，如高压灭菌的时间少于要求的时间，也会使培养基中的污染物生长，并得出错误的阳性结果。
在任务繁重的实验室里，这些问题可能更容易出现。
要检查高压蒸汽灭菌器的温度，温度过高会使必需的营养素变性甚至炭化，这可能使受到抑制的微生物达不到最佳复活。
潜在的假阳性结果带来的明显问题，是无法区别因疏忽大意而产生的培养基污染和直接来源于被检验样品的真正的污染。
Ⅳ。
无菌检验1991年10月11 日，药品管理局颁布了关于经无菌加工艺和最终灭菌生产药品的规定，并列出—份经无菌加工工艺生产的受污染或潜在污染而致收回的药品清单。
许多对收回药品的调查或检查都起因于初次无菌检验不合格。
FDA对生产厂的生产、质控、调查及其缺陷进行中核的结果以及最初无菌检验不合格的证据最终导致厂对这些药品的收回。
美国药典规定，用于进行无菌检验的设施与用于制药的设施应该是一致的，并规定，用于无菌检验的设施不应比无菌加工生产设施造成更多的微生物污染机率。
因此，合理的设计应该包括更衣区域和通过气闸。
环境的监测和着装应该与药品生产中相一致。
由于无菌检验中包括大量药品和培养基的操作，因此检查应包括对无菌检验的实地观察；即使有些公司曾借口实地检查可能会使分析人员紧张而试图阻止，检查组应注意此问题，使实地观察能以尽可能不干扰正常操作环境的方式进行。
然而这种考虑不足以成为放弃这部分检查的充分理由。
检查无菌检验程序最重要的方面之一就是审查元菌检验初次阳性结果的记录。
要求列出不合格的检验结果以便于审查生产、质控记录和调查报告，特别是对于风险性很大的无菌分装的药品，应审查无菌检验初次阳性结果和调查结果。
对于生产厂来说，如果无菌分装的产品在初次无菌检验时出现阳性结果，但无菌检验中的对照未发现异常，则很难决定是否发放该种产品。
应检查是否设立阴性对照品，设立阴性对照对高质量的无菌检验儿为重要；无菌检验规范的做法包括使用已知的最终灭菌或经辐射灭菌的样品作为系统对照，或者使用培养基模拟分装试验中分装的安瓿或西林瓶。
要特别注意生产无菌分装药品的厂家从未发现初次阳性无菌检验结果的情况，尽管这种情况可能发生，但毕竟少见。
曾经有一个案例，某生产厂的记录表明没有阳性结果，实际上他们的记录被改动了。
不存在初次阳性结果，还说明这项检查未通过验证来显示有没有来自于药品或防腐剂的残留抑制物的存在。
检查自动化无菌检验系统或无菌隔离技术，如用于无菌检验的La Calhene装置，这些装置保证了不需要人直接移取样品。
如果被检样品在此系统中检验发现初次检验不合格，很难根据—次复检决定药品是否发放，特别是检验对照是阴性的时候。
评价无菌检验样品的培养时间。
该问题已被阐明，美国药典提出样品至少需要培养7天，而—项提议要求美国药典将其改为14天；希望诸位能评价—项具体的分析过程和产品以确定其最适培养时间。
7天可能不够充分，特别是发现有生长过程缓慢的微生物存在时，培养基灌装、环境、无菌检验结果和其他数据应加以审查以确保没有发现生长缓慢的微生物。
此外，还应比较各种培养方法，以确定它们是否符合已被批准或将被批准的申请书中所列的方法。
V．方法和检验程序的验证确定检验程序的来源。
生产厂的检验程序有几个方面的来源，如美国药典、《细菌学分析手册》及其他微生物参考资料。
事实上对各种有害微生物进行的检验程序不可能得到全面验证，但是应确保样品中的抑制物被中和。
在检查过程中，包括批准前的检查中，要评价微生物检验的方法。
例如我们希望检验方法可以鉴别一些微生物的存在，如洋葱假单孢菌或其他有害的假单孢菌属。
在进行批准前检查时，要将正在使用的方法和申请中呈递的方法进行比较，同时证实实验室拥有必要的检验设备，这些设备在进行关键检验时确实可用并处于良好的运行状态。
美国药典规定，如果已经过正确的验证，另一种方法可以得出同样甚至更好的结果，它可以用来代替法定检验方法。
你也许会发现人们已使用脱水的培养基来制备培养基。
良好的操作包括定期用极少量微生物去测试制备好的培养基，包括使用美国药典规定的指示微生物和正常菌群。
培养基促进生长的能力可能受到培养基制备程序、灭菌过程(过热)和储存的影响，它们是对任何实验室进行检查的重要指标，也是实现微生物实验室优良管理的重要考虑事项。
Ⅵ．数据储存评价检验结果是记录在实验记录本(logbooks)上还是记录在活页检验单上，由于有些生产厂可能不愿提供表明微生物检验结果的表格、总结或打印稿，应当审查数据来鉴别生产过程中潜在的微生物。
当得不到此数据的总结时，检验组应审查充分的数据来建立他们自己对于实验室结果和质量控制程序的总结。
一些实验室将事先印好的表格用于记录检验数据。
有些实验室还指出在检查中审查微生物检验数据的唯一途径是审查单批记录。
然而在大多数情况下，事先印出的表格都是一式几份，而第二，三份总是存在中心档案中。
一些公司以实验记录本记录资料，这些实验记录本也应该被查阅。
另外，很多生产厂装备厂自动化微生物系统，如用于鉴别微生物的Vitek装置。
这种检验的记录和对样品来源的鉴别对于识别生产过程中潜在的微土物问题具有同样价值。
鉴别微生物的自动化系统比较普遍地用于制造非肠道药品的生产厂，用于对从环境、水系统、验证过程和人员的分离物的常规鉴别。
我们巴尔的摩地区的微生物学家都是自动化微生物分析系统使用方面的专家。
他们最先在FDA实验室里使用这种设备并且对于验证这些设备的方法有丰富的经验。
关于该系统的情况或有关问题，可以向巴尔的摩地区实验室查询。
另外，大规模使用这类设备的应当接受来自巴尔的摩地区实验室专家的检查。
Ⅶ．管理审查微生物检验结果是数据评价及解释中一个较难的领域。
评价微生物检验结果需要受过微生物学的专业训练并具备经验。
要厂解检验的方法，而且更重要及更难的是要厂解检验的局限性。