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计算机化的实验室数据收集系统的验证和确认Validation and Qualification of Computerized Laboratory Data Acquisition SystemsPDA 技术报告No. 31Technical Report No. 31 PDA1. Objective 目的The purpose of this article is to provide guidance to laboratory scientists, technicians and managers responsible for the implementation, testing, control and usage of Laboratory Data Acquisition Systems (LDAS) used within a GMP, GLP, and GCP regulated environment.本文为实验室科学家、技术员以及管理者提供了在GMP, GLP和GCP 规范化环境下使用的LDAS的执行、测试、控制和用途方面的指南。
2. Scope 范围This article specifically addresses computerized LDAS within a regulated environment. This guidance is also applicable to systems considered critical to the operations of a company, department or function regardless of the system’s regulatory impact. The scope of this article excludes the typical Laboratory Information Management System (LIMS). The fundamental difference between a LIMS and an LDAS system is that the LDAS has a laboratory instrument as its primary focus, such as a computerized HPLC, whereas a LIMS, though instruments may be attached, has the management of data as its primary focus. The guiding key practices for testing and controlling an LDAS are similar to those for testing and controlling a LIMS;-1 the fundamental differences lie in the application of these key practices.本文仅涉及在规范化环境下的计算机化的LDAS。
PDA的技术报告目录PDA——Parenteral Drug Association，注射用药物协会（/）PDA技术报告目录／PDA PublicationsTechnical Methods Bulletin No.1 - Extractables from Elastomeric Closures: Analytical Procedures for Functional Group Characterization/IdentificationTechnical Methods Bulletin No.2 - Elastomeric Closures: Evaluation of Significant Performance and Identity CharacteristicsTechnical Methods Bulletin No.3 - Glass Containers for Small Volume Parenteral Products: Factors for Selection and Test Methods for IdentificationTechnical Information Bulletin No.2 - Generic Test Procedures for Elastomeric Closures Technical Information Bulletin No.4 - Aspects of Container/Closure IntegrityTechnical Report No.1 - Validation of Steam Sterilisation CyclesTechnical Report No.3 - Validation of Dry Heat Processes used for Sterilisation and DepyrogenationTechnical Report No.4 - Design Concepts for the Validation of a Water for Injection System Technical Report No.5 - Sterile Pharmaceutical Packaging: Compatibility and Stability Technical Report No.7 - DepyrogenationTechnical Report No.8 - Parametric Release of Parenteral Solutions Sterilized by Moist Heat Sterilization, 1987 (Please note: Technical Report No. 8 has been superseded by Technical Report No. 30 and is no longer available.)Technical Report No.9 - Review of Commercially Available Particulate Measurement Systems Technical Report No.10 - Parenteral Formulations of Proteins & Peptides: Stability and Stabilizers Technical Report No.11 - Sterilization of Parenterals by Gamma RadiationTechnical Report No.12 - Siliconization of Parenteral Drug Packaging ComponentsTechnical Report No.13 - Fundamentals of a Microbiological Environmental Monitoring Program Technical Report No.14 - Industry Perspective on the Validation of Column-Based Separation Processes for the Purification of ProteinsTechnical Report No.15 - Industry Perspective on Validation of T angential Flow Filtration in Bio-pharmaceutical ApplicationTechnical Report No.16 - Effect of Gamma Irradiation on Elastomeric ClosuresTechnical Report No.17 - Current Practices in the Validation of Aseptic Processing ? 1992 Technical Report No.18 - PDA Report on the Validation of Computer Related Systems Technical Report No.19 - Rapid/Automated ID Methods SurveyTechnical Report No.20 - Report on Survey of Current Industry Gowning PracticesTechnical Report No.21 - Bioburden Recovery ValidationTechnical Report No.22 - Process Simulation Testing for Aseptically Filled ProductsTechnical Report No.23 - Industry Survey on Current Sterile Filtration PracticesTechnical Report No.24 - Current Practices in the Validation of Aseptic Processing 1996 Technical Report No.25 - Blend Uniformity Analysis: Validation and In-Process Testing Technical Report No.26 - Sterilizing Filtration of LiquidsTechnical Report No.27 - Pharmaceutical Package IntegrityTechnical Report No.28 - Process Simulation Testing for Sterile Bulk Pharmaceutical Chemicals Technical Report No.29 - Points to consider for Cleaning ValidationTechnical Report No.30 - Parametric Release of Pharmaceuticals Terminally Sterilized by Moist HeatTechnical Report No.31 - Validation & Qualification of Computerized Laboratory Data Acquisition SystemsTechnical Report No.32 - Auditing of Suppliers Providing Computer Products and Services for Regulated Pharmaceutical OperationsTechnical Report No.33 - Evaluation, Validation & Implementation of New Microbiological Testing MethodsTechnical Report No.34 - Design and Validation of Isolator Systems for the Manufacturing and Testing of Health Care ProductsTechnical Report No.35 - A Proposed Training Model for the Microbiological Function in the Pharmaceutical IndustryTechnical Report No.36 - Current Practices in the Validation of Aseptic Processing - 2001 Technical Report No.39 - Cold Chain Guidance for Medicinal Products: Maintaining the Quality of Temperature-Sensitive Medicinal Products Through the Transportation EnvironmentTechnical Report No.40 - Sterilizing Filtration of GasesTechnical Report No.41 - Virus FiltrationTechnical Report No.42 - Process Validation of Protein ManufacturingTechnical Report No.43 - Identification and Classification of Nonconformities in Molded and Tubular Glass Containers for Pharmaceutical ManufacturingTechnical Report No.44 - Quality Risk Management for Aseptic ProcessesTechnical Report No.45 - Filtration of Liquids Using Cellulose-Based Depth Filters, 2008。
Evaluation, V alidation and Implementation of New Microbiological Testing Methods新微生物测试方法的评估、验证和执行Technical Report No. 33 第33号技术报告PDA目录第一部分: 新微生物方法的选择1.0 简介. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11.1 文件范围. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2 文件目的. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.3 文件结构概览. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22.0 技术概要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22.1微生物方法类型的一般说明. . . . . .. . . . . . . . . . .. . . . . . . . .. . 22.2 技术审核. . .. . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . .. . . . 32.3 基于生长的技术. . . . . . . .. . . . . . . . . . . . . . . . . . . . . .. . . .. .32.3.1 ATP生物发光. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. ..32.3.2 CO2生产的色度检测. . . . . . . . . . . . . . . . . . . . . . . .. . . .42.3.3 顶部空间压力变更的测量. . . .. . . . . . . . . . . . . .. .. . . .. .42.3.4阻抗. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .42.3.5 生物化学分析. . .. . . . . .. . . . . . .. . . . . . . . . . . . . . . .42.4 基于活性的技术. . .. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . 62.4.1 固相血细胞计数. . . . . .. . . . . . . . . . . . . . . . . . . . . . . . 62.4.2 流式荧光细胞计. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .62.5 人工技术. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.1 脂肪酸概要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.2 质谱学. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.3 ELISA酶联免疫吸附测定. . . . . . . . . . . . . . . . . .. . . . . . 72.5.4 荧光探针检测. . . .. . . . . . . . . . . . . . . . . . . .. . . . . . . . 72.5.5 细菌内毒素-鲎变形细胞溶解物测试. . . . . . . . . . . . . ..72.6 基于核酸的技术. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. (8)2.6.1 探针. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .82.6.2 核糖型/分子型. . .. . . . . . . . . . . . . . . . . . . . . . . . . . . 82.6.3 聚合酶链反应. . .. . . . . .. . . . . . . . . . . . . . . . . . . . . .93.0监管审查. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .103.1微生物测试总分类. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.1 加工过程中测试. . .. . . . . . . . . . . . . . . . . . . . . . (10)3.1.2 产品测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.3 定性测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.1.4 定量测试. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (10)3.2 药典微生物测试方法参考文件. . .. . . . . . . . . . . . . . . . . . . . 103.2.1 制药用水. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ..103.2.2 抗菌效力测试. . . . .. . . . . . . . . . . . . . . . . . . . . . . (10)3.2.3 微生物限度测试. . . . . .. . . . . . . . . . . . . . . . . . (11)3.2.4 无菌测试. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . (11)3.2.5 环境监测. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (11)3.2.6 微生物鉴定. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .113.3 改变微生物测试方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.3.1 新微生物测试方法介绍的监管观点 . . . . . . . . . . . . . ..113.3.2 药典观点. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (12)3.4 药典变更. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (13)3.5 新微生物方法的管理者评估. . . . . . . . . . . . . . . . . . . . .13 第二部分: 如何验证并执行新微生物测试方法4.0 验证流程. . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . .154.1 设备确认模型. . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . .164.1.1 供应商/说明要求. . . . . . . . . . . . . . . . . . . . . . . . . . 164.1.1.1 测试方法选择. . . . . . . . . . . . . . . . . . . . .174.1.1.2 供应商选择. . . . . . . . . . . . . . . . . . . . . . .174.1.2 验证方法设计. . . . . . . . . . . . . . . . . . . . . . . . . . (18)4.1.3 安装确认. . . . . . . . . . . . . . . . . . . . . . . . . . . . (18)4.1.4 运行确认. . . . . . . . . . . . . . . . . . . . . . . . . . . . (20)4.1.5 性能确认. . . . . . . . . . . . . . . . . . . . . . . . . . ....... (20)4.2验证标准. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (21)4.2.1测试样品制备. . .. . . . . . . . . . . . . . . . . ....... . . . (23)4.2.2 微生物方法的活性：特别注意. . . . . . . . . . . . . . . . 254.2.2.1 样品分布错误. . .. . . . . . . . . . . . . . . (25)4.2.2.2 细胞形态. . . . . . . . . . . . . . . . . . . . . . . . 254.2.2.3 代谢活动. . .. . . . . . . . . . . . . . . . . . . . . 254.2.3使用推荐验证标准的方案设计. .. . . . . . . . . . . . . . .264.3微生物方法验证的特殊考虑. . . . . . . . . . . . . . . . . . . . . . . 294.3.1在实验室和公司内使用多件相同设备. . . . . . . . . . . . 294.3.2微生物设备的独特测试要求. .. . . . . . . . . . . . . . . . . 305.0 术语. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32第一部分新微生物测量方法的选择1.0简介1.1文件范围本文件的目的是对制药、生物技术和医药设备工业为确保产品质量进行成功评估、验证和实施所需新微生物方法提供指南。
PDA Technical Report No. 29Points to Consider forCleaning ValidationDRAFTMarch 30, 1998TR28_002.PDFPDA Pharmaceutical Cleaning Validation Task Force James P. Agalloco, Agalloco & AssociatesWill Brame, Rhone-Poulenc RorerBohdan Ferenc, Novartis Pharmaceuticals Corp.William E. Hall, Ph.D., Hall & AssociatesKevin Jenkins, Pharmacia & Uphohn, Inc.John T. LaMagna, Pfizer, Inc.Russell E. Madsen, PDA (Chairman)Michael V. Mullen, Ph.D., Eli Lilly and CompanyDietmar Wagenknecht, Fujisawa USA, Inc.Carmen M. Wagner, Ph.D., Wyeth-Lederle Vaccines & PediatricsPrefaceThis document provides guidance relative to the validation of cleaning for a broad range of processing systems and product types within the pharmaceutical industry. This effort commenced in 1991 in conjunction with individuals representing the biotechnology community. Early on it was agreed to separate the development of cleaning validation guidance into "biotechnology" and "pharmaceutical" segments. The committees worked in parallel for a number of years and shared early drafts to ensure that what would be produced by each committee would be compatible. The biotech effort culminated in PDA's 1995 publication of "Cleaning and Cleaning Validation: A Biotechnology Perspective". The "pharmaceutical" committee continued the development of its document after the publication of the biotech effort, and completed its stand alone guidance in the fall of 1997.Our goal had always been to outline cleaning validation practices across a range of equipment, process, and product applications and the inclusion of this flexibility was certainly a factor in the length of time it took to complete this effort. During the course of assembling this document, we recognized the commonality of certain themes, issues, and concerns relative to cleaning and cleaning validation across the industry. We also realized that in order to apply the principles in different operating settings, that some narrowing of the document would be necessary. As a result, we have included perspectives on the application of the guidance in various areas: finished pharmaceuticals, bulk pharmaceutical chemicals, biopharmaceuticals and clinical products. Inclusion of biopharmaceuticals in this effort is not intended to replace the more comprehensive coverage provided by our partner committee, but rather to provide greater insight regarding the broad application of the guidance provided herein.This Technical Report was disseminated in draft for public review and comment prior to publication. Many of the submitted comments have been included in the final document. We believe this approach accomplished the widest possible review of the document and ensures its suitability as a valuable guide to industry in the area of cleaning validation. This document should be considered as a guide; it is not intended to establish any mandatory or implied standard.Russell E. MadsenChairman, Pharmaceutical Cleaning Validation Task ForceTable of Contents1.Introduction (1)1.1 Background (1)1.2 Purpose (1)1.3 Scope (1)1.4 Report Organization (2)Finished Pharmaceuticals (2)Biopharmaceuticals (2)Bulk Pharmaceutical Chemicals (3)Clinical Products (4)2.The Cleaning Continuum (4)2.1 Use of the Cleaning Continuum (4)2.2 Cleaning Program Criteria (5)Automated Cleaning 76 Manual Cleaning (5)Clean-In-Place (CIP) 76 Clean-Out-of-Place (COP) (6)2.3 Equipment Characteristics / Materials of Construction (6)Dedicated 76 Non-Dedicated Manufacturing Equipment (6)Dedicated 76 Non-Dedicated Cleaning Equipment (6)Non-Product Contact 76 Product Contact Surfaces (7)Non-Critical Site 76 Critical Site (7)Minor Equipment 76 Major Equipment (7)Materials of Construction (8)2.4 Product Attributes (8)Low Risk 76 High Risk Drugs (8)Highly Characterized 76 Poorly Characterized (9)Non-Sterile 76 Sterile (9)2.5 Formulation Attributes (10)Solids 76 Liquids (10)Soluble (Active or Excipient) 76 Insoluble (Active or Excipient) (10)2.6 Operational Issues (10)Single Product Facility 76 Multiple Product Facility (10)Campaign Production 76 Batch Production (11)Simple Equipment Train 76 Complex Equipment Train (11)3.Cleaning Validation (11)3.1 The Cleaning Validation Program (11)3.2 Product Grouping (12)3.3 Manufacturing Equipment Grouping (13)3.4 Cleaning Method Groupings (14)3.5 Cleaning Agent Groupings (14)4.Residues and Residue Removal (14)4.1 Types of Residue (14)4.2 Cleaning Agents (14)4.3 Microbiological Contaminants (15)4.4 Other Contaminants to be Removed (15)4.5 Cleaning Agent and Surface Interactions (16)5.Cleaning of Equipment (16)5.1 Types of Cleaning Processes (16)Manual (16)Semi-Automated (16)Automated (17)5.2 Clean-in-Place (CIP) Systems (17)5.3 Clean-Out-of-Place (COP) Systems (18)5.4 Cleaning Porous Equipment (18)5.5 Disposable Equipment (18)5.6 Placebo Batches as a Cleaning Method (18)5.7 Residue Removal and Cleaning Methods (18)5.8 Equipment (19)5.9 Equipment Design Considerations (19)5.10 Cleaning Frequency (20)Between batches of different products (20)Between batches of the same product (campaign) (20)5.11 Post-Cleaning Equipment Storage (20)5.12 Monitoring Cleaning Cycles (21)6.Cycle Development (21)6.1 Cleaning Agent Selection (22)6.2 Product Considerations (22)6.3 Cleaning Parameter Selection (23)6.4 Standard Operating Procedures (23)6.5 Cleaning Records (24)6.6 Operator Training (24)7.Sampling Techniques and Analytical Methods (24)7.1 Sampling Techniques (25)Swabs and Wipes (25)Rinse Sampling (25)Coupon Sampling (26)Solvent Sampling (26)Product Sampling (27)Placebo Sampling (27)Direct Surface Monitoring (28)7.2 Visual Examination (28)7.3 Relationship of Analytical Method, Sampling Method, and Limit (29)iv7.4 Specific versus Non-Specific Testing (29)7.5 Analytical Methods (30)7.5.1 Direct Surface Analysis (30)7.5.2 pH (31)7.5.3 Conductivity (31)7.5.4 Total Organic Carbon (TOC) (31)7.5.5 Enzymatic (Bioluminescence) (32)7.5.6 Light Microscopy (32)7.5.7 Gravimetric Analysis (33)7.5.8 Titration (34)7.5.9 High Performance Liquid Chromatography (HPLC) (34)7.5.10 Thin Layer Chromatography (TLC) (34)7.5.11 Capillary Zone Electrophoresis (CZE) (35)7.5.12 Fourier Transform Infrared (FTIR) (35)7.5.13 Enzyme Linked Immunosorbant Assay (ELISA) (36)7.5.14 Atomic Absorption/Ion Chromatography (AA/IC) (36)7.5.15 Ultraviolet (UV) Spectrophotometry (36)7.6 Pass-Fail Testing Methods (37)7.7 Analytical Methods Validation (37)7.8 Microbial and Endotoxin Detection and Testing (37)8.Limits Determination (38)8.1 The Scientific Rationale for Cleaning (38)8.2 Contamination of the Next Product (38)8.3 Considerations for Developing Limits (38)8.4 Limits Based on Medical or Pharmacological Potency of the Product (38)8.5 The Basis for Quantitative Limits (39)8.6 Limits Based on the Toxicity of the Residue (40)8.7 Limits Based on the Analytical Limitations (41)8.8 The Meaning of “None Detected” (41)8.9 Dividing a Limit Among Various Pieces of Equipment (41)9.Ongoing Verification of Cleaning (42)9.1 Verification of Cleaning (42)9.2 Monitoring of Automatic and Manual Cleaning (42)10.Change Control (43)11.Appendices (44)11.1 Glossary of Terms (44)11.2 Suggested Reading (49)v1.Introduction1.1 BackgroundIn recent years, cleaning has achieved a position of increasing importance in the pharmaceutical industry. The current good manufacturing practices (CGMP) regulations recognize that cleaning is a critical issue to ensure product quality. Virtually every aspect of manufacturing involves cleaning, from the initial stages of bulk production to the final dosage form.The CGMPs in the United States, Europe and other parts of the world have provided the pharmaceutical industry with general guidance for cleaning requirements. For example, in the U.S., section 211.67 of part 21 of the Code of Federal Regulations (CFR) states that "Equipment and utensils shall be cleaned, maintained, and sanitized at appropriate intervals to prevent malfunctions or contamination that would alter the safety, identity, strength, quality, or purity of the drug product beyond the official or other established requirements." Section 211.182 of part 21 of the CFR identifies that cleaning procedures must be documented appropriately, and that a cleaning and use log should be established. In addition to CGMPs, various inspectional guideline documents published by the FDA contain expectations regarding cleaning in the pharmaceutical industry. Cleaning is also addressed in the PIC recommendations on cleaning validation and in the SFSTP Commission report "Validation des procédés de nettoyage."It has always been the responsibility of the regulated industry and the regulatory agencies to interpret the CGMPs and to create programs and policies which establish the general requirements as specific practices. Recognizing the importance of the relationship between cleaning and product quality, regulatory agencies are demanding greater evidence of cleaning effectiveness through validation or verification.1.2 PurposeThe purpose of this publication is to identify and discuss the many factors involved in the design, validation, implementation and control of cleaning programs for the pharmaceutical industry.The document does not attempt to interpret CGMPs but provides guidance for establishing a cleaning validation program. It identifies the many factors to be considered for all segments of the pharmaceutical industry. It also identifies specific points to be considered by dosage form manufacturers, manufacturers of clinical trial materials (CTMs) and manufacturers of bulk pharmaceutical chemicals and biochemicals. The report covers the different approaches which may be appropriate for the different stages of product development from the early research stages to the commercially marketed product.1.3 ScopeThis paper applies to biopharmaceutical, bulk pharmaceutical and finished dosage form operations; liquid, dry, solid and semi-solid dosage forms are covered in both sterile and non-sterile presentations. Both clinical and marketed product cleaning validation programs are identified.The manufacture of modern pharmaceuticals is a complex process involving highly technical personnel, complex equipment, sophisticated facilities and complicated processes. Individuals responsible for all aspects of the production, approval and validation of products, such as quality control, quality assurance, engineering, validation, production, research and development, contractors and vendors and regulatory affairs personnel may use this document as a resource for establishing or reviewing the cleaning programs within their facilities.The validation programs described herein assume that an overall validation program with appropriate controls is already in place for the facility, utilities, equipment and processes. The cleaning of the environment is not specifically covered, however many of the same concerns that are considered for the cleaning of process equipment also impact the cleaning of the environment. The monitoring of microbiological and endotoxin contamination and steps for their elimination are mentioned in several sections and should be part of the cleaning validation program. However this document is not intended to be a comprehensive treatise on microbiological control, or endotoxin limitation. Other documents have addressed microbiological programs and methods for the environmental monitoring which can be applied to cleaning.1.4 Report OrganizationEach of the major topics of this document starts with a general section which applies to all segments of the pharmaceutical industry. Points to be considered for specific industry segments such as biopharmaceuticals, bulk pharmaceutical chemicals, clinical products may vary, depending on the specific product type. A glossary is provided at the end of the report.Finished PharmaceuticalsFinished pharmaceuticals represent solid formulations, semi-solid formulations, liquid and aerosol formulations with various routes of administration. Over-the-counter and prescription pharmaceuticals for both human and veterinary use are included in this category.The common characteristics shared by finished pharmaceuticals are their manufacture by combining raw materials and active ingredients to create the final dosage form.Pharmaceutical manufacturers often make a large number of product types in one facility; often there are several different strengths prepared of the same product. The cleaning problems include the large number of processes and product types manufactured within one facility. The number of cleaning methods, assays and types of equipment to be tested are often staggering. This is complicated by the issues surrounding the use of non-dedicated equipment. Thus, the establishment of a cleaning validation policy which is applicable to all products is often very difficult. BiopharmaceuticalsBioprocess manufacturing, starting with microbial, animal or insect cells, or DNA derived host cells or other cells modified to make a specialized product, can be performed in several ways. Indeed, new methods for bioprocessing are constantly being developed. The most common method is through large scale fermentation (such as bacterial cell culture or mammalian cell culture) followed by highly specific purification steps. Other methods include the development of an antibody in host animals (such as ascites), cloning of cells or tissues, or transgenic generation of cellular components, namely,proteins. Many in the biopharmaceutical industry consider the stages of fermentation to be similar to other pharmaceutical industry processes. For example, the initial stages of the large scale fermentation have a striking similarity to bulk pharmaceutical chemical production. Later, harvest and purification steps find more in common with pharmaceutical processes. It is important to remember however, that other bioprocessing methods used in the biopharmaceutical industry differ greatly from traditional pharmaceutical processes.Cleaning for biopharmaceuticals presents special concerns due to the large number of impurities such as cellular debris, waste products of cellular metabolism, media constituents and buffer salts generated or added during manufacture which must be eliminated from the equipment. In the case of mammalian cell cultures, due to the nature of the source material, microbial contamination is of great concern. Identification of the residues is often quite difficult because they may vary from batch to batch. The large variety of proteinaceous materials present in the residue make differentiation of the contaminants from one another a challenge.Due to the nature of the biopharmaceutical production, multi-product facilities represent an area of regulatory concern. In order to control the production within a multi-product facility, it is necessary to ensure that special precautions are taken which preclude product to product carryover. Cleaning is an integral part of the strategies designed to ensure that there is no cross-contamination in these facilities. The terms cleaning and cleaning validation in multi-product facilities often include the facility itself, and therefore emphasis is placed on changeover validation.Cleaning for biotechnology products has been described in "Cleaning and Cleaning Validation: A Biotechnology Perspective," PDA, Bethesda, MD, 1996.Bulk Pharmaceutical ChemicalsBulk pharmaceutical chemical processes are typically biochemical or chemical syntheses carried out on a relatively large scale. The bulk pharmaceutical chemicals may be provided to pharmaceutical manufacturers as active or inactive ingredients for eventual inclusion in a finished dosage form pharmaceutical. The bulk pharmaceutical chemical manufacturing process for active ingredients is typically enclosed in large tanks with direct transfer of materials from tank to tank after a particular chemical reaction has occurred. The initial stages of the bulk pharmaceutical chemical drug development are reminiscent of the chemical industry. At some point during the process, the manufacturer must, in accordance with CGMPs have identified a process step after which the process will strictly comply with the CGMPs.Bulk pharmaceutical chemical production, due in large part to the scale of manufacture and its use of strong reagents and chemicals, is often performed in closed systems which may use automated or semi-automated Clean-In-Place technologies. The difficulties in the validation of cleaning processes often stem from the inaccessibility of many areas to direct sampling. The contaminants to be removed include precursor molecules, intermediates, byproducts, impurities or other physical forms such as isomers or polymorphs, which exist from early stages in the process.Clinical ProductsIn this document, clinical products identify those products which are currently registered as an investigational status due to their involvement in clinical trials. The Clinical Products category identifies the special care that must be taken with these products which may not be as fully characterized as marketed materials. Both pharmaceutical and biopharmaceutical drug products and drug substances are included in this category.Cleaning in a clinical manufacturing setting is often complicated by the use of small scale manually cleaned equipment. Clinical manufacturing may represent a period during which process improvements are made, and therefore the same equipment may not be used each time the product is made. Also, since clinical products are often manufactured in development facilities, the subsequent products may not be known. The next materials manufactured may be research products, development products, placebo products or other clinical products. Our intent is to address cleaning of equipment in Phase III and later, but it may be appropriate to consider the same approaches in earlier phases as well. Typically, assays for low level detection of the active ingredient and its excipients will need to be developed and validated. Verification of cleaning effectiveness, as opposed to traditional validation, is prevalent since information on the material is not readily available. 2.The Cleaning ContinuumThe subject of cleaning validation is one which the pharmaceutical and biotechnology industries have struggled with. Progress to a consensus in approach in the industry has been slowed by the number and complexity of issues surrounding the cleaning process and the variety of facilities, products and equipment in use. The development of a universal approach to cleaning validation is unlikely given these variations.2.1 Use of the Cleaning ContinuumThe intent of this section is to describe the limits of the cleaning continuum (see Table 1). These limits represent the extremes in the range of operating differences found within the industry which preclude a uniform approach. At each end of the continuum, the cleaning validation requisites are either simple or complex. Recognition that there are many of these coupled limits, and that each cleaning process has a unique place within each level of the continuum, explains why specific industry-wide approaches have been so difficult to develop.The cleaning continuum provides some of the primary points to consider in any cleaning validation program. The continuum helps firms to establish the parameters which are critical factors for individual products, thereby enabling them to set priorities, develop grouping philosophies and establish the "scientific rationale" which will govern the cleaning program. The continuum will assist in determining which processes, equipment and products represent the greatest concerns and may help to establish the criticality of cleaning limits and methods. The continuum should be used during the initial phases of defining a cleaning validation program or during new product development. The cleaning continuum includes: cleaning program criteria, equipment characteristics, quality attributes of equipment design, formulation/product attributes, analytical methodology and manufacturing/process attributes. All of the factors in the continuum directly affect the ability toclean; however, their relative importance and criticality may be different from one company to another.Table 1: The Cleaning Continuum Manual....................................................Automated Cleaning Clean-out-of-Place (COP)......................................Clean-in-Place (CIP) Dedicated Equipment....................................Non-Dedicated Equipment Product Contact Surfaces..............................Non-Product Contact Surfaces Non-Critical Site...................................................Critical Site Minor Equipment..............................................Major Equipment Low Risk Drugs................................................High Risk Drugs Highly Characterized.........................................Poorly Characterized Sterile............................................................Non-Sterile Solid Formulations...........................................Liquid Formulations Soluble.............................................................Insoluble Single Product Facility.....................................Multiple Product Facility Campaigned Production.................................Non-Campaigned Production Simple Equipment plex Equipment Train 2.2 Cleaning Program CriteriaWhen establishing a cleaning validation program, it is important to first characterize the types of cleaning that are used in the facility. The cleaning methods that are used in a facility can reveal important factors with regard to process control, process reproducibility, the best ways in which to challenge the process, the best ways in which to collect samples and the best ways in which to monitor cleaning effectiveness during routine cleaning.Automated Cleaning 76 Manual CleaningAutomated cleaning will usually provide reproducible results. Process control is inherent in automated systems and process monitoring is frequently integral with the control system.Automated systems may not adjust to present conditions. The validation of an automated system requires that the cycle is proven to be rugged and will provide reproducible results under a given range of operating conditions. Control system validation is a large part of the validation of an automated cleaning system.Manual cleaning is a universal practice within the pharmaceutical industry. There are many pieces of equipment and portions thereof for which construction and/or configuration make manual cleaning a necessity. The control of manual cleaning is accomplished by operator training, well defined cleaning procedures, visual examination of equipment after use and prior to the next use, and well-defined change control programs. It may be desirable to identify worst case cleaning situations (in terms of operator experience and/or cleaning methodology) for validation purposes. With manual cleaning, concern must also be given to the ruggedness of the method. Successful reproducibility is a function of strict adherence to written procedures.Clean-In-Place (CIP) 76 Clean-Out-of-Place (COP)The cleaning of large pieces of equipment may be performed in the equipment's permanent location, generally in a configuration very similar to that in which it is utilized for production.This procedure is widely known as Clean-In-Place (CIP). Smaller equipment items are frequently transported to a designated cleaning or wash area where the cleaning procedure is performed. This practice is known as Clean-Out-of-Place (COP), but the term is not as prevalent as its counterpart.The additional activities involved with transport of equipment to and from the wash room, component identification, the elimination of cross-contamination potential during transfer, and cleaning and storage prior to use make the validation of COP procedures somewhat more difficult than the comparable CIP activity. The need for manual manipulation is an integral part of many COP procedures and requires detailed procedures and training. The manual manipulation makes COP concerns similar to those of manual cleaning in place.The use of automated washing machines to COP smaller items is an important part of many COP systems. The use of these systems reduces the differences between CIP and COP significantly. These systems are considered to be highly reproducible in their cleaning performance and are gaining wide acceptance.2.3 Equipment Characteristics / Materials of ConstructionEquipment usage during production is another important aspect to consider in establishing a cleaning validation program. It is important to understand not only the range of products that are likely to come into contact with the various equipment surfaces, but also the role that the equipment plays in the production train. This will help to establish the contamination and cross-contamination potentials of the equipment.Equipment design characteristics, as established during product development, are often driven by equipment functionality and the requirements of the process. With the current emphasis on cleaning validation, it makes sense that "cleanability" be a key criterion in the design of equipment.Dedicated 76 Non-Dedicated Manufacturing EquipmentDedicated equipment is used solely for the production of a single product or product line.Concerns over cross-contamination with other products are markedly reduced. Dedicated equipment must be clearly identified with the restrictions of use in order to prevent potential errors during cleaning and preparation.Where the same piece of equipment is utilized for a range of product formulations, (i.e., non-dedicated equipment), the prevention of cross-contamination between products becomes the main objective in the cleaning validation effort. Clearly, cleaning non-dedicated equipment represents a more significant obstacle to overcome.Dedicated 76 Non-Dedicated Cleaning EquipmentThe issues of dedicated and non-dedicated equipment can also arise when considering the equipment used for cleaning. CIP systems, for example, are frequently used for manydifferent tanks in a single facility. Inherently, the design of CIP systems should preclude cross-contamination through appropriate valving and back-flow prevention. Care should be taken with shared devices which apply cleaning agents, such as spray balls or spray nozzles which, themselves, may require cleaning. Certainly any recirculation within the CIP system should be configured carefully during system design and monitored closely during routine operation.COP equipment, such as an ultrasonic sink, may also be used for multiple equipment loads. With cleaning apparatus such as the sink, the removal of potential contaminants from the sink, itself is a concern. Sinks and washers frequently use recirculation systems to economically remove residuals from surfaces without undue waste. The cleanliness of the recirculated materials should be evaluated during cleaning validation to ensure that contaminants are not being redeposited on the equipment to be cleaned.Non-Product Contact 76 Product Contact SurfacesTraditionally, the validation of cleaning has focused on product contact surfaces. Programs for the elimination of cross-contamination must address non-product contact surfaces if they are to be truly effective. In practice, cleaning validation requirements may change with non-product contact surfaces in accordance with the less critical nature of these areas. When establishing the requirements for non-product contact surfaces, it is important to review the possible interactions of that area with the process.Non-Critical Site 76 Critical SiteCritical sites are those locations in which a contaminant is in danger of affecting a single dose with a high level of contamination. Critical sites often require special cleaning emphasis. It may be appropriate to establish more intensive sampling schedules for critical sites, set tighter acceptance criteria for critical sites and ensure that enough detail is included in cleaning procedures to provide for reproducible cleaning of critical sites.Minor Equipment 76 Major EquipmentThe distinction between "major" and "minor" equipment is not a definitive one. The CGMPs make mention (211.105) of "major" equipment, but are silent on the subject of "minor" equipment except with regard to items described as utensils (211.67). Despite this failure within the CGMPs, it is necessary to identify those pieces of equipment (major) which are central to the production process and those pieces of equipment (minor) which perform a secondary role.Typically the cleaning of "major" equipment will be the subject of individual, highly specific SOPs. In contrast, "minor" equipment and "utensils" are often cleaned using broadly defined procedures which describe the methods to be used in general terms.。
Validation of Moist Heat Sterilization Processes: Cycle Design, Development, Qualification and Ongoing ControlTechnical Report N01 (Revised 2007) SupplementVol. 61, N0 S-1c 2007 by PDAP D AParenteral Drug Association湿热灭菌的验证：灭菌程序的设计，开发，确认和日常监控2007年增补，第一卷N0.S-1美国注射剂协会制药科学及技术杂志编写委员：中国医药设备工程协会技术委员会发行单位：中国医药设备工程协会湿热灭菌程序的验证：灭菌程序的设计、开发、确认以及日常监控目录1.0引言 (4)1.1范围 (4)2.0术语 (5)3.0灭菌科学 (11)3.1 灭菌模式 (12)3.1.1 耐热参数（D T） (13)3.1.2 温度系数（z值） (15)3.1.3 灭菌率（Lethal Rate）和累计杀灭时间（lethality，F） (16)3.2灭菌指示剂 (20)3.2.1 生物指示剂（BIs） (20)3.2.2 化学监测器 (21)3.3热力学和蒸汽质量 (22)3.3.1 温度和热量 (22)3.3.2 蒸汽 (28)3.3.3 纯蒸汽质量的测试 (28)4.0 灭菌程序的开发 (30)4.1 设计方法 (30)4.1.1 灭菌程序设计方法中残存曲线的应用 (31)4.2 装载类型 (34)4.2.1 多孔/坚硬装载的定义 (34)4.2.2 液体装载的定义 (34)4.3 灭菌程序 (35)4.3.1 饱和蒸汽灭菌程序 (35)4.3.2 空气加压程序 (38)4.4 灭菌程序的开发 (40)4.4.1 多孔/坚硬装载灭菌程序的开发 (40)4.4.2 液体装载灭菌程序的建立 (45)4.5 稳定性研究 (48)5.0 灭菌程序的性能确认 (48)5.1 物理确认 (49)5.1.1 热分布 (49)5.1.2 热穿透 (49)5.2 生物指示剂确认 (50)5.2.1 生物指示剂挑战系统 (50)5.2.2 生物指示剂的使用和放置 (51)5.3 灭菌程序性能确认合格标准 (52)5.4等效灭菌器 (53)5.5 分组法 (54)5.5.1 典型产品法 (54)5.5.2 典型容器规格/装量法 (54)5.5.3 典型物品法 (54)5.5.4 典型装载法 (54)6.0 日常工艺控制 (55)6.1 常规放行 (55)6.2 灭菌器系统的适用性试验 (55)6.3 变更控制 (55)6.4 灭菌程序定期再确认 (56)7.0 参考文献 (56)1.0引言PDA的原始技术报告第一期“蒸汽灭菌程序的验证”于1978年出版，它为一代制药科学家和工程师介绍了蒸汽灭菌的原理。
ACTIVE PHARMACEUTICAL INGREDIENTS COMMITTEE (APIC)GUIDANCE ON ASPECTS OF CLEANING VALIDATIONIN ACTIVE PHARMACEUTICAL INGREDIENT PLANTS原料药工厂中清洁验证指南Revision September 2016Table of Contents 目录1.0 FOREWORD 前言This guidance document was updated in 2014 by the APIC Cleaning Validation Task Force on behalf of the Active Pharmaceutical Ingredient Committee (APIC) of CEFIC.本指南文件于2014年由APIC清洁验证工作组代表CEFIC的APIC委员会进行了更新。
The Task Force members are:- 以下是工作组的成员―Annick Bonneure, APIC, Belgium―Tom Buggy, DSM Sinochem Pharmaceuticals, The Netherlands―Paul Clingan, MacFarlan Smith, UK―Anke Grootaert, Janssen Pharmaceutica, Belgium―Peter Mungenast, Merck KGaA, Germany.―Luisa Paulo, Hovione FarmaCiencia SA, Portugal―Filip Quintiens, Genzyme, Belgium―Claude Vandenbossche, Ajinomoto Omnichem, Belgium―Jos van der Ven, Aspen Oss B.V., The Netherlands―Stefan Wienken, BASF, Germany.With support and review from:- 以下为提供支持和进行审核的人员―Pieter van der Hoeven, APIC, Belgium―Anthony Storey, Pfizer, U.K.―Rainer Fendt, BASF, Germany.A further revision of the guidance document has now been done in 2016 to bring it in line with the European Medicines Agency Guidance on use of Health Based data to set acceptance criteria for cleaning. The main changes were introduced in Chapter 4, Acceptance Criteria.本指南文件进一步修订已于2016年完成，使其与EMA使用基于健康数据设定清洁可接受标准的指南保持一致。
Evaluation, V alidation and Implementationof New Microbiological Testing MethodsTechnical Report No. 33PDAMay/June 2000Vol.54, No. 3, May/June 2000, Supplement TR33 iTask Force MembersBrian Bauer, Ph.D., Merck & Co., Elkton, VirginiaMark Claerbout, Lilly Research Laboratories, Indianapolis, IndianaWarren Casey, Ph.D., GlaxoWellcome R&D, Research Triangle Park, North CarolinaAnthony M. Cundell, Ph.D., Wyeth-Ayerst Pharmaceuticals, Pearl River, New York (Chair)Martin Easter, Ph.D., Celsis Ltd., Cambridge, EnglandEdward Fitzgerald, Ph.D. Consultant, (USP Microbiology Subcommittee)Carol Gravens, BioMerieux, Inc., Hazelwood, MissouriDavid Hussong, Ph.D., CDER, FDA, Rockville, MarylandMichael Korcynzski, Ph.D., PDA Training Institute, Baltimore, Maryland (USP Microbiology Subcommittee) Robin Lerchen, American Pharmaceutical Partners, Melrose Park, IllinoisFrederic J. Marsik, Ph.D. CDER, FDA, Rockville, MarylandAmy Meszaros, StatProbe Inc, Ann Arbor, MichiganJeanne Moldenhauer, Ph.D., Jordan Pharmaceuticals, Inc., Elk Grove, IllinoisManju Sethi, Qualicon, Wilmington, DelawareScott Sutton, Ph.D., Alcon Laboratories, Fort Worth, Texas (USP Microbiology Subcommitee)Martin Tricarico, Chemunex (USA), Monmouth Junction, New JerseyAmanda Turton, Millipore Corp, Bedford, MassachusettsChristine V ojt, Johnson & Johnson Diagnostics Inc., Rochester, New YorkKirsty Wills, Celsis Ltd., Cambridge, EnglandJon Wuannlund, Becton Dickinson Microbiology Systems, Cockeysville, MarylandPDA TECHNICAL REPORT NO.33EVALUATION, VALIDATION AND IMPLEMENTATIONOF NEW MICROBIOLOGICAL TESTING METHODSTable of ContentsPart One: Selection of New Microbiological Methods1.0Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.1Scope of Document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2Purpose of Document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.3Overview of Document Structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22.0Technology Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22.1Generic Description of Types of Microbiological Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22.2Technology Review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32.3Growth-based Technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32.3.1ATP Bioluminescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32.3.2Colorimetric Detection of CO2 Production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42.3.3Measurement of Change in Head Space Pressure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42.3.4Impedance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42.3.5Biochemical Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42.4Viability-based Technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62.4.1Solid Phase Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62.4.2Flow Fluorescent Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62.5 Artifact-based Technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.1Fatty Acid Profiles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.2Mass Spectometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.3ELISA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.4Fluorescent Probe Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72.5.5Bacterial Endotoxin-Limulus Amebocyte Lysate Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .72.6 Nucleic Acid-based Technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.6.1DNA Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.6.2Ribotyping/Molecular Typing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.6.3Polymerase Chain Reaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93.0Regulatory Review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.1General Classification of Microbiological Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.1.1In-Process Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.1.2Product Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.1.3Qualitative Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.1.4Quantitative Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.2 Compendial Microbiological Test Method References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.2.1Water for Pharmaceutical Purposes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.2.2Antimicrobial Effectiveness Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103.2.3Microbial Limit Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.2.4Sterility Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.2.5Environmental Monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.2.6Microbial Identification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 Vol.54, No. 3, May/June 2000, Supplement TR33 iii3.3Changing a Microbiological Test Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113.3.1The Regulatory Perspective on the Introduction of New Microbiological Test Methods . . . . 113.3.2The Compendial Perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123.4Obtaining Compendial Change . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133.5 Regulators Assessment of New Microbiological Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Part Two: How to Validate and Implement a New Microbiological Test Method4.0The Validation Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .154.1The Equipment Qualification Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164.1.1Vendor/Specification Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164.1.1.1Test Method Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174.1.1.2Vendor Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174.1.2Validation Plan Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184.1.3Installation Qualification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184.1.4Operation Qualification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204.1.5Performance Qualification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 204.2Validation Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 214.2.1Preparation of Test Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 234.2.2Variability of Microbiological Methods: Special Note . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .254.2.2.1Sample Distribution Error . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254.2.2.2Cell Morphology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254.2.2.3Metabolic Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 254.2.3Protocol Design Using Recommended Validation Criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264.3Special Considerations for the Validation of Microbiological Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 294.3.1Use of Multiple Pieces of the Same Equipment within the Laboratory and Company . . . . . . . . . . . . . 294.3.2Unique Testing Requirements for Microbiological Equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 305.0Glossary of Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 326.0References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 iv PDA Journal of Pharmaceutical Science & TechnologyPART ONE:Selection of New Microbiological Methods 1.0 INTRODUCTION1.1 Scope of DocumentThis document is intended to provide guidance for the successful evaluation, validation, and implemen-tation of new microbiological methods needed by the pharmaceutical, biotechnology, and medical de-vice industries to assure product quality. Applica-tions for these methods include but are not limited to Microbial Limit Testing, Sterility Testing, Anti-microbial Effectiveness Testing, Microbiological Monitoring of Clean Rooms and Other Controlled Environments, Water for Pharmaceutical Purposes Monitoring, and Microbial Characterization and Identification.The intended audience of this report is microbiolo-gists who are responsible for the validation of the microbiological test methods used in the routine mi-crobiology testing laboratory. The document should be of interest to suppliers of testing equipment, mi-crobiology managers and supervisors, validation specialists, quality control personnel responsible for the approval of validation protocols and the re-lease of product, and regulatory agencies.1.2 Purpose of DocumentMicrobiological testing plays an ever increasing role in the pharmaceutical laboratory. In response to this, a variety of new methodologies have emerged in re-cent years which automate existing methods, make use of surrogate markers for growth, or are based on wholly new technologies. These new methodolo-gies offer significant improvements in terms of the speed, accuracy, precision, and specificity with which testing can be performed.The majority of testing performed today relies on century-old methods, based on the recovery and growth of microorganisms, using solid or liquid mi-crobiological growth media. This is true in part be-cause these methods can be very effective and have a long history of application in both industrial and clinical settings. However, they are often limited by slow microbial growth rates, the unintended selectiv-ity of microbiological culture, and the inherent vari-ability of microorganisms in their response to culture methods. In spite of the limitations of current culture methods, acceptance of new and potentially superior methods is often slow. We believe this is due in part to a lack of clear guidance regarding the demonstra-tion of their equivalence to existing methods accept-able to regulatory agencies and validation of the equip-ment associated with the new methods. This techni-cal report hopes to provide guidance to assist with the evaluation, validation, and implementation of the newer microbiological methods.Considerable guidance can be found regarding the validation of chemical methods. Examples include USP General Informational Chapter <1225> Valida-tion of Compendial Method s (1), and a recent publi-cation by the International Conference on Harmoni-zation (ICH) Validation of Analytical Methods (2). These publications provide very specific instruction regarding the demonstration of new analytical chem-istry methods and their equivalence to existing meth-ods. In contrast, virtually no guidance specific to mi-crobiological testing has been published. Possible exceptions are the ASM Cumitech publication V erifi-cation and Validation of Procedures in the Clinical Microbiology Laboratory (3), that addresses pathogen isolation and identification and antimicrobial suscep-tibility testing, and the new USP General Information Chapter <1227> Validation of Microbial Recovery from Pharmacopeial Articles (4). However, more guidance is needed. Because microbiological meth-ods are inherently different than chemical assays, this lack of agreed upon demonstration criteria can present serious obstacles to their implementation.When instrumentation is developed for existing mi-crobiological methods to automate sample handling, result reading, or data management or to miniaturize the test procedure, it is not difficult to demonstrate the equivalency of the alternate method using guidelines developed for chemical assays, because the test re-mains essentially the same. In a similar fashion, when a new technology continues to rely on the measure-ment of microbial growth (e.g., impedance, ATP bioluminescence or other metabolic changes in aVol.54, No. 3, May/June 2000, Supplement TR33 1microbial culture), equivalence can be readily dem-onstrated. However, when a new method is based on novel technology without direct ties to the ex-isting method (e.g., microbial identification by DNA amplification versus patterns of biochemical reac-tions, or counting fluorescent- labeled bacterial cells instead of colony-forming units on an agar plate), demonstration of equivalency may require a new application of the validation principles, although the method provides higher quality results.The application of these new technologies may be compared to the replacement of the Most Probable Number-multiple fermentation tubes with mem-brane filtration methods for counting coliforms in water, and the change from solely morphological features to physiological and biochemical charac-teristics to DNA-based methods for the identifica-tion of bacteria. For example, the ability to count individual, viable, fluorescent-labeled microbial cells by scanning a membrane with a laser should be superior to counting colony-forming units; the nucleic acid contained in a microorganism is con-servatively unique to that species, i.e., genotypic identification, hence is superior to patterns of bio-chemical reactions that are currently used to iden-tify bacteria, i.e., phenotypic identification.This document, was developed as a collaborative effort amongst representatives from test manufac-turers, the pharmaceutical and device industry, stan-dards organizations, and regulatory agencies. It is intended to provide a general approach to the intro-duction of new microbiology methods in a govern-ment-regulated environment. It is anticipated that by providing agreed upon performance standards, the development, demonstration and implementa-tion of superior methods will be greatly accelerated.1.3 Overview of Document StructureThis document was written to establish industry-wide criteria on what constitutes an acceptable mi-crobiology test and how to prove it to the satisfac-tion of a regulatory agency.The document is divided into two major sections. In the Method Selection section, a review of micro-biology testing methods is followed by an overview of compendial applications which make use of these tests. A discussion of requirements for regulatory acceptance and economic justification completes the section. In the Validation section, criteria used for validation and demonstration of equivalence are de-fined, and approaches to validation methods and documentation are described. Finally, a decision tree, glossary of commonly used terms, and a bibliogra-phy are provided to present an overview of the en-tire process.2.0 TECHNOLOGY OVERVIEW2.1 Generic Description of Types ofMicrobiological MethodsMicrobiological test methods can be divided into three general categories, based on their function: 1) detection of the presence or absence of micro-organisms in a test sample, 2) enumeration of mi-croorganisms present in a test sample, and 3) char-acterization and identification of microorganisms either present in test samples or from a pure cul-ture.Presence/absence tests may be designed to detect diverse types of microorganisms, as in sterility test methods, or may be intended for detection of specific microbial species or genera, as in the tests for Pharmacopeial Indicator Organisms. USP, Ph. Eur. and JP have the same set of indicator organ-isms, i.e., Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmo-nella species.The results obtained when testing samples by dif-ferent enumeration methods based on classical mi-crobiological techniques may vary greatly, de-pending on the test conditions, e.g., media type and incubation temperature, conditions, and du-ration.An example is the difference seen in plate counts from water samples using a low nutrient medium2 PDA Journal of Pharmaceutical Science & Technologysuch as R2A agar with a 5 to 7 day incubation period at ambient temperature, versus Soybean –Casein Digest agar incubated for 48 hours at 30 to 35°C. Significant differences in counts may also be seen when comparing non-growth based direct detection methods with classical methods, with higher counts obtained by the former due to suboptimal culture conditions in microbiological media.Many types of microbial characterization and identification test methods are used, ranging from classical Gram’s staining and simple manual tests for specific enzymes associated with bacterial cells such as catalase and cytochrome oxidase, to compendial methods using the appearance of microbial colonies on selective/differential growth media, to semi-automated biochemical profiling systems, to methods based on the de-tection of specific nucleic acid sequences or cell membrane fatty acid composition.The test methods vary greatly in their cost, ease of use, duration, and performance as measured by analytical parameters such as specificity, sen-sitivity, reproducibility, and ruggedness.2.2 Technology ReviewAs discussed earlier, the century-old methods de-veloped by the pioneering microbiologists Pasteur, Koch, Lister, and many others is the technology base for current compendial microbiological test meth-ods. These methods are based on providing condi-tions to allow microbial cells present in test samples to grow and replicate sufficiently to allow their de-tection, typically by visual examination of a plate or broth. Specific compendial tests will be discussed later in the section on Regulatory Review, so the focus of this section will be on newer technologies. New technology for microbiology quality assur-ance testing is a rapidly developing area. The Task Force has attempted to include all technologies with current application in the pharmaceutical in-dustry. However, exclusion of a technology or application of a technology is not intentional, and does not imply that the technology is not suitable. This review is simply the best available informa-tion at the time. Without doubt, further develop-ments will occur during the lifetime of this docu-ment, which will mean that new technologies or new applications of existing technologies will be brought to market. For convenience, the technologies are divided into 1) growth-based technologies, 2) vi-ability-based technologies, 3) cellular component or artifact-based technologies, and 4) nucleic acid-based technologies.Reference to representative suppliers (see Tables 1, 2, 3, and 4) is merely given as examples compiled by the Task Force members and does not infer commer-cial endorsement on the part of the Task Force or PDA.2.3 Growth-based TechnologiesThese methods are based on the measurement of biochemical or physiological parameters that reflect the growth of the microorganisms.2.3.1 ATP BioluminescenceThe presence of Adenosine Triphosphate (ATP) is a well-documented marker for cell viability. All cells store energy in the form of ATP. An increase in cell numbers results in an increase in ATP lev-els. ATP bioluminescence utilizes the luciferin-lu-ciferase reaction of the firefly to detect the pres-ence of ATP by measuring the light emitted. The presence or absence of microbial contamination in a sample can be determined by measuring the in-crease in ATP levels following incubation. ATP bi-oluminescence reduces the test time to approxi-mately one third of that taken by the traditional method. This is because it is a more sensitive end-point detection system, using sensitive chemistry and instrumentation, rather than relying on the hu-man eye. This means that identical principles are in use, i.e., multiplication of microorganisms, but the amount of replication required for detection is sig-nificantly reduced: 103 cells per mL as opposed to 107 or greater, to determine turbidity or colonies on a plate.Vol.54, No. 3, May/June 2000, Supplement TR33 32.3.2 Colorimetric Detection of COP roductionTest samples are placed in culture bottles for moni-toring. The samples are incubated, agitated, and monitored for the presence of microorganisms. These systems use colorimetric detection of CO production from the growth of organisms. Some systems will detect color change, flag a positive sample, and alert the user. These systems are often referred to as non-invasive microbial detection sys-tems and can accommodate a large number of samples. Although commonly used clinically for blood cultures, the method could be used for steril-ity testing.2.3.3 Measurement of Change in Head Space PressureThese systems are based on non-invasive, continu-ous, automated monitoring of microbial cultures. Electronic transducers are used to sense positive or negative pressure changes in the head space of each culture bottle. These changes are caused by micro-bial growth. If significant production and/ or con-sumption of gas is detected, samples are flagged as positive. Large quantities of samples can be placed into these instruments for testing with frequent monitoring of the head space pressure. Although commonly used clinically for blood cultures, the method could also be used for sterility testing.2.3.4 ImpedanceImpedance measures microbial activity by elec-trical methods. These instruments measure ionic changes occurring within the growth media as bacteria multiply. The impedance detection time is inversely proportional to the number of micro-organisms present at initial inoculation. Bacteria metabolize larger, weakly charged molecules and produce smaller highly charged by-products. For example, large weakly charged molecules such as proteins are hydrolyzed to many highly charged amino acids. Two electrodes are then used to measure these ionic changes in the broth or agar culture.2.3.5 Biochemical AssaysMicrobial cell suspensions of pure cultures are tested with a series of biochemical substrates. Mi-croorganisms are known to have particular reac-tions to these biochemicals, e.g., carbohydrate uti-lization. By matching the biochemical results with a database of corresponding results, one can determine the identification of the organism be-ing tested. Many of these methods are performed and recorded manually. High volume, automated instruments are also commercially available to read miniature cultures and identify the microor-ganisms from the pattern of reactions in the data-base.In the following tables, presence/absence, enu-meration and identification methods are designed as P, E, and I respectively.224 PDA Journal of Pharmaceutical Science & TechnologySteriScreen and MicroStar RapidMicrobiology and MicroCount Digital Systems (Millipore Corp, Bedford, MA & RapiScreen (Celsis, Evanston, IL)Bact/Alert (Organon-Teknika, Durham,NC) & ESP Microbial Detection System (AccuMed International, Detroit, MI)API Systems (bioMerieux, Hazelwood,MO), BIOLOG Systems (Biolog, San Diego, CA) & VITEK System (bioMerieux, Hazelwood, MO)Bactometer (bioMerieux, Hazelwood,MO) & Malthus Microbial Detection System (Malthus Diagnostics North Ridgeville, OH)Spiral Plating System (Spiral Biotech,Bethesda, MD)Iso-Grid (QA LifeScience, San Diego,CA)ConventionalConventionalConventionalRaw material & product screening, water monitor-ing, pre-sterile filtration bioburden monitoring (E)Sterility testing (P)Sterility testing (P)Microbial identification (I)Bioburden monitoring,antimicrobial effectiveness testing (E)Bioburden monitoring (E)Bioburden monitoring (E)Pathogen monitoring (E)Bioburden monitoring,water monitoring,antimicrobial effectiveness testing (E)Bioburden monitoring,water monitoring, D-Value analysis (E)Bioburden monitoring (E)Sterility testing (P),Water monitoring (E)ATPBioluminescenceColorimetric Detection of Carbon Dioxide Production Headspace Pressure Biochemical &Physiological ReactionsImpedance /ConductivitySpiral plating Hydrophobic Grid Membrane Filter MethodsPour Plate MethodMost Probable Number Multiple -Tube Method Membrane Filtration MethodTable 1: Growth-based Microbiological Methods Technology Representative Commercial Products Principal Applications Vol.54, No. 3, May /June 2000, Supplement TR33 5。
