Regulated Bioanalytical Laboratories

Michael Zhou, PhD, is Director of Bioanalytical Chemistry at Synta Pharmaceuticals Corporation and a highly regarded expert on analytical/bioanalytical operations, regulatory compliance, including Good Laboratory Practice (GLP), cGMP, GCP regulations, and ICH, GCLP, BMV guidelines. Dr. Zhou has given numerous presentations and workshops / short courses on the topics at national and international conferences and has authored over fifty research articles and two book chapters on analytical/bioanalytical chemistry

Preface xiii

Acknowledgment xvii

Contributors and Advisors xix

1 Introduction, Objectives, and Key Requirements for GLP Regulations 1

1.1 Introduction 1

1.1.1 Good Laboratory Practices 1

1.1.2 Bioanalytical Laboratories—Bioanalysis 4

1.1.3 Good Laboratory Practices Versus Bioanalytical Labs/ Bioanalysis 7

1.2 Objectives and Key Requirements for GLP Regulations 8

1.3 Fundamental Understanding of GLP Regulations and Principles 10

1.3.1 Elements of Good Laboratory Practices 11

1.4 Key Elements of Bioanalytical Methods Validation 16

1.4.1 Reference Standards 19

1.4.2 Method Development—Chemical/Chromatographic Assay 20

1.4.3 Calibration/Standard Curve 21

1.4.4 Stability 21

1.4.5 Reproducibility 23

1.4.6 Robustness or Ruggedness 23

1.5 Basic Principles of Bioanalytical Method Validation and Establishment 23

1.5.1 Specific Recommendations for Method Validation 24

1.5.2 Acceptance Criteria for Analytical Run 29

References 33

2 Historic Perspectives of GLP Regulations, Applicability, and Relation to Other Regulations 35

2.1 Historic Perspectives of GLP Regulations 35

2.1.1 Economic Assessment 39

2.1.2 Environmental Impact 40

2.2 Applicability and Relations to Other Regulations/Principles 42

2.2.1 GLP, GCP, GMP, and Part 11 42

2.2.2 General Terminologies and Definitions of GxPs (GLP, GCP, and cGMP) 47

2.3 Comparison of FDA GLP, EPA GLP Regulations, and OECD GLP Principles 47

2.3.1 US and OECD GLP Similarity and Differences 53

2.4 Applications of GLP to Multiple Site Studies 55

2.4.1 Roles and Responsibilities 57

2.4.2 Performance of the Studies 61

2.4.3 Applications of GLP to In Vitro Studies for Regulatory Submissions 64

2.5 21 CFR Part 11 in Relation to GLP Programs 66

2.5.1 A New Risk-Based Approach 67

2.5.2 Understanding Predicate Rule Requirements 67

2.5.3 21 CFR Part 11 Best Practices 68

2.5.4 Use of Electronic Signatures 71

2.6 GLP, cGMP, and ISO Applicabilities, Similarity, and Differences 74

2.6.1 GLPs, cGMPs, ISO 17025:2005: How Do They Differ? 74

2.6.2 GLPs Versus GMPs 74

2.6.3 GLPs Versus ISO/IEC 17025:2005 75

2.6.4 ISO Versus GLPs 76

2.7 Good Clinical Practices and Good Clinical Laboratory Practices 78

2.8 Gap and Current Initiatives on Regulating Laboratory Analysis in Support of Clinical Trials 80

References 84

3 GLP Quality System and Implementation 87

3.1 GLP Quality System 87

3.1.1 Regulatory Inspection for GLP Quality System 95

3.1.2 Good Laboratory Practice Inspections 99

3.1.3 GLP Quality System Objectives 103

3.2 Global GLP Regulations and Principles 106

3.2.1 General 106

3.2.2 Responsibilities and Compliance 107

3.2.3 Statement of Compliance in the Final Report 107

3.2.4 Protocol Approval 108

3.2.5 Assignment of Study Director 108

3.2.6 Laboratory Qualification/Certification 108

3.2.7 Authority Inspections 108

3.2.8 Archiving Requirements 108

3.3 Implementation of GLP Regulations and OECD Principles 109

3.3.1 Planning (Master Schedule) 114

3.3.2 Personnel Organization 115

3.3.3 Curriculum Vitae 115

3.3.4 Rules of the Conducts of Studies 116

3.3.5 Content of Study Protocol 116

3.3.6 Approval of Study Protocol 118

3.3.7 Distribution of Study Protocol 118

3.3.8 Protocol Amendment 118

3.3.9 Standard Operating Procedures 119

3.3.10 SOP System Overview 119

3.3.11 Characterization 121

3.3.12 Test Item/Article Control before Formulation 121

3.3.13 Preparation of the Dose Formulation 123

3.3.14 Sampling and Quality Control of Dose Formulation 125

3.4 Initiatives and Implementation of Bioanalytical Method Validation (Guidance for Industry BMV—May 2001) 126

3.4.1 Summary 127

References 128

4 Fundamental Elements and Structures for Regulated Bioanalytical Laboratories 131

4.1 Introduction 131

4.2 Fundamental Elements for Bioanalytical Laboratories 133

4.2.1 Document Retention and Archiving 136

4.3 Basic Requirements for GLP Infrastructure and Operations 139

4.4 GxP Quality Systems 143

4.4.1 Laboratory Instrument Qualification and Validation 149

4.4.2 Procedural Elements and Function that Maintain Bioanalytical Data Integrity for GLP Studies 150

References 166

5 Technical and Regulatory Aspects of Bioanalytical Laboratories 167

5.1 Fundamental Roles and Responsibilities of Bioanalytical Laboratories 167

5.1.1 Technical Functions of Bioanalytical Laboratories 168

5.1.2 Basic Processes in Bioanalytical Method Development, Validation, and Sample Analysis 173

5.2 Qualification of Personnel, Instrumentation, and Analytical Procedures 178

5.2.1 From Regulatory Perspectives: Personnel, Training, and Qualification 183

5.2.2 Facility Design and Qualifications 186

5.2.3 Equipment Design and Qualification 186

5.2.4 Analytical/Bioanalytical Method Qualification and Validation along with Related SOPs 197

5.3 Regulatory Compliance with GLP Within Bioanalytical Laboratories 204

5.4 Joint-Effort from Industries and Regulatory Agencies 206

5.4.1 Ligand-Binding Assays In-Study Acceptance Criteria 213

5.4.2 Determination of Metabolites during Drug Development 216

5.4.3 Incurred Sample Analysis 216

5.4.4 Documentation Issues 217

5.4.5 Analytical/Validation Reports 218

5.4.6 Source Data Documentation 218

5.4.7 Final Report Documentation 219

5.4.8 Stability Recommendation 219

5.4.9 Matrix Effects for Mass Spectrometric-Based Assays 221

5.4.10 System Suitability 222

5.4.11 Reference Standards 222

5.4.12 Validation Topics with No Consensus 222

5.4.13 Specific Criteria for Cross-Validation 223

5.4.14 Separate Stability Experiments Required at – 70°C if Stability Shown at – 20°C 223

5.4.15 Stability Criteria for Stock Solution Stability 224

5.4.16 Acceptance Criteria for Internal Standards 224

5.4.17 Summary 224

References 226

6 Competitiveness of Bioanalytical Laboratories—Technical and Regulatory Perspectives 229

6.1 Technical Aspect of Competitive Bioanalytical Laboratories 229

6.2 Bioanalytical Processes and Techniques 232

6.2.1 Sample Generation, Shipment, and Storage 232

6.2.2 Sample Preparation 233

6.3 Enhancing Throughput and Efficiency in Bioanalysis 243

6.3.1 Chromatographic Separation 244

6.3.2 Selective and Sensitive Detection 251

6.4 Technical Challenges and Issues on Regulated Bioanalysis 254

6.4.1 Matrix Effect 254

6.4.2 Method Validation and Critical Issues during Sample Analysis 256

6.4.3 Method Transfer 258

6.5 Regulatory Aspects of Competitive Bioanalytical Laboratories 264

6.5.1 General Consideration 264

6.5.2 Historical Perspective 265

6.5.3 Personnel—Training and Qualification 267

6.5.4 Facility—Design and Qualifications 269

6.5.5 Equipment Design and Qualification 270

6.5.6 Standard Operating Procedures 272

6.5.7 Laboratory/Facility Qualification Perspectives 272

6.6 Advanced/Competitive Bioanalytical Laboratories 277

6.6.1 Strategy Versus Tactics 278

6.6.2 Bioanalytical Laboratory Assessment 279

6.6.3 Capacity 279

6.6.4 Experience 280

6.6.5 Quality 281

6.6.6 Performance and Productivity Measures 281

6.6.7 Information Technology and Data Management 282

6.6.8 Communication 282

6.6.9 Financial Stability 283

6.6.10 Ease of Use 283

6.6.11 Contracting Bioanalytical Services 284

6.6.12 The Contracting Process 284

6.7 Applications and Advances in Biomarker and/or Ligand-Binding Assays within Bioanalytical Laboratories 286

References 290

7 Sponsor and FDA/Regulatory Agency GLP Inspections and Study Audits 297

7.1 GLP versus Biomedical Research Monitoring and Mutual Acceptance of Data for Global Regulations and Inspections 298

7.2 Purposes and Benefits of Regulatory Inspections/Audits 303

7.2.1 Criteria for Selecting Ongoing and Completed Studies 304

7.2.2 Areas of Expertise of the Facility 305

7.2.3 Establishment Inspections 305

7.2.4 Organization and Personnel (21 CFR 58.29, 58.31, 58.33) 305

7.2.5 Quality Assurance Unit (QAU; 21 CFR 58.35) 307

7.2.6 Facilities (21 CFR 58.41–58.51) 308

7.2.7 Equipment (21 CFR 58.61–58.63) 309

7.2.8 Testing Facility Operations (21 CFR 58.81) 310

7.2.9 Reagents and Solutions (21 CFR 58.83) 311

7.2.10 Animal Care (21 CFR 58.90) 311

7.2.11 Test and Control Articles (21 CFR 58.105–58.113) 312

7.2.12 Test and Control Article Handling (21 CFR 58.107) 313

7.2.13 Protocol and Conduct of Nonclinical Laboratory Study (21 CFR 58.120–58.130) 314

7.2.14 Study Protocol (21 CFR 58.120) 314

7.2.15 Test System Monitoring 314

7.2.16 Records and Reports (21 CFR 58.185–58.195) 314

7.2.17 Data Audit 316

7.2.18 General 316

7.2.19 Final Report Versus Raw Data 317

7.2.20 Specimens Versus Final Report 318

7.2.21 Refusal to Permit Inspection 318

7.2.22 Sealing of Research Records 318

7.2.23 Samples 319

7.3 Typical Inspections/Audits and Their Observations 320

7.4 Regulatory Challenges for Bioanalytical Laboratories 321

7.4.1 Introduction 321

7.4.2 Analysis of Current FDA Inspection Trends 324

7.4.3 Discussion and Analysis of Specific Potential FDA 483

Observation Issues 325

7.4.4 Method Validation Issues 325

7.4.5 Batch Runs Acceptance Criteria Issues 329

7.4.6 Events/Deviations Investigation/Resolution Issues 331

7.4.7 Test Specimen Accountability Issue 333

7.4.8 Recommendations to Support an Effective FDA Inspection Readiness Preparation 334

7.5 Handling and Facilitating GLP or GxP Audits/ Inspections 334

7.5.1 General Preparation for an Inspection 336

7.5.2 Why Are Audits/Inspections Needed and Conducted? 342

7.5.3 Written Policy in Place 342

7.5.4 Positions on Controversial Issues 343

7.5.5 The Inspection Coordinator 344

7.5.6 Follow-Up Procedures 348

7.5.7 Summary 349

References 351

8 Current Strategies and Future Trends 353

8.1 Strategies from General Laboratory and Regulatory Perspectives 354

8.2 Strategies from Technical and Operational Perspectives 356

8.3 Biological Sample Collection, Storage, and Preparation 360

8.3.1 Sample Collection and Storage 360

8.3.2 Sample Preparation Techniques 361

8.3.3 Off-Line Sample Extraction 364

8.3.4 On-Line Sample Extraction 364

8.4 Strategies for Enhancing Mass Spectrometric Detection 366

8.4.1 Enhanced Mass Resolution 368

8.4.2 Atmospheric Pressure Photoionization 369

8.4.3 High-Field Asymmetric Waveform Ion Mobility Spectrometry 370

8.4.4 Electron Capture Atmospheric Pressure Chemical Ionization 370

8.4.5 Mobile Phase Optimization for Improved Detection and Quantitation 371

8.4.6 Anionic and Cationic Adducts as Analytical Precursor Ions 372

8.4.7 Derivatization 372

8.5 Strategies for Enhancing Chromatography 374

8.5.1 Ultra-Performance Chromatography 375

8.5.2 Hydrophilic Interaction Chromatography for Polar Analytes 376

8.5.3 Specialized Reversed-Phase Columns for Polar Analytes 377

8.5.4 Ion-Pair Reversed-Phase Chromatography for Polar Analytes 378

8.6 Potential Pitfalls in LC–MS/MS Bioanalysis 378

8.6.1 Interference from Metabolites or Prodrugs due to In-Source Conversion to Drug 378

8.6.2 Interference from Metabolites or Prodrugs due to Simultaneous M + H+ and M + NH4+ Formation or Arising from Isotopic Distribution 379

8.6.3 Pitfall in Analysis of Two Interconverting Analytes due to Inappropriate Method Design 383

8.6.4 Matrix Effect 383

8.7 Trends in High-Throughput Quantitation 386

8.7.1 System Throughput 386

8.7.2 High-Speed HPLC 386

8.8 Trends in Hybrid Coupling Detection Techniques 388

8.9 Trends in Internal R&D and External Outsourcing 388

8.10 Trends in Ligand-Binding Assays and LC–MS/MS for Biomarker Assay Applications 397

8.11 Trends in Study Design and Evaluation Relating to Bioanalysis 399

8.12 Trends in Applying GLP to In Vitro Studies in Support of Regulatory Submissions 403

8.13 Trends in Global R&D Operations 404

8.14 Trends in Regulatory Implementations 407

8.14.1 Calibration Range and Quality Control Samples 407

8.14.2 Incurred Sample Reproducibility (Duplicate Sample Analysis) 408

8.14.3 LIMS and Electronic Data Handling, Security, Archiving, and Submission 409

8.15 Trends in Global Regulations and Quality Standards 412

8.16 Trends in Compliance with 21 CFR Part 11 414

8.16.1 21 CFR Part 11 Software Requirements 415

8.16.2 Building a Roadmap for Compliance with 21 CFR Part 11 415

8.16.3 Low Hanging Fruits in the Roadmap for Compliance with 21 CFR Part 11 416

8.17 Summary 419

References 421

9 General Terminologies of GxP and Bioanalytical Laboratories 431

9.1 General Terminologies for GxP and Bioanalytical Laboratories 431

9.2 GLP Basic Concepts and Implementation 469

9.2.1 The Study Protocol 470

9.2.2 Raw Data 471

9.2.3 The GLP Archive and the Archivist 472

9.2.4 Expansion of GLP Scope 473

9.2.5 OECD GLP 473

9.3 GLP Guidance Documents 474

9.3.1 FDA Guidance for Industry on Bioanalytical Method Validation 474

9.3.2 OECD GLP Guidance Documents 474

9.3.3 Swiss GLP Guidance Documents 475

References and Sources for Above Terminologies 475

Appendix A Generic Checklist for GLP/GXP Inspections/Audits 479

Appendix B General Template for SOP 489

Appendix C Typical SOPs for GLP/Regulated Bioanalytical Laboratory 493

Quality Assurance—GLP 493

Bioanalytical—GLP Laboratories 494

Appendix D Basic Equipment/Apparatus for Bioanalytical Laboratory 497

Appendix E Website Linkages for Regulated Bioanalysis 499

Index 503