HANDBOOK OF LC-MS BIOANALYSIS -- CONTENTS -- PREFACE -- CONTRIBUTORS -- ABBREVIATIONS -- PART I OVERVIEW OF LC-MS BIOANALYSIS -- 1 ROLES OF LC-MS BIOANALYSIS IN DRUG DISCOVERY, DEVELOPMENT, AND THERAPEUTIC DRUG MONITORING -- 1.1 INTRODUCTION -- 1.2 LC-MS BIOANALYSIS IN DRUG DISCOVERY -- 1.2.1 Structure-Activity Relationships from High-Throughput Screening -- 1.2.2 Structure-PK-PD Relationships -- 1.2.3 Candidate Selection -- 1.3 LC-MS BIOANALYSIS IN PRECLINICAL DEVELOPMENT OF DRUGS -- 1.3.1 Toxicokinetics -- 1.3.2 Preclinical ADME and Tissue Distribution Studies in Animals -- 1.4 LC-MS BIOANALYSIS IN CLINICAL DEVELOPMENT OF DRUGS -- 1.4.1 First-in-Human Studies -- 1.4.2 Human ADME Studies -- 1.4.3 Human Drug-Drug Interaction Studies -- 1.4.4 Renal Impaired and Hepatic Impaired Studies in Human -- 1.4.5 Phase II and Phase III Studies -- 1.4.6 "Fit-for-Purpose" Biomarker Measurement Using LC-MS in Clinical Samples -- 1.4.7 Other LC-MS Assays Needed for Clinical Development of Drugs -- 1.4.8 LC-MS Bioanalysis in Postapproval Studies (Phase IV) of Drugs -- 1.4.9 LC-MS Bioanalysis in BE and BA Studies for Generic Drugs -- 1.4.10 LC-MS Bioanalysis in Therapeutic Drug Monitoring -- 1.5 LC-MS BIOANALYSIS OF LARGE MOLECULE DRUGS AND BIOPHARMACEUTICALS -- 1.6 GUIDANCE AND REGULATIONS FOR LC-MS BIOANALYSIS -- 1.7 GENERAL CONSIDERATIONS OF A ROBUST LC-MS BIOANALYTICAL METHOD -- 1.8 CONCLUSIONS -- REFERENCES -- 2 OVERVIEW: FUNDAMENTALS OF A BIOANALYTICAL LABORATORY -- 2.1 INTRODUCTION -- 2.2 KEY ELEMENTS OF A BA LABORATORY -- 2.2.1 Facility -- 2.2.2 Infrastructure -- 2.2.3 Compliance -- 2.2.4 Documentation -- 2.3 QUALITY ASSURANCE -- 2.4 SUPPORTING FUNCTIONS -- 2.4.1 Quality Control -- 2.4.2 Sample Management -- 2.4.3 Archivist -- 2.4.4 Technical Support -- 2.4.5 Planning -- 2.4.6 Report Generation -- 2.5 CRO MONITORING -- 2.5.1 Personnel.

2.5.2 Outsource Process -- 2.6 CONCLUSIONS -- REFERENCES -- 3 INTERNATIONAL REGULATIONS AND QUALITY STANDARDS OF BIOANALYSIS -- 3.1 INTRODUCTION -- 3.2 GLOBAL BIOANALYTICAL GUIDANCE -- 3.3 BIOANALYTICAL QUALITY -- 3.3.1 Bioanalytical Quality Strategies -- 3.3.2 Quantitative Assessment -- 3.3.3 Qualitative Assessment -- 3.4 SCIENCE, QUALITY, AND REGULATION -- ACKNOWLEDGMENTS -- REFERENCES -- PART II CURRENT UNDERSTANDING OF LC-MS BIOANALYSIS-RELATED REGULATIONS -- 4 CURRENT REGULATIONS FOR BIOANALYTICAL METHOD VALIDATIONS -- 4.1 INTRODUCTION -- 4.2 CONTEXT OF THE REGULATORY ENVIRONMENT -- 4.3 VALIDATIONS OF METHODS -- 4.3.1 Matrix Requirements -- 4.3.2 Reference Standards -- 4.3.3 Robustness Testing -- 4.3.4 Sensitivity -- 4.3.5 Selectivity -- 4.3.6 Recovery -- 4.3.7 Matrix Effects -- 4.3.8 Calibration Curves -- 4.3.9 QC Samples -- 4.3.10 Stability in Matrix -- 4.3.11 Stock Solution Stability -- 4.3.12 System Suitability -- 4.3.13 Carryover -- 4.3.14 Determination of Metabolites -- 4.3.15 Incurred Sample Reanalysis -- 4.3.16 Run Size -- 4.3.17 Reporting -- 4.3.18 Other Topics -- 4.4 CONCLUSION -- REFERENCES -- 5 CURRENT UNDERSTANDING OF BIOANALYTICAL ASSAY REPRODUCIBILITY: INCURRED SAMPLE REANALYSIS, INCURRED SAMPLE STABILITY, AND INCURRED SAMPLE ACCURACY -- 5.1 INTRODUCTION -- 5.2 INCURRED SAMPLE REANALYSIS -- 5.2.1 Principles and Practice of ISR -- 5.2.2 Statistical Approaches for Assessment of ISR -- 5.2.3 Case Studies on ISR -- 5.2.4 Dried Blood Spot ISR -- 5.2.5 Approach for Efficient Conduct of ISR -- 5.2.6 Recommendations for ISR Investigation -- 5.3 INCURRED SAMPLE STABILITY -- 5.4 INCURRED SAMPLE ACCURACY -- 5.4.1 Methodology for ISA Assessment -- 5.4.2 ISA Data Interpretation -- 5.4.3 Reflections on the Use of ISA in Bioanalytical Method Validation -- 5.5 SUMMARY -- REFERENCES -- 6 LC-MS BIOANALYTICAL METHOD TRANSFER -- 6.1 INTRODUCTION.

6.2 PREPARATION FOR METHOD TRANSFER -- 6.3 CURRENT UNDERSTANDING ON REGULATORY REQUIREMENTS FOR BIOANALYTICAL METHOD TRANSFER -- 6.4 METHOD TRANSFER -- 6.4.1 Partial/Cross-validation versus Full Validation -- 6.4.2 Method Modification -- 6.4.3 Acceptance Criteria -- 6.5 COMMON CAUSES OF BIOANALYTICAL METHOD TRANSFER FAILURE -- 6.6 INVESTIGATION OF A METHOD TRANSFER FAILURE -- 6.7 SUMMARY -- REFERENCES -- 7 METABOLITES IN SAFETY TESTING -- 7.1 INTRODUCTION -- 7.2 TIMING OF ADME STUDIES WITH RADIOLABELED MATERIALS -- 7.3 FIH STUDIES -- 7.4 STANDARD FREE QUANTIFICATION AND ITS LIMITATIONS -- 7.5 TIERED OPTIONS FOR DETERMINATION OF HUMAN METABOLITE EXPOSURES AND RESPECTIVE LIMITATIONS -- 7.5.1 Tier 1-Metabolite Profiling Assay -- 7.5.2 Tier 2-Standard Free Quantification/Response Factor Determination -- 7.5.3 Tier 3-Qualified Assays -- 7.5.4 Tier 4-Validated Assays -- 7.6 SUMMARY -- REFERENCES -- 8 A COMPARISON OF FDA, EMA, ANVISA, AND OTHERS ON BIOANALYSIS IN SUPPORT OF BIOEQUIVALENCE/ BIOAVAILABILITY STUDIES -- 8.1 INTRODUCTION TO BIOAVAILABILITY/ BIOEQUIVALENCY STUDIES -- 8.2 REGULATIONS FROM THE US FDA -- 8.2.1 History and Validation -- 8.2.2 Conduct of the Bioanalysis -- 8.3 REGULATIONS FROM THE EUROPEAN MEDICINES AGENCY -- 8.3.1 The Bioanalytical Method -- 8.3.2 Bioanalysis of Enantiomers -- 8.4 REGULATIONS FROM THE BRAZILIAN SANITARY SURVEILLANCE AGENCY (ANVISA) -- 8.4.1 Method Validation -- 8.4.2 Conduct of Bioanalysis -- 8.5 OTHER INTERNATIONAL GUIDELINES -- 8.5.1 Indian Department of Health -- 8.5.2 Health Canada -- 8.5.3 China State FDA -- 8.6 CONCLUSION -- REFERENCES -- 9 A COMPARISON OF THE GUIDANCE OF FDA, OECD, EPA, AND OTHERS ON GOOD LABORATORY PRACTICE -- 9.1 INTRODUCTION -- 9.2 FDA VERSUS EPA ON GLP -- 9.3 FDA GLP VERSUS OECD GLP PRINCIPLES -- 9.4 SOME COUNTRY SPECIFIC REQUIREMENTS ON GLP -- 9.5 GLP INSPECTION -- 9.6 SUMMARY.

REFERENCES -- 10 CURRENT UNDERSTANDING OF BIOANALYSIS DATA MANAGEMENT AND TREND OF REGULATIONS ON DATA MANAGEMENT -- 10.1 INTRODUCTION -- 10.1.1 Historical Overview -- 10.1.2 The Need for Regulated Bioanalysis Data Management -- 10.2 BIOANALYTICAL WORKFLOW AND DATA MANAGEMENT -- 10.2.1 Bioanalytical Workflow and Data Flow -- 10.2.2 Quality Data Management Software Systems -- 10.3 COMPUTER SYSTEMS VALIDATION -- 10.3.1 Overview -- 10.3.2 Outline of Validation Procedures -- 10.3.3 Future Direction of Computer Systems Validation -- 10.3.4 Trend of the Regulations on Data Management -- 10.4 CHALLENGES FOR BIOANALYTICAL DATA INTEGRITY AND SECURITY -- 10.5 FUTURE PERSPECTIVES -- 10.6 CONCLUSIONS -- REFERENCES -- 11 REGULATORY INSPECTION TRENDS AND FINDINGS OF BIOANALYTICAL LABORATORIES -- 11.1 INTRODUCTION -- 11.2 CURRENT REGULATORY INSPECTION TRENDS -- 11.3 INADEQUATE INVESTIGATION-THE MDS CASE -- 11.4 DATA INTEGRITY CONCERN-THE CETERO CASE -- 11.5 DISCUSSION AND ANALYSIS OF SPECIFIC REGULATORY INSPECTION FINDINGS -- 11.5.1 Data Integrity/Electronic Records Issues -- 11.5.2 Method Validation Issues -- 11.5.3 Batch Runs Acceptance Criteria Issues -- 11.5.4 Events/Deviations Investigation/Resolution Issues -- 11.5.5 Test Specimen Accountability Issue -- 11.6 RECOMMENDATIONS TO SUPPORT AN EFFECTIVE FDA INSPECTION-READINESS PREPARATION -- REFERENCES -- PART III BEST PRACTICE IN LC-MS BIOANALYSIS -- 12 ASSESSMENT OF WHOLE BLOOD STABILITY AND BLOOD/PLASMA DISTRIBUTION OF DRUGS -- 12.1 ASSESSMENT OF WHOLE BLOOD STABILITY OF DRUGS -- 12.1.1 Factors Affecting the Stability of Drugs in Blood -- 12.1.2 Considerations of Experimental Design for Whole Blood Stability Assessment of Drugs -- 12.1.3 The Choice of Analytical Matrix for Blood Stability Assessment -- 12.1.4 Stability of Metabolites -- 12.1.5 Statistical Methods for Blood Stability Assessment.

12.2 BLOOD/PLASMA DISTRIBUTION OF DRUGS -- 12.2.1 Mechanisms of Blood/Plasma Distribution/Partitioning of Drugs -- 12.2.2 Measurement of the Blood-to-Plasma Partition Ratio -- 12.2.3 Factors to be Considered in Blood/Plasma Partition Assessment of Drugs -- 12.3 SUMMARY -- ACKNOWLEDGMENT -- REFERENCES -- 13 BEST PRACTICE IN BIOLOGICAL SAMPLE COLLECTION, PROCESSING, AND STORAGE FOR LC-MS IN BIOANALYSIS OF DRUGS -- 13.1 INTRODUCTION -- 13.2 SAMPLE COLLECTION -- 13.2.1 In vivo Sample Collection -- 13.2.2 Choice of Anticoagulant -- 13.2.3 Other Sample Tube Additives -- 13.2.4 Preparing Plasma from Blood -- 13.2.5 Minimizing Clot Formation -- 13.2.6 Difference between Plasma and Serum -- 13.2.7 Collection of Urine Samples -- 13.2.8 Collection of Tissue Samples -- 13.2.9 Collection of Fecal Samples -- 13.2.10 Best Practice for Use of Centrifuges -- 13.2.11 Time Factors -- 13.2.12 Working on Ice -- 13.3 DOCUMENTATION -- 13.3.1 Procedures for Nonclinical Sample Collection -- 13.3.2 Procedures for Clinical Sample Collection -- 13.3.3 Sample Identification and Randomization -- 13.4 REDUCING ADSORPTION OF ANALYTE TO CONTAINERS -- 13.5 SAMPLE COLLECTION FOR PEPTIDES AND PROTEINS -- 13.6 STABILIZING SAMPLES -- 13.6.1 Physical Factors -- 13.6.2 Chemical Factors -- 13.6.3 Stability and Dried Blood Spots -- 13.7 BIOLOGICALLY HAZARDOUS SAMPLES -- 13.8 SAMPLE STORAGE -- 13.8.1 Types of Container for Sample Storage -- 13.8.2 Material -- 13.8.3 Temperature of Samples Storage -- 13.8.4 Disaster Recovery Plan -- 13.9 TRANSPORT AND SHIPPING OF SAMPLES -- 13.9.1 Practicalities of Transporting Samples -- 13.9.2 Packaging -- 13.9.3 Documentation -- 13.9.4 Monitoring Samples in Transit -- 13.9.5 Receipt of Samples -- 13.10 COMPLIANCE CHECKLIST -- 13.11 SUMMARY -- ACKNOWLEDGMENT -- REFERENCES -- 14 BEST PRACTICES IN BIOLOGICAL SAMPLE PREPARATION FOR LC-MS BIOANALYSIS.

14.1 WHY DO SAMPLE PREPARATION?.