HPLC Method Development for Pharmaceuticals (eBook)

Front Cover 1
HPLC METHOD DEVELOPMENT FOR PHARMACEUTICALS 4
Copyright Page 5
CONTENTS 6
PREFACE 14
CONTRIBUTORS 16
Chapter 1. Overview of HPLC Method Development for Pharmaceuticals 20
I. Introduction 20
II. Theoretical Considerations 21
III. HPLC Columns and Column Packings 21
IV. Column Selection 22
V. Chiral Separations 22
VI. Contemporary HPLC 23
VII. Hyphenated Methods 24
VIII. Sample Preparation 24
IX. Instrument Qualification and Software Validation 25
X. Pharmaceutical Development 25
XI. Drug Discovery 26
XII. Early Phase Method Development 27
XIII. Late Phase Development 27
XIV. In-Process Testing 28
XV. Method Development for Biomolecules 28
XVI. Method Validation 29
XVII. Troubleshooting 29
XVIII. Molecularly Imprinted Polymers 29
References 30
Chapter 2. HPLC Theory 32
Abstract 32
I. Introduction 32
II. Basic Chromatographic Descriptors 33
III. Efficiency 34
IV. Resolution 39
V. Main Components of the Retention Mechanism 41
VI. General Column Mass Balance 43
VII. Partitioning Model 46
VIII. Adsorption Model 47
IX. Void Volume Considerations 48
X. Thermodynamic Relationships 50
XI. Secondary Equilibria 54
XII. Salt Effect 56
XIII. Effect of Different Counteranions 57
XIV. Inclusion of Secondary Equilibria in the Mass Balance 59
XV. Conclusions 62
References 62
Chapter 3. HPLC Columns and Packings 64
Abstract 64
I. Introduction 65
II. Column Chemistry 66
III. Speed and Resolution 87
IV. Specialty Columns from Nanobore to Preparative Chromatography 96
V. Summary 99
References 99
Chapter 4. Column Characterization and Selection 104
Abstract 104
I. Introduction 105
II. Characteristics of RP-HPLC Columns and Chromatographic Tests 109
III. Column Classification and Selection 120
IV. Conclusions 123
Acknowledgment 124
References 124
Chapter 5. Chiral Separations 130
Abstract 130
I. Introduction 131
II. Separation of Enantiomers on HPLC Chiral Stationary Phases 134
III. Practical Guidelines to Chiral HPLC of Pharmaceuticals 154
IV. Conclusions 156
References 157
Chapter 6. Contemporary Liquid Chromatographic Systems for Method Development 164
Abstract 164
I. Introduction 165
II. Traditional Instrumentation for HPLC Method Development 166
III. Contemporary Method Development Systems 172
IV. Migrating Methods from HPLC to UPLC 188
V. UPLC Applications in Pharmaceutical Analysis 193
VI. Summary and Conclusions 202
Acknowledgments 203
References 204
Further Reading 205
Chapter 7. Hyphenated Techniques 208
Abstract 208
I. Introduction and Background 209
II. Combined Liquid Chromatography/Mass Spectrometry 210
III. Combined Liquid Chromatography/Nuclear Magnetic Resonance Spectroscopy 237
IV. Conclusions 250
Acknowledgments 250
References 251
Chapter 8. HPLC Sample Preparation 256
Abstract 256
I. Introduction 257
II. Fundamentals of Extraction and Chemical Equilibrium 258
III. Choice of Samples Preparation 264
IV. Direct Injection 265
V. Liquid–Liquid Extraction 266
VI. Solid-Phase Extraction 267
VII. Solid Samples 273
VIII. Additional Sample Preparation Methods 275
IX. Conclusions 282
References 282
Chapter 9. Instrument Qualification and Software Validation 288
Abstract 289
I. Introduction 289
II. Definitions 290
III. Qualification Model 297
IV. Discussion: Case Study HPLC 310
V. Summary and Conclusions 313
Acknowledgments 314
References 314
Chapter 10. Pharmaceutical Development: From Pre-Clinical to Post Approval 316
Abstract 316
I. Introduction 317
II. The Role of HPLC in Drug Discovery 319
III. The Role of HPLC in Pre-clinical Development 322
IV. The Role of HPLC in Clinical Development 326
V. Post Approval 331
VI. Conclusions 334
References 334
Chapter 11 . HPLC Method Development for Drug Discovery LC-MSAssays in Rapid PK Applications 336
Abstract 337
I. Introduction 337
II. Tandem MS Selected Reaction Monitoring (SRM) Development 338
III. HPLC Method Development 341
IV. Sample Preparation 343
V. Matrix Effects 348
VI. Background Interference: Enhanced Mass Resolution Strategy 353
VII. Limit of Quantitation, Dynamic Range and Linearity 359
VIII. Assay Samples and Generate Pharmacokinetic Reports 363
IX. Conclusions 364
Acknowledgment 364
References 364
Chapter 12. HPLC Method Development in Early Phase Pharmaceutical Development 372
Abstract 372
I. Introduction 373
II. Forced Decomposition and Impurity Profiling 374
III. Orthogonal Screening 376
IV. Method Selection and Optimization 381
V. Additional Methods 385
VI. Setting the Stage for Late Phase HPLC Method Developments 386
VII. Conclusions 388
Acknowledgments 388
References 388
Chapter 13. HPLC Method Development in Late Phase Pharmaceutical Development 392
Abstract 392
I. Introduction 393
II. Goals of Development 394
III. Planning Phase 396
IV. Method Development Phase 401
V. Method Evaluation Phase 413
VI. Method Transfer Phase 417
VII. Method Performance Monitoring and Feedback 419
VIII. Method Development Cycle Times and Documentation 422
IX. Conclusions 422
References 423
Chapter 14. Use of HPLC for In-process Testing 426
Abstract 426
I. Introduction 426
II. Various Examples of In-Process Testing in Drug Substance Development 428
III. Fast HPLC 438
IV. In-Process Testing in Chemical Production 439
V. In-Process Testing in Pharmaceutical Production 441
VI. Conclusions and Future Directions 442
Acknowledgments 442
References 442
Chapter 15. Method Development for Biomolecules 444
Abstract 444
I. Introduction 445
II. Protein and Peptide Separations by HPLC 449
III. Case Studies 453
IV. Summary and Conclusions 456
Abbreviations 457
References 457
Chapter 16. Method Validation 460
Abstract 460
I. Introduction 461
II. Validation Process 463
III. Validation Characteristics 463
IV. Additional Validation Characteristics 469
V. Robustness Testing 471
VI. Conclusions 476
References 476
Chapter 17. Troubleshooting in High Performance Liquid Chromatography 478
Abstract 478
I. Introduction 479
II. Problems Observed in Chromatograms 479
III. Operating Parameters 487
IV. Leaks 491
V. Pressure Problems 493
VI. Troubleshooting Action Plan 494
VII. Conclusions 496
References 496
Internet Resources 496
Chapter 18. Molecularly Imprinted Polymers as Sorbents for Separations and Extractions 498
Abstract 498
I. Introduction 498
II. MIP Theory 500
III. Synthesis 506
IV. Use of Imprinted Polymers in Separation Science 510
V. Pharmaceutical Application for MIPS 512
VI. Pros and Cons of MIP Sorbents 514
VII. Conclusions and Future Outlook 515
References 516
Index 524