Hplc and Uhplc for Practicing Scientists

A concise yet comprehensive reference guide on HPLC/UHPLC that focuses on its fundamentals, latest developments, and best practices in the pharmaceutical and biotechnology industries
 
Written for practitioners by an expert practitioner, this new edition of HPLC and UHPLC for Practicing Scientists adds numerous updates to its coverage of high-performance liquid chromatography, including comprehensive information on UHPLC (ultra-high-pressure liquid chromatography) and the continuing migration of HPLC to UHPLC, the modern standard platform. In addition to introducing readers to HPLC's fundamentals, applications, and developments, the book describes basic theory and terminology for the novice, and reviews relevant concepts, best practices, and modern trends for the experienced practitioner.
 
HPLC and UHPLC for Practicing Scientists, Second Edition offers three new chapters. One is a standalone chapter on UHPLC, covering concepts, benefits, practices, and potential issues. Another examines liquid chromatography/mass spectrometry (LC/MS). The third reviews at the analysis of recombinant biologics, particularly monoclonal antibodies (mAbs), used as therapeutics. While all chapters are revised in the new edition, five chapters are essentially rewritten (HPLC columns, instrumentation, pharmaceutical analysis, method development, and regulatory aspects). The book also includes problem and answer sections at the end of each chapter.
* Overviews fundamentals of HPLC to UHPLC, including theories, columns, and instruments with an abundance of tables, figures, and key references
* Features brand new chapters on UHPLC, LC/MS, and analysis of recombinant biologics
* Presents updated information on the best practices in method development, validation, operation, troubleshooting, and maintaining regulatory compliance for both HPLC and UHPLC
* Contains major revisions to all chapters of the first edition and substantial rewrites of chapters on HPLC columns, instrumentation, pharmaceutical analysis, method development, and regulatory aspects
* Includes end-of-chapter quizzes as assessment and learning aids
* Offers a reference guide to graduate students and practicing scientists in pharmaceutical, biotechnology, and other industries
 
Filled with intuitive explanations, case studies, and clear figures, HPLC and UHPLC for Practicing Scientists, Second Edition is an essential resource for practitioners of all levels who need to understand and utilize this versatile analytical technology. It will be a great benefit to every busy laboratory analyst and researcher.

List of contents

Author's Biography xvii
 
Biographies of Contributors xix
 
Preface xxi
 
Foreword xxiii
 
Acknowledgments xxv
 
1 Introduction 1
 
1.1 Introduction 1
 
1.1.1 Scope 1
 
1.1.2 What Is HPLC? 2
 
1.1.3 A Brief History 3
 
1.1.4 Advantages and Limitations 4
 
1.1.5 Ultra-High-Pressure Liquid Chromatography (UHPLC) 4
 
1.2 Primary Modes of HPLC 4
 
1.2.1 Normal-Phase Chromatography (NPC) 5
 
1.2.2 Reversed-Phase Chromatography (RPC) 5
 
1.2.3 Ion-Exchange Chromatography (IEC) 6
 
1.2.4 Size-Exclusion Chromatography (SEC) 8
 
1.2.5 Other Separation Modes 8
 
1.3 Some Common-Sense Corollaries 10
 
1.4 How to Get More Information 11
 
1.5 Summary 11
 
1.6 Quizzes 11
 
1.6.1 Bonus Quiz 12
 
References 12
 
2 Basic Terms and Concepts 15
 
2.1 Scope 15
 
2.2 Basic Terms and Concepts 16
 
2.2.1 Retention Time (tR), Void Time (tM), Peak Height (h), and Peak Width (wb) 16
 
2.2.2 Retention Volume (VR), Void Volume (VM), and Peak Volume 16
 
2.2.3 Retention Factor (k) 18
 
2.2.4 Separation Factor (alpha) 19
 
2.2.5 Column Efficiency and Plate Number (N) 20
 
2.2.6 Peak Volume 20
 
2.2.7 Height Equivalent to a Theoretical Plate or Plate Height (HETP or H) 21
 
2.2.8 Resolution (Rs) 21
 
2.2.9 Peak Symmetry:Asymmetry Factor (As) and Tailing Factor (Tf) 23
 
2.3 Mobile Phase 24
 
2.3.1 General Requirements 24
 
2.3.2 Solvent Strength and Selectivity 25
 
2.3.3 pH Modifiers and Buffers 27
 
2.3.4 Acidic Mobile Phases 28
 
2.3.5 Ion-Pairing Reagents and Chaotropic Agents 29
 
2.3.6 High-pH Mobile Phases 29
 
2.3.7 Other Operating Parameters: Flow Rate (F) and Column Temperature (T) 30
 
2.4 The Resolution Equation 31
 
2.5 The Van Deemter Equation 33
 
2.6 Isocratic vs. Gradient Analysis 35
 
2.6.1 Peak Capacity (n) 35
 
2.6.2 Gradient Parameters (Initial and Final Solvent Strength, Gradient Time (tG), and Flow Rate) 36
 
2.6.3 The 0.25 DeltatG Rule: When Is Isocratic Analysis More Appropriate? 37
 
2.7 The Concept of Orthogonality and Selectivity Tuning 38
 
2.8 Sample Capacity 41
 
2.9 Glossary of HPLC Terms 41
 
2.10 Summary and Conclusion 42
 
2.11 Quizzes 42
 
2.11.1 Bonus Quiz 44
 
References 44
 
3 HPLC Columns and Trends 45
 
3.1 Scope 45
 
3.1.1 Glossary and Abbreviations 45
 
3.2 General Column Description and Characteristics 46
 
3.2.1 Column Hardware - Standard vs. Cartridge Format 47
 
3.3 Column Type 47
 
3.3.1 Types Based on Chromatographic Mode 48
 
3.3.2 Column Types Based on Dimension 48
 
3.3.3 Column Length (L) 48
 
3.4 Column Packing Characteristics 50
 
3.4.1 Support Type 50
 
3.4.2 Particle Size (dp) 51
 
3.4.3 Surface Area and Pore Size (dpore) 51
 
3.4.4 Bonding Chemistries 52
 
3.5 Modern HPLC Column Trends 54
 
3.5.1 Silica Support Material 54
 
3.5.2 Hybrid Particles 55
 
3.5.3 Novel Bonding Chemistries 58
 
3.5.4 Shorter and Narrower Columns Packed with Small Particles 61
 
3.5.5 Micro-LC and Nano-LC 62
 
3.5.6 Monoliths 64
 
3.5.7 Superficially Porous Particles (SPP) 65
 
3.5.8 Micropillar Array Chromatography (muPAC) 67
 
3.6 Guard Columns 69
 
3.7 Specialty Columns 69
 
3.7.1 Bioseparations Columns 69
 
3.7.2 Chiral Col

About the author

MICHAEL W. DONG, PHD, is the Principal of MWD Consulting, providing expert training and consulting service in HPLC/UHPLC, and pharmaceutical analysis. He is a former senior scientist, for GENENTECH, INC, small molecule analytical chemistry and quality control department. Dr. Dong conducts short courses on HPLC/UHPLC, drug development process, and drug quality at national meetings such as Pittcon, ACS, HPLC, and EAS. He also provides consulting services on CMC, HPLC method development, and solving drug quality issues. He holds a Ph.D. degree in analytical chemistry and has authored 120+ journal articles and three books.

Summary

A concise yet comprehensive reference guide on HPLC/UHPLC that focuses on its fundamentals, latest developments, and best practices in the pharmaceutical and biotechnology industries

Written for practitioners by an expert practitioner, this new edition of HPLC and UHPLC for Practicing Scientists adds numerous updates to its coverage of high-performance liquid chromatography, including comprehensive information on UHPLC (ultra-high-pressure liquid chromatography) and the continuing migration of HPLC to UHPLC, the modern standard platform. In addition to introducing readers to HPLC's fundamentals, applications, and developments, the book describes basic theory and terminology for the novice, and reviews relevant concepts, best practices, and modern trends for the experienced practitioner.

HPLC and UHPLC for Practicing Scientists, Second Edition offers three new chapters. One is a standalone chapter on UHPLC, covering concepts, benefits, practices, and potential issues. Another examines liquid chromatography/mass spectrometry (LC/MS). The third reviews at the analysis of recombinant biologics, particularly monoclonal antibodies (mAbs), used as therapeutics. While all chapters are revised in the new edition, five chapters are essentially rewritten (HPLC columns, instrumentation, pharmaceutical analysis, method development, and regulatory aspects). The book also includes problem and answer sections at the end of each chapter.
* Overviews fundamentals of HPLC to UHPLC, including theories, columns, and instruments with an abundance of tables, figures, and key references
* Features brand new chapters on UHPLC, LC/MS, and analysis of recombinant biologics
* Presents updated information on the best practices in method development, validation, operation, troubleshooting, and maintaining regulatory compliance for both HPLC and UHPLC
* Contains major revisions to all chapters of the first edition and substantial rewrites of chapters on HPLC columns, instrumentation, pharmaceutical analysis, method development, and regulatory aspects
* Includes end-of-chapter quizzes as assessment and learning aids
* Offers a reference guide to graduate students and practicing scientists in pharmaceutical, biotechnology, and other industries

Filled with intuitive explanations, case studies, and clear figures, HPLC and UHPLC for Practicing Scientists, Second Edition is an essential resource for practitioners of all levels who need to understand and utilize this versatile analytical technology. It will be a great benefit to every busy laboratory analyst and researcher.

Product details

• Authors: Michael W Dong, Michael W. Dong, Michael W. (Synomics Pharma Dong, MW Dong, Dong Michael W.
• Publisher: Wiley, John and Sons Ltd
• Languages: English
• Product format: Paperback / Softback
• Released: 31.08.2019
• EAN: 9781119313762
• ISBN: 978-1-119-31376-2
• No. of pages: 416
• Subjects: Natural sciences, medicine, IT, technology > Chemistry > Theoretical chemistry; Chemie, Analytische Chemie, Chromatographie, Life Sciences, Biowissenschaften, biotechnology, chemistry, Analytical Chemistry, Biotechnologie i. d. Biowissenschaften, Pharmaceutical & Medicinal Chemistry, Pharmazeutische u. Medizinische Chemie