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Introduction to Modern Liquid Chromatography
The latest edition of the authoritative reference to HPLC

High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample. Snyder and Kirkland's Introduction to Modern Liquid Chromatography has long represented the premier reference to HPLC. This Third Edition, with John Dolan as added coauthor, addresses important improvements in columns and equipment, as well as major advances in our understanding of HPLC separation, our ability to solve problems that were troublesome in the past, and the application of HPLC for new kinds of samples.

This carefully considered Third Edition maintains the strengths of the previous edition while significantly modifying its organization in light of recent research and experience. The text begins by introducing the reader to HPLC, its use in relation to other modern separation techniques, and its history, then leads into such specific topics as:

  • The basis of HPLC separation and the general effects of different experimental conditions
  • Equipment and detection
  • The column—the "heart" of the HPLC system
  • Reversed-phase separation, normal-phase chromatography, gradient elution, two-dimensional separation, and other techniques
  • Computer simulation, qualitative and quantitative analysis, and method validation and quality control
  • The separation of large molecules, including both biological and synthetic polymers
  • Chiral separations, preparative separations, and sample preparation
  • Systematic development of HPLC separations—new to this edition
  • Troubleshooting tricks, techniques, and case studies for both equipment and chromatograms
Designed to fulfill the needs of the full range of HPLC users, from novices to experts, Introduction to Modern Liquid Chromatography, Third Edition offers the most up-to-date, comprehensive, and accessible survey of HPLC methods and applications available.
1100173327
Introduction to Modern Liquid Chromatography
The latest edition of the authoritative reference to HPLC

High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample. Snyder and Kirkland's Introduction to Modern Liquid Chromatography has long represented the premier reference to HPLC. This Third Edition, with John Dolan as added coauthor, addresses important improvements in columns and equipment, as well as major advances in our understanding of HPLC separation, our ability to solve problems that were troublesome in the past, and the application of HPLC for new kinds of samples.

This carefully considered Third Edition maintains the strengths of the previous edition while significantly modifying its organization in light of recent research and experience. The text begins by introducing the reader to HPLC, its use in relation to other modern separation techniques, and its history, then leads into such specific topics as:

  • The basis of HPLC separation and the general effects of different experimental conditions
  • Equipment and detection
  • The column—the "heart" of the HPLC system
  • Reversed-phase separation, normal-phase chromatography, gradient elution, two-dimensional separation, and other techniques
  • Computer simulation, qualitative and quantitative analysis, and method validation and quality control
  • The separation of large molecules, including both biological and synthetic polymers
  • Chiral separations, preparative separations, and sample preparation
  • Systematic development of HPLC separations—new to this edition
  • Troubleshooting tricks, techniques, and case studies for both equipment and chromatograms
Designed to fulfill the needs of the full range of HPLC users, from novices to experts, Introduction to Modern Liquid Chromatography, Third Edition offers the most up-to-date, comprehensive, and accessible survey of HPLC methods and applications available.
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Introduction to Modern Liquid Chromatography

Introduction to Modern Liquid Chromatography

Introduction to Modern Liquid Chromatography
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Introduction to Modern Liquid Chromatography

Introduction to Modern Liquid Chromatography

Overview

The latest edition of the authoritative reference to HPLC

High-performance liquid chromatography (HPLC) is today the leading technique for chemical analysis and related applications, with an ability to separate, analyze, and/or purify virtually any sample. Snyder and Kirkland's Introduction to Modern Liquid Chromatography has long represented the premier reference to HPLC. This Third Edition, with John Dolan as added coauthor, addresses important improvements in columns and equipment, as well as major advances in our understanding of HPLC separation, our ability to solve problems that were troublesome in the past, and the application of HPLC for new kinds of samples.

This carefully considered Third Edition maintains the strengths of the previous edition while significantly modifying its organization in light of recent research and experience. The text begins by introducing the reader to HPLC, its use in relation to other modern separation techniques, and its history, then leads into such specific topics as:

  • The basis of HPLC separation and the general effects of different experimental conditions
  • Equipment and detection
  • The column—the "heart" of the HPLC system
  • Reversed-phase separation, normal-phase chromatography, gradient elution, two-dimensional separation, and other techniques
  • Computer simulation, qualitative and quantitative analysis, and method validation and quality control
  • The separation of large molecules, including both biological and synthetic polymers
  • Chiral separations, preparative separations, and sample preparation
  • Systematic development of HPLC separations—new to this edition
  • Troubleshooting tricks, techniques, and case studies for both equipment and chromatograms
Designed to fulfill the needs of the full range of HPLC users, from novices to experts, Introduction to Modern Liquid Chromatography, Third Edition offers the most up-to-date, comprehensive, and accessible survey of HPLC methods and applications available.

Product Details

ISBN-13: 9781118210390
Publisher: Wiley
Publication date: 09/20/2011
Sold by: JOHN WILEY & SONS
Format: eBook
Pages: 960
File size: 25 MB
Note: This product may take a few minutes to download.

About the Author

Lloyd R. Snyder, PhD, is a Principal at LC Resources in Walnut Creek, California. He is the author or coauthor of several books including An Introduction to Separation Science, Introduction to Modern Liquid Chromatography, Second Edition, the bestselling Practical HPLC Method Development, Second Edition, and the comprehensive High-Performance Gradient Elution, all published by Wiley.

Joseph J. Kirkland, PhD, is Vice President of Research and Development for Advanced Materials Technology, Inc., and coauthor of Introduction to Modern Liquid Chromatography, Second Edition, Practical HPLC Method Development, Second Edition, and Modern Size-Exclusion Liquid Chromatography, Second Edition, all published by Wiley.

John W. Dolan, PhD, is a Principal at LC Resources. He is author of the popular "LC Troubleshooting" column in LCGC magazine and coauthor with Lloyd Snyder of Troubleshooting LC Systems and High-Performance Gradient Elution.

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Table of Contents

Preface xxxi

Glossary of Symbols and Abbreviations xxxv

1 Introduction 1

1.1 Background Information 2

1.2 A Short History of HPLC 6

1.3 Some Alternatives to HPLC 8

1.4 Other Sources of HPLC Information 12

References 15

2 Basic Concepts and the Control of Separation 19

2.1 Introduction 20

2.2 The Chromatographic Process 20

2.3 Retention 24

2.4 Peak Width and the Column Plate Number N 35

2.5 Resolution and Method Development 54

2.6 Sample Size Effects 69

2.7 Related Topics 74

References 83

3 Equipment 87

3.1 Introduction 88

3.2 Reservoirs and Solvent Filtration 89

3.3 Mobile-Phase Degassing 92

3.4 Tubing and Fittings 96

3.5 Pumping Systems 104

3.6 Autosamplers 113

3.7 Column Ovens 125

3.8 Data Systems 127

3.9 Extra-Column Effects 131

3.10 Maintenance 131

References 144

4 Detection 147

4.1 Introduction 148

4.2 Detector Characteristics 149

4.3 Introduction to Individual Detectors 160

4.4 UV-Visible Detectors 160

4.5 Fluorescence Detectors 167

4.6 Electrochemical (Amperometric) Detectors 170

4.7 Radioactivity Detectors 172

4.8 Conductivity Detectors 174

4.9 Chemiluminescent Nitrogen Detector 174

4.10 Chiral Detectors 175

4.11 Refractive Index Detectors 177

4.12 Light-Scattering Detectors 180

4.13 Corona-Discharge Detector (CAD) 184

4.14 Mass Spectral Detectors (MS) 185

4.15 Other Hyphenated Detectors 191

4.16 Sample Derivatization and Reaction Detectors 194

References 196

5 The Column 199

5.1 Introduction 200

5.2 Column Supports 200

5.3 Stationary Phases 217

5.4 Column Selectivity 227

 

5.5 Column Hardware 238

5.6 Column-Packing Methods 240

5.7 Column Specifications 244

5.8 Column Handling 246

References 250

6 Reversed-phase Chromatography for Neutral Sam- Ples 253

6.1 Introduction 254

6.2 Retention 256

6.3 Selectivity 263

6.4 Method Development and Strategies for Optimizing Selectivity 284

6.5 Nonaqueous Reversed-Phase Chromatography (narp) 295

6.6 Special Problems 297

References 298

7 Ionic Samples: Reversed-phase Ion-pair and Ion- Exchange Chromatography 303

7.1 Introduction 304

7.2 Acid–Base Equilibria and Reversed-Phase Retention 304

7.3 Separation of Ionic Samples by Reversed-Phase Chromatography (RPC) 319

7.4 Ion-Pair Chromatography (IPC) 331

7.5 Ion-Exchange Chromatography (IEC) 349

References 357

8 Normal-phase Chromatography 361

8.1 Introduction 362

8.2 Retention 363

8.3 Selectivity 376

8.4 Method-Development Summary 385

8.5 Problems in the Use of NPC 392

8.6 Hydrophilic Interaction Chromatography (HILIC) 395

References 401

9 Gradient Elution 403

9.1 Introduction 404

9.2 Experimental Conditions and Their Effects on Separation 412

9.3 Method Development 434

9.4 Large-Molecule Separations 464

9.5 Other Separation Modes 465

9.6 Problems 470

References 471

10 Computer-assisted Method Development 475

10.1 Introduction 475

10.2 Computer-Simulation Software 481

10.3 Other Method-Development Software 491

10.4 Computer Simulation and Method Development 492

References 497

11 Qualitative and Quantitative Analysis 499

11.1 Introduction 499

11.2 Signal Measurement 500

11.3 Qualitative Analysis 516

11.4 Quantitative Analysis 520

11.5 Summary 529

References 529

12 Method Validation 531
with Michael Swartz

12.1 Introduction 532

12.2 Terms and Definitions 534

12.3 System Suitability 542

12.4 Documentation 543

12.5 Validation for Different Pharmaceutical-Method Types 546

12.6 Bioanalytical Methods 548

12.7 Analytical Method Transfer (AMT) 554

12.8 Method Adjustment or Method Modification 561

12.9 Quality Control and Quality Assurance 564

12.10 Summary 565

References 566

13 Biochemical and Synthetic Polymer Separations 569
with Timothy Wehr, Carl Scandella, and Peter Schoenmakers

13.1 Biomacromolecules 570

13.2 Molecular Structure and Conformation 571

13.3 Special Considerations for Biomolecule HPLC 579

13.4 Separation of Peptides and Proteins 584

13.5 Separation of Nucleic Acids 618

13.6 Separation of Carbohydrates 625

13.7 Separation of Viruses 630

13.8 Size-Exclusion Chromatography (SEC) 631

13.9 Large-Scale Purification of Large Biomolecules 641

13.10 Synthetic Polymers 648

References 658

14 Enantiomer Separations 665
with Michael Lämmerhofer, Norbert M. Maier and Wolfgang Lindner

14.1 Introduction 666

14.2 Background and Definitions 666

14.3 Indirect Method 670

14.4 Direct Method 675

14.5 Peak Dispersion and Tailing 681

14.6 Chiral Stationary Phases and Their Characteristics 681

14.7 Thermodynamic Considerations 715

References 718

15 Preparative Separations 725
with Geoff Cox

15.1 Introduction 726

15.2 Equipment for Prep-LC Separation 730

15.3 Isocratic Elution 736

15.4 Severely Overloaded Separation 748

15.5 Gradient Elution 751

15.6 Production-Scale Separation 754

References 755

16 Sample Preparation 757
with Ronald Majors 757

16.1 Introduction 758

16.2 Types of Samples 759

16.3 Preliminary Processing of Solid and Semi-Solid Samples 760

16.4 Sample Preparation for Liquid Samples 764

16.5 Liquid–Liquid Extraction 764

16.6 Solid-Phase Extraction (SPE) 771

16.7 Membrane Techniques in Sample Preparation 790

16.8 Sample Preparation Methods for Solid Samples 791

16.9 Column-Switching 796

16.10 Sample Preparation for Biochromatography 797

16.11 Sample Preparation for LC-MS 800

16.12 Derivatization in HPLC 802

References 805

17 Troubleshooting 809

Quick Fix 809

17.1 Introduction 810

17.2 Prevention of Problems 811

17.3 Problem-Isolation Strategies 819

17.4 Common Symptoms of HPLC Problems 821

17.5 Troubleshooting Tables 865

References 876

Appendix I. Properties of HPLC Solvents 879

I.1 Solvent-Detector Compatibility 879

I. 2 Solvent Polarity and Selectivity 882

I. 3 Solvent Safety 885

References 886

Appendix II. Preparing Buffered Mobile Phases 887

II.1 Sequence of Operations 887

II.2 Recipes for Some Commonly Used Buffers 888

Reference 890

Index 891

What People are Saying About This

From the Publisher

"In summary, I would strongly recommend this book for anyone working with LC and LC/MS. It is a finely crafted introduction that adequately covers nearly every aspect of the science. The emphasis on basic principles and practical aspects ensures that it will be a useful reference for many years to come." (J Am Soc Mass Spectrom, 2011)

"This third edition is highly cross-referenced, so as to allow the reader to follow up on topics of special interest, or to clarify questions that may arise during reading. The third edition of Introduction to Modern Liquid Chromatography will continue to be the HPLC reference book for thousands of readers, either experienced workers who may wish to explore topics of his/her choice, or find an answer to specific problems, or beginners who would like to understand and know the possibilities offered by the technique." (Chemistry Journals, 11 April 2011)

"This classic text on liquid chromatography has been thoroughly updated through the addition of information on modern instrumentation, columns, and troubleshooting. It is a valuable resource for practicing chromatographers at all levels.." (Anal Bioanal Chem, 2011)

"In summary, I would strongly recommend this book for anyone working with LC and LC/MS. It is a finely crafted introduction that adequately covers nearly every aspect of the science. The emphasis on basic principles and practical aspects ensures that it will be a useful reference for any years to come." (American Society for Mass Spectrometry, 21 Januay 2011)

"The text is illustrated with many figures and tables originating from authors' own work or taken from the literature . . . both groups of readers will find in this book plenty of information and inspiration." (Journal of Separation Science, 1 June 2010)

"Following their highly successful second edition (1979), which appeared more than 30 years ago, Snyder (LC Resources) and Kirkland (Advanced Materials Technology) have teamed with Dolan (LC Resources) and additional collaborators to provide an excellent update of their earlier work." (CHOICE, July 2010)

"It is difficult in these times to ever call a scientific book a "bargain", but this truly is. The wealth of information contained in these almost 1000 pages is invaluable. The book is comprehensive, eminently readable and approachable, and highly useful for both the skilled chromatographer and those new to the technique. Everyone using a liquid chromatographic instrument would benefit from owning a copy." (JACS, 2010)

"It's current, clearly-indexed, well-referenced, and comprehensive. The inclusion of John Dolan as author has significantly expanded the sections on troubleshooting, and additional expert contributors have enhanced the coverage of the specialist chapters." - Bruce Hamilton posted on the Chromatography Forum

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