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Chromatography. Principles and Instrumentation

  • ID: 3644039
  • Book
  • 280 Pages
  • John Wiley and Sons Ltd
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Provide students and practitioners with a solid grounding in the theory of chromatography and the metrics used in its applications

In this book, you will learn the fundamental principles underpinning chromatography and be able to connect those principles to the design and use of chromatographic systems.

Chromatography is the most widely used technique in modern analytical chemistry. Chromatography: Principles and Instrumentation, designed as a textbook, is focused on the current theory and the modern practice of chromatography.  It is intended for use in both quantitative analysis courses and instrumental methods courses.

Chromatography: Principles and Instrumentation features:

  • Numerous figures and problems that help the reader visualize these processes and identify the key variables in any given chromatography problem
  • Introduction to the most recent developments in chromatography
  • Case studies that illustrate the range of applications addressed by chromatography such as the analysis of performance–enhancing drugs in sports and the detection of deliberate contamination of food
  • Problems embedded throughout the chapters as well as at the end of each chapter so that students can check their understanding before continuing on to new sections
  • Clear and concise writing intended for undergraduate students and analysts just entering the field of chromatography.

The book, split into three chapters, covers Fundamentals of Chromatography, Gas Chromatography, and Liquid Chromatography. Chapter 1 provides an in–depth description of the processes governing chromatography and includes a systematic development of band broadening. Chapter 2 focuses on gas chromatography (GC). It introduces the components of GC instruments in the context of the function they accomplish and discusses the theory behind instrumentation, instrument design, and practical aspects. The final chapter addresses liquid chromatography. It emphasizes reversed–phase liquid chromatography (RPLC) because of its prominence, but you will also find descriptions of all of the common modes of LC.

The text of each section includes numerous headings and subheadings, making it easy for faculty and students to refer to and use the information within each chapter selectively. Students in analytical chemistry and biochemistry courses focused on chromatography will find this book a valuable resource in providing them with the knowledge needed to operate in modern laboratories.

Mark F. Vitha is currently a Professor at Drake University.  He received his Ph.D. from the University of Minnesota.  He is the series editor for The Chemical Analysis Series (Wiley) and is a co–editor of the book High Throughput Analysis for Food Safety (Wiley, 2014). He has been named as a Levitt Teacher of the Year, a Windsor Professor of Science, and a Ronald D. Troyer Research Fellow at Drake.

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PREFACE ix

1. Fundamentals of Chromatography 1

1.1 Theory 1

1.1.1 Component Separation 3

1.1.2 Retention Factor 6

1.1.3 Separation 11

1.1.4 Resolution and Theoretical Plates 13

1.2 Band Broadening 20

1.2.1 Diffusion 21

1.2.2 Linear Velocity 23

1.2.3 Broadening in Open Tubes with No Stationary Phase and No Retention 24

1.2.4 Broadening in Open Tubes with a Stationary Phase 28

1.2.5 Broadening in a Packed Column 34

1.2.6 Putting It All Together 43

1.2.7 Practical Consequences of Broadening Theory 45

1.3 General Resolution Equation 47

1.4 Peak Symmetry 51

1.5 Key Operating Variables 51

1.6 Instrumentation 53

1.7 Practice of The Technique 53

1.7.1 Quantitation 53

1.7.2 Internal Standards and the Method of Standard Additions 55

1.8 Emerging Trends and Applications 55

1.9 Summary 55

Problems 56

References 59

Further Reading 59

2. Gas Chromatography 61

2.1 Theory of Gas Chromatographic Separations 61

2.1.1 GC Columns and Partitioning 64

2.2 Key Operating Variables that Control Retention 64

2.2.1 Adjusting Retention Time: Temperature 65

2.2.2 Adjusting Retention Time: Temperature Programming 67

2.2.3 Adjusting Retention Time: Mobile Phase Flow Rate 69

2.2.4 Adjusting Retention Time: The Column and the Stationary Phase 72

2.2.5 Adjusting Retention Time: Summary 78

2.2.6 Measures of Retention 78

2.3 Gas Chromatography Instrumentation 82

2.3.1 Carrier Gas Supply 83

2.3.2 The Injection Port and the Solute Injection Process 83

2.3.3 Oven/Column Compartment 97

2.3.4 Detectors 98

2.4 A More Detailed Look at Stationary Phase Chemistry: Kovats Indices and Mcreynolds Constants 111

2.4.1 Kovats Retention Indices 111

2.4.2 Stationary Phase Selection 120

2.5 Gas Chromatography in Practice 124

2.5.1 Syringe Washing 124

2.5.2 Controls and Blanks/Ghost Peaks 124

2.5.3 Autosamplers 125

2.5.4 GC Septa 125

2.5.5 Qualitative Analysis 126

2.5.6 Quantitative Analysis 126

2.5.7 Derivatization 128

2.5.8 High–Speed GC 128

2.5.9 Tandem GC 129

2.5.10 Microfabricated GC 129

2.6 A Real–World Application of Gas Chromatography 131

2.6.1 GC and International Oil Trading 131

2.7 Summary 136

Problems 137

References 143

Further Reading 144

3. Liquid Chromatography 145

3.1 Examples of Liquid Chromatography Analyses 145

3.2 Scope of Liquid Chromatography 147

3.3 History of LC 148

3.3.1 Modern Packing Materials 149

3.4 Modes of Liquid Chromatography 152

3.4.1 Normal Phase Liquid Chromatography (NPLC) 152

3.4.2 Reversed–Phase Liquid Chromatography (RPLC) 154

3.4.3 Ion–Exchange Chromatography (IEX) 165

3.4.4 Hydrophilic Interaction Chromatography (HILIC) 173

3.4.5 Size Exclusion Chromatography (SEC) 175

3.4.6 Affinity Chromatography 178

3.5 HPLC Instrumentation 180

3.5.1 The Proportioning Valve 181

3.5.2 Mixing Chamber 181

3.5.3 Pumps 181

3.5.4 Injection 183

3.5.5 The Column and Particles 185

3.5.6 Guard Columns 187

3.5.7 Detectors 188

3.6 Specific Uses of and Advances in Liquid Chromatography 201

3.6.1 Chiral Separations 202

3.6.2 Preparative–Scale Chromatography 207

3.6.3 Ultra–High Performance Liquid Chromatography (UHPLC) for High–Speed Separations 212

3.6.4 Tandem–Column Liquid Chromatography 216

3.6.5 Two–Dimensional Liquid Chromatography (2D–LC) 218

3.7 Application of LC Analysis of Pharmaceutical Compounds in Groundwater 224

3.7.1 Sampling 225

3.7.2 Analysis Method for 21 Antibiotics Sample Pretreatment 225

3.7.3 Use of Internal Standards and Other Quality Assurance Issues 227

3.7.4 LC Analyses 228

3.7.5 Mass Spectrometric Selected Ion Monitoring Detection 228

3.7.6 Results 229

3.8 Summary 230

Problems 230

References 232

SOLUTIONS 237

Index 263

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Mark F. Vitha
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