Additional Information
Book Details
Abstract
Capillary electrophoresis–mass spectrometry (CE-MS) has become a very useful analytical technique for the profiling of highly polar and charged metabolites in biological samples. In this book, the unique features of CE-MS for metabolomics studies are highlighted including CE separation modes, capillary coatings and practical aspects of CE-MS coupling alongside a comprehensive overview of recent technological developments and applications.
CE-MS can be considered a relatively new technique in the field of metabolomics and it is therefore important to inform the scientific community about the possibilities of advanced CE-MS approaches for metabolomics studies. This book outlines the potential of this technique for researchers working in metabolomics, bioanalytics and biomarker analysis.
Strongly believing in the power of analytical technology to contribute to a better understanding of biochemical mechanisms underlying diseases, Rawi Ramautar studied both Pharmacochemistry and Analytical Sciences at Vrije Universiteit of Amsterdam in order to have the right background for a PhD training in this direction. In 2010, he completed his PhD on the development of capillary electrophoresis-mass spectrometry methods for metabolomics at the Utrecht University. Intrigued by metabolomics for disease prediction and diagnosis, Rawi switched to the Leiden University Medical Center to broaden his horizon on this topic. In 2013 and 2017, Rawi Ramautar received the highly prestigious Veni and Vidi personal grants, respectively, from the Netherlands Organisation for Scientific Research. Currently, he is a Principal Investigator (tenured) at the Leiden Academic Center for Drug Research of the Leiden University where his group is developing microscale analytical workflows for volume-restricted biomedical problems.
Table of Contents
| Section Title | Page | Action | Price |
|---|---|---|---|
| Front Cover | Cover | ||
| Capillary Electrophoresis–Mass Spectrometry for Metabolomics | i | ||
| Preface | v | ||
| About the Editor | vii | ||
| Contents | ix | ||
| Chapter\r1 - Capillary Electrophoresis–Mass Spectrometry for Metabolomics – From Metabolite Analysis to Metabolic Profiling | 1 | ||
| 1.1 Introduction | 1 | ||
| 1.2 Capillary Electrophoresis for Metabolite Analysis | 4 | ||
| 1.3 Capillary Electrophoresis–Mass Spectrometry for Metabolomics | 11 | ||
| 1.4 Conclusions | 15 | ||
| Acknowledgements | 16 | ||
| References | 16 | ||
| Chapter\r2 - CE-MS Workflows for Metabolomics: A Focus on Sample Preparation | 21 | ||
| 2.1 Introduction | 21 | ||
| 2.2 Sample Preparation: General Aspects | 24 | ||
| 2.3 Sample Preparation for Body Fluids | 25 | ||
| 2.3.1 Urine | 25 | ||
| 2.3.2 Blood, Plasma and Serum | 29 | ||
| 2.3.3 Other Biofluids | 36 | ||
| 2.4 Sample Preparation for Mammalian Cultured Cells | 38 | ||
| 2.5 Sample Preparation for Solid Samples: Tissues and Plants | 40 | ||
| 2.6 Sample Preparation for Microbial Cells, Parasites and Microbiota | 44 | ||
| 2.7 Conclusions | 46 | ||
| Acknowledgements | 46 | ||
| References | 46 | ||
| Chapter\r3 - Capillary Electrophoresis–Mass Spectrometry Using Non-covalently Coated Capillaries for Metabolic Profiling of Biological Samples | 53 | ||
| 3.1 Introduction | 53 | ||
| 3.2 Capillary Coatings in CE-MS | 55 | ||
| 3.3 Non-covalently Coated Capillaries for CE-MS-based Metabolomics | 56 | ||
| 3.4 PB–PVS and PB–DS–PB Coated Capillaries: Practical Aspects | 60 | ||
| 3.5 Conclusions | 62 | ||
| Acknowledgements | 63 | ||
| References | 63 | ||
| Chapter\r4 - Capillary Electrophoresis–Mass Spectrometry for Metabolomics Using New Interfacing Designs | 66 | ||
| 4.1 Introduction | 66 | ||
| 4.2 Low-flow Sheath-liquid Interfacing Designs | 68 | ||
| 4.3 Sheathless Interfacing Designs | 74 | ||
| 4.4 Conclusions | 79 | ||
| Acknowledgements | 79 | ||
| References | 80 | ||
| Chapter\r5 - Stacking or On-line Sample Concentration in CE-MS for Metabolomics | 83 | ||
| 5.1 Introduction | 83 | ||
| 5.2 Stacking Techniques – an Overview | 85 | ||
| 5.2.1 Field Amplification or Enhancement | 86 | ||
| 5.2.2 Transient Isotachophoresis (tITP) | 88 | ||
| 5.2.3 Dynamic pH Junctions | 89 | ||
| 5.2.4 Sweeping | 90 | ||
| 5.2.5 Analyte Focusing by Micelle Collapse (AFMC) | 91 | ||
| 5.2.6 Micelle to Solvent Stacking (MSS) | 92 | ||
| 5.2.7 Hyphenated Stacking | 93 | ||
| 5.3 Considerations for Stacking in CE-MS-based Metabolomics Studies | 94 | ||
| 5.3.1 Stacking with MS-compatible Reagents | 95 | ||
| 5.3.2 Stacking with Non-volatile Reagents | 96 | ||
| 5.4 Applications of Stacking in CE-MS-based Metabolomics Studies | 97 | ||
| 5.4.1 Applications of Field Amplification/Enhancement | 97 | ||
| 5.4.2 Applications of tITP | 102 | ||
| 5.4.3 Applications of Dynamic pH Junctions | 103 | ||
| 5.4.4 Applications of Sweeping, AFMC and MSS | 104 | ||
| 5.4.5 Applications of Hyphenated Stacking | 106 | ||
| 5.5 Conclusion | 109 | ||
| Acknowledgements | 109 | ||
| References | 109 | ||
| Chapter 6 - On-line Solid-phase Extraction Capillary Electrophoresis–Mass Spectrometry and Data Analysis Approaches for Metabolomics | 113 | ||
| 6.1 Introduction | 113 | ||
| 6.2 Experimental Procedures | 116 | ||
| 6.2.1 Mouse Blood Plasma Extraction | 116 | ||
| 6.2.2 Sample Preparation | 116 | ||
| 6.2.3 Apparatus and Procedures: SPE-CE-MS | 117 | ||
| 6.2.3.1 Construction of an SPE-CE Microcartridge | 117 | ||
| 6.2.3.2 SPE-CE-MS Methodology | 117 | ||
| 6.2.4 Multivariate Data Analysis: MCR-ALS and PLS-DA | 119 | ||
| 6.3 Results and Discussion | 120 | ||
| 6.3.1 Plasma Pretreatment | 120 | ||
| 6.3.2 Analysis of Plasma Samples by C18-SPE-CE-MS | 123 | ||
| 6.3.3 Multivariate Data Analysis Tools | 123 | ||
| 6.3.3.1 Feature Detection | 123 | ||
| 6.3.3.2 Statistical Analysis | 125 | ||
| 6.3.4 Metabolite Identification | 127 | ||
| 6.4 Conclusions | 127 | ||
| Acknowledgements | 131 | ||
| References | 131 | ||
| Chapter\r7 - CE-MS for Anionic and Cationic Metabolic Profiling: System Optimization and Applications | 134 | ||
| 7.1 Introduction | 134 | ||
| 7.2 CE-MS for Metabolome Analysis | 136 | ||
| 7.2.1 Cationic Metabolite Analysis | 137 | ||
| 7.2.2 Anionic Metabolite Analysis | 139 | ||
| 7.2.3 Interface | 140 | ||
| 7.2.4 MS Instrument | 142 | ||
| 7.3 Application | 144 | ||
| 7.3.1 Serum/Plasma | 144 | ||
| 7.3.2 Urine | 148 | ||
| 7.3.3 Tissue | 149 | ||
| 7.3.4 Cultured Cells | 151 | ||
| 7.3.5 Other Samples | 153 | ||
| 7.4 Conclusions | 155 | ||
| Acknowledgements | 156 | ||
| References | 156 | ||
| Chapter\r8 - CE-MS for Metabolomics: A Comparison with Other Techniques | 161 | ||
| 8.1 Introduction | 161 | ||
| 8.2 Design of the Experiment | 164 | ||
| 8.3 Sample Selection and Collection | 164 | ||
| 8.4 Sample Preparation | 165 | ||
| 8.5 Analysis | 166 | ||
| 8.6 Data Reprocessing | 175 | ||
| 8.7 Data Treatment | 176 | ||
| 8.8 Identification | 177 | ||
| 8.9 Pathway Analysis and Data Interpretation | 179 | ||
| 8.10 Conclusions | 180 | ||
| References | 180 | ||
| Chapter\r9 - Potential of CE-MS for Chiral Metabolic Profiling | 184 | ||
| 9.1 Introduction to Chiral Analysis | 184 | ||
| 9.1.1 Chirality and Its Relevance | 185 | ||
| 9.1.2 Chiral Recognition by Separation Techniques | 186 | ||
| 9.1.3 Possibilities of CE in Chiral Separations | 188 | ||
| 9.1.4 Chiral CE-MS | 190 | ||
| 9.2 Chiral Metabolic Profiling | 191 | ||
| 9.2.1 State-of-the-Art of Chiral Metabolic Profiling | 191 | ||
| 9.2.2 Applications of Chiral Metabolic Profiling by CE-MS | 192 | ||
| 9.2.2.1 Study of the Enantioselective Metabolism of Drugs | 192 | ||
| 9.2.2.2 Study of the Enantioselective Metabolism of Amino Acids | 198 | ||
| 9.2.2.3 d-Amino Acids as Diagnostic Markers of Diseases | 199 | ||
| 9.2.2.4 d-Amino Acids as Indicators of the State of a System | 199 | ||
| 9.2.3 CE-MS Methods to Potentially Study Chiral Profiles for Amino Acids and/or Amines | 202 | ||
| 9.3 Concluding Remarks and Future Trends | 205 | ||
| Acknowledgements | 206 | ||
| References | 206 | ||
| Chapter\r10 - Single-cell Metabolomics with Capillary Electrophoresis–Mass Spectrometry | 209 | ||
| 10.1 Introduction and Overview | 209 | ||
| 10.2 Sampling of Single Cells | 211 | ||
| 10.3 Metabolite Extraction | 214 | ||
| 10.4 Detection by Capillary Electrophoresis–Mass Spectrometry | 216 | ||
| 10.5 Data Processing | 218 | ||
| 10.6 Conclusions and Perspectives | 220 | ||
| Acknowledgements | 221 | ||
| References | 221 | ||
| Chapter\r11 - Capillary Electrophoresis–Mass Spectrometry for Lipid Analysis | 225 | ||
| 11.1 Introduction | 225 | ||
| 11.1.1 Lipids | 225 | ||
| 11.1.2 Lipid Analysis | 226 | ||
| 11.2 CE-MS of Lipids | 228 | ||
| 11.2.1 Phospholipids | 228 | ||
| 11.2.2 Glycolipids | 234 | ||
| 11.2.2.1 Lipopolysaccharides | 234 | ||
| 11.2.2.2 Gangliosides | 234 | ||
| 11.2.3 Acylcarnitines | 237 | ||
| 11.2.4 Lipidic Surfactants | 238 | ||
| 11.2.5 Long-chain Fatty Acids | 242 | ||
| 11.3 CE-MS Analysis of Lipids Using Coated Capillaries | 244 | ||
| 11.4 Concluding Remarks | 250 | ||
| Acknowledgements | 251 | ||
| References | 252 | ||
| Chapter\r12 - Multisegment Injection–Capillary Electrophoresis–Mass Spectrometry: A Robust Platform for High Throughput Metabolite Profiling with Quality Assurance | 255 | ||
| 12.1 Introduction | 255 | ||
| 12.2 CE-MS in Metabolomics: Overcoming Poor Robustness | 257 | ||
| 12.3 Capillary Preparation and Conditioning: Buffer Compatibility Issues in CE-MS | 258 | ||
| 12.4 Versatile Serial Injection Formats with Temporal Signal Pattern Recognition | 262 | ||
| 12.5 Reliable Quantification with Quality Assurance and Batch Correction Adjustment | 268 | ||
| 12.6 Applications of MSI-CE-MS: Biomarker Discovery in Clinical Metabolomic Studies | 273 | ||
| 12.7 Conclusions | 277 | ||
| Acknowledgements | 278 | ||
| References | 278 | ||
| Subject Index | 283 |