Chromatography of Lipids in Biomedical Research and Clinical Diagnosis (eBook)

Front Cover 1
Chromatography of Lipids in Biomedical Research and Clinical Diagnosis 4
Copyright Page 5
Contents 6
Journal of Chromatography Library 12
List of Abbreviations 16
Preface 20
Acknowledgments 21
List of Contributors 22
Chapter 1: General strategies for practical chromatographic analysis of lipids 24
1.1 Introduction 24
1.2 Chemical structure and chromatographic utility of lipids 24
1.3 Selection, extraction, storage and stability of samples 29
1.4 Purification, preliminary segregation and derivatization 34
1.5 Resolution and quantitation of solutes 40
1.6 Identification and quantitation of unresolved components 52
1.7 Validation of quantitation 59
1.8 Summary and conclusions 64
1.9 Acknowledgments 65
References 65
Chapter 2: Polar capillary GLC of intact natural diacyl and triacylglycerols 71
2.1 Introduction 71
2.2 Rules of resolution 71
2.3 Preparation of samples 74
2.4 Analytical methodology 75
2.5 Applications 79
2.6 Comparison with other Methods 91
2.7 Summary and conclusions 96
2.8 Acknowledgments 97
References 97
Chapter 3: High pressure liquid chromatography of arachidonic acid metabolites involved in inflammation 99
3.1 Introduction 99
3.2 Chromatographic properties of arachidonic acid metabolites 101
3.3 Reparation of samples for HPLC 103
3.4 Norinal-phase HPLC of arachidonic acid metabolites 105
3.5 Argentation HPLC of arachidonic acid metabolites 109
3.6 Reversed-phase HPLC of arachidonic acid metabolites 113
3.7 Quantitation of arachidonic acid metabolites by HPLC 123
3.8 Summary and conclusions 127
3.9 Acknowledgments 128
References 128
Chapter 4: Application of GC/MS techniques to the analysis of prostaglandins and related substances 130
4.1 Introduction 130
4.2 Pathways and chemical structures 131
4.3 Collection of samples 133
4.4 Analytical methodologies 135
4.5 Application of GC/MS with NICI to biological problems 142
4.6 LCMS 147
4.7 MSMS 147
4.8 Acknowledgments 149
References 149
Chapter 5: The GLC of plasma intact lipids in clinical research 151
5.1 Introduction 151
5.2 Historical survey 151
5.3 GLC analysis 152
5.4 Recommended procedure in the analysis of biological samples 171
5.5 Application of the intact lipid profile 179
5.6 Conclusions 183
References 184
Chapter 6: HPLC of the arachidonoyl molecular species of glycerophospholipids in alveolar macrophages and immune responses 186
6.1 Introduction 186
6.2 Separation of the molecular species of diacyl, alkylacyl and alkenylacyl analogues derived from glycerophospholipids of rabbit alveolar macrophages by HPLC 188
6.3 Composition of molecular species of glycerophospholipids 194
6.4 Synthesis of arachidonoyl molecular species 196
6.5 Release of 20:4 from glycerophospholipids in opsonized zymosan-stimulated alveolar macrophages 202
6.6 Summary and conclusions 209
6.7 Acknowledgments 210
References 211
Chapter 7: HPLC of diacylglycerol and phospholipase C sensitive glycerolipids in microsomes of normal tissues and dystrophic muscle (Type Duchenne) 214
7.1 Introduction 214
7.2 Materials and methods 214
7.3 General aspects of the determination of the species pattern of microsomal DG and PC 223
7.4 Species pattern of DG and PC in subcellular preparations of normal and dystrophic human muscle 235
7.5 Summary and conclusions 244
References 245
Chapter 8: Chromatographic analysis of phosphoinositides and their breakdown products in activated blood platelets / neutrophils 248
8.1 General introduction 248
8.2 Extraction of phosphoinositides and their breakdown products 250
8.3 Chromatographic analysis of phosphoinositides 252
8.4 Analytical methods used in the quantitation of inositol phospholipids 262
8.5 Analysis of molecular species of phosphatidylinositol 264
8.6 Chromatographic separation and identification and quantitation of inositol phosphates 268
8.7 Chromatographic analysis of diacylglycerols. monoacylglycerols and free fatty acids 275
8.8 Chromatographic procedures employed in the analysis of phosphatidic acid and lysophosphatidic acid 281
8.9 Conclusions 285
8.10 Acknowledgements 285
References 286
Chapter 9: TLC and HPTLC of phospholipids and glycolipids in health and disease 289
9.1 Introduction 289
9.2 Materials and principal methods for TLC and HPTLC 290
9.3 Application of HPTLC 314
9.4 Summary and conclusions 328
References 328
Chapter 10: HPLC of molecular species of glycerophospholipids in studies of lipoproteins and lipid transport 334
10.1 Introduction 334
10.2 HPLC analysis of diacylphospholipids 335
10.3 Application of molecular species analysis of phosphatidylcholine to biological systems 352
10.4 Future directions of research 366
10.5 Acknowledgments 369
References 369
Chapter 11: High performance liquid chromatoyraphy of glycophingolipids in brain disease 371
11.1 Introduction 371
11.2 Extraction and isolation of glycosphingolipids 372
11.3 General HPLC techniques 373
11.4 Separation of native glycosphingolipids 376
11.5 HPLC analysis of perbenzoylated derivatives 377
11.6 p-Nitrobenzoyl derivatives of neutral glycosphingolipids 385
11.7 Comments 385
11.8 Polyglycosylceramides 386
11.9 Gangliosides 386
11.10 Molecular species separation by reversed-phase HPLC 389
11.11 Liquid chromatography-mass spectrometry (LC/MS) 391
11.12 Applications 395
References 398
Chapter 12: GC/MS of molecular species of glycerophospholipids 401
12.1 Introduction 401
12.2 Brain glycerophospholipids 402
12.3 Platelet-activating factor 411
12.4 Summary and conclusions 421
12.5 Acknowledgments 423
References 424
Chapter 13: LC/MS of natural glycerolipids, sterols and steryl esters 426
13.1 Introduction 426
13.2 Instrumentation and general methodology 426
13.3 Identification of molecular species 430
13.4 Quantitation of molecular species 446
13.5 Metabolic studies with deuterated lipids 452
13.6 Sumnary and conclusions 460
13.7 Acknowledgments 461
References 461
Subject Index 464