Liquid Chromatography in Biomedical Analysis (eBook)

Front Cover 1
Liquid Chromatography in Biomedical Analysis 4
Copyright Page 5
Contents 6
List of Contributors 10
Preface 12
Chapter 1. Liquid chromatography in biomedical analysis : basic approach 14
1. Introduction 14
2. Sample collection or sampling 14
3. Sample preparation 16
4. High-performance liquid chromatography 25
References 32
Chapter 2. Optimization of liquid chromatography for biomedically important compounds 34
1. Introduction 34
2. Relationships between retention and hydrophobicity of aromatic acids 35
3. Liquid chromatography of nitrogen-containing compounds 41
4. Chromatographic behaviour of aromatic acids on ion-exchangers 42
5. Liquid chromatography of bilogical fluids 44
6. Advanced optimization techniques in liquid chromatography 52
7. Conclusion 59
References 59
Chapter 3. Amino acids 60
1. Introduction 60
2. Post-column derivatization using ninhydrin 60
3. Post-column derivatization using OPA 65
4. Reactor 75
5. Pre-column derivatization 76
6. Related topics 86
7. Clinical applications 88
References 92
Chapter 4. Bile acids 94
1. Introduction 94
2. Extraction of bile acids in biological fluids 95
3. Separation of bile acids 100
4. Determination of bile acids 111
References 120
Chapter 5. Carbohydrates 122
1. Introduction 122
2. Principle of the liquid chromatography of carbohydrates 123
3. Liquid chromatography of monosaccharides 128
4. Liquid chromatography of oligosaccharides 141
5. Liquid chromatography in structural studies of carbohydrates 144
6. Liquid chromatographic assay of the activities of enzymes related to carbohydrates 148
7. Automated analysis of carbohydrates and its clinical applications 149
References 151
Chapter 6. Catecholamines 154
1. Introduction 154
2. Post-column fluorescence derivatization 155
3. Pre-column fluorescence derivatization 173
4. Amperometric electrochemical detector for HPLC 176
5. Coulometric detector 176
6. Conclusion 178
References 178
Chapter 7. Fatty acids 182
1. Introduction 182
2. Pretreatment of fatty acids in biological samples for HPLC analysis 185
3. Separation and determination of fatty acids by HPLC using refractive index detection 186
4. Separation and determination of fatty acids by HPLC using UV detec tion 187
5. Various fluorescence-labelling techniques for the determination of fatty acids by HPLC 192
References 201
Chapter 8. Nucleotides 204
1. Introduction 204
2. High-performance liquid chromatography 204
3. Extraction of nucleotides from cells and tissues 215
4. HPLC detectors for nucleotides 218
5. Quantification of nucleotides 219
6. Peak identification 219
References 220
Chapter 9. Porphyrins 222
1. Introduction 222
2. High-performance liquid chromatography of porphyrins 223
3. High-performance liquid chromatography of porphyrinogens 232
4. HPLC detectors for porphyrins and porphyrinogens 235
5. HPLC quantification of porphyrins 235
6. Sample preparation for HPLC of porphyrins 235
7. Biomedical applications 238
8. Assay of enzymes of the haem biosynthetic pathway by HPLC 241
References 243
Chapter 10. Prostaglandins 246
1. Introduction 246
2. Direct UV detection in the separation and determination of PGs, TXB2 and LTs by HPLC 250
3. UV-labelling techniques for the determination of PGs and their metabolites by HPLC 254
4. Fluorescent-labelling techniques for the determination of PGs and TXB2 by HPLC 255
5. Simultaneous separation of several closely related prostanoids by HPLC 256
6. Application of HPLC and GC-MS to the determination of prostanoids in biological samples 260
7. Application of HPLC and RIA to the determination of prostanoids in biological samples 263
References 265
Chapter 11. Steroid hormones 268
1. Introduction 268
2. Simultaneous determination of pregnenolone (.5 -pathway) and progesterone (.4 -pathway) metabolites by HPLC 5 272
3. HPLC determination of C19 steroids 276
4. HPLC determination of corticosteroids 280
5. Determination of estrogens in biological fluids by HPLC 285
6. Application of HPLC and RIA to the determination of steroid hormones in biological samples 288
References 292
Chapter 12. Miscellaneous 296
1. Introduction 296
2. Catecholamine metabolites 296
3. Indoleacetic acid 298
References 299
Subject Index 302
Journal o f Chromatography Library (other volumes in the series) 306