Carbohydrate analysis: high performance liquid chromatography and capillary electrophoresis

Z. El Rassi0444899812, 9780444899811

Carbohydrates and glycoconjugates play an important role in several life processes. The wide variety of carbohydrate species and their inherent polydispersity and heterogeneity require separation techniques of high resolving power and high selectivity such as high performance liquid chromatography (HPLC) and capillary electrophoresis (HPCE). In the last decade HPLC, and recently HPCE methods have been developed for the high resolution and reproducible quantitation of carbohydrates. Despite the importance of these two column separation technologies in the area of carbohydrates, no previous book describes specialized methods for the separation, purification and detection of carbohydrates and glycoconjugates by HPLC and HPCE.
Therefore, the objective of the present book is to provide a comprehensive review of carbohydrate analysis by HPLC and HPCE by covering analytical and preparative separation techniques for all classes of carbohydrates including mono- and disaccharides; linear and cyclic oligosaccharides; branched heterooligosaccharides (e.g., glycans, plant-derived oligosaccharides); glycoconjugates (e.g., glycolipids, glycoproteins); carbohydrates in food and beverage; compositional carbohydrates of polysaccharides; carbohydrates in biomass degradation; etc.
The book will be of interest to a wide audience, including analytical chemists and biochemists, carbohydrate, glycoprotein and glycolipid chemists, molecular biologists, biotechnologists, etc. It will also be a useful reference work for both the experienced analyst and the newcomer as well as for users of HPLC and HPCE, graduates and postdoctoral students.

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Table of contents :
CarbohydrateIm Analysis: High Performance Liquid Chromatography and Capillary Electrophoresis……Page 4
Copyright Page……Page 5
Preface……Page 6
List of Contributors……Page 8
Contents……Page 10
Part I: The Solute……Page 22
1.1 Introduction……Page 24
1.2 Sample purification……Page 25
1.3 Release of poly- or oligosaccharides from the sample……Page 38
1.4 Further degradation for structural study……Page 41
1.5 Exchanging solvents……Page 47
1.6 Sources for reference compounds……Page 50
1.7 References……Page 56
Part II: Analytical and Preparative Separations……Page 60
2.1 Introduction……Page 62
2.2 Fundamentals……Page 63
2.3 Stationary phases……Page 68
2.4 Mobile phases……Page 77
2.5 Separation methodologies and selected applications……Page 82
2.6 References……Page 115
3.1 Introduction……Page 124
3.2 The chromatographic system……Page 125
3.3 Applications of hydrophilic interaction chromatography to carbohydrates……Page 135
3.4 Examples of applications of hydrophilic interaction chromatography to carbohy- drates……Page 155
3.5 Conclusions……Page 158
3.6 References……Page 163
4.2 Cation-exchange HPLC of carbohydrates……Page 168
4.3 Anion-exchange HPLC of carbohydrates……Page 185
4.5 References……Page 198
5.2 Monosaccharide analysis……Page 202
5.3 Oligosaccharide analysis……Page 209
5.4 In-line and off-line desalting after HPAEC separations……Page 220
5.5 Other important applications……Page 222
5.6 Abbreviations……Page 225
5.7 References……Page 226
6.2 Use of high performance liquid chromatography……Page 232
6.3 Use of high performance capillary electrophoresis……Page 246
6.4 References……Page 251
7.1 Introduction……Page 254
7.2 The chromatographic system……Page 258
7.3 Applications of size-exclusion chromatography in the carbohydrate field……Page 268
7.4 Conclusions……Page 282
7.5 References……Page 283
8.1 Introduction……Page 288
8.2 Overview of the fundamentals of capillary electrophoresis……Page 289
8.3 The electrophoretic system……Page 306
8.4 HPCE methodologies and applications……Page 332
8.6 References……Page 376
9.2 Equipment……Page 382
9.3 Stationary phases……Page 387
9.4 General guidelines for preparative HPLC of carbohydrates……Page 392
9.5 Specific preparative HPLC techniques for various classes of carbohydrates……Page 394
9.6 Bibliographic information……Page 402
9.7 References……Page 404
Part III: The Detection……Page 410
10.1 Introduction……Page 412
10.2 Voltammetric basis of pulsed electrochemical detection……Page 414
10.3 Design and optimization of PED waveforms……Page 418
10.4 Representative LC-PED results for carbohydrates……Page 424
10.5 Other applications of LC-PED……Page 434
10.6 Future improvements……Page 440
10.8 Acknowledgements……Page 443
10.9 Appendices……Page 444
10.10 References……Page 448
11.1 Introduction……Page 452
11.2 Refractive index of mixtures……Page 453
11.3 Instrumentation……Page 454
11.4 Concluding remarks
……Page 465
11.5 References……Page 467
12.1 Introduction……Page 468
12.2 Techniques and strategies……Page 470
12.3 Protein glycosylation……Page 480
12.4 Glycosphingolipids……Page 512
12.5 Lipo-peptido-glycans and miscellaneous classes……Page 514
12.6 Conclusions and future challenges……Page 525
12.8 Appendix……Page 526
12.9 References……Page 528
13.1 Introduction……Page 536
13.2 Evaporative light scattering detector – principle, detector technology and char- acteristics……Page 537
13.3 Analysis of carbohydrates and carbohydrate derivatives……Page 546
13.4 Conclusion and future prospects……Page 557
13.5 References……Page 560
14.1 Introduction……Page 562
14.2 History of development……Page 563
14.3 Bases behind chiroptical detectors……Page 564
14.4 Instrumentation……Page 566
14.5 Applications……Page 569
14.7 References……Page 574
15.1 Introduction and scope……Page 576
15.2 Carbonyl derivatization……Page 579
15.3 Hydroxy derivatization……Page 585
15.4 Miscellaneous……Page 587
15.5 Convertible derivatization……Page 589
15.6 Post-column derivatization……Page 591
15.8 References……Page 592
16.2 Immobilization of enzymes in reactors……Page 598
16.3 Applications of post-column enzyme reactors in liquid chromatography of carbohydrates……Page 602
16.4 Reactor kinetics……Page 620
16.5 References……Page 624
17.2 UV detection in HPLC and HPCE……Page 628
17.3 Fluorescence detection in HPCE……Page 639
17.4 Other miscellaneous detection in HPLC and HPCE……Page 648
17.5 Conclusions……Page 657
17.7 References……Page 658
Subject Index……Page 662
Journal of Chromatography Library……Page 690