Carbohydrate Analysis by Modern Chromatography and Electrophoresis: El-Rassi: 9780444500618: Book: : eCampus.com

This book is an updated and expanded edition of Carbohydrate Analysis, High Performance Liquid Chromatography and Capillary Electrophoresis and is concerned with the analysis of carbohydrates by modern chromatography and electrophoresis including analytical and preparative high performance liquid chromatography (HPLC), thin layer chromatography (TLC), field flow fractionation (FFF), capillary electrophoresis (CE), capillary electrochromatography (CEC), polyacrylamide gel electrophoresis (PAGE), gas chromatography (GC) and supercritical fluid chromatography (SFC).

Thirty-one chapters cover: various modes of HPLC, CE, CEC, FFF, GC and SFC that are currently applied to the analysis of carbohydrates; discussions on analytical and preparative separations; descriptions of the principles of detection and quantitative determination of carbohydrates by the various separation techniques; reviews of sample preparations; and information on important applications. Furthermore, the book describes in detail the different direct and indirect detection methods that have been introduced for the sensitive detection of carbohydrates.

This title is useful for a wide audience including separation scientists; analytical chemists and biochemists; carbohydrate chemists; glycoprotein and glycolipid chemists; molecular biologists; and biotechnologists. The book is also a useful reference for both the experienced analyst and the newcomer and for users of modern chromatography and electrophoresis.

� Contains 31 chapters covering all aspects of carbohydrate analysis by modern chromatography and electrophoresis
� Each chapter discusses the basic principles, advantages and limitations, and applications of the particular detection technique
� Useful reference for both the experienced analyst and the newcomer

Preface

List of Contributors

Part I The Solute

Preparation of Carbohydrates for Analysis by Modern Chromatography and Electrophoresis

(38)

Andrew J. Mort

Margaret L. Pierce

Introduction

(1)

Strategy for sample preparation

(3)

Factors to consider

(1)

Examples from our current projects

(1)

Working with small amounts of sample

(1)

Optimizing sample preparation

(1)

Sample purification

(16)

Glycoproteins

(6)

Proteoglycans

(1)

Glycosylphosphatidylinositol anchors

(1)

Glycolipids

(2)

Lipopolysaccharides, lipo-oligosaccharides, and lipoarabinomannans

(3)

Polysaccharides

(1)

Bacterial exo-polysaccharides

(1)

Plant cell walls

(3)

Mono- and oligosaccharides

(1)

Release of poly- or oligosaccharides from the sample

(5)

Glycoproteins and proteoglycans

(1)

Enzymic cleavage of the carbohydrate portion from glycoproteins and proteoglycans

(1)

Chemical cleavage of the carbohydrate portion from glycoproteins and proteoglycans

(1)

Cleanup of oligosaccharides released from glycoproteins

(1)

Cleavage of glycolipids

(1)

Cleavage of lipopolysaccharides

(1)

Further degradation for structural study

(6)

Enzymic methods

(2)

Chemical methods

(1)

Chemical fragmentation of poly- and oligosaccharides

(1)

Degradations directed to particular functional groups

(3)

Exchanging solvents

(1)

Acknowledgements

(1)

References

(6)

Part II Modern Liquid Chromatography: Fundamentals, Methodologies and Applications

Reversed-Phase and Hydrophobic Interaction Chromatography of Carbohydrates and Glycoconjugates

(62)

Ziad EI Rassi

Introduction

(1)

Basic theoretical concepts

(3)

Chromatographic systems, methodologies and applications

(49)

Reversed-phase chromatography

(1)

Stationary phase

(6)

Mobile phase

(2)

Some general aspects of RPC of carbohydrates and selected applications

(28)

Ion-pair reversed-phase chromatography

(7)

Hydrophobic interaction chromatography

(1)

Stationary phase

(3)

Mobile phase

(2)

Some general aspects of HIC of glycoproteins and selected applications

(1)

Acknowledgements

(1)

References

(9)

HPLC of Carbohydrates on Graphitized Carbon Columns

103

(18)

Kyoko Koizumi

Introduction

103

(1)

The characteristics of GCC

104

(1)

Retention mechanism

104

(1)

Stationary phases

104

(1)

Mobile phases

105

(1)

HPLC of carbohydrates on GCC

105

(12)

Monosaccharides

105

(2)

Oligosaccharides

107

(1)

Disaccharides

107

(4)

N-linked oligosaccharides

111

(1)

Chito-oligosaccharides

111

(1)

Sulfated oligosaccharides

111

(1)

Oligosaccharide alditols

112

(1)

Cyclodextrins (CDs)

113

(1)

CDs and mono-branched CDs

113

(2)

Multibranched CDs

115

(1)

Glycopeptides

116

(1)

Conclusion

117

(1)

Acknowledgements

118

(1)

References

118

(3)

High Performance Hydrophilic Interaction Chromatography of Carbohydrates with Polar Sorbents

121

(44)

Shirley C. Churms

Introduction

121

(1)

The chromatographic system

122

(10)

Polar sorbents used in HILIC

122

(1)

Microparticulate silica

122

(1)

Bonded-phase packings based on silica

123

(2)

Polymer-based packings for HILIC

125

(2)

Mobile phases

127

(1)

Operating variables

128

(1)

Detection systems

129

(3)

Applications of hydrophilic interaction chromatography to carbohydrates

132

(20)

Adsorption and partition chromatography on silica

132

(5)

Partition chromatography on amine-modified silica

137

(2)

Partition chromatography on silica carrying bonded polar phases

139

(1)

Amino phases

139

(5)

Amino-cyano, polyamine and amide phases

144

(2)

Hydroxylic phases

146

(1)

Novel polar phases

146

(3)

Partition chromatography on polymer-based packings

149

(3)

Examples of applications of hydrophilic interaction chromatography to carbohydrates

152

(6)

Conclusions

158

(2)

References

160

(5)

HPLC of Carbohydrates with Cation- and Anion-Exchange Silica and Resin-Based Stationary Phases

165

(42)

Christian W. Huck

Christian G. Huber

Gunther K. Bonn

Introduction

165

(1)

Separation of carbohydrates on cation-exchange stationary phases

165

(17)

Separation mechanisms

165

(1)

Ion exchange

166

(1)

Ion exclusion

166

(1)

Size exclusion

166

(1)

Ligand exchange

167

(2)

Ion-moderated partition chromatography

169

(1)

Interactions with sulfonate groups

170

(1)

Interactions with the support matrix

171

(1)

Silica-based cation-exchange stationary phases

171

(5)

Polymer-based cation-exchange stationary phases

176

(1)

Some basic characteristics of polymer-based cation-exchange stationary phases

176

(3)

Coupling of cation-exchange columns with different counter-ions

179

(2)

Variables affecting the liquid chromatographic separation of carbohydrates on cation exchangers

181

(1)

Particle size

181

(1)

Cross-linking

181

(1)

Column temperature

181

(1)

Eluent composition

182

(1)

Separation of carbohydrates on anion-exchange stationary phases

182

(19)

Separation mechanisms

183

(1)

Anion-exchange chromatography at high pH

183

(1)

Use of borate complexation in carbohydrate chromatography

183

(1)

Silica-based anion-exchange stationary phases

184

(7)

Polymer-based anion-exchange stationary phases

191

(1)

Anion-exchange chromatography at high pH

191

(7)

Anion-exchange chromatography of carbohydrates as borate complexes

198

(3)

Applications

201

(1)

References

202

(5)

High-Performance Anion-Exchange Chromatography of Carbohydrates on Pellicular Resin Columns

207

(44)

Ye Zhang

Yuan C. Lee

Introduction

207

(2)

Instrumentation and operation

209

(12)

General considerations of HPAEC at high pH

209

(1)

The columns

209

(1)

The CarboPac PA-1, PA-10 and PA-100 columns

209

(1)

The Carbopac MA-1 column

210

(1)

The trap columns

210

(1)

Mobile phase and elutions

210

(1)

The NaOH/NaOAc eluent system

210

(1)

The nitrate eluent system

211

(2)

The alkaline-earth ion spiked eluent system

213

(1)

Other eluent systems

214

(1)

Neutral and mildly acidic elution conditions

215

(1)

Detection

216

(1)

HPAEC--PAD

216

(1)

HPAEC--RD

217

(1)

HPAEC--FD/UVD and HPAEC--CD

217

(1)

Desalting

218

(2)

Influence of temperature on retention

220

(1)

Separation mechanisms

221

(6)

Effective charges

221

(1)

pKa values

222

(1)

Steric effects

222

(2)

Hydrophobic interactions

224

(1)

Charge-to-mass ratios

225

(2)

Applications

227

(17)

Monosaccharide analysis

227

(1)

N-glycan analysis

228

(3)

O-glycans and milk- or glycolipid-derived oligosaccharides

231

(2)

Acidic saccharides

233

(1)

Sialic acids

233

(1)

Sulfated and phosphorylated saccharides

233

(1)

Other acidic saccharides

234

(1)

Glucans

234

(1)

Linear and branched glucans

234

(1)

Cyclic glucans

235

(1)

Chain-length distribution analysis

236

(1)

Examination of hydrolytic conditions

237

(1)

Examination of enzymatic reactions

238

(1)

Examination of other reactions and protocols

239

(1)

Carbohydrate analysis in cell cultures

240

(2)

Preparative HPAEC for MS/NMR

242

(1)

Carbohydrate analysis of glycoproteins on PVDF membranes

243

(1)

Acknowledgements

244

(1)

References

244

(7)

Lectin-Affinity Chromatography of Carbohydrates

251

(16)

Tamao Endo

Introduction

251

(1)

Preparation of immobilized lectins

251

(1)

Carbohydrate-binding specificities of various immobilized lectins

252

(7)

Concanavalin A (Con A)

252

(1)

Aleuria aurantia lectin (AAL)

253

(1)

Lens culinaris agglutinin (Lentil lectin, LCA)

253

(1)

Datura stramonium agglutinin (DSA)

253

(2)

Erythroagglutinin (E4-PHA)

255

(1)

Leukoagglutinin (L4-PHA)

256

(1)

Lectins with specificities directed to the peripheral portions of outer chains

256

(3)

Serial lectin column chromatography for the separation of N-glycans

259

(2)

Perspectives

261

(3)

References

264

(3)

Modern Size-Exclusion Chromatography of Carbohydrates and Glycoconjugates

267

(38)

Shirley C. Churms

Introduction

267

(4)

Development of modern size-exclusion chromatography

268

(3)

The chromatographic system

271

(10)

Types of column packing for modern SEC

271

(1)

The mobile phase

271

(4)

Operating variables

275

(1)

Column variables

275

(1)

Sample variables

276

(1)

Flow rate and temperature

277

(1)

Detection methods

278

(3)

Applications of size-exclusion chromatography in the carbohydrate field

281

(17)

Analysis of oligosaccharide mixtures

282

(1)

Fractionation of polysaccharides and glycoconjugates

283

(12)

Molar-mass distribution analysis

295

(3)

Conclusion

298

(1)

References

299

(6)

Molar Mass Determination of Complex Bioglycopolymers by Size Exclusion Chromatography and Light Scattering Detection

305

(42)

Kornelia Jumel

Introduction

305

(2)

Theory

307

(4)

Light scattering

307

(3)

Size exclusion chromatography

310

(1)

Instrumentation

311

(8)

Low-angle laser light scattering (LALLS)

312

(1)

Multi-angle light scattering (MALLS)

313

(2)

Right angle instrument

315

(4)

Specific refractive index increment

319

(1)

Starch

319

(3)

Cellulose and cellulose derivatives

322

(1)

Galactomannans

323

(2)

Pectin

325

(1)

Fructans

325

(1)

Marine polysaccharides

326

(5)

Carrageenan

326

(1)

Toxicity of carrageenans

327

(2)

Gelation mechanism

329

(1)

Alginates

330

(1)

Agars

330

(1)

Chitosan

331

(1)

Exopolysaccharides

331

(5)

Xanthan

332

(1)

Scleroglucan

333

(3)

Glycoproteins

336

(3)

Hyaluronic acid

339

(1)

Heparin

340

(1)

Conclusions

341

(1)

References

341

(6)

Multi-Dimensional Mapping of N-Linked Glycans by HPLC

347

(40)

Noriko Takahashi

Introduction

347

(1)

Code numbers for the glycans

348

(1)

Multi-dimensional mapping technique

348

(3)

Outline of the method

348

(1)

2-D mapping technique for neutral N-glycans

349

(2)

Extension to 3-D mapping technique for sialyl N-glycans

351

(1)

Contribution of component monosaccharides to the coordinates of neutral and sialyl pyridylaminated N-glycans on a 2-D map

351

(6)

Parameterization of contribution of component monosaccharides to elution volumes of PA-glycans on ODS- and amide-silica columns

353

(1)

Calculation of the unit contribution values

353

(1)

Application of the UC value diagrams together with the multi-dimensional map

354

(3)

Application of multi-dimensional mapping to various N-glycans

357

(27)

N-Glycan structures of human integrin α5β1 (35 different neutral, mono-, di-, tri- and tetra-sialyl N-glycans) (integrin)

357

(1)

Separation of N-glycans by three successive HPLC steps

357

(1)

Identification of N-glycan structures on the 2-D map

357

(8)

N-Glycans of murine neuropsin produced insect cells (containing di-fucosyl tri-mannosyl core)

365

(1)

PA-N-glycan profile of neuropsin glycoprotein and the structural characterization using 2-D mapping technique

366

(1)

Identification of glycans A-1, C-2, and E on the 2-D map

367

(4)

N-Glycans of human urinary kallidinogenase (containing GalNAc residues at the non-reducing ends)

371

(1)

Preparation of PA-glycans

372

(1)

Structural characterization of PA-glycans by a combination of HPLC and exoglycosidase digestion

372

(2)

Monosaccharide analysis in the outer chains of N-glycans b--v

374

(1)

H-NMR analyses of N-glycans m, p, g, i, v and r

375

(2)

Methylation analyses of N-glycans p, m and v

377

(1)

Structural analysis of each N-glycan by 2-D sugar mapping

377

(7)

References

384

(3)

HPLC Analysis of Oligosaccharides Derived from Glycosaminoglycans in Biological Materials

387

(16)

Toshio Imanari

Toshihiko Toida

Ichiro Koshiishi

Hidenao Toyoda

Introduction

387

(2)

Analysis of glycosaminoglycans as their unsaturated disaccharides

389

(8)

Determination of hyaluronan, chondroitin sulfate and dermatan sulfate

389

(1)

Determination of heparin and heparan sulfate

390

(1)

GAG analyzer by HPLC

391

(1)

Apparatus

392

(1)

HPLC conditions

392

(2)

Preparation of unsaturated disaccharides from GAGs in biological materials

394

(1)

Urine

394

(1)

Blood cells and plasma

395

(1)

Tissues

396

(1)

Analysis of di- and oligosaccharides from GAGs

397

(3)

Determination of HA, CS and DS as their disaccharides by solvolysis

397

(1)

Determination of keratan sulfate

397

(2)

Sequencing analysis of oligosaccharides derived from GAGs

399

(1)

Exosequence analysis of oligosaccharides from CS

399

(1)

Exosequence analysis of oligosaccharides from HP/HS

400

(1)

Determination of intact GAGs

400

(1)

Conclusion

401

(1)

References

401

(2)

Starch Chain Length Analysis by Using an Anion-Exchange Chromatography System Equipped with an Enzyme Reactor and a PAD Detector

403

(20)

Kit-Sum Wong

Jay-Lin Jane

Introduction

403

(2)

HPAEC-ENZ-PAD for carbohydrate analysis

405

(6)

Separation of amylodextrins by HPAEC

405

(3)

Detection of amylodextrin by PAD

408

(2)

Detection of amylodextrin by PAD with a postcolumn enzyme reactor

410

(1)

HPAEC-ENZ-PAD for starch chain length analysis

411

(5)

Advantages and disadvantages

416

(2)

Acknowledgements

418

(1)

References

419

(4)

TLC and HPLC of Glycosphingolipids

423

(60)

Johannes Muthing

Introduction

423

(6)

General remarks

423

(1)

Physiological functions of GSLs

424

(2)

Role of GSLs in disease

426

(3)

Structural features, biodistribution, and membranous organization of GSLs

429

(4)

Chemical structure and nomenclature

429

(3)

Subcellular distribution

432

(1)

Supramolecular organization

432

(1)

Fundamentals of isolation and structural characterization

433

(1)

Thin-layer chromatography of GSLs

434

(27)

Materials and methods

435

(1)

TLC plates

435

(1)

Sample application, solvents, and development of chromatograms

435

(1)

Staining and quantification

436

(1)

Continuous and multiple development

437

(1)

Short-bed continuous development

437

(2)

Multiple development

439

(1)

Automated multiple development

440

(1)

Two-dimensional TLC

441

(1)

Preparative TLC

442

(4)

Combined TLC and mass spectrometry

446

(1)

Overlay technique

447

(1)

General procedure

448

(1)

Antibodies

448

(4)

Enzymatic modification

452

(2)

Toxins

454

(3)

Lectins

457

(1)

Other proteins and related compounds

457

(1)

Viruses

458

(1)

Bacteria

459

(1)

Cells

460

(1)

Conclusion and perspectives

460

(1)

High-performance liquid chromatography (HPLC) of GSLs

461

(9)

HPLC of derivatized GSLs

461

(1)

HPLC of underivatized GSLs

462

(1)

Preparative anion-exchange HPLC of gangliosides

463

(1)

Materials and methods

463

(1)

HPLC of mouse brain ganglio-series gangliosides on TMAE-Fractogel

464

(1)

HPLC of granulocyte neolacto-series gangliosides on TMAE-Fractogel

465

(3)

HPLC of GM3(Neu5Ac) and GM3(Neu5Gc) on TMAE-Fractogel

468

(2)

Acknowledgements

470

(1)

References

470

(13)

Analysis of Carbohydrates in Food and Beverages by HPLC and CE

483

(22)

Tomoyoshi Soga

Introduction

483

(1)

HPLC

484

(3)

Separation mechanisms

484

(1)

Hydrophilic interaction

484

(1)

Ligand exchange

484

(1)

Borate complex anion exchange

485

(1)

Size exclusion

485

(1)

High-performance anion exchange

485

(1)

Detection systems

485

(1)

Refractive index detection

486

(1)

Pulsed amperometric detection

486

(1)

487

(2)

CE systems

487

(1)

Derivatized mono- and oligosaccharide methods

487

(1)

Indirect UV detection methods for carbohydrates at high pH

487

(1)

Carbohydrate-borate complexes

488

(1)

Applications

489

(8)

HPLC systems

489

(1)

Mono- and oligosaccharides

489

(5)

Organic acids and carbohydrates

494

(1)

CE systems

494

(1)

Mono- and oligosaccharides

494

(2)

Inorganic anions, organic acids, amino acids and carbohydrates

496

(1)

Conclusions

497

(3)

Acknowledgements

500

(1)

References

500

(5)

Part III Preparative Liquid Chromatography

Preparative HPLC of Carbohydrates

505

(30)

Kevin B. Hicks

Arland T. Hotchkiss Jr.

Introduction

505

(1)

Equipment

506

(5)

Solvent delivery systems

506

(1)

Pre- and guard columns

506

(1)

Injection systems

506

(1)

Column hardware

507

(1)

Equipment for commercial large-scale production of carbohydrates

507

(1)

Equipment for packing columns

507

(1)

Detection systems

508

(1)

Fraction collectors

509

(1)

Equipment and techniques for desalting and/or neutralizing fractions from preparative HPAEC

509

(2)

Stationary phases

511

(5)

Normal phases

511

(1)

Reversed phases

512

(1)

Cation-exchange resins

513

(2)

Anion-exchange phases

515

(1)

Miscellaneous phases

516

(1)

General guidelines for preparative HPLC of carbohydrates

516

(2)

Obtaining adequate resolution (R)

517

(1)

Selection of column size

517

(1)

Selection of column flow rate

518

(1)

Specific preparative HPLC techniques for various classes of carbohydrates

518

(9)

Neutral monosaccharides and derivatives

518

(1)

Acidic and basic monosaccharides and derivatives

519

(1)

Neutral oligosaccharides

520

(3)

Acidic and basic oligosaccharides

523

(4)

Glycopeptides and other glycoconjugates

527

(1)

Bibliographic information

527

(2)

References

529

(6)

Preparative Purification and Utilization of Tyrosinamide N-Glycan Libraries

535

(38)

Kevin G. Rice

V. Hayden Thomas

Wendy T. Collard

Introduction

535

(1)

Release of N-glycans from glycoproteins

536

(2)

Tyrosinamide derivatization of N-glycans

538

(14)

Enzymatic remodeling of tyrosinamide N-glycans

552

(9)

Tyrosinamide N-glycan aglycon transformation

561

(7)

Conclusions

568

(1)

Acknowledgements

569

(1)

References

569

(4)

Part IV Field Flow Fractionation

Field-Flow Fractionation for Molar Mass Characterization of Polysaccharides

573

(24)

Karl-Gustav Wahlund

Introduction

573

(1)

Basics of field-flow fractionation

574

(1)

Molecular-size fractionation by FFF

575

(2)

Sedimentation field-flow fractionation

576

(1)

Thermal field-flow fractionation

576

(1)

Flow field-flow fractionation

577

(1)

From retention time to molar mass

577

(5)

Indirect methods

577

(2)

Direct methods

579

(3)

FFF-MALS

582

(1)

Applications to various polysaccharides

582

(12)

Dextran

582

(1)

Pullulan

583

(4)

Cellulose derivatives

587

(2)

Starch (amylopectin and amylose)

589

(2)

Starch derivatives

591

(1)

κ-Carrageenan

591

(1)

Xanthan

592

(1)

Gum arabic

592

(1)

Ficoll™

593

(1)

Pectin

593

(1)

Hyaluronate

593

(1)

Polysulfonated polysaccharide

593

(1)

References

594

(3)

Part V Electrophoresis and Electrochromatography: Fundamentals, Methodologies and Applications

Capillary Electrophoresis and Electrochromatography of Carbohydrates

597

(80)

Ziad El Rassi

Introduction

597

(1)

Electrolyte systems

598

(7)

Borate-based electrolytes

598

(4)

Highly alkaline pH electrolyte systems

602

(2)

Metal cation-based electrolyte systems

604

(1)

Detection systems and pre-column derivatization

605

(28)

Detection of underivatized carbohydrates

605

(1)

Direct UV detection

605

(1)

Indirect UV and indirect fluorescence detection of underivatized carbohydrates

606

(7)

Electrochemical detection

613

(5)

Mass spectrometry detection

618

(1)

Refractive index detection

619

(1)

Spectrophotometric determination of glucose via its enzyme degradation products

619

(1)

Detection of labeled carbohydrates

620

(1)

Dynamically labeled carbohydrates

620

(1)

Pre-column derivatization

621

(12)

Separation approaches and selected applications

633

(34)

Monosaccharides

633

(1)

Underivatized monosaccharides

633

(2)

Derivatized monosaccharides

635

(11)

Oligosaccharides

646

(1)

Underivatized oligosaccharides

646

(1)

Derivatized oligosaccharide

647

(10)

Polysaccharides

657

(1)

Underivatized polysaccharide

657

(4)

Dynamically labeled and derivatized polysaccharides

661

(2)

Glycolipids

663

(4)

Capillary electrochromatography

667

(5)

Conclusions

672

(1)

Acknowledgements

672

(1)

References

672

(5)

Studies on Carbohydrate--Protein Interaction by High Performance Capillary Affinity Electrophoresis

677

(14)

Susumu Honda

Atsushi Taga

Introduction

677

(1)

Principle of association/dissociation constant estimation by capillary affinity electrophoresis

677

(2)

Normal system (protein as sample/carbohydrate ligand as buffer additive)

679

(4)

Using carbohydrate ligand having electric charge

679

(1)

Using carbohydrate ligand not having electric charge

680

(1)

Chemical introduction of a strongly negative charge

680

(1)

Competitive binding

681

(2)

Reverse system (carbohydrate ligand as sample/protein as buffer additive)

683

(6)

Single carbohydrate system

683

(1)

Multiple carbohydrate system

684

(3)

Comparative study of glycoprotein glycoform--protein interaction

687

(2)

Conclusion

689

(1)

References

690

(1)

Analysis of Glycoproteins and their Glycopeptide and Glycan Fragments by Electrophoresis and Capillary Electrophoresis

691

(96)

Nguyet Thuy Tran

Marion Cabanes-Macheteau

Myriam Taverna

Introduction

691

(2)

Glycoform analyses

693

(49)

Conventional electrophoresis

693

(1)

One-dimensional gel electrophoresis

693

(6)

Two-dimensional gel electrophoresis

699

(1)

Detection of glycoproteins

699

(5)

Capillary electrophoresis

704

(1)

CZE

704

(17)

MEKC or CZE in the presence of surfactants

721

(5)

CIEF

726

(8)

Other separation modes: CGE and CEC

734

(4)

Detection of glycoproteins separated by CE

738

(3)

Electrophoresis in miniaturized devices

741

(1)

Glycopeptide mapping

742

(16)

Peptide mapping by CZE

744

(4)

Peptide mapping by CEC and electrically assisted capillary liquid chromatography (cLC)

748

(2)

Peptide mapping by CIEF

750

(2)

Combination of HPLC and CE

752

(1)

Detection of glycopeptides

753

(5)

Glycan mapping

758

(19)

Release of oligosaccharide from glycoproteins

758

(1)

Enzymatic release

759

(1)

Chemical release

760

(1)

Oligosaccharide labelling

760

(2)

Oligosaccharide mapping

762

(1)

762

(10)

CEC

772

(1)

Fluorophore-assisted carbohydrate electrophoresis

773

(4)

References

777

(10)

Fluorphore-Assisted Carbohydrate Electrophoresis (FACE®): Rapid Enzymatic Analysis of Oligosaccharide Mixtures from Recombinant Glycoproteins for Batch Comparability

787

(12)

C. Hague

I. Masada

B.K. Brandley

Introduction

787

(1)

Materials

788

(1)

Oligosaccharide profiling: aryl sulfatase B

789

(1)

Enzymatic sequencing of N-linked oligosaccharides from FACE® bands

790

(2)

Rapid enzymatic analysis of oligosaccharide mixtures from recombinant iduronidase for batch comparability

792

(5)

Conclusions

797

(1)

Acknowledgements

797

(1)

References

797

(2)

Capillary Electrophoresis of Intact and Depolymerized Glycosaminoglycans and Proteoglycans

799

(30)

Nikos K. Karamanos

Anders Hjerpe

Biological significance of GAGs and PGs

799

(6)

Structure of GAGs

801

(4)

Functional oligomeric structures of GAGs

805

(1)

CE-based methodology for characterizing GAGs

805

(6)

Detection of GAGs in CE analysis

808

(1)

Detection of underivatized GAGs

808

(1)

Precolumn derivatization of GAGs and detection

809

(2)

Strategies for analyzing and elucidating the GAG/PG structure

811

(12)

CE analysis of intact and oligomeric GAGs

812

(1)

Hyaluronan

812

(1)

Galactosaminoglycans

812

(2)

HS and heparin

814

(1)

Analysis of depolymerized GAGs

814

(1)

Hyaluronan

814

(4)

Galactosaminoglycans

818

(2)

HS and heparin

820

(3)

Applications

823

(1)

Abbreviations

824

(1)

References

824

(5)

Part VI Gas Chromatography

A Current Perspective on Analysis of Sugar Monomers Using GC-MS and GC-MS/MS

829

(16)

Alvin Fox

Introduction

829

(2)

General discussion on sample preparation for GC analysis

831

(2)

Automated preparation of alditol acetate derivatives of sugars present in complex matrices

833

(3)

GC and GC-MS analysis of alditol acetate derivatives

836

(6)

Conclusions

842

(1)

References

842

(3)

Gas Chromatography--Mass Spectrometric Analysis of Monosaccharides after Methanolysis and Trimethylsilylation. Potential for the Characterization of Substances of Vegetal Origin: Application to the Study of Museum Objects

845

(60)

Philippe Mejanelle

Jean Bleton

Alain Tchapla

Serge Goursaud

Introduction

845

(3)

Experimental

848

(1)

Sample preparation

848

(1)

Gas chromatography--mass spectrometry

848

(1)

Methanolysis of standard monosaccharides

848

(15)

Methanolysis of aldoses and uronic acids

848

(1)

Structures of derivatives formed from the different sugar classes

848

(3)

Variation of composition with temperature

851

(8)

Methanolysis of ketohexoses

859

(4)

Identification of monosaccharides by gas chromatography

863

(5)

Identification of monosaccharides by electron impact--mass spectrometry

868

(20)

General features of mass spectra

868

(4)

Fragmentation reactions under EI ionization

872

(1)

General fragmentation pattern

872

(3)

Influence of the ring size

875

(1)

Specific fragmentation of uronic acids and furanurono-6,3-lactones

876

(1)

Specific fragmentations of ketohexoses

877

(1)

Specific fragmentations of 1,6-anhydrosugars

878

(1)

Identification of sugar classes

879

(3)

Differentiation of stereoisomeric methyl glycosides

882

(1)

Differentiation of hexopyranosides

883

(1)

Differentiation of pentopyranosides

883

(2)

Differentiation of 6-deoxyaldopyranosides

885

(1)

Differentiation of hexuronic acids

885

(3)

Applications

888

(12)

Analysis of reference samples

889

(3)

Analysis of manuscript inks

892

(2)

Analysis of the materials and substances of a mummy cartonnage

894

(3)

Analysis of coatings from objects of ethnographic interest

897

(3)

Conclusion

900

(1)

References

901

(4)

Part VII Detection

Pulsed Electrochemical Detection of Carbohydrates at Noble Metal Electrodes following Liquid Chromatographic and Electrophoretic Separation

905

(42)

William R. LaCourse

Introduction

905

(2)

Voltammetric basis of pulsed electrochemical detection

907

(5)

Residual response at Au and Pt electrodes

907

(2)

Glucose response at Au electrodes

909

(2)

Mechanistic aspects of PED response

911

(1)

Design and optimization of PED waveforms

912

(7)

Historical significance of pulsed potential cleaning

912

(1)

General waveform design

913

(1)

Waveform optimization

914

(4)

Carbohydrate response revisited

918

(1)

Representative LC-PED results for Mode I detections

919

(10)

Carbohydrates

919

(1)

Oligosaccharides and gradient elution

920

(4)

Enzymatic post-column reactors

924

(1)

Alcohols

925

(1)

Amino alcohols

925

(2)

Aminosugars and glycoconjugates

927

(2)

Other considerations

929

(1)

Integrated pulsed amperometric detection

929

(5)

Voltammetry

929

(1)

Chronoamperometry

930

(1)

The IPAD waveform

931

(1)

IPAD waveform optimization

932

(2)

Representative LC-PED results for Mode II detections

934

(4)

Amines and amino acids

934

(2)

Peptides

936

(1)

Sulfur compounds

937

(1)

Future improvements

938

(2)

Faster waveforms

938

(1)

Capillary electrophoresis

938

(1)

Indirect detection

939

(1)

Conclusions

940

(2)

Acknowledgements

942

(3)

Appendix A. selected applications of LC--PED to mixtures of alditols and carbohydrates

942

(1)

References cited in Appendix A

943

(1)

Appendix B. Selected applications of LC--PED in pharmaceutical and life sciences

943

(1)

References cited in Appendix B

944

(1)

References

945

(2)

Electrochemical Detection of Carbohydrates at Constant Potential after HPLC and CE Separations

947

(14)

Richard P. Baldwin

Introduction

947

(2)

Overview

949

(4)

Electrode material

949

(1)

Separation considerations

950

(1)

Instrumentation

950

(3)

Applications

953

(2)

Conclusions

955

(3)

Constant potential vs. pulsed detection

957

(1)

EC detection in general

957

(1)

Acknowledgements

958

(1)

References

958

(3)

Mass Spectrometry of Oligosaccharides

961

(82)

Mark Stahl

Alexander von Brocke

Ernst Bayer

Introduction

961

(1)

Sample preparation for mass spectrometry

962

(2)

Mass spectrometry of oligosaccharides

964

(18)

Mass spectrometers

964

(1)

Magnetic sector field mass spectrometer (EB)

964

(1)

Quadrupole mass spectrometer (Q)

965

(1)

Time-of-flight (TOF) mass spectrometer

965

(1)

Quadrupole-time of flight (Q-TOF) mass spectrometer

966

(1)

Ion trap (IT) mass spectrometer

966

(1)

Ion cyclotron resonance Fourier transform mass spectrometer (ICR--FT--MS)

967

(1)

Ionization methods

967

(1)

Electron impact (EI) ionization mass spectrometry

967

(1)

Chemical ionization (CI) mass spectrometry

968

(2)

Fast atom bombardment (FAB) mass spectrometry

970

(1)

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry

971

(4)

Electrospray ionization (ESI) mass spectrometry

975

(7)

Summary

982

(1)

Fragmentation

982

(33)

Fundamental fragmentation reactions and nomenclature of fragments

982

(1)

Fragmentation of the glycosidic bond

983

(1)

Cross-ring fragmentation

984

(1)

Branched oligosaccharides

984

(1)

Internal fragmentation

985

(1)

Derivatized oligosaccharides

986

(1)

Fragmentation during fast atom bombardment ionization

987

(2)

Fragmentation induced by MALDI mass spectrometry

989

(6)

Fragmentation induced by ESI and CID

995

(20)

Sequencing of oligosaccharides by partial degradation in combination with MS''

1015

(2)

Coupling of separation methods with MS

1017

(21)

GC/MS

1017

(6)

LC/MS

1023

(2)

Applications of HPLC/MS

1025

(7)

CE/ESI--MS

1032

(3)

Sample preconcentration for CE/MS

1035

(3)

Conclusion

1038

(1)

Acknowledgements

1039

(1)

References

1039

(4)

Pre- and Post-Column Detection-Oriented Derivatization Techniques in HPLC of Carbohydrates

1043

(28)

Sumihiro Hase

Introduction and scope

1043

(3)

Carbonyl derivatization

1046

(15)

Formation of amines by reductive amination

1046

(7)

Convertible derivatization

1053

(1)

Tritium labeling

1054

(1)

Derivatization of glycamines

1054

(1)

Carbonyl derivatization into imines (Schiff bases) or glycosylamines

1055

(4)

Miscellaneous

1059

(2)

Hydroxy and amino derivatization

1061

(2)

Post-column derivatization

1063

(1)

References

1064

(7)

Enzyme Reactors and Enzyme Electrodes for the Liquid Chromatographic Determination of Carbohydrates

1071

(30)

Luc J. Nagels

Introduction

1071

(1)

Immobilization of enzymes in reactors

1072

(2)

Applications of post-column enzyme reactors in liquid chromatography of carbohydrates

1074

(17)

Detection methods suited for combination with post-column enzyme reactors

1074

(2)

Detectors combined with oxidase reactors

1076

(4)

Detectors combined with dehydrogenase reactors

1080

(1)

Chromatographic determination of mono- and disaccharides using dehydrogenase-based enzyme reactors

1080

(3)

Chromatographic determination of mono- and disaccharides using oxidase-based enzyme reactors

1083

(3)

Determination of oligo- and polysaccharides

1086

(5)

Determination of carbohydrate related molecules

1091

(1)

Reactor kinetics

1091

(4)

Enzyme electrodes for the HPLC determination of carbohydrates

1095

(2)

References

1097

(4)

Carbohydrate Analysis by LC and SFC using Evaporative Light Scattering Detection

1101

(34)

Michel Lafosse

Bernard Herbreteau

Introduction

1101

(1)

Evaporative light scattering detector: principle, detector technology and characteristics

1102

(11)

Nebulization

1103

(1)

LC nebulizer

1103

(3)

SFC nebulizer

1106

(1)

Vaporization

1107

(1)

Measurement of the scattered light intensity: quantitative analysis

1108

(3)

Characteristic properties of the detector

1111

(1)

Which detector should be used with the chromatographic analysis of a given mixture?

1112

(1)

LC analysis of carbohydrates and carbohydrate derivatives

1113

(15)

Separation-detection dependency

1113

(1)

Eluents incompatible with ELSD characteristics

1113

(1)

Volatile eluents

1113

(1)

ELSD quality of a chromatographic eluent

1113

(1)

Stability of the stationary phase

1114

(1)

Automatic sugar analysis in a complex mixture

1115

(2)

Isocratic and gradient LC with polar and apolar stationary phases

1117

(1)

Diol and some other polar stationary phases

1117

(1)

Aminopropyl-bonded stationary phases

1118

(1)

Apolar stationary phases

1119

(1)

Quantitative determination

1120

(4)

Carbohydrate derivatives

1124

(4)

Packed column SFC of carbohydrates and derivatives

1128

(3)

Conclusions and future prospect

1131

(1)

References

1132

(3)

Chiroptical Detectors for HPLC of Carbohydrates

1135

(16)

Neil Purdie

Introduction

1135

(1)

History of development

1136

(1)

Bases behind chiroptical detectors

1137

(2)

Instrumentation

1139

(3)

General descriptions

1139

(1)

Polarimeters

1139

(1)

Spectropolarimeters

1139

(1)

Modifications for chromatographic detection

1140

(1)

Calibrations

1141

(1)

Instruments

1141

(1)

Compounds

1142

(1)

Applications

1142

(6)

Bulk systems

1143

(1)

Bulk systems with induced chirality

1143

(2)

LC systems

1145

(2)

LC systems with induced chirality

1147

(1)

Conclusion

1148

(1)

References

1148

(3)

Author Index

1151

(46)

Subject Index

1197


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