Aqueous Size-Exclusion Chromatography (eBook)

Front Cover 1
Aqueous Size–Exclusion Chromatography 4
Copyright Page 5
Contents 6
Preface 14
List of Contributors 18
Part I: Separation Mechanisms 20
Chapter 1. Size exclusion parameters 22
1. Introduction 22
2. Solute retention mechanism in SEC 23
3. Acknowledgements 39
References 39
Chapter 2. Partitioning: Hydrophobic interactions 42
1. Introduction 42
2. Hydrophobic nature of sugars as an origin of hydrophobic properties of polysaccharide gels 43
3. Hydrophobic interactions in hydrophilic gels 60
4. Conclusion 71
References 72
Chapter 3. Electrostatic effects 74
1. Introduction 74
2. SEC of low MW salts 75
3. SEC of polyelectrolytes (polyanions) 79
4. SEC of polycations 82
5. Electrostatic effects in SEC of proteins 85
6. Models for electrostatic effects 89
References 92
Chapter 4. Exclusion chromatography of inorganic compounds 96
1. Introduction 96
2. Distribution processes of inorganic salts 96
3. Inorganic polymers 117
4. Metal complexes 123
References 130
Part II: Characterization of Stationary Phases 136
Chapter 5. Pore size distributions 138
1. Introduction 138
2. Pore structure of supports for size exclusion chromatography 139
3. Characterization of pore size 143
4. Impact of pore dimensions on size exclusion parameters 165
Acknowledgements 169
References 169
Chapter 6. Structural analysis of porous materials by measurement of size exclusion 176
1. Introduction - Principle and general view of chromatographic analysis for solid-solute interaction 176
2. Size exclusion chromatography for porous structure analysis 177
3. Concluding remarks 188
References 188
Chapter 7. Column efficiency 190
1. Introduction 190
2. Definition of column efficiency 192
3. Measurement of column efficiency 195
4. Operational variables of column efficiency 197
5. Problems associated with aqueous SEC 203
References 208
Part III: New Packings 210
Chapter 8. Native and bonded silicas in aqueous SEC 212
1. Introduction 212
2. The physical properties of support and column and their role in SEC separations 213
3. The surface chemistry of silica SEC packings 227
4. Commercial native and bonded silica SEC packings and columns and their operation 238
5. Conclusion 250
References 251
Chapter 9. Rigid polymer gels for SEC and their application to biopolymers 254
1. Introduction 254
2. Currently available size exclusion chromatographic columns 255
3. General separation data on SEC columns 265
4. Application of SEC to biological systems 270
5. Acknowledgment 282
References 282
Part IV: Biopolymers 286
Chapter 10. Biopolymers. I. Protein chromatography in denaturing and non-denaturing solvents 288
1. Introduction 288
2. Packings for SEC of proteins 289
3. Eluant systems 297
4. Sample preparation 311
5. Column geometry 314
References 314
Chapter 11. Biopolymers. II. Serum lipoproteins 316
1. Introduction 316
2. Analytical methods of serum lipoproteins by HPLC 318
3. Separation of serum lipoproteins by HPLC 323
4. Analysis of serum lipoproteins by HPLC 328
5. Quantitation of lipids in major lipoprotein classes 334
6. Precision and sensitivity of the HPLC method 336
7. Correlation of lipoprotein analysis between HPLC and ultracentrifugation 337
8. Application of the HPLC method 338
9. Conclusion 344
References 344
Chapter 12. Application of SEC/LALLS to biopolymer assemblies 346
1. Introduction 346
2. Low angle laser light scattering 346
3. Outline of low angle laser light scattering 347
4. Arrangement of instruments in SEC/LALLS system 348
5. Application of SEC/LALLS technique to simple proteins 349
6. Application of SEC/LALLS technique to the study of molecular weight of native membrane proteins 352
7. Conclusion 360
Acknowledgements 360
References 361
Part V: Associating Systems 362
Chapter 13. Measurement of equilibrium constants by exclusion methods 364
1. Introduction 364
2. Description of the interacting system 366
3. Static equilibrium methods 368
4. Dynamic methods 375
5. Effects of Donnan equilibrium on binding studies 388
References 390
Chapter 14. Frontal boundary analysis in size exclusion chromatography of self-associating proteins 394
1. Introduction 394
2. Theory of large zone SEC 398
3. Experimental approach 401
4. Specific examples of protein systems studied by large zone SEC 406
5. Conclusions 415
References 415
Chapter 15. Exclusion chromatography of micelles 418
1. Introduction 418
2. Thermodynamics of micellar solution 420
3. Exclusion chromatography of micelles 427
References 455
Subject Index 460
Journal of Chromatography Library (other volumes in the series) 470