Preparative chromatography /
Preface xv
About the Editors xvii
List of Abbreviations xix
Notation xxiii
1 Introduction 1
Henner Schmidt-Traub and Reinhard Ditz
1.1 Chromatography, Development, and Future Trends 1
1.2 Focus of the Book 4
1.3 Suggestions on How to Read this Book 4
References 6
2 Fundamentals and General Terminology 9
Andreas Seidel-Morgenstern
2.1 Principles and Features of Chromatography 9
2.2 Analysis and Description of Chromatograms 13
2.2.1 Voidage and Porosity 13
2.2.2 Retention Times and Capacity Factors 16
2.2.3 Efficiency of Chromatographic Separations 17
2.2.4 Resolution 20
2.2.5 Pressure Drop 23
2.3 Mass Transfer and Fluid Dynamics 25
2.3.1 Principles of Mass Transfer 25
2.3.2 Fluid Distribution in the Column 27
2.3.3 Packing Nonidealities 28
2.3.4 Extra-Column Effects 29
2.4 Equilibrium Thermodynamics 29
2.4.1 Definition of Isotherms 29
2.4.2 Models of Isotherms 31
2.4.2.1 Single-Component Isotherms 31
2.4.2.2 Multicomponent Isotherms Based on the Langmuir Model 33
2.4.2.3 Competitive Isotherms Based on the Ideal Adsorbed Solution Theory 34
2.4.2.4 Steric Mass Action Isotherms 37
2.4.3 Relation Between Isotherms and Band Shapes 38
2.5 Column Overloading and Operating Modes 44
2.5.1 Overloading Strategies 44
2.5.2 Beyond Isocratic Batch Elution 45
References 46
3 Stationary Phases 49
Michael Schulte
3.1 Survey of Packings and Stationary Phases 49
3.2 Inorganic Sorbents 50
3.2.1 Activated Carbons 50
3.2.2 Synthetic Zeolites 54
3.2.3 Porous Oxides: Silica, Activated Alumina, Titania, Zirconia, and Magnesia 54
3.2.4 Silica 55
3.2.4.1 Surface Chemistry 57
3.2.4.2 Mass Loadability 59
3.2.5 Diatomaceous Earth 59
3.2.6 Reversed Phase Silicas 60
3.2.6.1 Silanization of the Silica Surface 60
3.2.6.2 Silanization 60
3.2.6.3 Starting Silanes 61
3.2.6.4 Parent Porous Silica 61
3.2.6.5 Reaction and Reaction Conditions 62
3.2.6.6 Endcapping 62
3.2.6.7 Chromatographic Characterization of Reversed Phase Silicas 63
3.2.6.8 Chromatographic Performance 63
3.2.6.9 Hydrophobic Properties Retention Factor (Amount of Organic Solvent for Elution), Selectivity 65
3.2.6.10 Shape Selectivity 65
3.2.6.11 Silanol Activity 67
3.2.6.12 Purity 68
3.2.6.13 Improved pH Stability Silica 68
3.2.7 Aluminum Oxide 69
3.2.8 Titanium Dioxide 70
3.2.9 Other Oxides 71
3.2.9.1 Magnesium Oxide 71
3.2.9.2 Zirconium Dioxide 71
3.2.10 Porous Glasses 72
3.3 Cross-Linked Organic Polymers 73
3.3.1 General Aspects 74
3.3.2 Hydrophobic Polymer Stationary Phases 77
3.3.3 Hydrophilic Polymer Stationary Phases 78
3.3.4 Ion Exchange (IEX) 79
3.3.4.1 Optimization of Ion-Exchange Resins 81
3.3.5 Mixed Mode 88
3.3.6 Hydroxyapatite 88
3.3.7 Designed Adsorbents 91
3.3.7.1 Protein A Affinity Sorbents 91
3.3.7.2 Other IgG Receptor Proteins: Protein G and Protein L 96
3.3.7.3 Sorbents for Derivatized/Tagged Compounds: Immobilized Metal Affinity Chromatography (IMAC) 96
3.3.7.4 Other Tag-Based Affinity Sorbents 101
3.3.8 Customized Adsorbents 102
3.3.8.1 Low Molecular Weight Ligands 105
3.3.8.2 Natural Polymers (Proteins, Polynucleotides) 108
3.3.8.3 Artificial Polymers 111
3.4 Advective Chromatographic Materials 111
3.4.1 Adsorptive Membranes and Grafted-Polymer Membranes 114
3.4.2 Adsorptive Nonwovens 115
3.4.3 Fiber/Particle Composites 117
3.4.4 Area-Enhanced Fibers 117
3.4.5 Monolith 118
3.4.6 Chromatographic Materials for Larger Molecules 121
3.5 Chiral Stationary Phases 121
3.5.1 Cellulose- and Amylose-Based CSP 122
3.5.2 Antibiotic CSP 128
3.5.3 Cyclofructan-Based CSP 128
3.5.4 Synthetic Polymers 128
3.5.5 Targeted Selector Design 130
3.5.6 Further Developments 132
3.6 Properties of Packings and Their Relevance to Chromatographic Performance 132
3.6.1 Chemical and Physical Bulk Properties 132
3.6.2 Morphology 133
3.6.3 Particulate Adsorbents: Particle Size and Size Distribution 133
3.6.4 Pore Texture 134
3.6.5 Pore Structural Parameters 137
3.6.6 Comparative Rating of Columns 137
3.7 Sorbent Maintenance and Regeneration 138
3.7.1 Cleaning in Place (CIP) 138
3.7.2 CIP for IEX 140
3.7.3 CIP of Protein A Sorbents 140
3.7.4 Conditioning of Silica Surfaces 143
3.7.5 Sanitization in Place (SIP) 145
3.7.6 Column and Adsorbent Storage 145
References 146
4 Selection of Chromatographic Systems 159
Michael Schulte
4.1 Definition of the Task 164
4.2 Mobile Phases for Liquid Chromatography 167
4.2.1 Stability 168
4.2.2 Safety Concerns 172
4.2.3 Operating Conditions 172
4.2.4 Aqueous Buffer Systems 176
4.3 Adsorbent and Phase Systems 178
4.3.1 Choice of Phase System Dependent on Solubility 178
4.3.2 Improving Loadability for Poor Solubilities 180
4.3.3 Dependency of Solubility on Sample Purity 183
4.3.4 Generic Gradients for Fast Separations 184
4.4 Criteria for Choosing Normal Phase Systems 184
4.4.1 Retention in NP Systems 186
4.4.2 Solvent Strength in Liquid-Solid Chromatography 188
4.4.3 Pilot Technique Thin-Layer Chromatography Using the PRISMA Model 190
4.4.3.1 Step (1): Solvent Strength Adjustment 199
4.4.3.2 Step (2): Optimization of Selectivity 199
4.4.3.3 Step (3): Final Optimization of the Solvent Strength 200
4.4.3.4 Step (4): Determination of the Optimum Mobile Phase Composition 200
4.4.4 Strategy for an Industrial Preparative Chromatography Laboratory 202
4.4.4.1 Standard Gradient Elution Method on Silica 203
4.4.4.2 Simplified Procedure 204
4.5 Criteria for Choosing Reversed Phase Systems 206
4.5.1 Retention and Selectivity in RP Systems 208
4.5.2 Gradient Elution for Small Amounts of Product on RP Columns 212
4.5.3 Rigorous Optimization for Isocratic Runs 213
4.5.4 Rigorous Optimization for Gradient Runs 217
4.5.5 Practical Recommendations 222
4.6 Criteria for Choosing CSP Systems 223
4.6.1 Suitability of Preparative CSP 223
4.6.2 Development of Enantioselectivity 224
4.6.3 Optimization of Separation Conditions 226
4.6.3.1 Determination of Racemate Solubility 226
4.6.3.2 Selection of Elution Order 226
4.6.3.3 Optimization of Mobile/Stationary Phase Composition, Including Temperature 226
4.6.3.4 Determination of Optimum Separation Step 227
4.6.4 Practical Recommendations 227
4.7 Downstream Processing of mAbs Using Protein A and IEX 231
4.8 Size-Exclusion Chromatography (SEC) 236
4.9 Overall Chromatographic System Optimization 237
4.9.1 Conflicts During Optimization of Chromatographic Systems 237
4.9.2 Stationary Phase Gradients 241
References 246
5 Process Concepts 251
Malte Kaspereit and Henner Schmidt-Traub
5.1 Discontinuous Processes 252
5.1.1 Isocratic Operation 252
5.1.2 Gradient Chromatography 253
5.1.3 Closed-Loop Recycling Chromatography 256
5.1.4 Steady-State Recycling Chromatography (SSRC) 258
5.1.5 Flip-Flop Chromatography 259
9783527816330 352781633X 9783527816347 3527816348 9783527816316 3527816313
9783527816330 Wiley
GBC032111 bnb
019727223 Uk
Preparative layer chromatography.
Chromatographic analysis.
Chromatographic analysis.
Preparative layer chromatography.
Electronic books.
QD79.P74
543.8