A Practical Handbook of Preparative HPLC

Donald A Wellings1856174662, 9781856174664, 9780080458854

A distillation of over twenty years of the author’s practical experience of this process, this book deliberately steers clear of complex theoretical aspects and concentrates more on the ‘black art’ elements of the techniques involved. As a result is a book for frequent use in the laboratory, especially of interest to those in the pharmaceutical, agrochemical and biotechnology industries and to anyone involved in the analysis and purification of drugs, small molecules, proteins and DNA.

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Table of contents :
Cover Page……Page 1
Don Wellings……Page 3
Title Page: A Practical Handbook of Preparative HPLC……Page 4
ISBN 1856174662……Page 5
Contents (with page links)……Page 6
Preface……Page 8
Foreword……Page 10
Abbreviations……Page 12
1 The history and development of preparative HPLC……Page 14
A Colourful origin!……Page 16
The saviour of many a frustrated chemist!……Page 17
Confused?……Page 27
2 Fluid dynamics, mass transport and friction……Page 30
Food for thought!……Page 32
Why plates?……Page 33
Picture this?……Page 36
Did you know?……Page 37
Think of the bigger picture!……Page 39
3 Modes of chromatographic separation……Page 42
The most commonly utilized modes………Page 44
Particle size……Page 45
Remember the forest?……Page 46
3.1 Normal Phase Chromatography……Page 48
A sophisticated and complex Ball Mill!……Page 49
3.2 Reversed Phase Chromatography……Page 50
3.3 Chiral Separations……Page 54
Chirality……Page 55
Optical purity……Page 56
Separation of enantiomers……Page 57
3.4 Ion Exchange Chromatography……Page 60
Invest in a conductivity meter!……Page 63
3.5 Exclusion Chromatography……Page 66
3.6 Affinity Chromatography……Page 67
Have you realized?……Page 69
4 How to get started……Page 70
4.1 Packing a Column……Page 72
Slurry concentration……Page 76
Slurry and column preparation……Page 77
Column packing protocol for column type 2 (Figure 4.4)……Page 78
4.2 What and Where!……Page 79
4.3 Product Recovery……Page 82
Ready packaged!……Page 84
4.4 Productivity……Page 85
A sledgehammer to crack a nut!……Page 88
5 Process development and optimization……Page 90
5.1 Sample Self Displacement for Purification of a Peptide……Page 92
Analytical HPLC……Page 103
Gradient selection……Page 104
Loading study……Page 105
Flow rate optimization……Page 106
5.2 Boxcar Injections for Chiral Separations……Page 108
6 Documentation and record keeping……Page 114
6.1 Equipment Qualification……Page 116
6.2 Process Documentation……Page 118
Does the development have to be cGMP compliant?……Page 119
Quality checks on consumables!……Page 121
Don’t lose a sheet!……Page 122
Appendices……Page 124
PROCESS INSTRUCTIONS……Page 126
CONTENTS……Page 128
DEVIATION SHEET……Page 130
1.03 Raw Materials……Page 132
1.04 Equipment……Page 134
Analytical HPLC Strength Method……Page 136
1.07 Ancillary SOPs……Page 138
1.08 Cleaning……Page 140
1.09 Personnel and Responsibilities……Page 142
1.10 Safety……Page 144
Safety Precautions……Page 145
OPERATION 2 PRE-PURIFICATION COLUMN PREPARATION……Page 148
OPERATION 3 PURIFICATION……Page 150
PURIFICATION DEVIATION SHEET……Page 158
EXAMPLE OF FRACTION RECONCILIATION DOCUMENT……Page 162
OPERATION 1 SUMMARY……Page 164
References……Page 172
CHAPTER 1……Page 174
CHAPTER 3……Page 176
CHAPTER 4……Page 178
CHAPTER 6……Page 179
Index (with page links)……Page 180
B……Page 182
C……Page 183
D……Page 184
E……Page 185
G……Page 186
I……Page 187
M……Page 188
P……Page 189
S……Page 191
V……Page 193