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Principles and Practice of Bioanalysis

Name: Principles and Practice of Bioanalysis
Brand: Taylor & Francis Ltd
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By: Richard F. Venn (Editor)

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Paperback | 22 June 2000 | Edition Number 2

At a Glance

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382 Pages

Edition Type
Revised

Dimensions(cm)
25.4 x 17.8 x 1.91

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Published: 25th February 2008

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Principles and Practice of Bioanalysis provides a guide to the methods available and the techniques currently used in this field. It provides up to the minute information and guidance on the methods and strategy used in developing and running ultra-trace analyses for drugs, metabolites and other substances. The authors writes in an informal and didactic style, offering a logical path through the problems of small molecule (bio)analysis and enables readers to choose appropriate methods of analysis for their needs. Principles and Practice of Bioanalysis provides an overview of analytical methods for analytical scientists within the pharmaceutical industry, research and development, the agrochemical industry, and scientists in the health service, biology and biochemistry. It also gives postgraduate students a useful reference for their research methods.

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Non-Fiction Science Biology, Life Sciences Biochemistry Medicine Nursing & Ancillary Services Pharmacy & Dispensing Pre-Clinical Medicine & Basic Sciences Other Branches of Medicine Pharmacology Engineering & Technology Biochemical Engineering

Biotechnology

List of contributors	p. xvi
Preface	p. xvii
Physico-chemical properties of drugs and metabolites and their extraction from biological material	p. 1
Introduction	p. 1
Metabolite isolation	p. 1
Bioanalysis	p. 1
Enrichment of drugs and metabolites	p. 1
Differences between metabolite isolation and drug analysis	p. 2
Physico-chemical properties of drugs and solvents	p. 2
Energy changes on solution	p. 2
Molecular phenomena behind solubility/miscibility	p. 3
Water miscibility and water immiscibility	p. 8
Partition	p. 9
Extraction efficiency	p. 9
Ionisation and its effect on the extraction of drugs	p. 11
Ionisation, pH and pK	p. 11
Titration curves	p. 12
Henderson-Hasselbach equation	p. 13
Buffers	p. 16
Distribution coefficient	p. 17
Solvent extraction	p. 18
Choice of solvent	p. 18
Mixed solvents	p. 20
Dealing with plasma proteins and emulsions	p. 21
Choice of pH for solvent extraction	p. 21
Artefacts arising during the extraction of drugs and metabolites	p. 21
Modification and derivatisation of drugs and metabolites	p. 24
Ion-pair extraction	p. 26
Recoveries	p. 26
The 'first law of drug metabolism'	p. 26
Bibliography	p. 27
Solid-phase extraction	p. 28
Introduction	p. 28
General properties of bonded silica sorbents	p. 30
Sorbent/analyte interactions	p. 30
Solvation	p. 30
Non-polar	p. 31
Polar	p. 32
Ion exchange	p. 32
Covalent	p. 33
Mixed-mode interactions	p. 34
Polymeric sorbents	p. 35
Miscellaneous	p. 36
Sample pretreatment of different biological matrices	p. 36
Liquid samples	p. 36
Protein binding	p. 36
Solid samples	p. 37
Developing SPE methods	p. 37
Example of an SPE method	p. 38
Disc cartridges	p. 38
Potential advantages	p. 39
Disadvantages	p. 40
96-Well format (e.g. Porvair Microsep system)	p. 40
Direct injection of plasma	p. 41
Other new developments	p. 41
Fines	p. 41
A cartridge in a pipette tip?	p. 41
Conclusions and future perspectives	p. 42
Bibliography	p. 42
Basic HPLC theory and practice	p. 44
Origins	p. 44
Applications	p. 44
Apparatus	p. 45
Column	p. 45
Plumbing	p. 47
Pumps	p. 48
Injectors	p. 49
Column ovens	p. 50
Detectors	p. 50
The chromatographic process	p. 50
Basic principles	p. 50
Molecular forces	p. 51
Distribution	p. 51
Theoretical plates	p. 52
The chromatogram	p. 54
Retention	p. 54
Resolution	p. 54
Peak shape	p. 58
Effect of temperature	p. 61
Effect of flow rate and linear velocity	p. 62
Effect of sample volume	p. 64
Separation mode	p. 64
Normal phase	p. 64
Reverse phase	p. 65
Gradient reverse phase	p. 68
Ion suppression and ion pairing	p. 69
Ion exchange	p. 72
Others	p. 72
Column care	p. 73
Bibliography	p. 74
HPLC optimisation	p. 75
Objective	p. 75
System parameters	p. 75
Reverse-phase HPLC	p. 76
Ion-pair HPLC	p. 82
Ion-exchange HPLC	p. 85
Normal-phase HPLC	p. 87
Chiral HPLC	p. 90
Chiral columns	p. 91
Diastereoisomers	p. 94
Chiral complexing agents	p. 96
Chiral summary	p. 96
Column switching in HPLC	p. 97
Gradient reverse-phase HPLC	p. 100
Column conditions	p. 101
Computerised optimisation of HPLC	p. 103
Conclusions	p. 104
Glossary	p. 104
References	p. 105
HPLC detectors	p. 106
Introduction	p. 106
Principles of detection	p. 107
Solute-property detectors	p. 107
Bulk-property detectors	p. 108
Selectivity in detectors	p. 108
Detector response	p. 109
Linearity	p. 109
Time constant	p. 110
Detector types	p. 111
UV-visible detectors	p. 111
Fluorescence detectors	p. 115
Electrochemical detectors	p. 120
Multifunctional detectors	p. 122
Radiochemical detectors	p. 123
Other detectors	p. 124
Sensitivity considerations	p. 126
Irradiation	p. 126
Pre-column derivatisation	p. 126
Post-column derivatisation	p. 127
Selectivity	p. 127
Detector problems	p. 128
Noise due to bubbles	p. 128
Spurious peaks	p. 128
Baseline instability	p. 128
Appendix	p. 129
Buying a detector	p. 129
Which detector to use?	p. 129
Bibliography	p. 129
Gas chromatography: what it is and how we use it	p. 131
Why gas chromatography works	p. 131
Factors that affect the chromatography	p. 132
Choices in GC	p. 133
Stationary phase	p. 133
Mobile phase	p. 135
Column length	p. 136
Column diameter	p. 136
Film thickness	p. 136
Flow rate	p. 137
Temperature	p. 137
Some rules of thumb	p. 138
GC hardware	p. 138
Pneumatics	p. 139
Sample introduction	p. 140
Detectors	p. 145
Derivatisation for GC	p. 147
A GC strategy for bioanalysis	p. 148
Bibliography	p. 148
Thin-layer chromatography	p. 149
Introduction	p. 149
Uses of TLC	p. 150
Preparative TLC	p. 150
Metabolic profiling	p. 151
'Rules of thumb'	p. 155
Some recommended solvent systems	p. 158
Detection of compounds on TLC plates	p. 158
Bibliography	p. 159
Capillary electrophoresis: an introduction	p. 160
Introduction	p. 160
How capillary electrophoresis works	p. 160
Why capillary electrophoresis works	p. 162
Electro-osomotic flow	p. 162
Free-solution capillary electrophoresis	p. 162
Micellar electrokinetic capillary chromatography	p. 164
Electrochromatography (electrically driven HPLC)	p. 165
CE hardware	p. 166
The capillary	p. 166
Sample introduction	p. 166
Detectors in CE	p. 168
Sensitivity in CE	p. 169
Use in bioanalysis	p. 169
Bibliography	p. 170
Immunoassay techniques	p. 171
Introduction	p. 171
Definitions	p. 171
Theory	p. 172
Mass action	p. 172
Competitive assays	p. 173
Non-competitive assays	p. 174
Requirements for immunoassay	p. 175
Antibody	p. 175
Label	p. 175
Separation	p. 177
Practical aspects	p. 178
Preparation of hapten-carrier protein conjugates	p. 178
Immunisation	p. 179
Antibody detection	p. 180
Antibody titres	p. 180
Calibration curves	p. 180
Matrix effects	p. 182
Data handling	p. 182
Standard curves	p. 182
Fitting	p. 182
Precision profile	p. 182
Advantages of immunoassay	p. 184
Sensitivity	p. 184
Throughput	p. 184
Selectivity	p. 184
Ease	p. 184
Automation	p. 184
Disadvantages of immunoassays	p. 185
Time: how long does it take?	p. 185
Selectivity	p. 185
Matrix effects	p. 185
What can go wrong?	p. 185
Matrix effects	p. 185
Concentration effects	p. 187
Immunoassay strategy	p. 187
Example	p. 187
Sampatrilat	p. 187
Affinity chromatography	p. 187
Immobilisation techniques and media	p. 190
Elution techniques	p. 191
Re-use/reconditioning	p. 192
The interface between affinity chromatography and analysis	p. 193
The future	p. 193
Phage libraries for antibodies	p. 193
Monoclonal antibodies	p. 193
Molecular imprinting	p. 193
Non-competitive assays for small molecules	p. 194
Use of low-specificity immunoassay for discovery compounds	p. 194
Indwelling optical fibre probes	p. 194
Summary	p. 194
Bibliography	p. 194
Automation of sample preparation	p. 196
Introduction	p. 196
Approaches to automation	p. 197
SPE	p. 197
Protein precipitation methods	p. 198
Multi-well plate technology	p. 198
Liquid-handling procedures	p. 198
Avoiding evaporation	p. 199
Simple automation	p. 199
Column switching	p. 200
Prospekt and Merck OSP-2	p. 202
Benchtop instruments - sequential sample processing	p. 202
Zymark BenchMate	p. 203
Gilson ASPEC XL	p. 203
Hamilton MicroLab	p. 204
Benchtop instruments - parallel sample processing	p. 205
Zymark RapidTrace	p. 205
Gilson ASPEC 4	p. 205
Multiple probe liquid-handling robots	p. 205
Gilson ASTED	p. 206
Full robotic systems	p. 207
When to automate?	p. 207
Example methods	p. 208
Conclusions and future perspectives	p. 208
Bibliography	p. 209
Fundamental aspects of mass spectrometry: overview of terminology	p. 211
Introduction	p. 211
Inlets	p. 211
Septum inlet	p. 211
Direct probe inlet	p. 212
GC inlets	p. 212
LC inlets	p. 213
Ion sources	p. 216
Introduction	p. 216
Electron impact ionisation	p. 216
Chemical ionisation	p. 218
Atmospheric-pressure chemical ionisation	p. 219
Fast atom bombardment	p. 220
Thermospray	p. 221
Electrospray	p. 223
Other desorption techniques	p. 225
Analysers	p. 226
Single-focusing magnetic instruments	p. 226
Double-focusing instruments	p. 228
Quadrupole analysers	p. 229
Time of flight (ToF) analysers	p. 231
Ion-trap mass analysers	p. 231
Detectors	p. 233
Electron multipliers	p. 233
Negative-ion detection	p. 234
Data acquisition and processing	p. 234
Instrument control	p. 234
Data acquisition/preliminary data processing	p. 234
Secondary data processing/data presentation	p. 235
Bibliography	p. 239
Applications of mass spectrometry: quantitative mass spectrometry	p. 240
Quantification	p. 240
Gas chromatography-mass spectrometry (GC-MS)	p. 240
Liquid chromatography-mass spectrometry (LC-MS)	p. 241
Quantitative API LC-MS and its contribution to the drug development process	p. 241
Internal standardisation	p. 242
Data acquisition	p. 243
Selected ion versus mass chromatogram	p. 243
Mass analysis	p. 243
Calculation of the mass of the selected ion	p. 244
Data storage and processing	p. 245
Developing a quantitative method	p. 245
Analysis of prostanoids by GC-MS	p. 246
An example of thermospray LC-MS	p. 249
Examples of API LC-MS	p. 250
The future	p. 253
Bibliography	p. 254
Mass spectrometric identification of metabolites	p. 255
Objectives	p. 255
Introduction	p. 255
Tandem mass spectrometry (MS-MS)	p. 256
Theory	p. 256
Instrumentation	p. 256
MS-MS scans and their application to metabolite identification	p. 258
Isotopically labelled compounds in metabolite identification	p. 265
Practical aspects for the identification of metabolites by mass spectrometry	p. 266
Introduction	p. 266
Electron impact ionisation and chemical ionisation	p. 268
Fast atom bombardment	p. 270
Thermospray LC-MS	p. 271
Electrospray LC-MS	p. 273
Ion-trap mass spectrometry coupled to external atmospheric-pressure ionisation sources	p. 274
Summary	p. 275
Overall comments	p. 276
Bibliography	p. 277
Nuclear magnetic resonance in drug metabolism	p. 278
Introduction	p. 278
Basic theory of the NMR phenomenon	p. 278
Parameters of the NMR spectrum	p. 280
Chemical shift	p. 280
Spin-spin coupling	p. 281
Intensity	p. 286
Practical considerations	p. 286
Types of spectrometer	p. 286
Sample preparation	p. 287
NMR applications in drug development	p. 288
No sample preparation	p. 288
Solid-phase extraction sample preparation	p. 288
HPLC fractions	p. 291
Fluorinated compounds	p. 291
Stable isotope labelling	p. 293
Plasma metabolites	p. 294
Biochemical changes	p. 294
Summary	p. 294
Appendix: fourier transform and some multi-pulse techniques	p. 294
Why use pulse NMR?	p. 294
The pulse	p. 296
Time and frequency	p. 296
Multi-pulse experiments	p. 296
Conclusion	p. 301
Bibliography	p. 301
Strategy in metabolite isolation and identification	p. 302
Stage 1: radiochemical synthesis	p. 302
Choice of label	p. 302
Position of [superscript 14]C label	p. 303
Stage 2: animal experiments	p. 303
Routes of excretion	p. 304
Formulation and route of administration	p. 304
Collection of urine and bile	p. 304
Stage 3: metabolite isolation and characterisation	p. 304
Enrichment	p. 304
Analysis	p. 306
Separation	p. 307
Purification	p. 315
Characterisation	p. 315
Stage 4: identification of metabolites	p. 321
Mass spectrometry	p. 323
NMR	p. 325
Degradation, derivatisation and comparison with authentic material	p. 327
Ambiguities	p. 330
Stage 5: quantitative aspects of metabolism	p. 330
Quantification of excretion balance studies	p. 330
Quantitative aspects of metabolite isolation	p. 331
Quantitative measurement of metabolic profiles	p. 331
In vitro studies	p. 333
Isolation of metabolites from in vitro incubations	p. 334
Cross-species comparisons of metabolic profiles	p. 336
Mechanistic studies	p. 337
Identification of plasma metabolities	p. 337
Good laboratory practice	p. 339
Conclusions	p. 341
Strategy for the development of quantitative analytical procedures	p. 342
Introduction	p. 342
Preliminary requirements	p. 343
Detection	p. 345
Separation	p. 348
Sample preparation	p. 349
Solid-phase extraction	p. 349
Extraction sequence	p. 350
Liquid/liquid extraction	p. 352
Quantification	p. 354
Rule of one and two	p. 354
Standardisation	p. 354
Peak height and area	p. 355
Calibration check	p. 355
Validation	p. 356
Support work	p. 356
Matrix substitution	p. 356
Stability	p. 357
Metabolites	p. 358
Conclusions	p. 358
Index	p. 359
Table of Contents provided by Syndetics. All Rights Reserved.

Principles and Practice of Bioanalysis

At a Glance

New Edition

Paperback

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How to return your order

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