Sample Preparation for Hyphenated Analytical Techniques

Sample Preparation for Hyphenated Analytical Techniques

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English
240 Pages

Description

Linking “standard” but often mutually incompatible analytical techniques – so called hyphenation – generally leads to enhanced analytical performance, so hyphenated techniques are widely used in areas where samples are presented in complex matrices, eg environmental, pharmaceutical and biochemical analysis. With these hyphenated techniques, sample preparation is often the most time-consuming step in analysis, particularly where compounds are present in low concentration, and it has a huge influence on the quality of the analytical results. Sample preparation is still not given the importance it deserves, however.

The purpose of this book is to demonstrate the sample preparation chemistry that has enabled the present sensitivity, specificity and high throughput available across a range of hyphenated analytical techniques, and to illustrate the successful utilization of existing sample preparation methodologies in various analytical applications. It identifies the problems in biology, environmental science and pharmaceutical chemistry that require new ideas in chemistry and provides considered opinion on those newer techniques that may address these problems. By dealing with wider issues than is generally found in review papers, this book will provide analytical chemists with insights that are not available by searching the literature for papers on a specific topic.

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Published 12 February 2009
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EAN13 9781405148030
License: All rights reserved
Language English

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Contents
Contributors
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2
3
Introduction: current techniques and issues in sample preparation JACK M. ROSENFELD References
Molecular pathology: applications of genomic analyses to diagnosis of genetic diseases JACK H. LICHY and QI LIANG 2.1 Overview 2.2 Molecular diagnostics: types of assay offered and clinical examples 2.2.1 Properties of nucleic acids relevant to clinical diagnosis 2.2.2 Two clinical examples 2.3 Methods of molecular pathology 2.3.1 Methods for isolation of nucleic acids from body fluids and solid tissues 2.3.2 Methods for amplification 2.3.3 Methods of separation and detection 2.4 More extensive hyphenation of molecular diagnostics assays 2.4.1 Dedicated robotic methods 2.5 Conclusion References
Measurement of oxidative DNA damage by gas chromatographymass spectrometry and liquid chromatographymass spectrometry MIRAL DIZDAROGLU 3.1 Introduction 3.2 Measurement using mass spectrometric techniques 3.3 Gas chromatographymass spectrometry 3.3.1 Materials 3.3.2 Sample preparation 3.3.3 Instrumentation 3.3.4 Measurement 3.4 Liquid chromatographymass spectrometry 3.4.1 Materials 3.4.2 Sample preparation
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10 10 12 14
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3.4.3 Instrumentation 3.4.4 Measurement 3.5 Artifacts Disclaimer References
CONTENTS
Utility of chemical derivatization schemes for peptide mass fingerprinting JONATHAN A. KARTY, TAEYOUNG KIM and JAMES P. REILLY 4.1 Introduction 4.2 Experimental methodology 4.2.1 Band destaining and tryptic digestion prior to derivatizations 4.2.2 Guanidination 4.2.3 Esterification 4.2.4 MALDI 4.2.5 Database searching algorithms 4.2.6 Random spectra generation and statistical analysis 4.3 Results 4.3.1 Identification of a strong gel band 4.3.2 Identification of a weak gel band 4.3.3 More complex gel bands 4.3.4 Homologous proteins 4.3.5 Advantages of esterification 4.3.6 Inadequacy of peptide mass mapping 4.3.7 Sample preparation issues 4.3.8 Correlation between theoretical and experimental data 4.4 Discussion 4.4.1 Overview of peptide mass mapping data 4.4.2 Problems with esterification for routine digest derivatizations 4.4.3 Sources of chemical noise 4.4.4 Multiple proteins in a gel spot 4.4.5 Effect of guanidination and esterification on sensitivity 4.4.6 Peptide ion fragmentation 4.4.7 Comparison of Prodigiesto other mass mapping algorithms 4.5 Conclusions Acknowledgements References
Oligosaccharides HÉLÈNEPERREAULT 5.1 Introduction 5.2 Oligosaccharides from glycoproteins
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55 55 56 56 56 58 58 59 63 66 66 69 71 72 73 74 74
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CONTENTS
5.2.1 General aspects 5.2.2 Glycoprotein purification and molecular mass measurement 5.2.3NGlycosylation 5.2.4OGlycosylation 5.2.5 Quantitative aspects 5.3 Oligosaccharides from other biological sources 5.3.1 General aspects 5.3.2 Milk oligosaccharides 5.3.3 Glycolipids 5.3.4 Oligosaccharides as degradation products 5.4 Methods 5.4.1 General 5.4.2 Gel electrophoretic separations 5.4.3 Capillary electrophoresis separations 5.4.4 Anion exchange chromatography 5.4.5 Affinity chromatography 5.4.6 HPLC and HPLC/ESIMS 5.4.7 Offline HPLC/MALDIMS 5.5 Summary and general considerations Acknowledgements References
Hyphenated techniques in drug discovery: purity assessment, purification, quantitative analysis and metabolite identification JENNY KINGSTON, DESMOND OCONNOR, TIM SPAREY and STEVEN THOMAS 6.1 Challenges associated with sample analysis and purification in support of early drug discovery 6.1.1 Introduction 6.1.2 Analysis in support of discovery chemistry 6.1.3 Purification of reaction mixtures 6.1.4 Conclusion 6.2 Quantitative bioanalysis 6.2.1 Introduction 6.2.2In vitrosamples 6.2.3In vivosamples 6.2.4 LCMS/MS 6.2.5 Mass spectrometer robustness 6.2.6 Traditional sample preparation techniques 6.2.7 Automated preparation ofin vitroandin vivosamples 6.2.8 Conclusion 6.3 Metabolite identification 6.3.1 Introduction 6.3.2 Sample generation
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86 89 92 93 94 94 95 96 99 99 99 100 100 103 103 104 106 106 108 108
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114 114 114 118 123 123 123 124 125 125 126 128 131 137 137 137 139
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6.3.3 Sample purification 6.3.4 Selective analytical 6.4 Conclusion References
CONTENTS
and concentration techniques
Environmental organic analytes MIRAPETROVICandDAMIÀBARCELÓ 7.1 Introduction 7.2 Initial considerations 7.2.1 Sampling of aqueous and solid environmental samples 7.2.2 Sample storage, homogenisation and preservation 7.3 Extraction of target compounds 7.3.1 Extraction from aqueous samples 7.3.2 Extraction from solid matrices samples (soil, sediment and sludge) 7.3.3 Biota 7.4 Extract cleanup 7.5 Sample preparation for specific compound classes 7.5.1 Polychlorinated and polybrominated compounds 7.5.2 Surfactants 7.5.3 Hormones and drugs 7.5.4 Volatile organic compounds (VOCs) 7.6 Conclusions and future trends Acknowledgements References
From cells to instrumental analysis HOSSEIN AHMADZADEH and EDGAR A. ARRIAGA 8.1 Introduction 8.2 Flow cytometry 8.3 Capillary electrophoresis (CE) 8.3.1 Whole single cell analysis 8.3.2 Singlecell injector 8.3.3 Laserinduced fluorescence (LIF) detection 8.4 Single cell gel electrophoresis (comet assay) 8.5 Sampling 8.5.1 Sampling acoustically levitated single cells 8.5.2 High throughput single cell sampling and analysis 8.5.3 Probing the cytoplasm of large cells 8.6 Mass spectrometry for single cells and tissues 8.6.1 Classical hyphenated techniques 8.6.2 Laser capture microdissection 8.7 Direct analysis of subcellular compartments by CE
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158 159 160 160 160 162 163 166 166 167 167
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173 174 176 176 178 179 181 181 181 182 183 185 185 186 187
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Acknowledgements References
CONTENTS
Studies on animal to instrument hyphenation: development of separationbased sensors for near realtime monitoring of drugs and neurotransmitters MALONNE DAVIES, BRYAN HUYNH, SARA LOGAN, KATHLEEN HEPPERT and SUSAN LUNTE 9.1 Introduction 9.2 Obtainingin vivobiological samples by classical techniques: limitations, challenges and other considerations 9.2.1 Animal considerations 9.2.2 Classical methods of blood sampling 9.2.3 Classical methods of tissue sampling 9.3 Modernin vivosampling techniques 9.3.1 Automated blood sampling 9.3.2 Ultrafiltration (UF) 9.3.3 Microdialysis sampling 9.4 Analytical methodologies for online microdialysis sampling 9.4.1 Nonseparationbased sensors 9.4.2 Separationbased sensors 9.5 Conclusions Acknowledgements References
Index
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188 188
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191 192 194 194 195 195 197 198 203 203 205 217 217 217
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