LIQUID CHROMATOGRAPHY– MASS …

1 LIQUID chromatography mass spectrometry :AN INTRODUCTIONL iquidChromatography ArdreyCopyright 2003 JohnWiley & Sons, :0-471-49799-1(HB);0-471-49801-7(PB)Anal ytical Techniques in the Sciences (AnTS)Series Editor: David J. Ando, Consultant, Dartford, Kent, UKA series of open learning/distance learning books which covers all of the majoranalytical techniques and their application in the most important areas of physical,life and materials Available in the SeriesAnalytical Instrumentation: Performance Characteristics and QualityGraham Currell, University of the West of England, Bristol, UKFundamentals of Electroanalytical ChemistryPaul Monk, Manchester Metropolitan University, Manchester, UKIntroduction to Environmental AnalysisRoger N.

2 Reeve, University of Sunderland, UKPolymer AnalysisBarbara H. Stuart, University of Technology, Sydney, AustraliaChemical Sensors and BiosensorsBrian R. Eggins, University of Ulster at Jordanstown, Northern Ireland, UKMethods for Environmental Trace AnalysisJohn R. Dean, Northumbria University, Newcastle, UKLiquid chromatography mass spectrometry : An IntroductionRobert E. Ardrey, University of Huddersfield, Huddersfield, UKForthcoming TitlesAnalysis of Controlled SubstancesMichael D. Cole, Anglia Polytechnic University, Cambridge, UKLIQUID chromatography mass spectrometry :AN INTRODUCTIONR obert E.

3 ArdreyUniversity of Huddersfield, Huddersfield, UKCopyright 2003 John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester,West Sussex PO19 8SQ, EnglandTelephone (+44) 1243 779777 Email (for orders and customer service enquiries): our Home Page on or Rights Reserved. No part of this publication may be reproduced, stored in a retrieval system ortransmitted in any form or by any means, electronic, mechanical, photocopying, recording,scanning or otherwise, except under the terms of the Copyright, Designs and Patents Act 1988 orunder the terms of a licence issued by the Copyright Licensing Agency Ltd, 90 Tottenham CourtRoad, London W1T 4LP, UK, without the permissionin writing of the Publisher.

4 Requests to thePublisher should be addressed to the Permissions Department, John Wiley & Sons Ltd, TheAtrium, Southern Gate, Chichester, WestSussex PO19 8SQ, England, or emailed or faxed to (+44) 1243 publication is designed to provide accurateand authoritative information in regard to thesubject matter covered. It is sold on the understanding that the Publisher is not engaged inrendering professional services. If professional advice or other expert assistance is required, theservices of a competent professional should be Wiley Editorial OfficesJohn Wiley & Sons Inc.

5 , 111 River Street, Hoboken, NJ 07030, USAJ ossey-Bass, 989 Market Street, San Francisco, CA 94103-1741, USAW iley-VCH Verlag GmbH, Boschstr. 12, D-69469 Weinheim, GermanyJohn Wiley & Sons Australia Ltd, 33 Park Road, Milton, Queensland 4064, AustraliaJohn Wiley & Sons (Asia) Pte Ltd, 2 Clementi Loop #02-01, Jin Xing Distripark, Singapore 129809 John Wiley & Sons Canada Ltd, 22 Worcester Road, Etobicoke, Ontario, Canada M9W 1L1 Wiley also publishes its books in a variety of electronic formats. Some content that appearsin print may not be available in electronic of Congress Cataloging-in-Publication DataArdrey, chromatography - mass spectrometry : an introduction / Robert E.

6 Cm. (Analytical techniques in the sciences)Includes bibliographical references and 0-471-49799-1 (cloth : alk. paper) ISBN 0-471-49801-7 (pbk. : alk. paper)1. LIQUID chromatography . 2. mass spectrometry . I. Title. II. A73 2003543 .0894 dc212002028084 British Library Cataloguing in Publication DataA catalogue record for this book is available from the British LibraryISBN 0-471-49799-1 (Cloth)ISBN 0-471-49801-7 (Paper)Typeset in 10/12pt Times by Laserwords Private Limited, Chennai, IndiaPrinted and bound in Great Britain by Antony Rowe Ltd, Chippenham, WiltshireThis book is printed on acid-free paper responsibly manufactured from sustainable forestryin which at least two trees are planted for each one used for paper PrefaceixPrefacexiAcknowledgementsxiiiAb breviations, Acronyms and SymbolsxvAbout the Authorxix1 What are the Advantages of Linking High PerformanceLiquid chromatography with mass spectrometry ?

7 What Capabilities are Required of the Combination? What Problems, if Any, Have to be Addressed to Allowthe LC MS Combination to Function, and Function Effectively?4 References52 LIQUID High Performance LIQUID Sample introduction (Injector) Mobile Stationary Chromatographic Identification Using High Performance LIQUID Chromatography21viLiquid chromatography mass Quantitation Using High Performance LIQUID The Need for High Performance LIQUID chromatography mass Spectrometry30 References313 mass Ionization Electron Chemical Fast-Atom Matrix-Assisted Laser Desorption Negative Ion The Quadrupole mass The (Quadrupole)

8 Ion-Trap mass The Double-Focusing and Tri-Sector mass The Time-of-Flight mass Tandem mass spectrometry (MS MS) Data Processing of mass Spectral The Total-Ion-Current Qualitative Quantitative The Use of Tandem mass Spectrometry71 References734 Interface The Moving-Belt The Direct- LIQUID - introduction The Continuous-Flow/Frit(Dynamic) Fast-Atom-Bombardment The Particle-Beam The Thermospray The Electrospray The Mechanism of Electrospray Sample The Appearance of the Electrospray Structural Informationfrom Electrospray The Atmospheric-Pressure Chemical Ionization The Mechanism of Atmospheric-Pressure ChemicalIonization123 References1265 Applications of High Performance LIQUID chromatography mass Method The Use of Experimental Design for The Choice of Electrospray or The Molecular Weight Determination of

9 Electrospray Spectra of Co-Eluting The Use of Selected-Ion Monitoring to Examinethe Number of Terminal Galactose Moieties on The Effect of Mobile-Phase Additives Structure Determination of Amino Acid Sequencing of The Use of Enzymes for Amino Acid The mass Spectral Fragmentation of Confirmation of Amino Acid Sequence Usingthe Analysis of LC MS Data from an EnzymeDigest of a Determination of the Amino Acid Sequence of a NovelProtein Using LC MS Data from an Enzyme Amino Acid Sequencing of PolypeptidesGenerated by Enzyme Digestion Using MS The Location of Post-Translational ModificationsUsing LC MS Data from an Enzyme The Location of Post-Translational ModificationsUsing MS The Analysis of Polysaccharides Presentin Glycosylated Location of the Position of Attachment ofa Glycan on the Polypeptide Backbone of a Molecular Weight Determination of Small (<1000 Da)

10 The Use of Fast-LC MS in Combinatorial Chemistry185viiiLiquid chromatography mass Structure Determination of Method Development for Structural The Use of Target-Compound Analysis andLC MS MS for the Identification of Drug The Use of High-Accuracy mass Measurementsin Combination with LC MS for the StructureDetermination of Drug The Use of Cone-VoltageFragmentation in Conjunctionwith High-Accuracy mass Measurements and LC MSfor Metabolite The Use of LC MSnfor the Identificationof Drug Requirements of a Quantitative MethodInvolving LC Quantitative Matrix Effects in LC The Method of Standard Additions to OvercomeMatrix The Quantitative Determination of DNAO xidation The Use of MS MS for Quantitative Determinations224 References233 Responses to Self-Assessment Questions235 Bibliography241 Glossary of Terms247SI Units and Physical Constants257 Periodic Table261 Index263 Series PrefaceThere has been a rapid expansion in the provision of further education in recentyears.