Hplc: A Practical User's Guide

HPLC HPLC A Practical User’s Guide SECOND EDITION Marvin C. McMaster WILEY-INTERSCIENCE A John Wiley & Sons, Inc., Publication Copyright © 2007 by John Wiley & Sons, Inc. All right reserved. Published by John Wiley & Sons, Inc., Hoboken, New Jersey. Published simultaneously in Canada. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, except as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization through payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750-8400, fax (978) 750-4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, (201) 748-6011, fax (201) 748-6008, or online at http://www.wiley.com/go/permission. Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives or written sales materials. The advice and strategies contained herein may not be suitable for your situation. You should consult with a professional where appropriate. Neither the publisher nor author shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages. For general information on our other products and services or for technical support, please contact our Customer Care Department within the United States at (800) 762-2974, outside the United States at (317) 572-3993 or fax (317) 572-4002. Wiley also publishes its books in a variety of electronic formats. Some content that appears in print may not be available in electronic formats. For more information about Wiley products, visit our web site at www.wiley.com. Library of Congress Cataloging-in-Publication Data: McMaster, Marvin C. HPLC, a practical user’s guide / Marvin C. McMaster. – 2nd ed. p. cm. Includes bibliographical references and index. ISBN-13: 978-0-471-75401-5 (cloth) ISBN-10: 0-471-75401-3 (cloth) 1. High performance liquid chromatography. I. Title. QD79.C454M36 2007 543′.84–dc22 2006040640 Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 CONTENTS PREFACE xi I HPLC PRIMER 1 1 Advantages and Disadvantages of HPLC 1.1 1.2 3 How It Works / 4 1.1.1 A Separation Model of the Column / 5 1.1.2 Basic Hardware: A Quick, First Look / 7 1.1.3 Use of Solvent Gradients / 8 1.1.4 Ranges of Compounds / 9 Other Ways to Make My Separation / 9 1.2.1 FPLC—Fast Protein Liquid Chromatography / 10 1.2.2 LC—Traditional Liquid Chromatography / 10 1.2.3 GLC—Gas Liquid Chromatography / 11 1.2.4 SFC—Supercritical Fluid Chromatography / 11 1.2.5 TLC—Thi