Liquid Chromatography In Clinical Analysis

462 Pages·1981·42.151 MB·English

by  Stephen R. Bakalyar (auth.), Pokar M. Kabra, Laurence J. Marton (eds.)

Liq u di Ch romatog raphy
ni Clinical Analysis
Biological Methods
Liquid Chromatography in clinical Analysis,
edited by Pokar .M Kabra dna Laurence .J
Marten, 1891
Metal Carcinogenesis Testing: Principles
and In Vitro Methods, yb xaM Costa, 0891
diuqiL
Chromatography
nI Clinical Analysis
detidE yb
Pokar M. Kabra
dna
Laurence J. Marton
University of California School of Medicine
San Francisco, California
The Humana Press Inc. • Clifton, New Jersey
Dedication
This volume si dedicated to George Brecher, M.D. for a lifetime of contributions
and devotion to Laboratory Medicine and for having the wisdom to encourage us to
establish our LC laboratory.
Library of Congress Cataloging in Publication Data
Main entry under title:
Liquid chromatography in clinical analyses.
(Biological methods)
Includes bibliographical references and index.
.1 Liquid chromatography. .2 Chemistry, Clinical-Technique. I. Marton,
Laurence J. II. Kabra, Pokar M. III. Series. [DNLM: .1 Chromatography, Liquid.
QD .97
C454 L765]
QP519.9. 55L L54 616.07'5'028 77392-08
ISBN 0-89603-026-1
Crescent Manor
P.O. Box 2148
Clifton, NJ 07015
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted, in
any form or by any means, electronic, mechanical, photocopying, microfilming,
recording, or otherwise, without written permission from the Publisher.
Printed in the United States of America.
ecaferP
Liquid chromatography si widely used in clinical laboratories for
monitoring a variety of therapeutic agents. In addition to its usefulness
in the areas of pharmacokinetics and toxicology, liquid chroma-
tography si currently being developed for the routine analysis of a
number of endogenous constituents. The present book si designed to
serve as a reference for, and stimulus to, scientists involved in patient
care monitoring.
In most instances, the authors review the fundamental concepts
underlying their respective approaches to the use of liquid
chromatography, and continue with detailed presentations of the
specifics of a particular method. This si done so that readers may gain
insight into the potential problems facing them in any application area,
based upon the cumulative experience of individuals who have been
pioneers in the field. The general concepts and approaches described
here change only slowly, and their proper understanding will serve the
biomedical scientist well even as specific methodology changes rapidly.
Liquid yhpargotamorhC ni lacinilC Analysis si an outgrowth of a
course sponsored by the Department of Laboratory Medicine of the
University of California in conjunction with the University's Extended
Programs in Medical Education.
We sincerely thank the contributors to this volume for their
dedication to quality, Mr. William Kerr, an outstanding hospital
administrator, for his willingness to explore new techniques, and our
wives and children for their support and understanding.
San Francisco Pokar M. Kabra
February, 1891 Laurence J. Marton
Contents
Chapter 1
Introduction to Liquid Chromatography
STEPHEN R. BAKALYAR
.I Introduction ........................................ 3
A. Overview of HPLC ............................... 3
B. Basic Facts of the HPLC System ................. 5
.1I Nature of Resolution ............................... 6
A. Zone Separation vs Zone Spreading ..... ........ 6
B. Stationary Phase Selectivity ..................... 9
C. Mobile Phase Selectivity ........................ 10
D. Column Efficiency ............................. 11
II1. Achieving the Separation .......................... 12
A. The Three Factors of Resolution ............ .... 12
B. Retention ...................................... 13
C. Efficiency ...................................... 14
D. Selectivity ..................................... 17
IV. Control and Monitoring Parameters ................ 17
A. Pressure and Flow-Rate ........................ 17
B. Temperature ................................... 18
V. Future Trends ...................................... 18
References ........................................ 19
vii
viii CONTENTS
Chapter 2
Instrumentation for Liquid Chromatography
DRAHCIR A. HENRY DNA GENRIKH YKSVONIROVIS
.I Introduction ....................................... 12
.1I Pumps and Reservoirs ............................. 22
II1. Sample Injectors .................................. 13
IV. Columns .......................................... 34
V. Detectors ......................................... 35
VI. Data Processing ................................... 43
VII. The Liquid Chromatograph as a System ............ 47
References ........................................ 47
Suggested Additional Literature .................... 48
Chapter 3
Liquid Chromatography Column Technology
RONALD E. MAJORS
.I Introduction ....................................... 15
.1I Types and Differences in Packings ................. 52
II1. Techniques for Packing LC Columns ............... 54
IV. Prepacked Columns for HPLC ..................... 55
.V Preparative Columns .............................. 56
VI. Protecting Microparticulate Columns ............... 57
VII. Modes of Liquid Chromatography .................. 59
A. Liquid-Solid (Adsorption) Chromatography (LSC)59
.B Bonded-Phase Chromatography (BPC) .......... 59
C. Ion Exchange Chromatography (IEC) ........... 60
.D Exclusion Chromatography (EC) ................ 60
VIII. Selection of the LC Mode .......................... 60
IX. Selection of Type of Column Packing .............. 63
X. Columns for Bonded-phase Chromatography ....... 63
A. Preparation of Bonded Phases .................. 64
.B Bonded-Phase Coverage and Stability ........... 65
C. Columns for Reverse-Phase Chromatography ... 66
XI. Columns for Adsorption and Normal Bonded-
Phase Chromatography ............................ 76
A. Liquid-Solid (Adsorption) Chromatography (LSC)76
.B Normal Bonded Phases ........................ 78
STNETNOC ix
Xll. Columns for Ion Exchange Chromatography ........ 18
XlII. Columns for Exclusion Chromatography ............ 86
XlV. Future Developments in Columns and Column
Technology ....................................... 89
XV. Conclusions .............................. • ........ 92
Acknowledgment .................................. 92
References ........................................ 92
Part II
Therapeutic Drug Monitoring dna Toxicology
Chapter 4
Why Measure Drug Levels?
SIWEL B. SHEINER
.I Introduction ....................................... 97
.1I Well-Accepted Uses of Drug Levels ................ 97
A. Overdosage .................................... 97
B. Failure of Regimen ............................. 98
II1. A Conceptual Model For Drug Use ................. 99
IV. Drug Levels for Therapeutic Monitoring ........... 100
A. Diagnosing Toxicity or Efficacy ................ 100
B. Rationale for Target Level Strategy ............ 101
C. Sources of Pharmacokinetic Variability ......... 104
D. Use and Misuse of Drug Levels ................ 105
.E Empirical Results of Using Drug Levels for
Therapeutic Monitoring ....................... 108
References ................................... 109
Chapter 5
Anticonvulsants
POKAR M. KABRA, BRIAN E. STAFFORD, DONNA M. MCDONALD,
DNA LAURENCE J. MARTON
.I Introduction ...................................... 111
.1I Collection and Preparation of Samples ............ 115
II1. Chromatography ................................. 117
IV. Detection and Quantitation ....................... 123
x CONTENTS
V. Stability of Columns .............................. 125
VI. Metabolites ...................................... 125
VII. Recent Developments and New Horizons .......... 136
Acknowledgments ................................ 136
References ....................................... 136
Chapter 6
Theophylline and Antiarrhythmics
F. L. VANDEMARK
.I Introduction ...................................... 139
.1I Analysis of Antiasthmatic Drugs ................... 139
A. Sample Pretreatment .......................... 140
B. Chromatography .............................. 143
II1. Antiarrythmics ................................... 147
A. Lidocaine and Procainamide ................... 147
B. Propranolol ................................... 150
C. Quinidine ..................................... 152
D. Disopyramide ................................. 157
IV. Summary ........................................ 157
Acknowledgement ................................ 159
References ....................................... 159
Editor's Note ..................................... 161
retpahC
7
Antibiotics
JOHN P. ANHALT
.I Introduction ...................................... 163
A. Case Histories ................................ 163
B. Need for Specificity ........................... 163
II. Efficient Utilization of Resources .................. 165
A. Reasons to Monitor ........................... 165
B. Mayo Clinic Experience ....................... 167
II1. Current Scope of Liquid Chromatographic Assays . 167
A. /3-Lactam Antimicrobics ....................... 168
B. Aminocyclitol Antimicrobics ................... 170
STNETNOC xi
C. Vancomycin .................................. 178
D. Chloramphenicol .............................. 180
IV. Conclusions ..................................... 183
References ....................................... 183
Chapter 8
Tricyclic Antidepressants
GARY J. SCHMIDT
.I Introduction ...................................... 187
.1I The Tricyclics .................................... 188
II1. Determination of Tricyclics in Physiological
Samples ......................................... 189
A. Sample Pretreatment .......................... 189
B. Chromatography .............................. 192
IV. Use of High pH Mobile Phases .................... 199
V. Determination of Hydroxy Metabolites ............. 203
VI. Conclusions ..................................... 206
Acknowledgments ................................ 208
References ....................................... 208
Chapter 9
Antineoplastic Drugs
GNAGFLOW SAD#E DNA YRSUOY DUOMHAM EL DEYAS
.I Drug Level Monitoring in Cancer Chemotherapy... 112
A. Investigational Clinical Trials .................. 211
B. Routine Therapeutic Applications: Methotrexate 212
.1I Analytical Procedures ............................ 213
A. Review of Liquid Chromatographic Analysis
of Antineoplastic Agents ...................... 213
B. Liquid Chromatographic Analysis of Selected
Drugs ......................................... 216
II1. Trends in Liquid Chromatographic Analysis of
Anti neoplastic Agents ............................ 219
References ....................................... 220