Table Of ContentTECHNIQUES AND INSTRUMENTATION IN ANALYTICAL CHEMISTRY
Volume 1 Evaluation and Optimization of Laboratory Methods and Analytical
Procedures. A Survey of Statistical and Mathemathecal Techniques
by D.L. Massart, A. Dijkstra and L. Kaufman
Volume 2 Handbook of Laboratory Distillation
by E. Krell
Volume 3 Pyrolysis Mass Spectrometry of Recent and Fossil Biomaterials.
Compendium and Atlas
by H.L.C. Meuzelaar, J. Haverkamp and F.D. Hileman
Volume 4 Evaluation of Analytical Methods in Biological Systems
Part A. Analysis of Biogenic Amines
edited by G.B. Baker and R.T. Coutts
Part B. Hazardous Metals in Human Toxicology
edited by A. Vercruysse
Part C. Determination of Beta-Blockers in Biological Material
edited by V. Marko
Volume 5 Atomic Absorption Spectrometry
edited by J.E. Cantle
Volume 6 Analysis of Neuropeptides by Liquid Chromatography and Mass
Spectrometry
by D.M. Desiderio
Volume 7 Electroanalysis. Theory and Applications in Aqueous and Non-Aqueous
Media and in Automated Chemical Control
by E.A.M.F. Dahmen
Volume 8 Nuclear Analytical Techniques in Medicine
edited by R. Cesareo
Volume 9 Automatic Methods of Analysis
by M. Valcarcel and M.D. Luque de Castro
TECHNIQUES AND INSTRUMENTATION IN ANALYTICAL CHEMISTRY — VOLUME 4
EVALUATION
OF ANALYTICAL METHODS
IN BIOLOGICAL SYSTEMS
general editor: R.A. de Zeeuw
PARTC
DETERMINATION OF BETA-
BLOCKERS IN BIOLOGICAL
MATERIAL
edited by
V. Marko
Institute of Experimental Pharmacology, Centre of Physiological Sciences,
Slovak Academy of Sciences, CS-84216 Bratislava, Czechoslovakia
ELSEVIER, Amsterdam — Oxford — New York — Tokyo 1989
ELSEVIER SCIENCE PUBLISHERS B.V.
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ISBN 0-444-87305-8
ISBN 0-444-41744-3 (Series)
© Elsevier Science Publishers B.V., 1989
All rights reserved. No part of this publication may be reproduced, stored in a retrieval system or
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No responsibility is assumed by the Publisher for any injury and/or damage to persons or property
as a matter of products liability, negligence or otherwise, or from any use or operation of any meth
ods, products, instructions or ideas contained in the material herein. Because of rapid advances in
the medical sciences, the Publisher recommends that independent verification of diagnoses and
drug dosages should be made.
Although all advertising material is expected to conform to ethical (medical) standards, inclusion in
this publication does not constitute a guarantee or endorsement of the quality or value of such
product or of the claims made of it by its manufacturer.
Printed in The Netherlands
XI
List of contributors
M. Ahnoff, Bioanalytical Chemistry, AB-Hässle, S-43183 Molndal,
Sweden.
J.G. Barnhill, Division of Clinical Pharmacology, Departments of
Psychiatry and Medicine, Tufts University School of Medicine
and New England Medical Centre, Boston, MA, 02111 USA.
D.J. Greenblatt, Division of Clinical Pharmacology, Departments of
Psychiatry and Medicine, Tufts University School of Medicine
and New England Medical Centre, Boston, MA, 02111 USA.
Z. Kâllay, Institute of Experimental Pharmacology, Centre of Phy
siological Sciences, Slovak Academy of Sciences, CS-84216
Bratislava, Czechoslovakia.
K. Kawashima, Department of Pharmacology, Kyoritsu College of
Pharmacy, 1-5-30 Minato-ku, Tokyo 105, Japan.
V. Marko, Institute of Experimental Pharmacology, Centre of Phy
siological Sciences, Slovak Academy of Sciences, CS-84216
Bratislava, Czechoslovakia.
E. Mutschler, Department of Pharmacology, Faculty of Pharmacy,
Biochemistry and Food Chemistry, University of Frankfurt,
D-6000 Frankfurt/M., FRG.
M.A. Peat, CompuChem Corporation, Research Triangle Park, NC,
27709, USA.
M. Schäfer-Körting, Department of Pharmacology, Faculty of Phar
macy, Biochemistry and Food Chemistry, University of
Frankfurt, D-6000 Frankfurt/M., FRG.
T. Trnovec, Institute of Experimental Pharmacology, Centre of Phy
siological Sciences, Slovak Academy of Sciences, CS-84216
Bratislava, Czechoslovakia.
XII
W.-R. Stenzel, Department of Physiology, Section of Veterinary
Medicine and Animal Production, Humboldt University, DDR-1040
Berlin, GDR.
T. Walle, Department of Cell and Molecular Pharmacology and Expe
rimental Therapeutics, Medical University of South Carolina,
Charleston, SC, 29425 USA.
U.K. Walle, Department of Cell and Molecular Pharmacology and
Experimental Therapeutics, Medical University of South
Carolina, Charleston, SC, 29425 USA.
A. Wellstein, Zentrum der Pharmakologie, Klinikum der J.W. Goethe-
-Universität, D-6000 Frankfurt/M. FRG (Present address:
National Institute of Health, NCI, Bethesda, MA, 2082 USA).
XIII
PREFACE
Evaluation of Analytical Methods in Biological Systems is a
problem-oriented series and the keyword associated with it is
"evaluation". For the evaluation of such a multi-dimensional field
as the determination of about fifty drugs by nine methods (spec-
trophotometry, spectrofluorometry, gas-liquid chromatography,
high-performance liquid chromatography, thin-layer chromato
graphy, radioimmunoassay, enzyme immunoassay, fluoroimmunoassay
and radioreceptor assay), complete information is inevitably nece
ssary. Therefore the main aim of the book is co provide a compre
hensive coverage of the theory and practice of the determination
of beta-blockers in biological material. If this aim has been
achieved, the reader will have available a valuable source of
information for the evaluation of the methods from the point of
view of his or her actual needs - the drug and the analytical
problem. In addition the author(s) of each chapter were requested
to approach the problem very critically in evaluating the possibi
lities of each method for the determination of beta-blockers in
biological material.
The book considers fifty beta-blockers (some more and some
less closely) and nine methods. The methods are divided into three
well known groups: (a) optical methods, (b) Chromatographie me
thods and (c) saturation methods. However, not only methods them
selves are treated. The beginning of almost all determinations is
connected with the pre-analytical manipulation of samples, and
these problems are dealt with at the beginning of the book. The
output of every determination presents information relevant to the
user - the pharmacologist, toxicologist or physician, and the
quality of this information is closely connected with the under
standing of the user s needs. These needs, in turn, should reflect
the possibilities of the analyst and the methods available. For a
better understanding, one chapter is devoted also to the pharmaco
logist and one to the pharmacokineticist.
Recent evidence points to the increased importance of distin
guishing optical isomers of drugs, especially beta-blockers, from
the point of view of their pharmacodynamics in addition to their
pharmacokinetics. In the light of this trend, one chapter on the
XIV
determination of optical isomers of beta-blockers in biological
material is included.
The supplements, which conclude the book, contain a list of
retention indices and a list of structures of beta-blockers.
It is hoped that analysts working on the determination of
beta-blockers, whatever their level of experience, will find some
thing of value in this book.
The Editor gratefully acknowledges the efforts of the contri
butors. He is indebted to Professor Rokus A. de Zeeuw for his
confidence and for the help given during the preparation of the
book and to Dr. M. Koufilovâ, whose contribution to the book was
much broader than the primary task of supervision of the English.
Bratislava Vladimir Marko
September, 1988
1
INTRODUCTION
V. MARKO
Institute of Experimental Pharmacology, Centre of Physiological
Sciences, Slovak Academy of Sciences, 84216 Bratislava (Czechoslo
vakia)
Beta-adrenoceptor antagonists, or more familiar by beta-
-blockers, have undoubtely belonged to the most widely studied
drugs over the past two decades. During this period, several
thousand papers have been written on various aspects of their
chemistry, biochemistry, pharmacology and clinical value. In 1982,
the 20th anniversary of the discovery of the first beta-blocker,
pronethanol, there were 23 beta-blockers on the market, 23 under
clinical trials and 60 under experimental investigation (ref. 1).
Beta-blockers marketed in 1987 are shown in Table 1. Their clini
cal uses range from angina pectoris, hypertension and tachycardia
to the control af migraine, chronic alcoholism, schizophrenia,
essential tremor and the cardiac effects of cocaine overdose, to
mention only a few.
TABLE 1
Recently marketed beta-blockers
Acebutolol Carteolol Pindolol
Alprenolol Indenolol Propranolol
Atenolol Labetalol Sotalol
Befunolol Levobunolol Timolol
Betaxolol Mepindolol
Bisoprolol Metipranolol
Bucumolol Metoprolol
Bufetolol Nadolol
Bunitrolol Nifenalol
Bupranolol Oxprenolol
Carazolol Penbutolol
2
The development of beta-blockers can be traced back to 1905,
when Langley was the first to suggest various inhibitory and
excitatory receptor substances (ref. 2.). In addition, there are
five other years that could be regarded as milestones in the
history of beta-blockers. The first is 1906, when Dale applied
the receptor concept to the adrenergic system (ref. 3). In 1948,
Ahlquist published his classical paper describing the effects of
six sympathetic stimulating compouds on a variety of adrenergic
responses, and thus introduced the alpha- and beta-receptor con
cept (ref. 4). Ten years after Ahlquist*s paper, in 1958 Moran and
Perkins (ref. 5) and Powell and Slater (ref. 6) discovered and
classified dichloroisoprenaline as the first beta-blocker. The
first commercially available beta-adrenoceptor blocking drug was
pronethanol, introduced by Black and Stephenson in 1962 (ref. 7).
The last year in this series is 1964, when pronethanol was re
placed with propranolol and when this drug was found to be effec
tive against angina pectoris, cardiac rhythm disturbances and
hypertension.
The above six milestones may be completed, from the point of
view of this book, by a seventh, i.e. 1965, when the first papers
reporting methods for the determination of beta-blockers appeared.
This occured three years after the discovery of pronethanol, and
these methods concerned pronethanol itself, propranolol (ref. 8)
and toliprolol (ref. 9).
Of the thousands of papers on beta-blockers that have
appeared since that time, about four hundred have dealt with
methods for the determination of these drugs in biological mate
rial. They have described the determination of about 50 beta-
-blockers by nine methods - UV-VIS spectrophotometry, fluorescence
spectrophotometry, gas-liquid chromatography, liquid or high-
-performance liquid chromatography, thin-layer chromatography,
radioimmunoassay, enzyme immunoassay, fluorescence immunoassay and
radioreceptor assay. Similarly to the other drugs (ref. 10), also
in the determination of beta-blockers the greatest attention has
been paid to Chromatographie methods (about three quarters of all
papers) with a prevalence of GLC and HPLC.
The historical development of methods for the determination
of beta-blockers has been closely connected with the development
of the analytical techniques themselves. This is illustrated in
Fig. 1, where this development is expressed as the number of
papers devoted to the different methods for the determination of
3
beta-blockers from 1965 to the mid 1980s. The first studies uti
lized the optical properties of the drugs for their analysis, with
specrophotometry and spectrofluorometry being the main methods.
The 1970s were the years of GLC for beta-blockers and the 1980s
are characterized by a spectacular rise in the high-performance
liquid Chromatographie method.
60 h
I
1 OPTICAL METHODS
H CHR0MAÎ0GRAPHIC METHODS
| SATURATION METH00S
- GLC
-- HPIC
I J I J I i l J ll
1965 66 1967-68 1-70 1971-72 1973 74 1975 76 197778 1979-80 198182 1983 84 1985 86
YEARS
Fig. 1. Historical development of methods for the determination of
beta-blockers expressed as the number of papers devoted to these
methods.
The preceding pages are the best answer to the question - Why
this book?
The second question which could arise is - Why now? The data
in Fig. 1 might best answer this question. The decreasing number
of papers indicates that the development of the methods is almost
finished and no substantial changes are to be expected. The future
will be a time of utilizing the developed methods. Hence now is
the best time to summarize the knowledge on this topic.
