Table Of ContentTECHNIQUES AND INSTRUMENTATION IN ANALYTICAL CHEMISTRY
Volume 1 Evaluation and Optimization of Laboratory Methods and Analytical
Procedures. A Survey of Statistical and Mathematical Techniques
by D.L. Massart, A. Dijkstra and L. Kaufman
Volume 2 Handbook of Laboratory Distillation
by E. Krell
Volume 3 Pyrolysis Maas 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
a
EVALU A T1O N
OF ANALYTICAL METHODS
IN BIOLOGICAL SYSTEMS
general editor: R.A. de Zeeuw
PART A
ANALYSIS
OF BIOGENIC AMINES
edited by
Glen 6. Baker and Ronald T. Coutts
Neurochemical Research Unit, Department of Aychiatry and Faculty of Pharmacy and
Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada
E LSEVlE R SClENTlF IC PUBLISHING COMPANY
- -
Amsterdam Oxford New York 1982
ELSEVIER SCIENTIFIC PUBLISHING COMPANY
Molenwerf 1
P.O. Box 21 1,1000 AE Amsterdam, The Netherlands
Distributors for the United States and Caneda:
ELSEVIER SCIENCE PUBLISHING COMPANY INC.
52, Vanderbilt Avenue
New York, NY 10017
Library of Congress Cataloging in Publication Data
Main entry under title:
Evaluation of analytical methods in biological systems.
(Techniques and instrumentation in analytical chemistry
chemistry ; v. 4)
Bibliography: p.
Includes index.
Contents: pt. A. Analysis of biogenic mines.
1. !nines--Analysis. 2. Amines in the body.
3. Biological chemistry--Technique. I. Baker, Glen B.,
.
1947- 11. Coutts, Ronald Thomson. 111. Series.
QP8Ol .A48E86 1982 574.19'285 82-11553
ISBN 0-444-42110-6 (v. 1)
ISBN 04444211 06 (VOl. 4)
ISBN 044441744-3 (Series)
0 Elsevier Scientific Publishing Company, 1982
All rights reserved. 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 or other-
wise, without the prior written permission of the publisher, Elsevier Scientific Publishing Company,
lo00 AH Amsterdam, The Netherlands
Printed in The Netherlands
XI11
Contributors
Baker, Glen B. Ferkany, John W.
Neurochemical Research Unit Departments of Neuroscience,
Department of Psychiatry Pharmacology and Psychiatry
University of Alberta Johns Hopkins University School
Edmonton, Alberta T6G 263, Canada of Medicine
Baltimore, Maryland 21205, USA
Baker, Judith M.
Neurochemical Research Unit Gelpi, Emilio
Department of Psychiatry Analytical Neurochemistry Unit
University of Alberta Instituto de Quimica Bio-Organica
Edmonton, Alberta T6G 263, Canada Consejo Superior de Investigaciones
Cie ntif ic a s
Candy, John M.
Jorge Girona Salgado S/N
MRC Neuroendocrinology Unit
Barcelona-34, Spain
Newcastle General Hospital
Newcastle-upon-Tyne NE4 6BE, U.K. Godse, Damodar D.
Clarke Institute of Psychiatry
Chiu, Andrew S.
University of Toronto
Clarke Institute of Psychiatry
Toronto, Ontario M5T 1R8, Canada
University of Toronto
Toronto, Ontario M5T 1R8, Canada Hubbard, John W.
Faculty of Pharmacy
Coutts, Ronald T.
University of Manitoba
Neurochemical Research Unit
Winnipeg, Manitoba R3T 2N2, Canada
Faculty of Pharmacy and
Pharmaceutical Sciences LeGatt, Donald F.
University of Alberta Neurochemical Research Unit
Edmonton, Alberta T6G 2N8, Canada Department of Psychiatry and
Faculty of Pharmacy and
Davis Bruce A.
Pharmaceutical Sciences
Psychiatric Research Division
University of A1 berta
University Hospital
Edmonton, Alberta T6G 2G3, Canada
Saskatoon, Saskatchewan S7N 0x0, Canada
Locock, R. Anthony
Dayton, Mark A.
Faculty of Pharmacy and
Medical Sciences Program
Pharmaceutical Sciences
Myers Hall
University of A1 berta
Indiana University
Edmonton, Alberta T6G 2N8, Canada
Bloomington, Indiana 47405, USA
Martin, Ian L.
Dewhurst, William G.
MRC Neurochemical Pharmacology Unit
Neurochemical Research Unit
Medical Research Council Centre
Department of Psychiatry
Medical School
University of Alberta
Cambridge CB2 2QD, U.K.
Edmonton, Alberta T6G 2G3, Canada
Midha, Kamal K.
Dryden, William F.
Col 1e ge of Pharmacy
Department of Pharmacology
University of Saskatchewan
University of Alberta
Saskatoon, Saskatchewan S7N OWO, Canada
Edmonton, Alberta T6G 2E1, Canada
Warsh, Jerry J.
Durden, David A.
' Clarke Institute of Psychiatry
Psychiatric Research Division
University of Toronto
University Hospital
Toronto, Ontario M5T 1R8, Canada
Saskatoon, Saskatchewan S7N 0x0, Canada
Wightman, R. Mark
Enna, S.J.
Department of Chemistry
Departments of Pharmacology and of
Indiana University
Neurobiology and Anatomy
Bloomington, Indiana 47405, USA
University of Texas Medical School
Houston, Texas 77025, USA
XV
PREFACE
Analytical chemistry has grown spectacularly over the last two decades.
As a result, it has become very difficult, if not impossible for many
scientists to keep abreast of the many developments, innovations and new
horizons. This does not only hold for specialists involved in the basic
aspects, but perhaps even more so for the non-specialists who wish to
apply analytical chemistry as a tool to provide answers to the problems
that they face in the course of their daily work, particularly in the area
of biomedicine and other life sciences.
A1 though the present analytical literature contains various excel lent
monographs and review series, appliers of analytical chemistry to biological
specimens often find that they focus too much on individual analytical
techniques and on their basic aspects, with little or no comparison of the
potentials and limitations of a particular technique as compared to others.
This leaves a need for a series which will evaluate the various analytical
techniques and approaches that can be used in a particular bioanalytical
field, taking into account the type of the problem, the type of the answer
needed and the impact of the biological matrix in which the measurements
are to be performed.
Thus, Evaluation of Analytical Systems in Biological Systems will be
problem-orientated, rather than technique-orientated with the emphasis on
evaluation in that the volumes in the Series will focus on a given bioana-
lytical problem and then provide a critical and comprehensive discussion
of the particular merits and pitfalls of the various techniques and approaches
available for solving that problem. This requires a team of expert authors
having not only an in-depth knowledge regarding the techniques as such but
also a broad and balanced overview over the entire domain. It is hoped that
in this way the reader will be provided the necessary information to select
the most suitable methodology for his particular application and also to
properly interpret the analytical answers produced by that methodology.
It is not surprising that the first volume deals with the analysis of
biogenic amines. There exists a heavy demand for reliable, sensitive,
selective and accurate analyses of these compounds in biological specimens.
On the other hand, major analytical innovations and applications have been
reported recently in this area, so that a thorough evaluation seems very
timely. We feel fortunate that Drs. Baker and Coutts have been willing to
accept the challenge to edit this volume and that they have been able to
find a team of outstanding authors to assist in this endeavour. The advice
and suggestions of Dr. Karel Macek, Prague, Czechoslovakia during the early
stages of this work are gratefully acknowledged.
XVI
Obviously, analysis in biological systems represents a rather broad field,
from which topics will be selected on the basis of their importance, time-
liness and feasibility. Books on the analysis of metals in human toxicology
and on benzodiazepine analysis are under preparation, whereas volumes on
alcohol analysis and on carbon monoxide are being set up. Additional topics
will not only be selected from the biomedical sciences, but also from other
areas such as environmental analysis, food and agricultural chemistry, etc.
Suggestions to this end as well as comments and suggestions for improvements
wi1 1 be greatly appreciated.
Rokus A. de Zeeuw
1
Chapter 1
AMINES OF BIOLOGICAL INTEREST AND THEIR ANALYSIS
GLEN B. BAKER AND RONALD T. COUTTS
Neurochemical Research Unit, Department of Psychiatry and Faculty of Pharmacy and
Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta T6G 2G3 (Canada)
1.1 INTRODUCTION
Biogenic or naturally-occurring amines have been the subject of a great deal of
research, particularly in the neurosciences. Much of this research has centered
around the catecholamines dopamine and noradrenaline and the indolalkylamine 5-
hydroxytryptamine (5-HT; serotonin). There is now a reasonable body of evidence
supporting the role of these amines as neurotransmitters, and the term "biogenic
amine" has become synonymous with the catecholamines and 5-HT. However, in recent
years, there has been increased interest in histamine and the 'trace' amines, and
in this book these are also included in the term biogenic amines. 'Trace' amines
are defined as a number of naturally-occurring amines which are present in the
central nervous system in much lower absolute quantities than the catecholamines
and 5-HT, and include 6-phenylethylamine, m- and p-tyramine, phenylethanolamine,
-m- and p-octopamine and tryptamine. The role of these substances in neurotrans-
mission is unclear, but it is known that their concentrations in brain are
affected markedly by a number of psychotropic drugs. They have also been implicated
in a variety of psychiatric and neurologic disorders, and this will be discussed
in further detail below.
There is now a voluminous literature on biogenic amines, and space does not
allow for a comprehensive review of the research which has been done on these sub-
stances in biological systems. Rather, it is the purpose of this introductory
chapter to highlight important aspects of the biogenic amines and to give a brief
description of the format of the remainder of the book.
1.2 CATECHOLAMINES AND THEIR 0-METHYLATED AMINE METABOLITES
The catecholamines (Fig. 1.1) have been the most extensively studied of the
biogenic amines because of interest in their role as neurohumors. A number of
excellent books and reviews are available describing the synthesis, metabolism
and localization of the catecholamines in nervous tissue (e.9. 1-14). It has been
proposed that one or more of the catecholamines are involved in the etiology and/
or symptomatology of a number of neurological and psychiatric disorders, includ-
ing migraine, schizophrenia, anxiety, depression, aggression, pheochromocytoma and
parkinsonism (1-3,7,9,14,15-22). In addition, a wide variety of psychotropic
2
HO
a b
HO
OH
C
Yb.,,,,
H3ch
HO
OH
d e
Fig. 1.1. Structures of the catecholamines and their 0-methylated amine
derivatives: a) dopamine, b) noradrenaline, c) adrenaline, d) 3-methoxytyramine,
e) normetanephrine and f) metanephrine.
3
and cardiac drugs are thought to interact with the synthesis, storage, metabolism
and/or receptor activity of the catecholamines (1-4,7,11,13,15,19-30). The
ability to form fluorescent derivatives of catecholamines with relative ease and
specificity has meant that much is now known about the localization of nerve tracts
containing these substances (see Chapter 5 of this volume). Electrolytic or
chemical lesions of these tracts as well as the administration of catecholamine-
depleting drugs have been used in animal models to study the involvement of the
catecholamines in a number of behavioural and physiological functions such as loco-
motion, exploratory activity, hunger, thirst and sexual activity (1-3,7-9,15,18,
21,26,31).
The distribution of the catecholamines dopamine (DA), noradrenaline (NA) and
adrenaline (A) and the ratios of each one to the others vary widely between the
peripheral and central nervous system, within the central nervous system itself
and amongst various species (1,2,6,7).
3-Methoxytyramine (3-MTA), normetanephrine (NMN) and metanephrine (MN) (Fig. 1.
are produced by the enzymatic action of catechol 0-methyl transferase (COMT) on DA
NA and A respectively. These 0-methylated amines have been identified in various
tissues and body fluids (32-39). Concentrations of these compounds in these loca-
tions can be altered dramatically in certain disease states and after administration
of a number of psychotropic drugs.
1.3 5-HYDROXYTRYPTAMINE
5-Hydroxytryptamine (Fig. 1.2), a substance which possesses strong vasocon-
strictive properties on smooth muscle, is found in many parts of the body, and is
located in nonneuronal sites (platelets, mast cells, enterochromaffin cells) as
well as in neurones (7,40,41). Although only 1-2% of this indolealkylamine in the
body is located in the brain (7), 5-HT has been of interest to neuroscientists for
some time because of its putative role as a neurotransmitter and its apparent
CH2CH2NH2
H
Fig. 1.2. Structure of 5-hydroxytryptamine.
4
involvement i n a number of psychological and physiological functions, including
emotion, sleep, hunger, thirst and sexual activity (5,7,8,15,26,31,40-45). As
with the catecholamines, a wide variety of psychotropic drugs affect the synthesis,
metabolism, storage and receptor activity of 5-HT (7,ll-13,15,21-25,30,40-45).
The localization of 5-HT-containing nerve tracts has also been investigated ex-
tensively (4,7,8,15,41-44). The involvement of 5-HT in a number of nervous dis-
orders, including migraine, depression and schizophrenia has been suggested (15,
16,21,22,41-49).
1.4 TRACE AMINES
The trace amines (Fig. 1.3) have not been investigated as thoroughly as the
catecholamines or 5-HT. The primary reason for this is the very low concentration
of these substances in the nervous system, which has necessitated the development
of highly sensitive and specific analytical techniques. The amount of literature
on these amines has increased enormously in recent years as these methods have
been applied to brain and body fluids.
Although they are present in the CNS in minute quantities, the turnover rate
of most of the trace amines is very rapid (50-53) and 8-phenylethylamine and
tryptamine can cross the blood-brain-barrier with ease (54); these two factors
may be very important from a physiological standpoint. Boulton (55) has pointed
out that the trace amines, like the catecholamines and 5-HT, are distributed hetero-
genously amongst brain areas.
Most of the trace amines exert rather strong effects on uptake and release of
the catecholamines and 5-HT (56-59) and metabolic conversions between some of the
trace amines and the catecholamines have been reported (60-63). Recent micro-
iontophoretic studies in brain have revealed that application of trace amines at
currents insufficient to affect baseline firing rates of neurones can alter drama-
tically the response of cells to DA, NA or 5-HT which are applied subsequently
(64,65). Although much yet remains to be known about the role of trace amines in
the CNS, the above findings and structural similarities to the putative neuro-
transmitter amines suggest that their function may be associated intimate y with
OA, NA and 5-HT.
1 .4.1 6-Phenyl ethyl ami ne
The neurochemistry, metabolism and. pharmacology of B-phenylethylamine PEA)
and its effects on animal behaviour have been reviewed extensively (66,67 . A
number of monoamine oxidase inhibitors are known to cause dramatic increases in
brain concentrations of PEA (68-72). One of the biochemical abnormalities result-
ing from phenylketonuria is an increased production of PEA and hence a greatly
elevated urine level of PEA and its major metabolite, phenylacetic acid (PAA)
(73,741.
