Table Of ContentANALYTICAL METHODS IN HUMAN
TOXICOLOGY
ANALYTICAL METHODS
IN
HUMAN TOXICOLOGY
Part 2
Edited by
A. S. CURRY
M
MACMILLAN
©The contributors 1986
Softcoverreprintofthehardcover1stedition1986
Allrights reserved. Noreproduction,copy or transmission
of this publicationmay bemade withoutwritten permission.
No paragraphofthis publication may be reproduced,copied
or transmitted savewithwritten permission orin accordance
with the provisions ofthe Copyright Act 1956 (asamended).
Any person who does any unauthorised act in relation to
this publication may beliable to criminal prosecution and
civilclaims for damages.
First published 1986
Published by
THE MACMILLANPRESS LTD
Houndmills,Basingstoke,Hampshire RG21 2XS
and London
Companies and representatives
throughout the world
Typeset by
TecSet Ltd, Wallington,Surrey
British Library Cataloguing inPublication Data
Analyticalmethodsinhuman toxicology.
Part 2.
1.Toxicology - Technique
I. Curry,A.S.
615.9'0028 RA1211
ISBN978-1-349-07246-0 ISBN978-1-349-07244-6(eBook)
DOI 10.1007/978-1-349-07244-6
Contents
TheContributors vi
Introduction ix
AREVIEW OFANALYTICALMETHODS D.Stevenson
andJ. W.Bridges
2 THEUSEOFHIGH PRESSURE LIQUIDCHROMATOGRAPHY
INHUMANTOXICOLOGY M.D. Osselton 35
3 FLUORESCENCE ANALYSIS L. A. KingandJ.N Miller 71
4 RADIOIMMUNOASSAYSINANALYTICALMETHODS IN
HUMANTOXICOLOGY G. W.Aherne 115
5 PHARMACOKINETICSANDPATIENT CARE G.P.Mould 131
6 DRUGANALYSES INADEPARTMENTOFFORENSIC
MEDICINE ANDTOXICOLOGY P.S.B.Minty 161
7 THEANALYSISOFNARCOTICS A. T.Holder 241
8 FIREGASES R.A.Anderson 289
9 THERAPEUTIC DRUGMONITORING P.A. Toseland,
D.J.BerryandJ. Williams 319
Index 345
The contributors
G.W.Aherne J. N.Miller
DepartmentofBiochemistry andRobens DepartmentofChemistry
InstituteofIndustrialandEnvironmental University ofTechnology
Health and Safety Loughborough
University ofSurrey LeicsLEll 3TU
Guildford
Surrey GU25XH
P.S.B.Minty
R.A.Anderson Departmentof Forensic Medicineand
DepartmentofForensicMedicineandScience Toxicology
TheUniversity ofGlasgow CharingCrossHospital MedicalSchool
GlasgowGl28QQ Fulham PalaceRoad
LondonW68RF
D.J. Berry
PoisonsUnit
Guy'sHospital G.P.Mould
LondonSEI 9RT ClinicalPharmacy Section
DepartmentofClinicalBiochemistry
J.W.Bridges St Luke'sHospital
Robens Instituteof Industrialand WarrenRoad
EnvironmentalHealth and Safety Guildford
University of Surrey Surrey GUI 3NT
Guildford
Surrey GU25XH
M.D.Osselton
Toxicology Division
A.T.Holder
Central Research Establishment
School ofChemistry
HomeOfficeForensicScienceService
Thames Polytechnic
Aldermaston
Woolwich
Reading
LondonSE18 6PF
BerksRG74PN
L.A.King
OperationalServicesDivision D.Stevenson
Central Research Establishment Robens Instituteof Industrialand
HomeOffice ForensicScienceService EnvironmentalHealth and Safety
Aldermaston University ofSurrey
Reading Guildford
BerksRG74PN Surrey GU25XH
The Contributors vii
P.A.Toseland J.Williams
DepartmentofClinicalChemistry DepartmentofPharmacology and
Guy'sHospital Therapeutics
LondonSEI 9RT WelshNational School ofMedicine
Cardiff
Introduction
In these two volumes on analytical methods in human toxicology twenty-six
authors have presented the latest experimental techniques and their application
to practical problems.
The individual chapters have been written by scientists who are in the fore
front of their science. They have not forgotten that the analyses have conse
quencesand many havecommentedon how vitalit isthat the results beaccurate
and precise.
The social scene today revolves very much around drugs and their use and
abuse,and there is a heavy responsibility on toxicologists to showwhere inthe
body a drug is localised and,if possible, how it isacting. Onlyby knowing such
fundamental facts canadvancesbemade in the future on arational basis.
Much of the work in these two volumes describes analytical methods using
'black boxes' - machines of high complexity and high cost but involvingvery
simple physicochemical principles. It isvital that the operator understands the
underlying principles if mistakes are not to be made,and it is refreshing to see
how many authors haverecognisedthis fact.
Unfortunately international quality control programs have demonstrated a
very poor performance when samples have been circulated on a 'known' basis
(let alone in 'blind trials'). The best equipment will only givethe best possible
results if it isoperated byscientists who areextremelyselfcritical,and although
this book (along with the previous volume) describes a host of experimental
techniques, it has to be read with care and should be used by experienced
personnel.
Human toxicology is not only a fascinating subject, but it also has a vast
potential for unravelling the solutions to diseases. To use an analogy, Erlich's
silver bullet has now to be fired and its position in tissue discovered to within
a few microns. The unlocking of molecular structure using monoclonal anti-
x Introduction
bodies coupled with advanced chemistry means that the key can now be made
with the knowledge of how the lock will respond. Medicalscienceis not only
knocking onthe door - it hasone foot insideaswell.
The editor has been mainly involvedwith forensic problems but it has been
refreshing for him to be able to have an overviewofthe wholesceneand to see
how many analytical techniques haveevolvedbecauseforensictoxicologistshave
required even more sensitivemethods of detection.Their criticalevaluationhas
added agreat dealto these two volumes.
It ishoped that these two volumeswillprovideknowledgethat willbeshared
and that willleadto evenfurther advances.
Reading,1986 A.S.C.
1
A Review of Analytical Methods
D. STEVENSON and J. W. BRIDGES
1.1 INTRODUCTION
For xenobiotics and their metabolites which form readily reversibleinteractions
with target sites for toxicity (e.g. receptors), the peak level of unbound com
pound present in the immediate vicinity of the target site should be directly
related to the biological response over a considerable concentration range. In
contrast, for chemicalswhich form ratherstable ligandsto their target, including
veryreactivexenobioticsandtheir metabolites,the amount ofcompound present
over a period of time isoftenmore important than the peak level.For anumber
of compounds, a direct correlation between total covalent binding to cellular
proteinsand theextentofhepatocellularnecrosishas been established.Similarly,
for several carcinogens, DNAalkylation has-been shown to bedirectly related to
their carcinogenic potency, although in some cases (e.g. dimethyl-nitrosamine)
such a correlation is not observed with total binding but rather with specific
adducts. There are a number of additional reasons why the concentration of a
xenobiotic and/or its metabolites that is achieved in a tissue may not correlate
with the biological response observed.Commoncausesare asfollows.
(a) The xenobiotic and/or its metabolites may be strongly bound to cellular
macromolecules which are not involved in the expression of biological
activity. For example, cadmium is very strongly bound in both liver and
kidney to the protein metallothionein;therefore, the measurement of total
cadmium levels does not correlate well with the extent of adverse change.
Organic compounds may also bind strongly but non-covalently to cellular
macromolecules, e.g. quinacrine is strongly boundin the liver,gentamicin is
slowly but strongly boundin the kidney.
(b) The toxic effect on the tissue may be mediated indirectly, e.g. at a site
which is distant from the observed site of biological activity, for example
the primary target may bethe nervous orendocrine system.
