Table Of ContentRecent Developments in
Mass Spectrometry in
Biochemistry and Medicine
----Volume 2----
Recent Developments in
Mass Spectrometry in
Biochemistry and Medicine
-----Volume 2 - - - - -
Edited by
Alberto Frigerio
"Mario Negri" Institute
Milan, Italy
Plenum Press· New York and London
Library of Congress Cataloging in Publication Data
International Symposium on Mass Spectrometry in Biochemistry and Medicine, 5th, Rimini,
1978.
Recent developments in mass spectrometry in biochemistry and medicine.
Organized by the Italian Group for Mass Spectrometry in Biochemistry and Medicine.
Includes index.
1. Mass spectrometry-Congresses. 2. Biological chemistry-Technique-Congresses. 3.
Drugs-Analysis-Congresses. I. Frigerio, Alberto. II. Italian Group for Mass Spectrometry in
Biochemistry and Medicine. III. Title. [DNLM: 1. Spectrum analysis, Mass-Congresses. Wl
RE106AM)
QP519.9M31571978 612'.01585 79-19982
ISBN-13: 978-1-4613-3020-2 e-ISBN-13: 978-1-4613-3018-9
001: 10.1007/978-1-4613-3018-9
Proceedings of the 5th Internati onal Symposium on Mass Spectrometry in
Biochemistry and Medicine, held at Rimini, Italy in June 1978
© 1979 Plenum Press, New York
Softcover reprint of the hardcover 1 st edition 1979
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N. Y. 10011
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
Preface
The papers collected in this volume were presented at the 5th
International Symposium on Mass Spectrometry in Biochemistry and
Medicine held at Rimini, Italy, in June 1978.
This meeting was organized by the Italian Group for Mass
Spectrometry in Biochemistry and Medicine, which was founded in
Milan, Italy, in 1975 by researchers working in different fields
(chemistry, pharmacology, medicine, biology).
I wish to thank Mrs. Anna Bernard, Mrs. Donata Castoldi and
Miss Vanna Pistotti for their invaluable technical assistance and
countless eff.orts in the preparation of this volume.
Alberto Frigerio
Milan,
March, 1979
v
Contents
The Place of Mass Spectrometry in Drug Metabolism
Studies .......................................... 1
A. Benakis
In Vitro and in vivo Studies on the Metabolism of
Dibenzo/c,f 7-/-1,2 7 Diazepine ••••••••.••••••••• 9
A. Frigerio, P~ Negrini, D~ Rotilio, A. De Pascale
and E. Rossi
Gas Chromatographic-Mass Spectrometric Identification
of Urinary Metabolites of Propildazine in Rat 27
L. Simonotti, R. Colombo and G. Pifferi
Mass Spectrometric Characterization of Sydnocarb ~
and its Major Metabolites in Rat ••••••••••••••••• 43
J. Tamas, M. Polgar, G. Czira and L. Vereczkey
Metabolism of an Aryloxy Beta Blocking Drug 53
G.L. Passetti, E. Grassi and A. Trebbi
Isolation and Identification of the Chloroform Soluble
Urinary Metabolites of Suloctidil in Man .••••..•• 69
R. Roncucci, W. Cautreels, M. Martens, C. Gillet,
K. Debast and G. Lambelin
Are Stable Sulfenic Acids Possible Metabolites of
Suloctidil? ...................................... 85
W. Cautreels, M. Martens, R. Roncucci, C. Gillet,
K. Debast and G. Lambelin
The Electron Impact Fragmentation of Products Related
to N-(Allyl)-3,3-Diphenylpropylamine, a
Metabolite from Thiopropamine ~ ••••••••••••••••• 97
V. Borzatta, M. Cristofori and A.G. Giumanini
vii
viii CONTENTS
Urinary Metabolites from (2-Ethyl-2,3-Dihydro-5-
Benzofuranyl) Acetic Acid .•••••••••••••••.•••••.• 107
C. Casalini, G. Cesarano, G. Mascellani,
G. Tamagnone and A.G. Giumanini
The Major Metabolites of Niclosamide: Identification
by Mass Spectrometry............................. 121
L.A. Griffiths and V. Facchini
The Metabolism of Deuterium-Labelled Analogues of
~1_,~6_, and ~7-Tetrahydrocannabinol and
the Use of Deuterium Labelling •••••••••••••••••.• 127
D.J. Harvey and W.D.M. Paton
Kinetics of Degenerate Nucleophilic Exchange of
C(3)-Hydroxy Group in l-Methyl-3-Hydroxy
-5-Phenyl-7-Cloro-2H-l,4-Benzodiazepin-
2-0ne ............................................ 149
D. Srzi~, L. Klasinc, B. Belin, F. Kajfez and
V. Sunji~
On the Analysis of Fermentation Broth During
Biosynthesis of Cephalosporin Antibiotics 155
M. Kac, J. Marsel and M. Pokorny
GC-MS Analysis by Electron Impact Ionization and
Chemical Ionization of Carcinogenic
2-Acetylaminofluorene and its Metabolites •.•••••• 163
C. Lallemant, G. Wood. M.F. Exilie, M. Chessebeuf,
P. Gambert, M. Hardy and P. Padieu
The Proximo-Distal Carcinogen of the K Region of
Benzo (a)Pyrene.... . . ... . .. . . . ... . . . . .. . . . . .. .... .. 179
G. Lhoest
Quantitative Determination of Vincamine in Human
Plasma by Gas Chromatography - Mass
Spectrometry ..................................... 191
P. Devaux, E. Godbille and R. Viennet
Mass Fragmentographic Quantification of Phosphonoacetic
Acid in the Blood of Monkeys Infected with
Herpesvirus Saimiri .............................. 205
J. Roboz. R. Suzuki. R. Hunt and J.G. Bekesi
The Determination of Blood Cyanide and Plasma Thiocyanate
by Gas Chromatography-Mass Spectrometry.......... 215
I. Thomson, R.A. Anderson and W.A. Harland
CONTENTS ix
Present State and Future Trends of Negative Ion
Mass Spectrometry in Toxicology, Forensic
and Environmental Chemistry...................... 227
H. Brandenberger
Mass Fragmentographic Determination of Polyamines
in the Urine of Premature Babies .••.••••..•....•• 257
H. Milon, I. Antener and S. Nordio
An Assay for Schizophrenia: Mass Spectrometric
Determination of Two Cadaverine Metabolites 269
H. Dolezalova, M. Stepita-Klauco, C. van der Velde
and V. Cassone
Analysis of the Urinary Steroids from a Pathological
Pregnancy by Liquid-Gel Chromatography and
Gas-Chromatography-Mass Spectrometry............. 283
H. Losty, R.J. Begue, M. Moriniere and P. Padieu
Identification of Atypical Bile Acids by Gas
Chromatography-Mass Spectrometry-Computer
Techniques ....•......•...........•...•........... 297
F. Stellaard and P.A. Szczepanik-Van Leeuwen
The Effects of Triisopropylacetic Acid on the
Metabolic Profile of Urinary Organic Acids 307
T. Kuhara, Y. Hirokata, S. Yamada, I. Matsumoto,
J.A. MacPhee and J.-E. Dubois
Application of Chemical Ionization MS to the Study
of Regional Brain Catecholamine Metabolism
in (S)-a-Methyldopa Treated Rats. ...•....•....... 317
D. Karashima, R.L. Cockerline, K.L. Melmon and
N. Castagno1i, Jr.
Concurrent Extraction and GC-MS Assay of Endogenous
TP, T, lAA, 5HT and SHlAA in Single Rat
Brain Samples .•.................•..............•. 331
F. Artigas and E. Gelpi
A New Method for Simplified Prostaglandin Profiling
by Selected Ion Monitoring ....... ...•....•....... 347
J. Rose116 and E. Gelpi
Analysis of Urinary Steroids by Liquid-Gel
Chromatography and Gas Chromatography -
Mass Spectrometry. Clinical Applications 355
R.J. Begue, M. Dumas, H. Losty, M. Moriniere,
C. Perrier and P. Padieu
x CONTENTS
A Micromethod for the Clinical Chemistry Routine
Analysis of Amino Acids in Blood and
Urine by Capillary Gas Liquid Chromatography
of Isobutyl Esters N(O)-Heptafluorobutyrate
Derivatives ......•....•.•••••••...••••••.•.....•. 377
J. Desgres, D. Boisson, F. Veyrac, M. Susse and
P. Padieu
Metabolic Profile of Ventricular Cerebrospinal Fluids 401
I. Matsumoto, N. Hisanaga, T. Shinka, T. Kuhara,
M. Yoshida, N. Kusano and S. Kuramoto
Energy and Metastable Characteristics in Peptides,
Electron Impact Appearance Potentials of
Sequence Ions in Tripeptides ••••.•...•.•....•...• 411
Z.V.I. Zaretskii and P. Dan
The Continuous Recording of Expired Cl60lBO by
Respiratory Mass Spectrometry After
Injection of H2lBO into the Vascular
System .•.•.......•..•...•••••.•.....•.•..•...••.. 427
K.-P. Pflug, K.-D. Schuster, H. Forstel and
J.P. Pichotka
A Continuous Mass Spectrometric Analysis of
Expired Physiological Gases: the Pulmonary
Gas Exchange ......•.•..•.•..•...••••...•••.....•. 435
S. Damato
Adaptation of Respiratory Mass Spectrometer to
Continuous Recording of Abundance Ratios
of Stable Oxygen Isotopes •..•.•...•...••..•.•.•.. 451
K.-D. Schuster, K.-P. Pflug, H. Forstel and
J.P. Pichotka
The Development of a Flexible Mass Spectrometer
Catheter •.....••.•...•••.•••••••••..•...•.•...... 463
T.D. Johnson, G.M. Watkins, J. Holsinger,
M.P. Roberts and D.D. Thomas
.........................................
Contributor Index 481
Index ...................................................... 483
THE PLACE OF MASS SPECTROMETRY IN DRUG METABOLISM STUDIES
A. Benakis
Laboratory of Drug Metabolism, Department of Pharmacology
University, Geneva, Switzerland
A vast fund of published information on the use of mass
spectrometry and associated techniques for the study of drug
metabolism has accumulated in recent years. The annual "Specialist
Periodical Reports" on mass spectrometry of the London Chemical
Society is particularly worthy of mention. This publication reviews
all the literature in the field. In the issue covering the two year
period from 1972 to 1974 (1), there are more than one hundred and
forty reviews of papers devoted to mass spectrometry applications
for drug metabolism. From 1974 to 1976 (2), there were a hundred
and twenty.
The papers presented at the annual international symposium on
mass spectrometry are published by Dr. Frigerio and greatly
appreciated (3-8). Other valuable publications in this field are
"Advances in Mass Spectrometry" put out by the London Institute
of Petroleum, and "Quantitative Mass Spectrometry in Life Sciences",
a report of the proceedings of the Ghent International Symposium
in 1976 (9).
Interesting mass spectrometric data can also be found in
numerous pharmacological journals, particularly in those specializing
in drug metabolism studies (10, 11, 12). The journal "Biomedical
mass spectrometry", in which papers on drug metabolism are well
represented, can also be mentioned (13).
Finally, new developments in mass spectrometry applied to drug
metabolism studies have been widely reviewed by other authors (14-
20).
The comments I shall be making in this paper will be those of
2 A. BENAK IS
one who habitually uses mass spectrometry techniques in drug
metabolism studies and pharmacokinetics and will concern these
fields rather than the technical and technological progress in mass
spectrometry.
During the years from 1958 to 1968, the study of drug metabolism
was relatively limited; it was conducted mainly by university
research teams in the context of fundamental studies and was
essentially concerned with the detoxication mechanism.
The first period could be called the "period of scientific
curiosity", during which some precise results were obtained and
certain techniques were devised and refined.
The period between 1968 and 1973. which I consider the second
period. was one of growing awareness and realization of the
importance of the problem.
Metabolic and pharmacokinetic studies became more generalized and
were performed not only in university laboratories but also and
especially by the pharmaceutical industry. During the seventies,
most of the drug metabolism laboratories of the pharmaceutical
industry. especially in the United States. centered their attention
on mass spectrometry and used the instruments of their company's
analytical laboratories. In just a few years. however. both indus
trial and university laboratories had acquired mass spe~trometers,
and frequently. GLC/MS as well.
Although this growing interest was certainly due in part to
the natural desire for a better understanding of the mode of action
of certain drugs by studying their biotransformation mechanisms,
it cannot be denied that the determining role in the development
of this field was played by the governmental authorities of an
ever-increasing number of countries who began to insist upon this
type of study.
It is my feeling that there is a need at present to better
define the objectives of the metabolic and pharmacokinetic studies
required by the authorities in many countries. I believe that this
problem is of interest to everyone performing studies of this kind.
Some controversial aspects are the ever-increasing demands of the
authorities with regard to results, and the consequent increasing
complexity and cost of the studies. This situation could be
disastrous in the long run since it is technically and economically
impossible to keep increasing, ad infinitum, the number of studies
required to introduce a new drug on the market.
The period from 1973 to the present has been very rich, both
in experimental results and perspectives.
By 1976, metabolic studies represented 18% of all the pharmaco-
