Modern Methods of Steroid Analysis

518 Pages·1973·9.243 MB·English

by  Erich Heftmann (Eds.)

Contributors
GUY Ε. ABRAHAM J. KARLE
G. ALBERS-SCHÖNBERG B. A. KNIGHTS
RODNEY C. BARKHURST AVINOAM KOWARSKI
FRANCIS BAYARD B. S. MlDDLEDITCH
INESE Z. BEITINS CLAUDE J. MIGEON
P. BELANGER K. NAKANISHI
GARY BROOKER MARGARET M. NOONE
C. J. W. BROOKS B. PLAZONNET
ALBERT W.XBURGSTAHLER DAVID A. SCHOOLEY
ARILD T. CHRISTENSEN B. SCHRÄDER
STANLEY J. CLARK W. ROY SLAUNWHITE, JR.
PIERRE CRABBE J. L. SMITH
JACK FISHMAN FELIKSA SNATZKE
JACEK K. GAWRONSKI GÜNTHER SNATZKE
C. C. HINCKLEY WOLFGANG STEIGEMANN
DONALD P. HOLLIS E. STEIGNER
ROGER W. JELLIFFE W. J. A. VANDENHEUVEL
DAVID F. JOHNSON PER VESTERGAARD
HERBERT H. WOTIZ
MODERN METHODS OF
STEROID ANALYSIS
Edited by ERICH HEFTMANN
Western Regional Research Laboratory
U.S. Department of Agriculture
Berkeley, California
1973
ACADEMIC PRESS New York and London
A Subsidiary of Harcourt Brace Jovanovich, Publishers
COPYRIGHT © 1973, BY ACADEMIC PRESS, INC.
ALL RIGHTS RESERVED.
NO PART OF THIS PUBLICATION MAY BE REPRODUCED OR
TRANSMITTED IN ANY FORM OR BY ANY MEANS, ELECTRONIC
OR MECHANICAL, INCLUDING PHOTOCOPY, RECORDING, OR ANY
INFORMATION STORAGE AND RETRIEVAL SYSTEM, WITHOUT
PERMISSION IN WRITING FROM THE PUBLISHER.
ACADEMIC PRESS, INC.
Ill Fifth Avenue, New York, New York 10003
United Kingdom Edition published by
ACADEMIC PRESS, INC. (LONDON) LTD.
24/28 Oval Road, London NW1
Library of Congress Cataloging in Publication Data
Heft mann, Erich.
Modern methods of steroid analysis.
Includes bibliographies.
1. Steroids-Analysis. I. Title [DNLM:
1. Steroids-Analysis. QU95 H461m 1973]
QD426.H43 547'.73 72-88330
ISBN 0-12-336640-2
PRINTED IN THE UNITED STATES OF AMERICA
List of Contributors
Numbers in parentheses indicate the pages on which the authors' contributions begin.
GUY E. ABRAHAM (451), Department of Obstetrics and Gynecology,
University of California at Los Angeles School of Medicine, Harbor
General Hospital Campus, Torrance, California
G. ALBERS-SCHÖNBERG (199), Merck Sharp and Dohme Research Labora-
tories, Rahway, New Jersey
RODNEY C. BARKHURST (349), Mathematics-Science Division, Haskell In-
dian Junior College, Lawrence, Kansas
FRANCIS BAYARD1 (471), Harriet Lane Service of the Children's Medical
and Surgical Center, The Johns Hopkins Hospital and University,
Baltimore, Maryland
INESE Z. BEITINS (471), Harriet Lane Service of the Children's Medical
and Surgical Center, The Johns Hopkins Hospital and University,
Baltimore, Maryland
P. BÉLANGER (199), Merck Sharp and Dohme Canada Limited, Pointe
Claire/Dorval, Quebec.
GARY BROOKER2 (437), Department of Medicine, University of Southern
California School of Medicine, Los Angeles, California
C. J. W. BROOKS (139), Chemistry Department, University of Glasgow,
Glasgow, Scotland
ALBERT W. BURGSTAHLER (349), Department of Chemistry, The Univer-
sity of Kansas, Lawrence, Kansas
ARILD T. CHRISTENSEN (281), Syntex Analytical Instruments, Cupertino,
California
STANLEY J. CLARK (71), Department of Biochemistry, Boston University
School of Medicine, Boston, Massachusetts
1 Present address : Centre Hospitalier Regional de Toulouse Hospital Purpan, Bloc
d'Urgence Médicale, Toulouse, France.
2 Present address : Department of Pharmacology, University of Virginia School of
Medicine, Charlottesville, Virginia.
xiii
xiv LIST OF CONTRIBUTORS
PIERRE CRABBÉ3 (331), Research Laboratories, Syntex S. Α., Mexico, D. F.,
Mexico
JACK FISHMAN (399), Institute for Steroid Research, Montefiore Hospital
and Medical Center, and Department of Biochemistry, Albert Ein-
stein College of Medicine, Bronx, New York
JACEK K. GAWRONSKI (349), Institute of Chemistry, A. Mickiewicz Uni-
versity, Poznan, Poland
C. C. HINCKLEY (265), Department of Chemistry and Biochemistry,
Southern Illinois University, Carbondale, Illinois
DONALD P. HOLLIS (245), Department of Physiological Chemistry, The
Johns Hopkins University School of Medicine, Baltimore, Maryland
ROGER W. JELLIFFE (437), Department of Medicine, University of South-
ern California School of Medicine, Los Angeles, California
DAVID F. JOHNSON (55), National Institute of Arthritis and Metabolic
Diseases, National Institutes of Health, Bethesda, Maryland
J. KARLE (293), Laboratory for the Structure of Matter, U.S. Naval Re-
search Laboratory, Washington, D.C.
B. A. KNIGHTS (103), Department of Botany, University of Glasgow,
Glasgow, Scotland
AVINOAM KOWARSKI (471), Harriet Lane Service of the Children's Medi-
cal and Surgical Center, The Johns Hopkins Hospital and University,
Baltimore, Maryland
B. S. MIDDLEDITCH4 (139), Chemistry Department, University of Glasgow,
Glasgow, Scotland
CLAUDE J. MIGEON (471), Harriet Lane Service of the Children's Medical
and Surgical Center, The Johns Hopkins Hospital and University,
Baltimore, Maryland
K. NAKANISHI ( 37 ), Department of Chemistry, Columbia University, New
York, New York
MARGARET M. NOONE (221), Sadtler Research Laboratories, Inc., Phila-
delphia, Pennsylvania
B. PLAZONNET (199), Merck Sharp and Dohme-Chibret Research Lab-
oratories, Clermont-Ferrand, France
DAVID A. SCHOOLEY ( 37 ), Biochemistry Department, Zoecon Corporation,
Palo Alto, California
3 Present Address: Labroatoire de Chimie Organique, C.E.R.M.O., Université
Scientifique et Médicale, Grenoble, France.
4 Present address: Institut for Lipid Research, Baylor College of Medicine, Houston,
Texas.
LIST OF CONTRIBUTORS XV
B. SCHRÄDER5 (231), Institut für Spektrochemie und Angewandte Spek-
troskopie, Dortmund, Germany
W. ROY SLAUNWHITE, JR. (419), Departments of Biochemistry and Pedi-
atrics, State University of New York at Buffalo, Buffalo, New York
J. L. SMITH (199), Merck Sharp and Dohme Research Laboratories, Rail-
way, New Jersey
FELIKSA SNATZKE (381), Institute of Organic Chemistry, Polish Academy
of Sciences, Warsaw, Poland
GÜNTHER SNATZKE (381), Institute of Organic Chemistry, University of
Bonn, Bonn, Germany
WOLFGANG STEIGEMANN (321), Max-Planck-Institut für Eiweiss- und
Lederforschung, Munich, Germany.
E. STEIGNER6 (231), Institut für Spektrochemie und Angewandte Spek-
troskopie, Dortmund, Germany
W. J. A. VANDENHEUVEL (199), Merck Sharp and Dohme Research Lab-
oratories, Rahway, New Jersey
PER VESTERGAARD ( 1 ), Research Center, Rockland State Hospital, Orange-
burg, New York
HERBERT H. WOTIZ (71), Department of Biochemistry, Boston University
School of Medicine, Boston, Massachusetts
5 Present address : Abteilung für Theoretische Organische Chemie, Universität
Dortmund, Dortmund-Hombruch, Germany.
c Present address : Internationale Dokumentationsgesellschaft für Chemie, Frankfurt
(Main), Germany.
Preface
When our first book on the "Biochemistry of Steroids" was published
(Reinhold, 1960), I was thoroughly familiar with the analysis of steroids
and found it easy to include as much on this subject as a biochemist
needed to know. Physical tests had only begun to replace the time-
honored "specific" color reactions, and they all could be performed in our
own laboratory. Structure elucidation was still an arcane art, requiring
months of labor and several hundred milligrams of sample. Although
paper chromatography had already opened large areas of steroid metab-
olism to intensive biochemical investigation, the analysis of minute
amounts of such important hormones as aldosterone presented great
difficulties.
Today the biochemist is faced with a bewildering array of instrumental
methods of analysis, most of them automated and computerized. The
steroid field seems to be a favorite playground of physical analysts, prob-
ably because it provides them with hundreds of analogous crystalline
model compounds with many possibilities of isomerism. Another reason is,
of course, that instrumental methods are so expensive that only com-
pounds with important biological properties, such as the steroid hor-
mones, can provide the justification for such large expenditures. How-
ever, instrumentation developed under the sponsorship of the biomedical
establishment is soon adopted by less affluent research fields in which it
supports the efforts of scientists in different disciplines.
My last book on "Steroid Biochemistry" (Academic Press, 1970) omits
sections on analytical chemistry. That field had grown so in the last
decade that it would have been impossible for me to include even the
most essential material in addition to covering the many important ad-
vances in steroid biosynthesis, metabolism, physiology, and pharmacology
without making the book too long.
When I began a review of the modern methods of steroid analysis, I
realized that to become an expert in all of the methods would be im-
possible. Therefore I invited prominent authorities in each of the seven
main subdivisions of instrumental analysis to contribute review chapters
on their specialties. These contributions comprise this volume.
xvii
xviii PREFACE
In editing "Modern Methods of Steroid Analysis," I have done my best
to eliminate duplication, to ask authors to fill lacunae, and to reconcile
individual styles. The treatment of the subject matter approaches the
level at which an investigator entering the steroid field or a steroid spe-
cialist contemplating the use of a new method will not only find an in-
troduction to it but guidance to further study as well. If some areas are
not as thoroughly covered as others, I must assume full responsibility.
I felt that timely publication of this book was as important as complete-
ness. I hope this volume will also be useful to investigators outside the
steroid field who are interested in methodology, since the methods dis-
cussed are largely applicable to other substances as well.
The section on chromatographic methods with special reference to the
new techniques of high-speed, high-resolution liquid chromatography
was placed at the beginning since most of the other techniques require
material that has been purified by chromatography. Gas chromatography
is presented with special reference to new detection and identification
methods. This is logically followed by a section on mass spectrometry,
since the combination of the gas Chromatograph with the mass spec-
trometer provides the modern analyst with one of his most powerful tools.
Sections III and IV present only the most recent developments in infrared
and Raman spectroscopy and nuclear magnetic resonance, respectively,
while the next section, on x-ray diffraction analysis, provides a comparison
of various crystallographic techniques. The sophistication in structure
analysis afforded by modern chiroptical methods is highlighted in Sec-
tion VI. The last section contains a number of radioisotope methods that
have provided the clinical investigator with the most sensitive and spe-
cific analytical tools devised so far.
My own contribution to this volume is that of an impresario, no more
and no less. Correspondence with the contributors has been a major
undertaking which was handled with patience, intelligence, and tact by
my wife, Brigitte. In addition to her duties as wife and mother, she has
spent many hours on this book, and deserves most of the credit for its
prompt appearance.
Erich Heftmann
List of Abbreviations
A 11-dehy drocorticoste- ft feet
rone GC-MS gas chromatography-
Aldo aldosterone mass spectrometry
atm atmospheres GLC gas-liquid chroma-
ATPase adenosinetriphos- tography
phatase 11-HA 11-hydroxyandroste-
Β corticosterone rone
BSA N,0-bis(trimethyl- 11-HE 11-hydroxyetiochola-
silyl)acetamide nolone
CD circular dichroism HMDS hexamethyldisilazane
Ci curie hr hour(s)
CM convolution molecule Hz hertz
cm centimeter, 10~2 m i.d. internal diameter
CMDMSi ( chloromethyl ) di- in. inch (es)
methylsilyl IR infrared
CPBA competitive protein 11-KA 11-ketoandrosterone
binding assay 11-KE 11-ketoetiocholanolone
cpm counts per minute LC liquid chromatography
DC direct current LLC liquid-liquid chroma-
DHA dehydroepiandroste- tography
rone LSC liquid-solid chroma-
DIDA double isotope deriva- tography
tive assay m meter
DOC deoxycorticosterone micron, 10"6 m
DSS dimethylsilapentane- mCi millicurie, 10~3 Ci
sulfonic acid mg milligram, 10-3 g
Ε cortisone microgram, 10~6 g
Εχ estrone MHz megahertz, 106 Hz
E estradiol ml milliliter, 10"3 1
2
E estriol \ microliter, 10~6 1
F 3 Cortisol mμ m millimeter, 10~3 m
FID flame-ionization de- MO methyl oxime
tector mp melting point
xix
XX LIST OF ABBREVIATIONS
MS mass spectrometry RI refractive index
ν frequency S Reichstein's Com-
ng nanogram, 10~9 g pound S, 11-deoxy-
nm nanometer, 10~9 m, πΐμ cortisol
ORD optical rotatory disper- SD standard deviation, σ
sion sq square
Ρ progesterone Τ testosterone
pg picogram, 10~12 g TFA trifluoroacetate
ppm parts per million TLC thin-layer chromatog-
psi pounds per square raphy
inch TMCS trimethylchlorosilane
Q deoxycorticosterone, TMSi trimethylsilyl
DOC TSIM trimethylsilylimidazole
rate of flow, relative UV ultraviolet
chromatographic
mobility