Advances in Tracer Methodology: Volume 2 A collection of papers presented at the Sixth, Seventh, and Eight Symposia on Tracer Methodology plus other papers selected by the editor

314 Pages·1995·12.068 MB·English

by  Martin D. Kamen (auth.), Seymour Rothchild (eds.)

Advances in
TRACER
METHODOLOGY
Volume 2
A Publication of the New England Nuclear Corporation
Advances in
TRACER
METHODOLOGY
Volume 2
A collection of papers presented at the Sixth, Seventh,
and Eighth Symposia on Tracer Methodology plus other
papers selected by the editor
Edited by
Seymour Rothchild
Distributed by
PLENUM PRESS
NEW YORK
1965
ISBN 978-1-4684-8624-7 ISBN 978-1-4684-8622-3 (eBook)
DOI 10.1007/978-1-4684-8622-3
Library of Congress Catalog Card Number 62-13475
©1965 New England Nuclear Corporation
Softcover reprint of the hardcover 1st edition 1965
575 Albany St., Boston, Massachusetts
All rights reserved
No part of this publication may be reproduced in any
form without written permission from the publisher
PREFACE
Advances in Tracer Methodology, Volume 2, records the pro
ceedings of the 6th, 7th, and 8th Symposia on Advances in Tracer
Methodology. These Symposia, which are part of a continuing
series sponsored by the New England Nuclear Corp. and the Packard
Instrument Company, Inc., are devoted to the entire isotope tracer
field: preparation and analysis of labeled compounds, applications
in the chemical, biochemical, and clinical fields, and health physics
considerations associated with tracer work.
The papers in this volume reflect certain trends which can be
noted in the tracer field: increasing reliance on biochemical methods
for labeling esoteric compounds, growing awareness of the problems
of radiation decomposition, the use of advanced instrumentation for
the analysis and detection of radioactive substrates, and the rapidly
growing application of tracer compounds to clinical diagnosis.
The three Symposia at which the papers were presented were
held on the following dates:
DATE LOCATION CHAIRMAN
11/16/62 New York City Daniel Steinberg, Chief
Laboratory of Ce1lular PhYSiology
and Metabolism,
National Heart Institute,
Bethesda, Maryland
3/8/63 Los Angeles Martin Kamen
University of California
San Diego, California
11/8/63 Chicago Joseph L. Rabinowitz
Veterans Administration Hospital
Philadelphia, Pennsylvania
Once again, it is the editor's pleasure to acknowledge the help
of his colleagues in planning and running the various Symposia, as
well as the excellent cooperation of the chairmen, who acquitted
themselves in their appointed tasks in a most efficient manner,
With humor and understanding.
S. R.
Boston, Mass.
December 4, 1964
CONTENTS
Early History of Carbon-14 .•.....•... 1
Martin D. Kamen
Methods for Labeling Compounds
Conditions for High Yield in the Labeling of Hydrocarbons
by Exchange • . . . • . • . . . . • • • . • . • . • . . . . . . • . . . 21
B. E. Gordon and J. J. Madison
C14-Labeling Using Carbene Insertion-Application to Satu-
rated Hydrocarbons . • . • . . . . . • . . • • . . • . . . . . . . . 39
M. A. Muhs, E. L. Bastin, and B. E. Gordon
Biosynthesis of Labeled Carbohydrates and Other Compounds
of Biochemical Interest. . . . . . . . . . . . . . . . . . . . • . . 49
S. Abraham
The Preparation of Labeled Albumins for Turnover Studies 61
Sheldon Margen and Harold Tarver
The Biosynthesis of C14_ and H3-Labeled Insulin. . . . 73
G. Eric Bauer, Arnold W. Lindall, and Arnold
Lazarow
Purification of High Specific Activity Acetic - H3 Anhydride 83
Herbert H. Henderson, Frank Crowley, and Leo
E. Gaudette
Special Analytical Techniques
Automatic Counting of Radioactivity on Two-Dimensional
Paper Chromatograms . . . . . . . . . . . • . • . . . . . . . . . 87
V. Moses
A Convenient Method for the Determination of Metabolically
Liberated C140Z • • • • • • • • • • • • • • • • • • • • • • • • • • • • 93
Eugene Roberts, Daisy G. Simonsen, and Betty
Sisken
Liquid Scintillation Counting of C14-Labeled Amino Acids on
Paper, Using Trinitrobenzene- I-Sulfonic Acid, and an
Improved Combustion Apparatus. . . . . . . . . . . . . . . . . 97
Claude F. Baxter and Ilse Senoner
vii
viii Contents
Zonal Scanning of Thin-Layer Chromatograms. 107
Fred Snyder
Continuous Scintillation Counting of Amino Acid Analyzer
Column Eluates. . . . . . . . . . . . . . . . . . . . . . . . . . .. 115
David H. Elwyn
Some Techniques of Radioactive Gas Chromatography for
Lipid Research . . . . . . . . . . . . . . . . . . . . . . . . . . .. 123
H. J. Dutton
The Microchemical Identification of Steroids from Biological
Media. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 135
David L. Berliner
Isotope Fractionation in the Absorption Chromatography of
Doubly-Labeled Compounds. . . . . . . . . . . . . . . . . . .. 145
Peter D. Klein
Quench Correction by Automatic External Standardization.. 155
Ariel G. Schrodt, James A. Gibbs, and Robert E.
Cavanaugh
Biochemical Applications
Analytical Studies Using Low Levels of Carbon-14 and
Tritium: A Method for Determining the Labeling Pattern
in myo-Inositol . . . . . . . . . . . . . . . . . • . . . . . . . . .. 163
Frank A. Loewus
Multiple Labeling to Determine Metabolic Pathways: Use of
Labeling Ratios. . . . . . . . . . . . . . . . . . . . . . . . . . .. 169
J. T. Van Bruggen and Paul Russell
Diffusion of Radioactively Labeled Molecules in Heart Muscle 179
Ernest Page
Use of Labeled Nonmetabolized Amino Acids in Biochemical
Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 183
Thomas R. Riggs and Halvor N. Christensen
The Use of DMO-2-C14 (5,5,-Dimethyl-2,4-0xazolidine
dione-2-C14) and Inulin-Carboxyl-C14 for Measurement of
Intracellular pH. . . . . . . . . . . . . . . . . . . . . . . . . . .. 189
Thomas C. Butler
Clinical Applications
AEC Licensing of Rad~oisotopes for Clinical Research. 193
George V. LeRoy
Contents ix
The Preparation of Radioactive Chemicals for Clinical Use 201
William H. Briner
Clinical Applications of Aldosterone .In Vivo Secretion Rate 213
Bernard Kliman
Steroid Secretory Mixtures in Man. • • . • . . . . • . . • • . . .. 221
Edward G. Biglieri
Isotopic Methods for Steroids in Human Blood . • . • . . . . .. 227
J. F. Tait, B. Little, S. A. S. Tait, A. Riondel,
C. Flood, E. Joachim, and M. Gut
Estimation of Alternative Metabolic Pathways of Catechol-
amines in Man. . . . . . . . . • . . . . . . . . . . . . . . . . . .. 237
Irwin J. ,Kopin
Use of Tracers in the Study of Biogenic Amine Compart-
ments. • • • ... . . . . . . . • . . • • • . • • . . . . . . . . . . . •. 243
Michael A. Beaven
Evaluation of Rates of Secretion and of Interconversion of
Steroid Hormones. . . • . • . . . . . . . • • • . . . . . . . . . .. 253
Erlio Gurpide and Jonah Mann
Diisopropylfluorophosphate-p32 (DFP_p32) as a Label for
Granulocytes. . . . . . . . . . • . . . . . . . . . . . . . . . . 261
Spencer O. Raab
Clinical Use of Liquid-Type Whole-Body Radioactivity De-
tectors .............. . 267
Hellmuth C. Heinrich
Paired Labeling with 1125 and 1131 • • • • • • • • • • • • • • • • • •• 277
David Pressman
The In Vivo Use of Doubly-Labeled Glucose to Determine
Cerebral Metabolism in Humans. . . . • . . . . . . . . . . .. 279
William Sacks
Carbon-14 Fat Oxidation Test: A New Method for Measuring
Fat Utilization in the Human. . . . . . . . . . . . . . . . 293
Susanne L. von Schuching and Arthur F. Abt
Health Physics
Coordinated Design of Radioisotope Laboratories . . . 303
Chih H. Wang, R. A. Adams, and Wyman K. Bear
Index ... 315
EARLY HISTORY OF CARBON-14
Martin D. Kamen*
The First College
University of California, La Jolla
When, how, and why was carbon-14 discovered? As T. S. Kuhn
has remarked [1], discovery is seldom a single event that can be
attributed wholly to a particular individual, time, or place. He notes
that some discoveries, such as those of the neutrino, radio waves,
and missing isotopes or elements, are predictable and present few
problems as far as establishment of priority is concerned. Others,
such as the discoveries of oxygen, x rays, and the electron, are
unpredictable .. These put the historian in a "bind" when he tries to
decide when, how, where, and by whom the discovery was made.
Much more rarely does he have a basis for an answer to the question
"Why?"
I propose in this account of the "prenatal" history of carbon-14
to provide the answers to my leading questions.t These make a
story which is a fragment of the whole record. That record must
be constructed by future historians who seek to probe the events of
a period in which there has been an unparalleled impact of intellec'
tual curiosity and scientific creativity on the structure of society.
The tremendous outburst of technology in the past half-century,
the result of the rise of nuclear science, has crowned man's quest
for the "philospher's stone" so successfully as to be hardly credible
even to the most optimistic alchemist. Tracer methodology, an
offspring of nuclear science, has provided essential support for the
ever-widening and deepening knowledge of structure and function in
biological systems, expressed as the dynamic science of molecular
biology.
*Dr. Kamen has been an active participant in the Advances in Tracer Methodology Sym
posia. He presented a paper at the Third Symposium and was Chairman of the Fourth
and Seventh Symposia. His fascinating account of the discovery of carbon-14 is of in
terest to everyone working with isotopes. This article was adapted from a paper which
he presented at a meeting of the American Chemical Society in Los Angeles in April 1963,
when he received the Society's 1963 Award for Nuclear Applications in Chemistry, and is
reprinted with permission, having appeared in J. Chern. Ed .. May 1963, and in Science
140:584 .. (1963). Copyright 1963 by the American Association for the Advancement of
Science.
tFor a brief account, concerned mainly with technical aspects, see [2].
2 Early History of Carbon-14
These developments have profound, but unknown, implications for
the future of our social structures. They obviously bring with them
an unexampled load of grist for the mills of cultural historians,
social scientists, and philosophers. Perhaps the novelists will dig
into the record of these exciting times for fresh insights into the
age-old drives of mankind.
Carbon-14, the long-lived carbon isotope, is the most important
single tool made available by tracer methodology, because carbon
occupies the central position in the chemistry of biological systems.
Thus it plays, and will continue to play, an essential role in the
elucidation of biochemical mechanisms, knowledge of which is essen
tial in the further development of molecular biology. Obviously, the
circumstances surrounding its discovery are valid objects of interest
for the historian.'"
INITIAL PHASES, 1934-1936
In the early 1930's, nuclear physics, immersed in the great
traditions of the Cambridge school led by Ernest Rutherford, was
concerned primarily with observations of processes associated with
the scattering of elementary nuclear particles by various atomic
nuclei. Reports in those times show painstaking determinations of
range-energy relations for the fundamental projectiles (protons,
deuterons, alpha particles). The energies used did not exceed
approximately 10 MeV, because of the limitations set by the rela
tively primitive accelerators and by the radiation characteristics
of the naturally radioactive materials that were available. The
rationale for such work, which often involved tedious attention to
detail and much labor, was that if enough precise facts were put
together, accurate binding energies for nuclei could be deduced.
From these energies, it was reasoned, there could be derived a
solid basis for further attack on the problem of the nature of nuclear
forces.
By 1933, such data-binding energies, angular distributions in
scattering experiments, and so on-had demonstrated that nuclear
forces could be described as analogous to saturation exchange
forces like those postulated previously for chemical bonding. The
so-called "alpha-particle" model of the nucleus already contained
the seeds of what was to be the full-fledged modern "shell" theory
of nuclei, to be developed later by Maria Mayer, Eugene Feenberg,
and others.
*1 wish to record a deeply felt personal indebtedness to two former associates. Dr. Franz
N. D. Kurie and Dr. Samuel Ruben. The interruption of Kurie's career by a debilitating
illness a few years ago deprived nuclear 'physics of one of its foremost investigators.
The untimely death of Ruben at the age of 29. while he was engaged in research on chemi
cal warfare in 1943. was an unmitigated catastrophe for modern biochemistry.
M.D. Kamen 3
As to my part in this, I was a young, eager student and had just
begun doctoral research, using the Wilson cloud chamber to study the
angular distribution of neutrons scattered in collisions with protons
and other nuclei. These researches were part of a general program
initiated in the laboratory of W. D. Harkins in the chemistry depart
ment at the University of Chicago [3). My deCision to work in this
field was largely a result of the influence of D. M. Gans, Harkins'
associate and an assistant professor in the department.*
Most significantly for this history, similar work was also under
way at Yale, where F. N. D. Kurie, investigating neutron-induced
disintegration of light elements, had obtained certain anomalous re
sults for the angular distributions of protons in collisions with
neutrons. In 1934 he proposed a radical interpretation [4) of certain
events he noted in the cloud chamber. When nitrogen was exposed
to fast neutrons, for instance, he noted that in some cases the ejected
nucleus produced a very long, thin track. This he ascribed to a
proton, rather than to an alpha particle. Thus, he supposed that the
usual reaction, N14(n,He4)Bl1, was accompanied by a less frequent
but readily observable reaction, N14(n, H1) C14. (As far as I am aware,
this is the first suggestion in the literature that C14 might exist.)
Kurie also suggested, however, that the tracks he was observing
might arise from H2, or even H3, and thus that the reactions
N14(n,H2)C13 and N14(n,H3)C12 were also possibilities. Infact, he felt
the reactions with emission of H2 and H3 were the more likely be
cause they resulted in nuclei of known stability.
What was radical about Kurie's suggestion was the idea that
something other than an alpha particle could emerge in a disinte
gration of a nucleus such as N14. But the physicists at the time
assumed from their everday experience that the alpha particle was
much the most likely nucleon to be formed in such a nuclear reac
tion. This belief found a ready basis in the relatively great stability
of the alpha particle, which was considered to exist as an entity in
all nuclei because of its relatively enormous binding energy per
nucleon, and because invariably in natural radioactivity it was the
only heavy nucleon ejected.
In the meantime, T. W. Bonner and W. M. Brubaker [li) published
observations on the energies of recoils induced by neutrons in in
elastic collisions with nitrogen nuclei. Assuming the usual reaction,
N14(n,He4)Bl1, they calculated from the mass values given by Hans
Bethe ~) that Q, the heat of reaction, was about 1.5 Mev. Most
significantly, however, Bonner and Brubaker [4.5] and W. Chadwick
and M. Goldhaber [7) independently reported that disintegration
of N14 occurred also with slow neutrons.
*1 am happy to record my great debt to Dr. Gans and also to Dr. Henry W. Newson. my
immediate predecessor in the research.