Automated Analysis of Amino Sugars using the Elson-Morgan Reaction

241 Pages·2015·10.82 MB·English

by  Lee Van Lenton

Yale University
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Yale Medicine Thesis Digital Library School of Medicine
Spring 5-31-1966
Automated Analysis of Amino Sugars using the
Elson-Morgan Reaction
Lee Van Lenten
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Van Lenten, Lee, "Automated Analysis of Amino Sugars using the Elson-Morgan Reaction" (1966).Yale Medicine Thesis Digital Library.
6.
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ABSTRACT
A combined chromatographic and analytical system is described
for the specific determination of amino sugars.
An adaptation of the Elson-Morgan reaction has been made using
the equipment of the Technicon AutoAnalyzer. The colorimetric
procedure is based partly on the work of Lewy and McAllan . The
Elson-Morgan reaction is a two-stage reaction which involves first
the reaction of the amino sugar with acetic anhydride or 2,4pentane-
dione and the formation of cyclic intermediates by heating in an al
kaline medium. In the second stage, the medium is acidifed and the
intermediates coupled with p_-dimethylaminobenzaldehyde to give a
colored product.
In the automated procedure developed, acetylation is performed
with an acetic acid - N,N-dimethylacetamide reagent and the cycliza-
tion is accomplished at elevated temperatures in a basic borate solu
tion. The reaction mixture is then acidified by the addition of an
acetic acid - formic acid - HC1 - water reagent and the p-dimethyl-
aminobenzaldehyde is added in an alcoholic solution. All reagents
are stable for several weeks.
Two chromatographic systems have been developed for the separation
of amino sugars without modification of the chromatographic section
of the Technicon Amino Acid Analyzer. These systems have distinctly
different elution patterns. It is possible to carry out simultaneous
ninhydrin and Elson-Morgan analyses of the chromatographic effluents.

AN AUTOMATED ANALYSIS OF AMINO SUGARS
USING THE ELSON-MORGAN REACTION
by
Lee Van Lenten
A.B., Colgate University, i960
A Thesis
Submitted to the Faculty
of the Yale University School of Medicine
in Candidacy for the Degree of
Doctor of Medicine
Department of Biochemistry
Yale University
1966
J 7 / 7
ACKNOWLEDGMENTS
This investigation was carried out in the laboratory of Dr. Fritz
Lipmann at the Rockefeller University under the guidance of Dr. John
D. Gregory.
I wish to thank Dr. Detlev W. Bronk, President of the Rockefeller
University, for the opportunities made available in the stimulating
atmosphere of the University and Dr. Lipmann for the opportunity of
working in his well-appointed laboratory. Dr. Gregory has been most
generous with his teaching, by precept and example, his encouragement
and friendship, and his willingness to let me wander down blind alleys
and make my own mistakes.
At Yale University Dr. Joseph S. Fruton has been kind enough to
sponsor this thesis and has given me skilled assistance in the
preparation of this manuscript.
My thanks also go to Mr. George D. Winter of the Technicon
Chromatography Corporation for his technical advice and assistance.
Last, but by no means least, I thank my wife, Beth, for her
moral support without which this investigation would never have been
completed, my sister, Ellen, for the art work in the figures, and
my parents, who are what parents should be.