Table Of ContentMethods in Carbohydrate Chemistry
Volume I : Analysis and Preparation of Sugars
Volume II : Reactions of Carbohydrates
Volume III : Cellulose
Volume IV: Starch
Volume V : General Polysaccharides
Volume VI : General Carbohydrate Methods
Volume VII : General Methods, Glycosaminoglycans,
and Glycoproteins
METHODS IN
Carbohydrate Chemistry
EDITORS
Roy L. Whistler James N. BeMiller
Department of Biochemistry Department of Chemistry
Purdue University and Biochemistry
Lafayette, Indiana Southern Illinois University
at Carbondale
Carbondale, Illinois
VOLUME VII
General Methods, Glycosaminoglycans, and Glycoproteins
ACADEMIC PRESS, INC.
(Harcourt Brace Jovanovich, Publishers)
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Toronto Montreal Sydney Tokyo Säo Paulo
Copyright © 1976, by Academic Press, Inc.
all rights reserved.
no part of this publication may be reproduced or
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ACADEMIC PRESS, INC.
Orlando, Florida 32887
United Kingdom Edition published by
ACADEMIC PRESS, INC. (LONDON) LTD.
24/28 Oval Road, London NW1 7DX
Library of Congress Cataloging in Publication Data
Whistler, Roy Lester.
Methods in carbohydrate chemistry.
Includes bibliographies.
CONTENTS: v. 1 Analysis and preparation of
sugars.—v.2. Reactions of carbohydrates.—v. 3.
Cellulose, [etc.]
1. Carbohydrates—Collected works. I. Wolfrom,
Melville Lawrence, (date) II. BeMiller,
James Ν. III. Title
QD321.W568 547'.78 61-18923
ISBN 0-12-746207-4 (v. 7)
PRINTED IN THE UNITED STATES OF AMERICA
84 85 86 87 98 7 6 5 4 3 2
Contributors to Volume VII
Article numbers are shown in parentheses following the names of the contributors.
CHARLES W. BAKER (26), Department of JOHN F. CODINGTON (39), Laboratory for
Cereal Chemistry and Technology, Carbohydrate Research, Departments
North Dakota State University, Fargo, of Biological Chemistry and Medicine,
North Dakota Harvard Medical School, and Massa-
chusetts General Hospital, Boston,
DAVID C. BAKER (1), Department of
Massachusetts
Chemistry, The Ohio State University,
Columbus, Ohio JAMES CONCHIE (34), Department of Car-
bohydrate Biochemistry, Rowett Re-
DONALD A. BAKER (12), ICN Pharma-
search Institute, Bucksburn, Aberdeen,
ceuticals, ICN Nucleic Acid Research
Scotland
Institute, Irvine, California
H. E. CONRAD (15, 25), Department of
JAMES N. BEMILLER (16), Department of
Biochemistry, University of Illinois,
Chemistry and Biochemistry, Southern
Urbana, Illinois
Illinois University at Carbondale, Car-
bondale, Illinois W. E. DICK, Jr. (3), Northern Regional
Research Laboratory, U.S. Department
L. J. BLACIK (20), Research-in-Aging
of Agriculture, Peoria, Illinois
Laboratory, Veterans Administration
Hospital, Downey, Illinois, and De- W. M. DOANE (8), Northern Regional
partment of Biochemistry, North- Research Laboratory, U.S. Department
western University Medical School, of Agriculture, Peoria, Illinois
Chicago, Illinois
F. DOWNS (35, 40, 41, 42), Department of
Μ ΑΧΙΝΈ S. BoRCHERDiNG (20), Research- Chemistry, H. Lehman College, Bronx,
in-Aging Laboratory, Veterans Admin- New York
istration Hospital, Downey, Illinois,
G. ENTLICHER (33), Department of Bio-
and Department of Biochemistry,
chemistry, Charles University, Prague,
Northwestern University Medical
Czechoslovakia
School Chicago, Illinois
JANA FURDOVÄ (5), Division of Biological
MOIRA BREEN (20), Research-in-Aging Sciences, National Research Council,
Laboratory, Veterans Administration
Ottawa, Canada
Hospital, Downey, Illinois, and De-
partment of Biochemistry, North- GARY R. GRAY (27), Department of
western University Medical School, Chemistry, University of Minnesota,
Chicago, Illinois Minneapolis, Minnesota
D. R. BUNDLE* (17), Department of S. HANESSIAN (11, 13), Department of
Chemistry, University of Alberta, Chemistry, University of Montreal,
Edmonton, Alberta, Canada Montreal, Quebec, Canada
J. A. CIFONELLI (23), Department of DONALD E. HOARD (4), P-L Biochemicals,
Pediatrics, The University of Chicago, Inc., Milwaukee, Wisconsin
Chicago, Illinois J. E. HODGE (3), Northern Regional
Research Laboratory, U.S. Department
of Agriculture, Peoria, Illinois
* Present address : Division of Bio-
logical Sciences, National Research MICHAEL J. HOLMAN (4), P-L Biochemi-
Council of Canada, Ottawa, Canada. cals, Inc., Milwaukee, Wisconsin
IX
χ CONTRIBUTORS ΤΟ VOLUME VII
DEREK HORTON (1, 14), Department of TAKASHI MURACHI* (31), Department of
Chemistry, The Ohio State University, Biochemistry, Nagoya City University,
Columbus, Ohio School of Medicine, Nagoya, Japan
SIDNEY P. KENT (43), Department of A. NEUBERGER (37), Department of Bio-
Pathology, University of Alabama in chemistry, Charing Cross Hospital
Birmingham, Birmingham, Alabama Medical School, London, England
J. KOCOUREK (33), Department of Bio- C. J. No1" (2), School of Chemistry,
chemistry, Charles University, Prague, University of New South Wales,
Czechoslovakia Kensington, New South Wales,
Australia
P. LAV ALLÉE (11), Department of Chem-
istry, University of Montreal, Montreal, ARTHUR S. PERLIN (19), Department
Quebec, Canada of Chemistry, McGill University,
Montreal, Quebec, Canada
R. U. LEMIEUX (17), Department of Chem-
istry, University of Alberta, Edmonton, MALCOLM B. PERRY (5, 6, 7, 10), Division
Alberta, Canada of Biological Sciences, National Re-
search Council, Ottawa, Canada
SU-CHEN LI (38), Delta Regional Primate
Research Center, Covington, Louisiana KERSTIN D. PHILIPS (14), Department of
Chemistry, The Ohio State University,
YU-TEH LI (38), Department of Bio-
Columbus, Ohio
chemistry, Tulane University, New
Orleans, Louisiana W. PIGMAN (35, 40, 41, 42), Department
of Biochemistry, New York Medical
BENGT LINDBERG (24), Department of
College, Valhalla, New York
Organic Chemistry, Arrhenius Labo-
ratory, University of Stockholm, THOMAS H. PLUMMER, Jr., (29), Division
Stockholm, Sweden of Laboratories and Research, New
York State Department of Health,
ALFRED LINKER (18), Veterans Adminis-
Albany, New York
tration Hospital, Salt Lake City, Utah
B. S. SHASHA (8), Northern Regional
KEYES B. LINSLEY (39), Laboratory for
Research Laboratory, U.S. Department
Carbohydrate Research, Departments
of Agriculture, Peoria, Illinois
of Biological Chemistry and Medicine,
Harvard Medical School, and Massa- VIRGINIA SHEPHERD (30), Department of
chusetts General Hospital, Boston, Biochemistry, University of Iowa, Iowa
Massachusetts City, Iowa
JÖRGEN LÖNNGREN (24), Department of J. E. SHIVELY (25), Department of Bio-
Organic Chemistry, Arrhenius Labora- chemistry, University of Illinois,
tory, University of Stockholm, Urbana, Illinois
Stockholm, Sweden
JACK M. SIEGEL (4), P-L Biochemicals,
R. D. MARSHALL (37), Department of Inc., Milwaukee, Wisconsin
Chemical Pathology, St. Mary's Hospi-
tal Medical School, London, England
MARTIN B. MATHEWS (21), Department of * Present address : Department of Clini-
Pediatrics, The University of Chicago, cal Science, Faculty of Medicine, Kyoto
Chicago, Illinois University, Kyoto, Japan.
REX MONTGOMERY (30), Department of f Present address : Arnotts Biscuits Pty.,
Biochemistry, University of Iowa, Iowa Ltd., Homebush, New South Wales,
City, Iowa Australia.
CONTRIBUTORS TO VOLUME VII xi
CYLA SILBER (39), Laboratory for Carbo- R. L. TAYLOR (25), Department of
hydrate Research, Departments of Bio- Biochemistry, University of Illinois,
logical Chemistry and Medicine, Urbana, Illinois
Harvard Medical School, and Massa-
CHARLES G. TINDALL, Jr. (1), Department
chusetts General Hospital, Boston,
of Chemistry, The Ohio State Univer-
Massachusetts
sity, Columbus, Ohio
R. A. SITTIG (20), Research-in-Aging
RICHARD L. TOLMAN* (12), ICN Pharma-
Laboratory, Veterans Administration
ceuticals, ICN Nucleic Acid Research
Hospital, Downey, Illinois, and Depart-
Institute, Irvine, California
ment of Biochemistry, Northwestern
University Medical School, Chicago, D. TRIMNELL (8, 9), Northern Regional
Illinois Research Laboratory, U.S. Department
of Agriculture, Peoria, Illinois
ROBERT G. SPIRO (28, 32, 36), Depart-
ments of Biological Chemistry and H. G. WEINSTEIN (20), Research-in-Aging
Medicine, Harvard Medical School, Laboratory, Veterans Administration
and the Elliott P. Joslin Research Hospital, Downey, Illinois, and De-
Laboratory, Boston, Massachusetts partment of Biochemistry, North-
western University Medical School,
A. P. A. STAUB (13), Department of
Chicago, Illinois
Chemistry, University of Montreal,
Montreal, Quebec, Canada ROY L. WHISTLER (26), Department of
Biochemistry, Purdue University,
J. D. STEVENS (2), School of Chemistry,
Lafayette, Indiana
University of New South Wales, Ken-
sington, New South Wales, Australia DAVID T. WILLIAMS (10), Division of Bio-
logical Sciences, National Research
AUDREY L. STONE (22), National Institute
Council, Ottawa, Canada
of Mental Health, Bethesda, Maryland
EUNICE S. WU (4), P-L Biochemicals, Inc.,
Ε. I. STOUT (8), Northern Regional Milwaukee, Wisconsin
Research Laboratory, U.S. Department
of Agriculture, Peoria, Illinois
MASON G. STOUT (4), ICN Pharmaceu-
tical, ICN Nucleic Acid Research Insti-
tute, Irvine, California
NORIKO TAKAHASHI (31), Department of * Present address : Merck Sharp &
Biochemistry, Nagoya City University, Dohme Research Laboratories, Rahway,
School of Medicine, Nagoya, Japan New Jersey.
Preface
In continuing to compile generally useful methods, the editors research
the literature with an awareness of the need for procedures that work.
Sometimes these needs suggest inclusion in the collection of procedures
published earlier in either this treatise or in primary journals, but substan-
tially improved since their original publication. More often, however, new
methods of preparation, analytical procedures, and reactions that continue
to form the fabric of the expanding body of carbohydrate chemistry are
selected in anticipation that they will aid those working with carbohydrates
in solving new or old problems.
There is a special emphasis in this volume on methods used with glyco-
proteins and glycosaminoglycans. Included are sixteen methods applicable
to glycoproteins and ten to glycosaminoglycans. Additional methods from
both these areas will appear in future volumes.
The remainder of the volume contains general methods. We have also
included a list of carbohydrate methods that can be found in other collec-
tions of methods but which have not been included in this treatise.
It has been a pleasure to work with the contributors to this volume. We
hope that carbohydrate chemists and biochemists will continue to make
suggestions for the contents of future volumes.
Roy L. Whistler
James N. BeMiller
xiii
Outline of Volume I
ANALYSIS AND PREPARATION OF SUGARS
Section I. General Methods
Carbohydrate Laboratory Techniques ; Chromatography
Section II. Monosaccharides
Trioses ; Tetroses ; Pentoses ; Hexoses; Heptoses; Deoxy Sugars; Amino
Sugars; Branched-Chain Sugars; C 14-Labeled Sugars; Inososes
Section III. Oligosaccharides
Reducing Oligosaccharides; Non-Reducing Oligosaccharides
Section IV. Analyses
Section V. Color Reactions of Carbohydrates
Section VI. Physical Measurements
Outline of Volume II
REACTIONS OF CARBOHYDRATES
Section I. General Considerations
Section II. Oxidation Products
Aldonic Acids; Uronic Acids; Aldaric Acids; Hexulosonic Acids;
Esterification
Section III. Reduction Products
Section IV. Derivatives of Nitrogen Bases
Glycosylamines ; Phenylhydrazine Derivatives
Section V. Etherification
Methyl Ethers; Benzyl Ethers; Triphenylmethyl Ethers; Sugar Anhy-
drides; Anhydroalditols; Diketohexose Dianhydrides; Dealkylation
Section VI. Esterification
Acetates; Acylglycosyl Halides; Benzoates; Carbanilates; Carbonates;
Sulfonates ; Nitrates ; Phosphates ; Sulfates
Section VII. Acetalation
Cyclic Acetal Derivatives ; Glycosidation ; Other Acetals
Section VIII. Unsaturated Sugars
Section IX. Aldosuloses (Osones)
xv
xvi OUTLINE OF VOLUMES I-VI
Section X. Acyclic Monosaccharides
Section XI. Thiosugars
Section XII. Configurational Inversion
Section XIII. The Oxo Reaction
Section XIV. Grignard and Friedel-Crafts Reactions
Section XV. Saccharinic Acids
Section XVI. Determination of Isotopic Carbon Distribution in Aldoses
Section XVII. Selected Methods in Carbohydrate Chemistry Found in
Other Collections
Outline of Volume III
CELLULOSE
Section I. Preparation of Cellulose
Section II. Chemical Analysis
Section III. Physical Analysis
Section IV. Degradation of Cellulose
Section V. Cellulose Esters: Preparation, Properties, Reactions, and
Analysis
Section VI. Cellulose Ethers: Preparation, Properties, Reactions, and
Analysis
Section VII. Microscopy
Section VIII. Laboratory Equipment
Section IX. Preparation and Analysis of C 14-Labeled Cellulose
Outline of Volume IV
STARCH
Section I. Preparation of Starch and Starch Fractions
Whole Starch; Starch Fractions
Section II. Chemical Analyses
Whole Starch and Modified Starches; Starch Fractions; Starch
Hydrolyzates
OUTLINE OF VOLUMES I-VI xvii
Section III. Physical Analyses
Whole Starch and Modified Starches; Starch Pastes; Starch Fractions;
Starch Hydrolyzates
Section IV. Microscopy
Section V. Starch Degradations
Section VI. Starch Derivatives and Modifications
Reactivity; Esters; Ethers; Oxidation
Outline of Volume V
GENERAL POLYSACCHARIDES
Section I. General Isolation Procedures
Section II. Polysaccharide Preparations
Section III. Chemical Analyses
Section IV. Physical Analyses
Section V. Molecular Weight Determinations
Section VI. Structural Methods
Section VII. Derivatives
Oxidation and Reduction; Esterification and Deacylation; Etherifica-
tion
Section VIII. Selected Methods Found in Other Collections
Outline of Volume VI
GENERAL CARBOHYDRATE METHODS
Section I. Separation and Analysis
Section II. Preparation of Mono- and Polysaccharides and Their Derivatives
Section III. Oxidation
Section IV. Acyclic Sugars
Section V. Etherification
