Table Of ContentSpringer Series in Wood Science
Editor: T. E. Timell
M. H. Zimmermann
Xylem Structure and the Ascent of Sap (1983)
1. F. Siau
Transport Processes in Wood (1984)
R.R. Archer
Growth Stresses and Strains in Trees (1986)
W.E. Hillis
Heartwood and Tree Exudates (1987)
S. Carlquist
Comparative Wood Anatomy (1988)
L. W. Roberts / P. B. Gahan / R. Aloni
Vascular Differentiation and Plant Growth Regulators (1988)
C. Skaar
Wood-Water Relations (1988)
1.M. Harris
Spiral Grain and Wave Phenomena in Wood Formation (1989)
B. 1. Zobel/ 1. P. van Buijtenen
Wood Variation (1989)
P. Hakkila
Utilization of Residual Forest Biomass (1989)
1.W. Rowe
Natural Products of Woody Plants (1989)
K.-E.L. Eriksson/R.A. Blanchette/P. Ander
Microbial and Enzymatic Degradation of Wood
and Wood Components (1990)
R. A. Blanchette / A. R. Biggs
Defense Mechanisms of Woody Plants Against Fungi (1992)
S. Y. Lin / C. W. Dence
Methods in Lignin Chemistry (1992)
G. Torgovnikov
Dielectric Properties of Wood and Wood-Based Materials (1993)
Stephen Y. Lin Carlton W. Dence (Eds.)
Methods
in Lignin Chemistry
With 190 Figures
Springer-Verlag
Berlin Heidelberg New York London Paris
Tokyo Hong Kong Barcelona Budapest
Dr. STEPHEN Y. LIN
LignoTech USA Inc.
100 Highway 51 South
Rothschild, WI 54474, USA
Professor Emeritus Dr. CARLTON W. DENCE
600 Black Walnut Court
Hendersonville, NC 28792, USA
Series Editor:
T.E. TIMELL
State University of New York
College of Environmental Science and Forestry
Syracuse, NY 13210, USA
Cover: Transverse section of Pinus lambertiana wood. Courtesy of Dr. Carl de Zeeuw,
SUNY College of Environmental Science and Forestry, Syracuse, New York
ISBN-13 :978-3-642-74067-1 e-ISBN-13:978-3-642-74065-7
DOT: 10.1007/978-3-642-74065-7
Library of Congress Cataloging-in-Publication Data. Methods in lignin chemistry / Stephen Y. Lin,
Carlton W. Dence (eds.). p. em. - (Springer series in wood science) Includes bibliographical
references and index. ISBN-13:978-3-642-74067-1 1. Lignin.
I. Lin, Stephen Y. II. Dence, Carlton w. III. Series. TS933.L5M47 1992 661'.802-dc20 92-6677
This work is subject to copyright. All rights are reserved, whether the whole or part of the material
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tained from Springer-Verlag. Violations are liable for prosecution under the German Copyright Law.
© Springer-Verlag Berlin Heidelberg 1992
Softcover reprint of the hardcover 1st edition 1992
The use of general descriptive names, registered names, trademarks, etc. in this publication does
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Typesetting: Best-set Typesetter Ltd., Hong Kong, and K + V Fotosatz GmbH, Beerfelden
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Dedication
As this book neared completion, we learned of the sudden and unex
pected death of Dr. Kyosti V. Sarkanen, Professor of Wood Chem
istry at the University of Washington. Just prior to his passing, Dr.
Sarkanen had reviewed several portions of this book and was in the
process of co-authoring the section on ozonolysis. Ironically, these
activities coincided with the culmination of a professional career that
spanned nearly five decades during which he established himself as
a dominant figure in the field of wood and, in particular, lignin
chemistry.
It is not our intention to echo here the numerous eUlogies marking
his passing or even to attempt to highlight his scientific accomplish
ments. It suffices to say that those of us who were fortunate to know
him personally as a teacher, researcher, adviser, colleague, or co
worker are fully aware of the enormous impact he made on the devel
opment of wood science; others who come to know him indirectly
through his published work have the opportunity to gauge his uni
queness for themselves. As co-editors, we deem it a privilege to
dedicate this book to the memory of Dr. Sarkanen as a fitting tribute
to a colleague whose research contributions and scientific insight pro
vided the basis for the shaping and refining of many of the concepts
expressed within its covers.
CARLTON W. DENCE and STEPHEN Y. LIN
Preface
The structural complexity of lignin has continually challenged the in
genuity of researchers to develop suitable methods for its charac
terization prior to and following a wide variety of chemical, biologi
cal, and physical treatments. Initially, activity along these lines was
fueled by a desire to interpret technical delignification (Le., pulping)
processes in terms of accompanying structural changes in the lignin.
Subsequently, increasingly wide ranging, in-depth investigations on
the structure and reactivity of lignin exposed the inadequacy of many
of the methods currently in use and underscored the ever-continuing
need to develop new methods capable of solving the unique analytical
problems associated with lignin. Characteristically, such methods
should be selective, sensitive, suitable for quantitative measurements,
and capable of being applied directly to, and without destruction of,
the lignin or lignocellulose sample. One notable example of the head
way being made in reaching this objective is the relatively recent devel
opment and refinement of methods based on the use of sophisticated
instrumentation, e.g., lH_ and 13C-NMR spectroscopy. Although the
utility of many of these and other recently developed methods de
scribed in this book has yet to be fully and satisfactorily exploited,
we believe that progress already made in this direction will continue
and most likely accelerate.
The decision to produce this book was prompted mainly by the
acknowledged need for an up-to-date, single source compilation of
lignin methodology. Hitherto, this need was, in part, satisfied by B. L.
Browning's two-volume set of books, Methods of Wood Chemistry,
which was published nearly 30 years ago. The recognition that these
books had, in part, become outdated was a major consideration in ar
riving at our decision to undertake to assemble a book reflecting the
current status of lignin methodology. While we envision Methods in
Lignin Chemistry as broadly serving the needs of professionals in
academia, the pulp and paper industry, and allied industries, we also
anticipate that the format adopted here for presenting the informa
tion will be particularly useful to those beginning a career in wood
and/or lignin chemistry and to those with no previous background in
or experience with lignin or lignocellulosics and whose interest is pro
jected to be on a one-time-only, short-term basis.
VIII Preface
The core element common to each chapter in this book is a de
tailed, step-by-step description of what, in the judgment of the
author(s), is the preferred method for conducting a specific analysis
or operation. This particular format was selected to enable the reader
to use the book essentially as a laboratory manual thereby obviating
the need to search for and evaluate methods scattered throughout the
literature. The methods included are a blend of the traditional and the
contemporary. In some instances, what appeared to be promising
analytical methods were bypassed in favor of more widely accepted
methods on the grounds that they had not been field-tested sufficient
ly to authenticate their reliability. The description of the preferred
method is augmented by supporting information including, for exam
ple, a survey of alternative methods, a discussion of the principle
underlying the preferred method, the advantages and disadvantages
of the method, the type and significance of information provided by
the analysis, and the presentation of typical analytical data.
From the onset of this undertaking it was obvious to us that the
range of topics we wished to include virtually necessitated our draw
ing on the expertise of a considerable number of individuals well
versed in the principles and applications of the selected methodology.
We were also aware that the adoption of a multi-author format entail
ed risks that might impact negatively on the uniformity, general
quality, and timetable for completion of the book. Without question,
some of these concerns did prove to be justified. On the other hand,
when this book is viewed in its totality, we believe that the overall
quality and usefulness of its contents vindicates our decision to pro
ceed as we did. With this in mind, we commend the international
team of authors for service well performed in helping to make
Methods in Lignin Chemistry a reality. Others who played important
roles in its creation are acknowledged below.
STEPHEN Y. LIN
CARLTON W. DENCE
Acknowledgments
We are indebted to the following persons who generously contributed
their time to review and recommend revision for several chapters in
this book: F. Agblevor, L. Amos, S. Black, J. Boxell, H. L. Chum,
R. Evans, D. A. I. Goring, M. Himmel, D. K. Johnson, L. Landucci,
J. Obst, S. Omori, M. G. Paice, K. V. Sarkanen, and T. P. Schultz. We
also acknowledge with gratitude the special contributions of
T. E. Timell, Series Editor, who, in addition to reviewing and editing
each manuscript, advised us on matters related specifically to organi
zation, language, and style conventions during the entire period in
which material for the book was being assembled.
Our sincere appreciation is also expressed to Sarah Remon for her
valued service in retyping promptly and accurately many of the edited
and revised manuscripts.
Finally, we individually wish to thank G. A. Rasmussen of
LignoTech USA, Inc. (SYL) for his support and encouragement in this
undertaking and the SUNY College of Environmental Science and
Forestry (CWD) for permission to discharge attendant editorial
responsibilities concurrently with customary academic duties.
Rib Mountain, WI STEPHEN Y. LIN
Hendersonville, NC CARLTON W. DENCE
August 1992
Contents
Chapter 1 Introduction ............................ .
1 Introduction ................................. . 3
C. W. DENeE and S. Y. LIN
1.1 General Structural Features of Lignin ............ 3
1.2 Variations in Concentration and Composition
of Lignin in Xylem ............................ 6
1.3 Isolation and Recovery of Lignin ................ 7
1.4 Physical Properties of Lignin ................... 8
1.5 Structure-Modifying Reactions .................. 9
1.5.1 Modification of the Aromatic Ring. . . . .. . . . . . . . . 9
1.5.1.1 Electrophilic Substitution ....................... 9
1.5.1.2 Conversion of Aromatic Rings to Nonaromatic
Cyclic Structures .............................. 11
1.5.1.3 Conversion of Cyclic to Acyclic Structures ........ 11
1.5.1.4 Ring Coupling and Condensation Reactions ...... 12
1.5.2 Modification of the Propanoid Side Chain ....... 12
1.5.2.1 Cleavage of Ether Bonds ....................... 12
1.5.2.2 Cleavage of Carbon-Carbon Bonds .............. 13
1.5.2.3 Substitution Reactions ......................... 14
1.5.2.4 Formation and Elimination of Multiple Bond
Functionalities ................................ 15
1.5.3 Future Analytical Needs ........................ 16
References ............................................ 17
Chapter 2 Detection and Determination ............. 21
2.1 The Detection of Lignin ...................... . 23
1. NAKANO and G. MESHITSUKA
2.1.1 Introduction ................................. . 23
2.1.2 Reagents for the Detection of Lignin ........... . 23
2.1.2.1 Aliphatic Compounds ......................... . 23
2.1.2.2 Phenols and Aromatic Amines ................. . 23
2.1.2.3 Heterocyclic Compounds ...................... . 24
XII Contents
2.1.2.4 Inorganic Reagents ........................... . 24
2.1.2.5 Unclassified Reagents ........................ . 25
2.1.3 Color-Forming Reaction Sequences ............. . 25
2.1.4 Procedures for the Detection of Lignin ......... . 28
2.1.4.1 Color Reaction with Phloroglucinol-Hydrochloric
Acid ....................................... . 29
2.1.4.2 The Maule Color Reaction .................... . 29
2.1.4.3 The Cross and Bevan Color Reaction ........... . 29
2.1.4.4 Color Reaction with Tosyl Chloride/Pyridine,
p-Nitrosodimethylaniline/Potassium Cyanide .... . 30
2.1.4.5 Color Reaction with Quinone Monochlorimide .. . 30
2.1.4.6 Color Reaction with Potassium Nitrosodisulfonate 31
2.1.4.7 Color Reaction Based on the Formation
of Nitrosophenols (Pearl-Benson Method) ....... . 31
References ............................................ 32
2.2 The Determination of Lignin 33
C.W. DENCE
2.2.1 Introduction ................................ . 33
2.2.2 Direct Methods .............................. . 33
2.2.2.1 Acid Hydrolysis of Wood and Pulp ............ . 33
2.2.2.2 Determination of Acid-Insoluble (Klason)
Lignin in Wood and Pulp ..................... . 34
2.2.2.3 Determination of Acid-Soluble Lignin in Wood
and Pulp ................................... . 39
2.2.2.4 Acid Hydrolysis of Annual Plants,
Developing Wood, and Forage ................. . 40
2.2.2.5 Determination of Acid-Insoluble Lignin in Annual
Plants, Developing Wood, and Forage .......... . 41
2.2.3 Indirect Methods ............................ . 42
2.2.3.1 Spectrophotometric Methods .................. . 42
2.2.3.2 Determination of Lignin in Wood and Pulp
by the Acetyl Bromide Method ................ . 44
2.2.3.3 Methods Based on Oxidant Consumption ....... . 48
2.2.3.4 Determination of the Kappa Number of Pulp ... . 50
2.2.4 Determination of Lignin in Solution ........... . 54
2.2.4.1 Determination of Dissolved Lignin by the Modified
Pearl-Benson (Nitrosation) Method ............. . 55
References ........................................... . 58
