2.

2 The Determination of Lignin

C.W. DENCE

2.2.1 Introduction

The determination of lignin is an analysis that is performed routinely for charac-

terizing lignocellulosic material, for assessing the effects of chemical, physical,
and biological treatments of wood and pulp, for monitoring effluents in wood
processing industries and, in the case of chemical pulps, for estimating bleach
chemical requirements. The general approaches followed in this determination,
together with their associated procedural variations, have been outlined and
thoroughly detailed by several authors (Brauns 1952, Brauns and Brauns 1960,
Browning 1967a, Pearl 1967). Therefore, the content of this section is confined
mainly to a description and discussion of the preferred and/or widely accepted
methods for determining lignin when present in a variety of representative
lignified materials.
In considering the various methods proposed for the determination of
lignin, it is important to bear in mind that none can be regarded as totally
satisfactory. This situation arises from the fact that lignin has yet to be isolated
in a pure or unmodified condition. As a result there is no definitive structural
formula of lignin currently available upon which to base the development of a
rational and quantitative measurement or to assess accurately the validity of
the methods currently available. In addition, many of the methods which are
commonly applied to the determination of lignin have fundamental defects
which seriously undermine their accuracy. For these reasons, values reported in
the literature may not always provide a reliable indication of the "true" lignin
content of a material.
In organizing the subject matter for this chapter, it was found expedient
to divide the presentation into two main subsections, one dealing with the
analytical methods applicable to lignin when initially present as a component of
a solid mixture and the other focusing on the determination of lignin in solution.

2.2.2 Direct Methods

2.2.2.1 Acid Hydrolysis of Wood and Pulp

A substantial proportion of the methods falling into this category consist of the
hydrolysis and solubilization of the carbohydrate component of the lignified
material, leaving the lignin as a residue which is determined gravimetrically.

Springer Series in Wood Science

Methods in Lignin Chemistry
(Edited by S.Y. Lin and C.W. Dence)
© Springer-Verlag Berlin Heidelberg 1992
34 c.w. DENCE

Interfering extraneous materials are removed prior to the hydrolysis step by

extraction with an appropriate solvent or combination of solvents. Hydrolysis
of the polysaccharides is catalyzed by a variety of strong mineral acids, applied
either individually or in combinations. The range of possibilities has been
surveyed by Browning (1967a) and Pearl (1967), who also included detailed
descriptions of many of the proposed methods.
The most widely applied variation of the acid hydrolysis approach employs
sulfuric acid to promote carbohydrate hydrolysis. The lignin isolated in this
treatment is referred to as acid-insoluble lignin, or more commonly as Klason
lignin, in recognition of the wood chemist who pioneered the development of
the isolation method. In its present modified form the method consists basically
of two steps: treatment of the lignified material with cold (1O-15°C) 72%
sulfuric acid for a specified time at 20°C followed by dilution of the acid to
3.0% and boiling to complete the hydrolysis. The procedure described below is
essentially the Official Test Method T222 om-83 of T APPI (Technical Associa-
tion of the Pulp and Paper Industry 1983) for the determination of lignin in
wood and pulp.

2.2.2.2 Determination of Acid-Insoluble (Klason) Lignin in Wood and Pulp

Reagents

Sulfuric acid, 72% H 2S0 4 solution, 24 ± O.lN, sp, gr. 1.6338 at 200W is
prepared by carefully pouring 665 ml of concentrated H 2S0 4 (95.5 to 96.5%, sp.
gr. 1.84) into 300 ml of water and, after cooling, diluting to 1000 ml. The
strength is adjusted to 24 ± 0.1 N after titration with standard alkali, or
me!lsurement of specific gravity. A variation of 0.1 % in the strength of acid
at this concentration causes a change of 0.0012 in specific gravity. The acid is
cooled to 1O-l5°C before use.
Ethanol-benzene mixture is prepared by mixing one volume of approxi-
mately 95% ethanol and two volumes of c.P. benzene.

Sample Preparation

The wood specimen is ground to pass a O.4mm (40-mesh) screen. A sample of

the wood meal equivalent to about 5 g on an oven-dry basis is placed in a coarse
porosity alundum or fritted glass extraction thimble and extracted successively in
a Soxhlet apparatus with ethanol-benzene for 6-8h and 95% ethanol for 4h.
Between extractions the wood meal is transferred to a Buchner funnel, where
the excess solvent is removed by suction and washing with ethanol.
At the conclusion of the extraction with 95% ethanol, the wood meal is
once again transferred to the Buchner funnel and the excess solvent removed by
suction and washing with distilled water. The wood is then transferred to a 1000-
ml Erlenmeyer flask, 500 ml of boiling distilled water is added and the flask is