Journal of Wood Chemistry and Technology

ISSN: 0277-3813 (Print) 1532-2319 (Online) Journal homepage: http://www.tandfonline.com/loi/lwct20

Relationships between Hemicellulose Composition

and Lignin Structure in Woods

Yu Huang, Linshan Wang, Yuesheng Chao, Deded Sarip Nawawi, Takuya

Akiyama, Tomoya Yokoyama & Yuji Matsumoto

To cite this article: Yu Huang, Linshan Wang, Yuesheng Chao, Deded Sarip Nawawi, Takuya
Akiyama, Tomoya Yokoyama & Yuji Matsumoto (2016) Relationships between Hemicellulose
Composition and Lignin Structure in Woods, Journal of Wood Chemistry and Technology, 36:1,
9-15, DOI: 10.1080/02773813.2015.1039543

To link to this article: http://dx.doi.org/10.1080/02773813.2015.1039543

Published online: 06 Oct 2015.

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 Journal of Wood Chemistry and Technology, 36:9–15, 2016
Copyright C Taylor & Francis Group, LLC
ISSN: 0277-3813 print / 1532-2319 online
DOI: 10.1080/02773813.2015.1039543

RELATIONSHIPS BETWEEN HEMICELLULOSE COMPOSITION AND LIGNIN

STRUCTURE IN WOODS

Yu Huang,1 Linshan Wang,2 Yuesheng Chao,2 Deded Sarip Nawawi,3 Takuya Akiyama,1
Tomoya Yokoyama,1 and Yuji Matsumoto1
1
Laboratory of Wood Chemistry, Department of Biomaterial Sciences, The University of Tokyo,
Tokyo, Japan
2
School of Sciences, Northeastern University, Shenyang, China
3
Department of Forest Products, Bogor Agricultural University, Bogor, Indonesia
Downloaded by [University of Nebraska, Lincoln] at 02:17 10 October 2015

The composition and absolute amount of neutral sugars were determined for 48 hardwood
species (including 17 hardwoods of genera Acacia, 14 hardwoods of genera Eucalyptus, and 17
hardwoods of other genera) and 14 softwood species by alditol-acetate method, and their rela-
tionships to the syringyl ratio (syringyl/(syringyl+guaiacyl)) of lignin, which was determined by
nitrobenzene oxidation, was investigated. In the hardwood species, an increase in the syringy
ratio of lignin correlated with a tendency toward increased xylose/glucose, rhamnose/glucose,
and arabinose/glucose ratios. However, the absolute amount of glucose in hardwood was
maintained in a small range (0.4–0.5 g in 1 g sample), independent of changes in the syringyl
ratio. In the softwood species, with increasing lignin content, the mannose/glucose ratio de-
creased, but the absolute amount of glucose remained almost constant. In both hardwood
and softwood species, a strong correlation was suggested between lignin, indicated by higher
syringy ratio, and hemicellulose, indicated by higher xylan/mannan ratio.

KEYWORDS. Hardwood, hemicellulose composition, lignin structure, softwood, syringyl ratio

INTRODUCTION lignin remain. In the lignin isolation processes,

whether MWL or CEL, isolated lignin still con-
Wood mainly consists of polysaccharides
tain a small amount of carbohydrate. [1,2] Thus,
(cellulose and hemicelluloses) and lignin. Lignin
it is generally assumed that lignin has connec-
is built up by oxidative coupling of three
tions with hemicellulosic polysaccharides. [3-9] If
major C6 C3 (phenypropane)-type precur-
lignin and hemicellulose cooperate with each
sors; namely, coniferyl alcohol, sinapyl alcohol,
other to build up physically and chemically
and 4-hydroxycinnamyl alcohol. Although the
strong wood cell walls, there should be rela-
chemical composition and structure of native
tionships between the structure of hemicellu-
wood cell walls is quite well known, the basic
lose and that of lignin.
nature of the interactions between these differ-
Softwood lignin consists only of guaiacyl-
ent components is still unclear. For example, in
type lignin. There is a relatively large amount
pulp fibers, although most of the lignin is re-
of condensed units. Hardwood lignin con-
moved, small amounts of structurally modified

Address correspondence to Yu Huang, Laboratory of Wood Chemistry, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo,
113-8657, Japan. E-mail: ad818187@yahoo.co.jp

9
 10
sists of both guaiacyl and syringyl types.
There is wide variation in the syringyl ratio
(syringyl/(syringyl+guaiacyl)), depending on the
wood species. Syringyl ratio is the most impor-
tant chemical characteristic of lignin, because it
is related to the bonding pattern of lignin units,
internally including its stereo structure, lignin
content[10,11] and chemical reactivity.[12] As re-
ported by Akiyama and Nawawi et al.,[10,11] with
an increase in the syringyl ratios in lignin aro-
matics, the number of β-O-4 units and the pro-
portion of erythro isomers increases, the pro-
portion of the erythro isomer in this structure is
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larger, and the lignin content is lower. A similar
tendency was also observed within the genera
such as Acacia and Eucalyptus. In addition, To-
moda et al.[12] found that delignification behav-
ior was quantitatively related to the chemical
structure of lignin by taking the syringyl ratio
as an index of lignin chemical structure. Thus,
the aromatic structure of lignin is an important
factor to determine not only the chemical char-
acteristics of lignin but also the chemical reac-
tivity of lignin during the biomass conversion
processes.
If there is also a relationship between the sy-
ringyl ratio of lignin and hemicellulose, this rela-
tionship could be seen through a correlation of
the hemicellulose composition with the syringy
ratio. Based on this consideration, 48 hardwood
species were analyzed to determine the com-
position and absolute amount of neutral sugars,
and to investigate the relationships of the sy-
ringyl ratio of lignin. Next, the relationship of
neutral sugars in 14 softwoods with lignin con-
tent was investigated. It is well known that hemi-
cellulose in native hardwood is mainly xylan,
whereas in softwood hemicellulose is mainly
glucomannans and some xylan. Thus, the xy-
lan/mannan ratio is an important chemical fea-
ture to discriminate hardwoods and softwoods,
as is the syringyl ratio. However, the relation-
ship between syringyl ratio and xylan/mannan
ratio in widely various wood species is still
unknown. To elucidate this, the relationship
between syringyl ratio and xylan/mannan ra-
tio was determined in 48 hardwoods and 14
softwoods.
Y. HUANG ET AL.
TABLE 1. Number of 62 woods examined
Sample Number of woods
Softwood 14
Hardwood 48
Genus Acacia 17
Genus Eucalyptus 14
Other genera 17
MATERIALS AND METHODS
Preparation of Wood Meals
The 62 wood species (48 hardwoods, in-
cluding 17 Acacia, 14 Eucalyptus, and 17
other different genera; and 14 softwoods;
see Table 1) were ground in a Wiley mill,
and the wood meals were extracted using
ethanol:benzene (1:2, v/v) for 8 h in a soxh-
let apparatus. The extracted Wiley wood was
dried using an air flow and subsequently in a
vacuum.
Determination of Lignin Content
Wood meal (1 g) was suspended in 72%
sulfuric acid (10 ml) at room temperature for
3 h. The solution was diluted with water un-
til the sulfuric acid concentration was 3% and
then autoclaved at 121◦ C for 30 min.[13,14] After
cooling, the insoluble fraction was separated by
filtration using a glass filter (G3 for softwoods,
G4 for hardwoods). The weight of Klason lignin
was measured after drying at 105◦ C for two days
and the acid-soluble lignin was determined by
measuring the UV absorption at 205 nm using
an extinction coefficient of 110 l g−1 cm−1.[15]
Alkaline Nitrobenzene Oxidation
Alkaline nitrobenzene oxidation analyses
were applied to the milled woods in accor-
dance with a common procedure.[16] Nitroben-
zene oxidation products were silylated with
N,O-bis(trimethylsilyl)acetamide at 100◦ C for
10 min and then analyzed by gas chromatog-
raphy using the following conditions: gas chro-
matograph, GC-17A with FID (Shimadzu Co.,
Kyoto, Japan); column, NB-1 (fused-silica cap-
illary column, 30 m, 0.25 mm i.d., GL Science
 HEMICELLULOSE COMPOSITION AND LIGNIN STRUCTURE
Inc., Tokyo, Japan); column program, kept for
15 min at 150◦ C, raised by 3◦ C/min to 210◦ C
and by 10◦ C/min to 280◦ C; injection tempera-
ture, 250◦ C; detector temperature, 280◦ C. Sy-
ringyl ratio [S/(S + V)] was calculated on the
basis of the formula
Syringyl ratio = S/(S + V) = (syringaldehyde
+syringicacid)/ × (vanillin + vanillicacid
+syringaldehyde + syringicacid)
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where syringaldehyde is 4-hydroxy-3,5-
dimethoxybenzaldehyde, syringic acid is
4-hydroxy-3,5-dimethoxybenzoic acid, vanillin
is 4-hydroxy-3-methoxybenzaldehyde, and
vanillic acid is 4-hydroxy-3-methoxybenzoic
acid.
Neutral Sugars Analysis
Neutral sugars in polysaccharides in the
samples were determined by the alditol-acetate
method.[17] Wood meal was placed in a vacuum
drying oven at 40◦ C, drying to constant weight.
A 1 ml aliquot of 72% sulfuric acid was added
to 50 mg of wood meal for acid hydrolysis at
room temperature for 4 h. Then, the mixture
was diluted to 4% sulfuric acid and hydrolyzed
at 120◦ C for 1 h. After cooling to room tem-
perature, the hydrolyzed solution was filtered
using a glass filter (G3 for softwoods, G4 for
hardwoods). The filtrate was adjusted to 100 ml
with water and used as the sample solution. A
1 ml aliquot of myo-inositol aqueous solution
(∼ 1 mg/ml) was added to 5 ml of sample so-
lution as internal standard solution. The mixed
solution was neutralized by barium hydroxide
to pH 5.5, and then centrifuged. About 20 mg
NaBH4 was added to the supernatant and kept
for 24 h. Excess NaBH4 was removed using
acetic acid. The mixture was evaporated to dry-
ness and then heated in an oven at 105◦ C for
15 min to ensure complete removal of water.
The dry residue was acetylated by 1 ml of acetic
anhydride at 120◦ C for 3 h, then analyzed using
gas chromatography under the following condi-
tions: gas chromatography: GC-14b with FID
11
(Shimadzu Co., Kyoto, Japan); column: TC-17
(fused-silica capillary column, 30 m, 0.25 mm
i.d) (GL Science Inc., Tokyo, Japan); column
program: 20 min at 220◦ C; injection tempera-
ture: 220◦ C; detector temperature: 230◦ C. The
amount of monosaccharide (g) in 1 g of ex-
tracted wood meal was calculated.
RESULTS AND DISCUSSION
The existence of chemical linkages be-
tween polysaccharides and lignin (LCC) has
been strongly suggested in spite of the ab-
sence of direct evidence. It is anticipated that
there are structural correlations between lignin
and polysaccharide, especially hemicellulose,
closely located with the LCC linkage. Therefore,
structural correlations of lignin and polysaccha-
rides were examined in this study.
When discussing polysaccharide from plant
cell walls, attention should be paid to the fol-
lowing. Almost all glucose is derived from cellu-
lose, and little derives from hemicellulose (glu-
curonoxylan and glucomannan in hardwoods,
glucomannan and galactoglucomannan in soft-
woods). Therefore, the amount of glucose can
be used as an indicator of the cellulose con-
tent. Xylose is the main component of xy-
lan (glucuronoxylan in hardwoods and ara-
binoglucuronoxylan in softwoods), and it may
be used as an index of xylan content. Simi-
larly, mannose can be taken as an index of
mannan (glucomannan in hardwoods, gluco-
mannan and galactoglucomannan in softwoods)
content.
Relationship Between Syringyl Ratio and
Absolute Amounts of Neutral Sugars in
48 Hardwoods
Hardwood lignin consists both of guaiacyl
and syringyl types. There is wide variation in
the syringyl content, depending on the wood
species. It was observed that for hardwoods in
the same genus, the syringyl ratios were con-
centrated in certain intervals. Syringyl ratios of
genus Acacia ranged from 0.4 to 0.7, whereas
those of genus Eucalyptus varied from 0.7 to
 12 Y. HUANG ET AL.
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FIGURE 1. Relationship between syringyl ratio and the absolute FIGURE 2. Relationship between syringyl ratio and xy-
amount of glucose in hardwoods. lose/glucose ratio in hardwoods.

ent wood species expressed different increas-

0.85. Genera were distinguished from each ing trends with syringyl ratio. Rhamnose/glucose
other by their syringyl ratios. ratios in genus Eucalyptus increased more
It was shown that clear, simple tendencies rapidly than those in genus Acacia. For the ma-
can be observed when structural characteristics jority of the 48 hardwoods, arabinose/glucose
and lignin content are expressed as a function of ratios increased with syringyl ratios (see Fig-
syringyl ratio.[10,11] Syringyl ratio is the most im- ure 5), but no clear trend could be elucidated
portant chemical characteristic of lignin. Thus, for galactose/glucose ratios (see Figure 6).
the relationship between syringyl ratio and neu- The results suggested that hemicellulose,
tral sugars in hardwoods was investigated. Re- not cellulose, changed in conjugation with
lationships between syringyl ratio and absolute lignin structure. The lignin content was lower in
amount of glucose are given in Figure 1 for all hardwood species with higher syringyl ratios.[10]
48 hardwoods. As shown in Figure 1, variations Accordingly, the total amount of neutral sugars
in the amount of glucose were quite limited, was higher with increasing syringyl ratios. On
ranging from 0.4 g to 0.5 g in 1 g samples. the whole, higher neutral sugar content seemed
Figure 2 shows the relationship between sy- to be attributed to an increase in xylose, rham-
ringyl ratio and xylose/glucose ratio. With an nose, and arabinose. Because wood cell walls
increase in the syringyl ratio of lignin, the xy-
lose/glucose ratio of 48 hardwoods showed an
increasing tendency, although quite wide vari-
ations were detected. A clearer upward trend
could be seen when comparisons were made
within the same genus. Figure 3 shows the re-
lationship between the syringyl ratio and man-
nose/glucose ratio. No clear trends were rec-
ognized among the 48 hardwoods. For Acacia
mangium wood, most mannose/glucose ratios
varied between 0.02 and 0.03. For Eucalyp-
tus woods, mannose/glucose ratios rose with
syringyl ratios.
Considering all 48 hardwoods, rham-
nose/glucose ratios increased with syringyl FIGURE 3. Relationship between syringyl ratio and man-
ratios, as shown in Figure 4. In addition, differ- nose/glucose ratio in hardwoods.
 HEMICELLULOSE COMPOSITION AND LIGNIN STRUCTURE
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FIGURE 4. Relationship between syringyl ratio and rham-
nose/glucose ratio in hardwoods.
contain large amounts of xylose and only a small
amount of rhamnose, higher neutral sugar con-
tent was mainly by glucuronoxylan.
RELATIONSHIPS BETWEEN LIGNIN
CONTENT AND THE ABSOLUTE
AMOUNT OF NEUTRAL SUGARS IN 14
SOFTWOODS
Softwoods with no syringyl-type lignin con-
tain only guaiacyl-type lignin; thus, the syringyl
ratio of all softwoods was zero. Considering the
negative correlation between lignin content and
syringyl ratio in hardwoods, the relationship be-
tween lignin content and neutral sugars was
studied for softwoods.
FIGURE 5. Relationship between syringyl ratio and arabi-
nose/glucose ratio in hardwoods.
13
FIGURE 6. Relationship between syringyl ratio and galac-
tose/glucose ratio in hardwoods.
As shown in Figure 7, the absolute amount
of glucose in 14 softwoods barely changed with
lignin contents. Figure 8 shows the relationship
between lignin contents and the ratios of each
neutral sugar to glucose in softwoods. The man-
nose/glucose ratio increased with decreasing
lignin content. For all other hemicellusic sugars,
the sugar/glucose ratios did not change much
with lignin content. The results suggested that a
lower lignin content was attributed to mannose
in softwoods.
Relationships Between Syringyl Ratio
and the Xylan/Mannan Ratio in 48
Hardwoods and 14 Softwoods
It is well known that not only lignin struc-
ture but also hemicellulose composition of
FIGURE 7. Relationship between lignin content and the absolute
amount of glucose in softwoods.
 14
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FIGURE 8. Relationship between lignin content and hemicellu-
losic sugars/glucose ratio in softwoods.
hardwoods are different from that of soft-
woods. Softwood hemicellulose has a higher
proportion of mannose, and hardwood hemi-
cellulose usually contains a higher propor-
tion of xylose. However, the relationship be-
tween xylose/mannose ratio and syringyl ra-
tio has not been studied. From this point of
view, the relationship between syringyl ratio
and xylose/mannose ratio in 48 hardwoods and
14 softwoods was investigated, as shown in
Figure 9. The range of xylose/mannose ratios in
softwoods was from 0.2 to 1.0, and it was from
2 to 29 in hardwoods. Thus, both syringyl ratio
and xylose/mannose ratio were able to distin-
guish hardwoods from softwoods. In the same
genus, the xylan/mannan ratio of genus Acacia
ranged from 5 to 12, whereas it ranged from
11 to 18 in genus Eucalyptus. Therefore, al-
though the xylose/mannose ratio in genus Aca-
cia and that in genus Eucalyptus were mostly dif-
ferent in range, different genera of hardwoods
cannot be distinguished completely because
of overlapping xylose/mannose ratios. More-
over, Figure 9 shows the relationship between
xylose/mannose ratio and syringyl ratio. With
the increase in syringyl ratio of lignin, the xy-
lose/mannose ratio of hardwoods seemed to
show an increasing trend. However, the xy-
lose/mannose ratio in the same genus, both
17 Acacia mangium woods and 14 Eucalyptus
woods, remained almost constant. This result
suggested that the xylan/mannan ratio is an im-
Y. HUANG ET AL.
FIGURE 9. Relationship between syringyl ratio and xy-
lose/mannose ratio in hardwoods.
portant chemical feature not only to discrim-
inate hardwoods and softwoods, but also to
characterize the difference of cell wall compo-
nent structures of different hardwoods.
CONCLUSIONS
In this study, the relationships between
the lignin structure and hemicellulose composi-
tion in different wood species were examined.
In both hardwoods and softwoods, hemicellu-
lose rather than cellulose changed in conju-
gation with lignin structure and amount. Ac-
cordingly, the diversity of hemicellulose in all
woods was closely related to the structure of
lignins. In the 48 hardwood species, when the
syringyl ratio was higher, the xylose/glucose,
rhamnose/glucose, and arabinose/glucose ratio
increased. In the case of the 14 softwoods,
the mannose/glucose ratio increased with a de-
crease in lignin content. For both hardwoods
and softwoods, affinity was suggested to exist
between lignin with a higher syringyl ratio and
hemicellulose with higher xylan/mannan ratio.
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