Industrial Crops and Products 31 (2010) 113–121

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Industrial Crops and Products

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Cell wall ultrastructure, anatomy, lignin distribution, and chemical composition

of Malaysian cultivated kenaf fiber
H.P.S. Abdul Khalil ∗ , A.F. Ireana Yusra, A.H. Bhat, M. Jawaid
Bioresource, Paper and Coating Division, School of Industrial Technology, Universiti Sains Malaysia, Minden, 11800 Penang, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: The basic characteristics and physical properties of kenaf (Hibiscus cannabinus L.) fibers cultivated in the
Received 17 June 2009 region of Penang, Malaysia were analyzed. For fundamental analysis, which includes nano-scale view-
Received in revised form ing for identification of kenaf cell wall ultra structure, fibers were viewed under transmission electron
10 September 2009
microscopy (TEM). Light microscopy (LM) was used to observe the physical characteristics, anatomy, and
Accepted 13 September 2009
lignin distribution. Anatomical study was further carried out by scanning electron microscopy (SEM).
Kenaf plants had a maximum height of 2.50 m, with a mean basal diameter of 1.74 cm. The wood (core)
is the most abundant tissue with proportions up to 78% in cross-section area and up to 68.5% in weight.
Keywords:
Cell wall ultrastructure
The mean dry density of stems was found to be 0.29 g/cm3 , while that of core was 0.21 g/cm3 along the
Anatomy stems. A combination of LM and image analysis techniques was used to measure cell wall thickness and
Lignin distribution fiber length of the bast and core fibers. The fiber length of the bast was found to be longest (3637 ␮m)
Fourier transform infrared spectroscopy as compared to the core (1100 ␮m). Vessel members average 284 ␮m in length and 72 ␮m in diameter.
The chemical composition was determined according to Technical Association of Pulp and Paper Industry
(TAPPI) methods. Fourier transform infrared (FT-IR) spectroscopy was used to determine the functional
groups present in the kenaf fiber samples.
© 2009 Elsevier B.V. All rights reserved.

1. Introduction producing panel products such as fiberboard and particleboard,

Kenaf (Hibiscus cannabinus L.) is a traditional, third world crop
after wood and bamboo that is poised to be introduced as a new
annually renewable source of industrial purpose in the so-called
developed economies. Kenaf is a warm-season annual fiber crop
growing in temperate and tropical areas. It is related to cotton, okra,
and hibiscus due to systematics. It is a fibrous plant, consisting of
an inner core fiber (75–60%), which produces low quality pulp, and
an outer bast fiber (25–40%), which produces high quality pulp, in
the stem. The plant grows to a height of 2.7–3.6 m and is harvested
for its stalks, from which the fiber is extracted. Kenaf requires less
than 6 months for attaining a size suitable for practical application.
Due to the fast growth and good fiber quality, since the 1960s, there
has been increasing interest in kenaf, primarily for its potential use
as a commercial fiber crop for the manufacture of newsprint and
other pulp and paper products, leading to collaborations between
R&D, economist, and market research.
Kenaf may be Malaysia’s next industrial crop based on research
findings about its technical and commercial potential. It was first
introduced in the early-1970s in Malaysia and was highlighted in
the late-1990s as an alternative and cheaper source of material for
∗ Corresponding author. Tel.: +60 04 6532200; fax: +60 04 6573678.
E-mail address: akhalilhps@gmail.com (H.P.S. Abdul Khalil).
textiles, and fuel. Malaysian kenaf is composed of two distinct
fibers, bast and core, with a makeup of about 35% and 65%, respec-
tively. Each fiber has its own usage; thus, separation of the fibers
produces higher monetary returns over whole-stalk kenaf. Major
factors involved in separation of kenaf into its two fractions include:
size and amount of each portion; type and number of separation
machinery; processing rate through separation machinery; mois-
ture content of whole-stalk kenaf; humidity of ambient air.
Kenaf is harvested early for production of high protein animal
feed or at maturity for its fiber and woody core. These raw materials
can be processed into pulp and paper, bio-composites for auto-
motive door trimmings and interior shelving, as well as building
materials (Natsuno et al., 2002; Nacos et al., 2006; Thi Bach et al.,
2003). Kenaf can be quickly and easily pulped and bleached with
the ecofriendly chemicals, and kenaf-based newsprint is strong, has
good ink retention, and does not easily yellow with age (Ververis
et al., 2004). Recognizing its immense potential and interest gener-
ated by its fiber potential for the wood-based sector, the Malaysian
government has pursued various measures to promote down-
stream value-added processing of kenaf as well as its cultivation
among smallholders and estate owners.
Kenaf production yields are favorable when compared to soft-
wood and the hardwood. Softwood is a generic term used in
woodworking and the lumber industries for wood from conifers
(needle-bearing trees from the order Pinales) while hardwood is

0926-6690/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.indcrop.2009.09.008
114 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
used to describe wood from broad-leaved angiosperm trees, mostly
deciduous, but not necessarily, in the case of tropical trees. With a
decrease in wood resources, interest has grown in using kenaf as
an alternative raw material for pulp because it has the excellent
advantages of being renewable, inexpensive, and easily grown even
under severe conditions such as low water supply and little fertil-
izer (Angelini et al., 1998; Ken-ichi et al., 2002). An understanding of
the chemical composition of kenaf lignin therefore gives an insight
into the application of kenaf as a raw material for pulp (Ken-ichi et
al., 2002).
With such abundance, relatively low capital investment, and a
renewable source of plant fibers, it is an advantage to explore the
vast economic performance potential of kenaf cultivation. The main
characteristics of the most important plant fibers are described
in terms of their microscopic features, chemical composition, and
physical properties. Basic properties of plant fibers can be related
to the properties of end products. Overall basic information about
cell wall structure and chemical composition of fiber is neces-
sary for industrial production. To date, there are reports evaluating
the chemical composition, thermal properties, anatomy, lignin dis-
tribution, microfibril angle, mechanical properties, and cell wall
structure of commodity plants in Malaysia for different types of
fibers.
Taking into consideration the potential use of plant fibers, it is
required that there should be more than a sufficient volume of agri-
cultural fiber available globally for new industrial products. The
extent to which this potential can be realized will depend upon
numerous factors, not least of which will be consumer demand,
the availability of suitable processing and product handling equip-
ment, and further development of existing and new technologies.
Hence, the knowledge from this basic study can be beneficial to
the food technologist, material scientist, and polymer chemist for
future applied research studies.
The researchers in different parts of the globe are focusing on
the utilization of kenaf stems (woody core and fibrous bast) for
panel manufacturing, pulp and paper (Voulgaridis et al., 2000). To
the best of our knowledge, there is no detailed study on the cell wall
ultrastructure, anatomy, and lignin distribution of kenaf core fibers
cultivated in Malaysian conditions. To fill this gap, we have recently
started a research program aiming to deepen the knowledge of the
cell wall properties and characteristics mentioned above for this
non-wood kenaf. However, available literature references contain a
few papers describing a kenaf experimental plantation established
as a secondary crop in sandy soils under mediterranean climate
conditions as a complementary raw material to the wood-based
panel industry (Alifragis et al., 1997). Furthermore, anatomical
characteristics and properties of kenaf stems have been studied
to assess the suitability of kenaf stems for the panel and pulp and
paper industry (Voulgaridis et al., 2000). The abundance and chem-
ical composition of kenaf lignin are the two important parameters
which provide an insight into the manufacture of kenaf-related
products (Natsuno et al., 2002). The characterization and quan-
tification of kenaf bast lignin at different harvesting time are very
important to use kenaf bast fibers widely as an industrial raw mate-
rial for pulps (Bibhuti et al., 2005). The purpose of this study is to
generate sets of data regarding the basic differences between the
bast and core fibers.
2. Experimental
2.1. Materials
Kenaf, sp.V36, was collected from a local plantation source,
Nibong Tebal Paper Mills (NTPM), Seberang Prai, Penang, Malaysia.
Stalks were harvested at the age of 4.5 months. The cultivation
conditions of this crop include a warm season with an average
temperature of 31 ◦ C and a relative humidity of 70–90% backed
by irrigation. For the measurement and properties studies, eight
kenaf stems were taken at a base position of 20 cm from the ground
level. For chemical analysis, kenaf fibers were separated into bast
and core manually. The chemicals used in this study were analyt-
ical grade and obtained from Mega Makmur Scientific Sdn. Bhd.,
Chembumi Sdn. Bhd. Fisher Scientific, Penang, Malaysia.
2.2. Methods
2.2.1. Morphology of kenaf fibers
The transverse sections of thickness 1 ␮m were cut using Sor-
vall Ultra microtome (MT 500) with a glass and diamond knife. For
anatomy and lignin distribution determination, embedded sam-
ples were stained with 1% toluidine blue and were viewed under
a polarized microscope (Olympus BX50) for transverse and longi-
tudinal section. For the ultra-thin sections (0.1 ␮m), samples were
stained with 2% uranyl acetate and lead citrate. The transverse sec-
tions were examined with a Phillips CM12 transmission electron
microscope (TEM).
The microscopic appearance of kenaf core and bast was exam-
ined with a scanning electron microscope (Leo Supra, 50 VP, Carl
Ziess, SMT, Germany, SEM) by using small samples of 2 mm long,
1–2 mm width and 1 mm thickness. Transverse surfaces of kenaf
material were smoothed by a microtome knife while radial and
tangential surface were split. Surfaces were coated with gold before
their examination by SEM.
2.2.2. Fiber analysis
The TEM images of the micro- and nano-scale fibers were used
to measure the thickness of the cell wall with the help of DIMAS
Expert software, using a two point measuring analysis. The lumen
diameter was also calculated using the same method. For each char-
acterization, approximately 100 measurements were recorded to
obtain average results.
The disc from each portion of the stem (top, middle, base) was
used for measurement of bast and core fiber length, vessel mem-
ber length and diameter and parenchyma cell length and width
and then discs were macerated according to a common labora-
tory method (Tsoumis, 1991). Measurements were carried out at
low magnification using DIMAS Expert software, using a two point
measuring analysis.
2.2.3. Characteristics and properties
The kenaf were harvested at ground level and measured the
stem height. Leaves and reproductive parts (flowers and flower
buds) were removed and stalks cut into 20 cm lengths from the
base. The stalks were cut into the discs from the base position in
order to characterize the kenaf stem, for each bast and core samples.
The sample discs were used for the determination of dry density of
the stem and the core by water displacement test using electronic
densimeter. The discs were also used to calculate the percentage
proportions (cross-sectional area and dry weight) of the whole
kenaf stem (bast and core). The samples were separated into bast
and core, kiln-dried and weighed and thereafter being oven dried
at 60 ◦ C for 45 min. Then, the weight proportion of bast and core
to total dry weight of stem was calculated. The acidity of the stem,
bast and core powder samples was calculated using pH meter.
2.2.4. Chemical characterization
The bottom parts of kenaf stalk were further hand-separated to
provide the core and bast samples and then air-dried. Dried samples
were pulverized in a grinding machine and ground to 40 mesh for
Soxhlet-extraction with ethanol–toluene (1:2, v/v) for 6 h. After air-
drying, the extractive-free samples were subjected to the chemical
 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
analysis. Moisture content was calculated based on oven dry weight
of the sample.
The composition (extractive, ␣-cellulose, hemicelluloses, lignin,
and ash contents) of the fibers was analyzed according to Techni-
cal Association of Pulp and Paper Industry (TAPPI) standards. The
holocellulose content (␣-cellulose + hemicelluloses) of the fibers
was determined by treating the fibers with a NaClO3 and NaOH
mixture solution. The ␣-cellulose content of the fibers was then
determined by further treating holocellulose with 17.5% NaOH to
remove the hemicelluloses. The difference between the values of
holocellulose and ␣-cellulose gives the hemicellulose content of
the fibers. The lignin content of the fibers was found by treating
them with a sulfuric acid solution based on TAPPI standard T222
om-83. The extractive-free samples were analyzed thrice for each
of the samples.
2.2.5. Fourier transform infrared (FT-IR) spectroscopy
FT-IR spectroscopy, Nicolet Avatar 360, (USA), was used to
examine the functional groups presents in the fibers. Perkin Elmer
spectrum 1000 was used to obtain the spectra of each sam-
ple. Two grams of powdered fiber samples (extractive-free) were
mixed with KBr (sample/KBr ratio 1/100). They were then pressed
into transparent thin pellets. FT-IR spectra of each sample were
obtained in the range of 4000–400 cm−1 . Spectral outputs were
recorded in the transmittance mode as a function of wave number.
3. Results and discussion
3.1. Cell wall ultrastructure
The transmission electron microscopy provides a better tool for
investigating cell wall ultrastructure. Figs. 1 and 2 show transverse
sections of kenaf cell wall fibers composed of intercellular layers:
Fig. 1. Transmission electron micrograph of ultra-thin section of kenaf fibers: (A) kenaf core fibers showing cell wall and lumen; (B) transverse section of core fibers; (C)
transverse section of kenaf bast fibers. Cell wall structure stained with uranyl acetate and lead citrate. Scale bars = 5000 nm in (A and C); 2000 nm in (B): CW: cell wall; L:
lumen; F: fiber; P: parenchyma.
115
the so-called primary wall (the first layer deposited during cell
development) and the secondary wall (S), which again is made up
of three layers (S1 , S2 , and S3 ). Kenaf core fibers showed great vari-
ability in size, shape, and structure of the cell wall fibers (Fig. 1B).
The cell wall fibers were round to polygonal in shape.
In all lignocellulosic fibers, these layers contain cellulose, hemi-
cellulose and lignin in varying amounts. The individual fibers are
bonded together by a lignin-rich region known as the middle
lamella. Cellulose attains its highest concentration in the S2 layer
(∼50%), and lignin is most concentrated in the middle lamella
(∼90%), which, in principle, is free of cellulose. The S2 layer was
usually by far the thickest layer and dominated the properties of
the fibers. This structure was similar to wood cell wall structure
that has been proposed by previous research (Abdul Khalil et al.,
2006, 2008). Cell wall structure for non-wood fiber has been found
to be different from the wood fiber (Abdul Khalil et al., 2006, 2008).
TEM observation in a prior study (Abdul Khalil et al., 2006)
showed that oil palm fibers contained primary and secondary wall
layers (S1 , S2 , and S3 ). This observation was similar to other fibers
such as flax, and jute. However, bamboo showed a different struc-
ture of cell wall layers. Bamboo culm fiber contained polylamellated
wall structure [middle lamella, primary and secondary wall (S0 ,
S1−1 , S2−t , Sn−l , Sn−t )]. A typical tertiary wall is not present, but
warts cover the innermost layer of the cell wall (Abdul Khalil et al.,
2006).
Compound middle lamella (CML) is a layer between the middle
lamella and both adjacent primary walls (Fromm et al., 2003). In
the S1 and S2 of the fibers, lignin was distributed homogeneously,
while its level increases in the CML and the cell corners. The lignin
particles occur in high concentrations throughout the CML and
intermingle with the microfibrils of the secondary wall layer, which
appears brighter, indicating a low lignin concentration. The transi-
tion between S1 and S2 was characterized by a change in microfibril
116 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
Fig. 2. Transmission electron micrograph of ultra-thin section of kenaf fibers: (A and B) transverse section of a multi-layered fiber wall at low and high magnification,
respectively, in core fibers; (C) transverse section of bast fibers. Scale bars = 2000 nm in (A and C); 1000 nm in (B): ML: middle lamella; CML: compound middle lamella; P:
primary wall; S1 , S2 , and S3 : secondary wall sub-layers; L: lumen.
direction and also by an increase in lignin concentration (Fromm et
al., 2003). The gap between cellulose microfibrils was filled with a
thin layer of the lignin–hemicellulose complex (Terashima, 2000).
3.1.1. Thickness of cell wall
The secondary wall of the cell wall and middle lamella clearly
appeared in ultra-thin transverse sections of kenaf fiber as shown in
Figs. 1 and 2. The primary wall of kenaf fiber is a thin solid bound-
ary at the outer layer of the cell. The cell wall was also found to
be composed of concentric layers of dense, lignified cells contain-
ing cellulose and lignin in their secondary walls. The dark staining
of middle lamella indicated that it was strongly lignified (Herbert
Morrison et al., 1999; Abdul Khalil et al., 2006). The middle lamella,
which glues the cells together, showed clear transition to the adja-
cent primary wall layers (Fig. 2). The primary cell wall is a thin
layer produced by cell division and the subsequent growth of xylem
mother cells.
The secondary wall is a thick layer situated inside the primary
wall in the inner part of the cell, consisting of an outer layer (S1 ),
a middle layer (S2 ), and an inner layer (S3 ). Cells with thick walls
contain a large S2 layer, whereas thin-wall cells have a small S2
layer. Previous reports suggested that the irregular thickness might
be better suited than uniform thickness to relieve the pressure of
the axial compression force on the tracheid wall.
The differences between the wall layers can be easily identi-
fied by the thickness of the cell wall. Boundaries between middle
lamella, and primary wall layer could not be exactly determined
because of the similarity in their chemical composition; therefore,
it was hard to distinguish between them. In addition, the forma-
tion of primary wall stopped at the maturity of the cell wall. The
S1 layer in kenaf core fiber was well-defined and could be distin-
guished from the adjoining S2 layer (Fig. 2B). Obviously, the S2 wall
layer occupies the largest area of the fiber wall layer in the cell wall
and is the thickest wall layer (primary or secondary). Random mea-
surements were taken on high magnification TEM micrographs, and
the thickness of the S2 layer in kenaf core fibers was found to be in
the range of 0.3–1.6 ␮m while for the kenaf bast fibers was in the
range of 1.5–2.5 ␮m.
A previous study (Abdul Khalil et al., 2008, 2006) stated that
secondary wall in oil palm fibers also clearly showed a typical three
layered structure, but the S3 and warty layer (inside the lumen)
could not be easily distinguished. The S3 layer plays a critical role
in some important processes such as pulping and paper making, but
it has received less attention because of difficulties in studying it. In
order to provide evidence, in this study, the presence of a distinct S3
layer in cell wall of kenaf fibers was observed, albeit with difficulty.
Kenaf fibers showed a thin layer thickness of S3 , which was not well
developed in some of the fibers.
The diameter of the cell wall lumen (Fig. 1) was another impor-
tant factor considered in pulp and paper processing. Higher lumen
diameter and lower fiber thickness gives better pulp mat forma-
tion in the paper making process. The average lumen diameter of
the kenaf core and bast fibers were 6.7 and 2.8 ␮m, respectively.
This showed that the core fibers had a higher lumen diameter with
lower cell wall thickness compared to the bast fibers. This indicates
kenaf core to be a potential fiber for replacing wood and other lig-
nocellulosic fibers as a new developed raw material in pulp and
paper industries.
The pit was a primary wall layer that was not fully covered by
the secondary wall layer. The secondary wall was formed on top
of the primary wall where the protoplasm occurred, except at the
plasmodesmata region, and was not formed on the membrane pit.
 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
Fig. 3. Transmission electron micrograph of ultra-thin section of kenaf fiber. Trans-
verse section of pit fiber wall. Scale bars = 1000 nm: CW: cell wall; PT: pit.
Plasmodesmata were soft cytoplast thread, from which links were
formed to adjacent cells (Eames and MacDaniels, 1974).
The pit structure can be viewed clearly in this kenaf core fiber
(Fig. 3). In some other fibers, pits are rarely seen, and they vary
greatly in dimension. They are considered to be weak points that
reduce the strength of wood pulp fibers. Pits in the kenaf fibers also
are shown in Fig. 6E at longitudinal section of the core fibers.
From observation, the type of pit in the kenaf fiber cell was a
bordered pit, as found in many plant fibers. Bordered pits were
found on the walls of hardwood vessels, tracheids, and fibers, and
in a more specialized form on the walls of conifer tracheids (Meylan
and Butterfield, 1972).
Fig. 4. Anatomical micrographs of kenaf by light microscopy illustrated by image analyzer: (A) transverse section of kenaf core fibers; (B) longitudinal section of kenaf core
fibers; (C) transverse section of kenaf bast fibers. Scale bars = 100 ␮m in (A–C): F: fiber; P: parenchyma; V: vessel.
117
3.2. Anatomical identification
The structure of kenaf anatomy was similar to wood anatomy,
where the vessel cells were grounded in the fibrous tissue. Illus-
tration by the image analyzer of kenaf anatomy showed that the
fibers are not uniform (Fig. 1B), but are rounded to polygonal in
shape (Fig. 4). The shape of a fibrous cell in the transverse section
was slightly different from the longitudinal section. The fibers and
vessels appeared more oval in longitudinal section compared to
the transverse section (Figs. 4 and 5). All these features can also be
viewed similarly under SEM as shown in Fig. 6A–D. In Fig. 6A and
B, clearly the different thickness of cell wall between the core and
the bast could be seen, where the bast fibers are thicker than the
core fibers.
The wood is diffuse-porous, consisting of fibers, vessel members
and typically rectangular ray parenchyma. In vessel member walls,
small pits are present and can be viewed in Fig. 6C at high magni-
fication in the SEM micrographs. Longitudinal section at tangential
surface of core with very small pits in the fibers and parenchyma-
tous cells is shown in Fig. 6C. From this section, the shape of long
vertical fibers with closed ends and the parenchymatous cells with
many inclusions can be observed. The rays are uniseriate or biseri-
ate (Fig. 6C). Fig. 6D indicates the long bast fibers in the longitudinal
section.
The major cell type was sclerenchyma, which had very thick
lignified cell walls and was often associated with conducting tissue
(xylem and phloem, which generally transport water up the plant
to the leaves and sugars away from the leaves) (Julian, 2000). Scle-
renchyma fibers have the cellulose microfibrils orientated along
their length and so are very stiff. From a botanical point of view,
fiber bundles of hemp, ramie, jute, kenaf, and Roselle are mainly
secondary phloem. Flax fiber bundles are mainly cortical, and sisal
and abaca (manila hemp) are more loosely associated with the vas-
118 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
cular tissue in stem and leaf. Sclerenchyma also occurs in leaves of
monocotyledons (grasses and lilies), giving New Zealand flax and
esparto (Julian, 2000).
3.3. Lignin distribution
Many methods exist for investigating the lignin distribution in
plant cell walls (Blanchette et al., 1987; Donaldson et al., 2001;
Fromm et al., 2003; Xu et al., 2006). In this study, the transverse
sections of kenaf fibers after toluidine blue reaction demonstrated
that the vessel wall and middle lamella were stained in deep blue,
which showed a high level of lignification.
The dark stain of toluidine blue in Fig. 5 shows cell wall to
be highly lignified in the middle lamella, which bonds the cells
together at wall layers. Lignin closely follows the cellulose microfib-
ril orientation in the secondary cell wall by using TEM as well as
SEM and concluded that the polymerization of monolignols was
affected by the arrangement of the polysaccharides that constitute
the cell wall (Fromm et al., 2003).
Toluidine blue is a polychromatic stain that has been widely
used to differentiate lignified tissues from non-lignified ones
(Abdul Khalil et al., 2006). Lignified cell walls generally stain bluish
in color. The fiber walls of many monocotyledons were character-
ized by successive lignifications, as cell walls of living fibers thicken
with increase in age. As a major component of the cell wall of tra-
cheids, vessels, and fibers, lignin contributes to the compression
strength of woody stems and to the waterproofing of conductive
elements within the xylem. Unfortunately, there is a lack of infor-
mation on lignin distribution in monocotyledons (Abdul Khalil et
al., 2006).
It has been reported that vessel cell walls must withstand
the large tensile forces resulting from the transpiration stream in
plants, and hence, an increased lignification of the cell wall would
Fig. 5. Light microscopic images of kenaf fibers: (A) transverse section of core fibers; (B) longitudinal section of core fibers; (C) transverse section of bast fibers; (D) longitudinal
section of bast fibers. Scale bars = 50 ␮m in (A–D): F: fiber; P: parenchyma; V: vessel.
be expected to increase the stiffness of the vessel wall and over-
come its tendency to collapse. As is well known, the high lignin
concentration present in the cell wall was beneficial for the fiber-
board industries but disadvantageous in pulp and paper industry.
3.4. Characteristics and properties
Some basic characteristics and properties of kenaf stems (core
and bast) were shown in Table 1. The stem height ranged between
1.45 and 2.50 m. The average diameters at the base were found to be
1.74 cm for the stem and 1.52 for the core part. The mean values of
dry density of stems studied (base position) was 0.29 g/cm3 while
that of core was 0.21 g/cm3 . Core is the major tissue of the stem with
cross-sectional area of 78.04% and a dry weight of 68.5% while in
case of bast, the proportions were 21.96% (in cross-sectional area)
and 32.2% for dry weight proportion. The differences in percent-
age of the proportion for the cross-sectional area and dry weight
between these two distinct fibers showed that the part that domi-
nates higher proportion (core) will produce more fibers. The pH of
core and the whole stem was found to be acidic, 5.21 and 5.87 but
neutral for bast, pH 7.13.
3.4.1. Cell dimensions
The mean value of cell dimensions in three positions (base, mid-
dle, top) of kenaf stems is shown in Table 2. The mean fiber length
of fiber bast (3637 ␮m) is about three times greater than the kenaf
core (1100 ␮m). Fiber length decreases from top to the base in case
of kenaf core while kenaf bast shows an increasing trend of the
fiber length from top to the middle and then a decreasing trend
from middle to the base as shown in Fig. 7. The mean vessel mem-
ber length and diameter are 284 and 72 ␮m, respectively, while the
average dimensions of parenchymatous cells are 74 ␮m (length)
and 42 ␮m (width). Vessel member length at the top (250 ␮m) was
 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121 119

Fig. 6. Scanning electron micrograph of kenaf fibers: (A) transverse section of core fibers; (B) transverse section of bast fibers; (C) longitudinal section of core fibers; (D)
longitudinal section of bast fibers.

Table 1 lower than that of the base (274 ␮m) and the middle (327 ␮m)
Characteristics and properties of kenaf stems.
of the stems. Vessel member diameter increases from top to the
Characteristics/properties Bast Core Stem base.
Dimension (cm)
Height (range) 145–250 3.5. Chemical composition
Diameter 1.52 (0.095) 1.74 (0.212)
Perimeter 5.73 (0.131) 6.60 (0.101)
The changes in chemical composition occur along the
Proportion (%) stalks/branches for the plant raw materials. There is a general ten-
Cross-section area 21.96 (2.03) 78.04 (2.51)
dency for a decrease in ␣-cellulose, lignin and ash content from
Weight proportion 32.2 68.5
the base of the stalks or branches to the top. This was expected
Density (g/cm3 ) 0.21 (0.038) 0.29 (0.044) because mature tissues (at the base) accumulate higher amounts
Acidity (pH) 7.13 5.21 5.87
of metabolic products than the younger parts at the top. However,
many researchers have noted (Ken-ichi et al., 2002; Ververis et al.,
2004) that the bast and core greatly differ not only in their morpho-

Table 2
Dimensions of kenaf bast and core stems.

Position from the top of plantsa Fiber dimensions, ␮m Average

A B C

Kenaf bast
Fiber length 3370 (211) 3980 (319) 3560 (286) 3637 (419)

Kenaf core
Fiber length 1360 (191) 1050 (112) 890 (93) 1100 (153)

Vessel members
Length 250 (84) 327 (92) 274 (79) 284 (104)
Diameter 62 (14.2) 72 (17.1) 83 (21.2) 72 (16.4)

Parenchymatous cells
Length 74 (16.3) 67 (14.5) 81 (19.1) 74 (17.6)
Width 41 (8.3) 39 (7.7) 46 (11.3) 42 (10.3)
a
A, B, C: correspond to the top, middle and base of the stem for both kenaf bast and core (standard deviations shown in parentheses).
120 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
Fig. 7. Vertical variation of fiber length for bast and core.
logic structure but also in their chemical composition (Ken-ichi et
al., 2002; Natsuno et al., 2002).
Kenaf bast and core were quite different in respect to their
chemical components. The chemical analysis data for the whole
kenaf (core and bast) and kenaf core and bast are illustrated in
Table 3. The result in this study showed that the percentages of
all the chemical constituents of kenaf (non-wood) are more or
less similar to the wood materials, except for the ash. Ash content
(carbonates, Ca, K, and some trace elements) in kenaf was greater
compared to the wood materials. This proved that the non-wood
fibers were high in silica, which was in a way known to be a dis-
advantage in mechanical strength properties for the end products
(Herbert Morrison et al., 1999; Ververis et al., 2004). The values
of ash content were 4%, 1.9%, and 5.4% for whole, core, and bast
fibers, respectively. The results for the chemical composition deter-
mined by this study were similar to the work done by Ashori et al.
(2006), except, for the ethanol–toluene extractive content which
was higher. Extractive was usually discarded and not used in the
production and manufacture.
From Table 3, the percentage of chemical composition in kenaf
fibers indicated that the kenaf core fibers were higher in holocellu-
lose and lignin, while kenaf bast fibers were higher in ␣-cellulose,
extractive, and ash content in comparison with both types of kenaf
fibers. ␣-Cellulose content in bast fiber, at 55%, was higher then
in the core, at 49%. High ␣-cellulose content is believed to provide
high strength in paper formation and other fiber end products. It
stated that during maturation of the kenaf plant the cellulose (bast
fiber 52–59%, core 44–46%) and lignin (bast fiber 9.3–13.2%, core
18.3–23.2%) contents increased significantly (Thi Bach et al., 2003).
Holocellulose contents were determined in this study to be 87.7%
for whole stalk, 87.2% for core and 86.8% for bast, all of which are
higher than according to the previous study (Ashori et al., 2006).
Kenaf lignin was composed of three lignin units, p-
hydroxyphenyl, guaiacyl, and syringyl, in varying ratios. The
whole kenaf stalk (core and bast) contained 21.2% lignin, higher
than results obtained by Ken-ichi et al. (2002). The core and bast
sample contained 19.2% and 14.7% lignin, respectively, which were
Table 3
Chemical composition of different fractions of kenaf fibers.
Kenaf whole (core + bast)
Extractive (%) 6.4
Holocellulose (%) 87.7
␣-Cellulose (%) 53.8
Lignin (%) 21.2
Ash (%) 4.0
a
Tsoumis (1991).
slightly lower than that of softwood (21–37%) and is advantageous
for pulping. Due to their lower lignin content (compared to wood),
non-wood plants can be pulped in one-third of the time needed
for softwoods and hardwoods. Pulping of non-wood fibers also
demands around 30% less chemical charge and reduced power
consumption in pulp refining. Bleaching of non-wood fibers was
also relatively easy. The above information means, in general, a
lower cost of production for some grades of paper from non-wood
plants (Ververis et al., 2004).
One of the advantages of kenaf pulping was its lignin content.
Relatively increased (>25%) lignin content may require additional
pulping time and chemical cost compared to those of other non-
wood raw materials. Analysis of samples at various heights/lengths
of the plant materials showed that lignin and cellulose content
depends on tissue maturity but does not change significantly within
each species. Paper strength also depends on the lignin and cel-
lulose content of raw plant materials; pulp mechanical strength
and especially tensile strength is directly proportional to cellulose
content, whereas lignin is an undesirable polymer and its removal
during pulping requires high amounts of energy and chemicals
(Ververis et al., 2004).
The core samples contained a higher amount of lignin, 19.2%,
compared to the bast samples, 14.7%, prepared from the same
location in the kenaf plant. This result concurs with the study
done (Ververis et al., 2004), but in a lower range. This discrepancy
may be attributed either to different methodologies or different
kenaf fiber varieties than those we have used in our analyses.
He also found that the bast samples have lower lignin content
than the inner bast and core samples and suggested that the
contents vary according to varieties, position and/or parts. For
the use of pulp, the low lignin content was expected (Herbert
Morrison et al., 1999; Ververis et al., 2004). In a detailed study
of maize (Zea mays L.) stalks, reported that lignin and cellulose
deposition increased with tissue maturation, with maximum rate
of lignin deposition following that of cellulose (Ververis et al.,
2004).
3.6. Fourier transform infrared spectroscopic (FT-IR) analysis
Fourier transform infrared (FT-IR) spectroscopy was used to
detect the presence of the functional groups that exists in kenaf
fibers. FT-IR spectroscopy is a powerful tool for studying the
physico-chemical and conformational properties of polysaccha-
rides. FT-IR spectra for the whole kenaf (core and bast), is shown
in Fig. 8. The spectra showed the presence of different functional
groups within the fiber sample.
Fig. 8 showed that the peaks for the whole kenaf (core and bast),
appeared at 3390 and 1190 cm−1 , corresponding to O–H stretching
and O–H bending frequencies, respectively, while the –CH2 stretch-
ing frequency appeared at 2910 cm−1 . In the spectrum of the kenaf
core as shown in Fig. 8, the peaks appeared at 3413 and 1047 cm−1
corresponding to the O–H stretching and O–H bending frequen-
cies, respectively, while the –CH2 stretching frequency in this case
appeared at 2910 cm−1 . In comparison, the peaks for the kenaf bast
shifts to the higher frequencies as compared to the whole kenaf and
Kenaf core Kenaf bast Softwooda Hardwooda
4.7 5.5 0.2–8.5 0.1–7.7
87.2 86.8 60–80 71–89
49.0 55.0 30–60 31–64
19.2 14.7 21–37 14–34
1.9 5.4 <1 <1
 H.P.S. Abdul Khalil et al. / Industrial Crops and Products 31 (2010) 113–121
Fig. 8. FT-IR spectrum of kenaf stem in the frequency range of 400–4000 cm−1 .
kenaf core. In this case the peaks appeared at 3430 cm−1 for O–H
stretching and 1028 cm−1 for O–H bending frequency. While the
CH2 stretching frequency in this case appeared at slightly higher
frequency and appeared at 2923 cm−1 . The blue shift in the O–H
group frequencies from the whole kenaf to the kenaf bast could be
due to the different hydroxyl group content in the fibers in the form
of cellulose, hemicelluloses, and lignin.
In the FT-IR spectra, the other prominent absorbance peaks were
in the range of 1739–1650 cm−1 . The peaks at 1739 and 1650 cm−1
(Fig. 8) represent carbonyl stretching (C O) for acetyl groups in
hemicelluloses and for aldehydic group present in the lignin. The
carbonyl chromophore does not occur in kenaf bast.
4. Conclusions
The above studies can be summarized as follows:
i) Transmission electron microscopy (TEM) has confirmed that
cell wall structure of all two types of kenaf core and bast fiber
consists of secondary wall (S1 , S2 , and S3 ) layers and middle
lamella.
ii) Core is the major tissue of the stem with larger cross-sectional
area and a dry weight as compared to the bast, thereby showed
that the dominant core portion will produce more fibers.
iii) The mean values of dry density of stem studied (base position)
was found to be higher than that of core. The pH of core and
the whole stem was found to be acidic but neutral for bast.
iv) Determinations of lignin distribution in kenaf fibers illustrate a
high level of lignification exhibited in the middle lamella. Kenaf
fibers showed positive staining with toluidine blue.
v) In general, chemical compositions of the two types of kenaf
fibers indicated that the kenaf core fibers were higher in holo-
cellulose and lignin, while kenaf bast fibers were higher in
␣-cellulose, extractive, and ash content. The chemical compo-
sitions of the whole kenaf stalk (core and bast) are still within
the range of the wood chemical composition except for the ash
content.
vi) Based on the anatomical characteristics and properties of kenaf
stems examined the potentials of using such material in wood
panel and pulp and paper manufacture are evident.
121
vii) The extractive content in the kenaf stem is relatively higher
and pH is acidic. On account of which it may affect the pulping
processes (consumption of chemicals, yield, etc.).
Acknowledgements
The researchers would like to thank the Ministry of Science,
Technology and Innovation (MOSTI) and the Universiti Sains
Malaysia, Penang for providing the research grant EScience Fund
(RM-9) 03-01-05-SF0334 or 305/PTEKIND/613323 that has made
this work possible.
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