WOOD RESEARCH

64 (3): 2019
411-422

THE CONTENTS OF PHENOLICS AND CELL WALL

COMPONENT OF EUCALYPTUS PELLITA F. MUELL
STEMWOOD AND BARK

Rizki Arisandi, Sri Nugroho Marsoem, Ganis Lukmandaru

Universitas Gadjah Mada, Faculty of Forestry
Department of Forest Products Technology
Yogyakarta, Indonesia

Tatsuya Ashitani, Koetsu Takahashi

Yamagata University, Faculty of Agriculture
Tsuruoka, Yamagata, Japan

(Received October 2018)

ABSTRACT

Eucalyptus pellita is the fast-growing species which is being developed for a raw material of
pulp and paper in Indonesia. The aim of this research was to evaluate the total phenolics (TPC)
and flavanols contents (TFC) in the stemwood and bark parts from four individual trees. Another
purpose was to determine its cell wall contents. Wood and bark materials in two vertical positions
(bottom and top) were successively extracted using dichloromethane, ethanol and hot water as
the solvents. Axial factor affected significantly in the values of hot water extract, TPC, and TFC
but no significantly affected the cell wall component contents. The ethanol extract levels in the
heartwood part was the significantly highest. It is noticed that the heartwood part had high levels
of the TPC and TFC and low level in lignin content. From this experiment, the comparatively
high levels of TPC and TFC in the heartwood indicate the potential antioxidative properties that
should be explored in the future. Further, the low content of Klason lignin in the heartwood part
would be an advantage for pulp production.

KEYWORDS: Extractive content, phenolics, cell wall components, pulp, E. pellita.

INTRODUCTION

Eucalyptus is a native genus from Australia and belongs to a family of Myrtaceae and
consists of 900 species (Brooker and Keing 2004). Eucalyptus species are important raw materials
for pulp, timber and charcoal industries, e.g. E. globulus, E. urophylla, E. grandis and various

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hybrids (Neiva et al. 2014, Sartori et al. 2016). One of the selected species for plantation forest of
Eucalyptus species is E. pellita. It has a natural distribution in Papua New Guinea (PNG), Irian
Jaya (Indonesia), and north Queensland (Australia) (Harwood et al. 1997).
E. pellita has shown great potential for plantation forest, where a commercially successful
plantation tree should include rapid growth under plantation conditions, straight stems with
limited branching, and decent wood quality for particular uses and products (Pinyopusarerk
et al. 1992, Dombro 2010). Nowadays, in Indonesia, E. pellita substituted Acacia mangium as an
industrial forest plantation (Hutan Tanaman Industri or HTI) as this species would be more
tolerant with a variety of soils and location conditions; distinct to be resistant to common pest and
diseases such as insect and fungi attacks (Harwood 1998, Dombro 2010).
Parts of the eucalyptus trees are known as a potent antioxidant activity such as terpenoids,
tannins, flavonoids, and phloroglucinol derivative (Boulekbache-Makhlouf et al. 2013). Many
of these compounds have therapeutic properties and are known for their anticarcinogenic,
antimutagenic, cardioprotective, anti-neurodegenerative, and antimicrobial activities (Babich
and Visioli 2003, Gursoy et al. 2009, Rababah et al. 2011). Some studies of Eucalyptus species
were reported that the woods contain biologically activity such as stump wood and bark parts of
E. globulus (Luís et al. 2014), 11 Eucalyptus species bark (E. botryoides, E. camaldulensis, E. globulus,
E. grandis, E. maculata, E. ovata, E. propinqua, E. resinifera, E. rudis, E. saligna, E. viminalis)
(Lima et al. 2017).
However, to our knowledge, investigations related to E. pellita are still limited, particularly
in phenolic contents. Only a few studies were conducted with regard to the chemical composition
of E. pellita wood which was from progeny trials in Indonesia (Fatimah et al. 2013, Lukmandaru
et al. 2016) and heart-rotted mature wood (Lukmandaru 2018). Hence, this study is to explore
the extractives, phenolic contents, and cell wall components on the axial and radial direction of
E. pellita stem wood and bark from natural forest.

MATERIALS AND METHODS

Wood specimen
E. pellita trees (4 individuals) were collected in May 2012 at the natural forest (tree age
was unknown) in Malind district (8.13o’S, 140.05°’E), Merauke, Papua. The amounts of total
diameter, heartwood proportion and total merchantable height ranged from 15.2 to 27 cm, 58.28
to 72.75%, 19 to 27 m, respectively. Each of E. pellita tree was cut at the bottom part (10% of total
height) and the top part (80% of total height) in disc form (± 5 cm in thickness) to obtain 8 discs
form. Each disc was divided into a bark, sapwood (± 0.5 cm from bark), and heartwood (± 1.0 cm
from the sapwood-heartwood border) to obtain 32 samples, which were taken from the cardinal
direction on the trunk to avoid radial variation, if any.
By visual inspection, the heartwood has a distinctive brown colour compared with lighter
sapwood colour. The bark and wood were separately milled to powder meal and were sieve-
screened (to pass a 1 mm sieve).

Extraction
The 5 g powder in 40-60 mesh size fractions was successively extraction by dichloromethane
and ethanol for 6 hours in a Soxhlet apparatus as well as hot water (refluxing in separated
extraction for 3 hours) (Morais and Pereira 2012). The temperature was set at 60°C for
dichloromethane, 90° for ethanol, and 110°C for hot water. Further, the solvent was evaporated

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by a rotary evaporator. Then, the extract was dried in an oven (103±2°C) and the extractive
content was quantified.

Chemicals
Aluminium chloride, sodium carbonate, sodium hydroxide, Folin-Ciocalteu’s phenol reagent
were purchased from Merck (Darmstadt, Germany) and standard components of gallic acid, and
(+)-catechin were purchased from Sigma-Aldrich (Chemie GmbH, USA) to analytical grade.

Determination of total phenolic content

Total phenolic content (TPC) was measured by Folin-Ciocalteu method (Singleton et al.
1999). Briefly, 2.5 ml of diluted Folin-Ciocalteu phenol reagent and an aqueous solution (1:9, v/v)
was mixed with 0.5 ml of the sample solution (0.25 mg.ml-1) in a 9 ml glass. After an interval
of 2 min, 2 ml of 7.5% aqueous sodium carbonate (Na 2CO3) was added to the glass, and the
mixture was allowed to stand for 30 min at ambient temperature. The absorbance for testing of
standard solutions was performed against the blank at 765 nm with an Ultraviolet (UV) / Visible
spectrophotometer (model UV-1800, Shimadzu, Tokyo, Japan). The standard curve was prepared
using 0.03125, 0.0625, 0.125, 0.25 mg.ml-1 solutions of gallic acid in methanolic (y = 0.0947
x - 0.0225; R 2 = 0.9991) and hot water (y = 0.1927 x -0.0085; R 2 = 0.9998). TPC was performed
as the mean ± standard deviation of four trees replication measurements and an expressed as
milligrams gallic acid equivalents (mg GAE/g dried extract).

Determination of total flavanol content

Total flavanol content (TFC) was quantified by vanillin HCl assay as described by Miranda
et al. (2016) with modification. To 0.5 ml (0.25 mg.ml-1) of the sample solution, 3 ml vanillin
reagent (4% vanillin methanol) and 1.5 ml HCl were added and the reaction is performed for
15 min at ambient temperature. The blank solution was prepared with the same procedure without
vanillin. The absorbance for testing of standard solutions was performed against the blank at
500 nm with an Ultraviolet (UV) / Visible spectrophotometer (model UV-1800, Shimadzu,
Tokyo, Japan). A set of reference standard solutions of (+)-catechin (0.03125, 0.0625, 0.125,
0.25 mg.ml-1) with (+)-catechin in methanol (y = 0.3591x - 0.0141; R 2 = 0.9995) and hot water
(y = 0.3426 x - 0.0079; R 2 = 0.9998). TFC were performed as the mean ± standard deviation of
the four trees replication measurements and an expressed in (+)-catechin equivalents (CE) (mg
CE/g dried extract).

Cell wall component in the heartwood and sapwood

The determination of wood of holocellulose and alpha-cellulose content was done with
chlorite acid modification of Wise method (Browning 1967). Further, Klason lignin content was
measured by standards of T 222 os - 1978 (TAPPI Standard 1992).

Statistical analysis
The data were statistically handled using the SPSS program (version 16 IBM, New York,
USA). Analysis of variance (ANOVA) was carried out and statistically significant differences
were set at a 95% confidence level. Two-way ANOVA was applied to determine the effect of
axial (bottom and top) and radial (bark, sapwood, and heartwood) direction on extractive content,
TPC, and TFC, and cell wall components. Further, Duncan test was performed to determine
specific differences between pairs of means. The analysis performed as the mean ± standard
deviation of the four tree replications measurements.

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RESULT AND DISCUSSION

Extractive content
By observing the colours, ethanol and hot water extracts showed dark and reddish colours.
Theoretically, those polar solvents will dissolve the phenolic compounds (Fengel and Wegener
1989, Sjöström 1993). Average of dichloromethane, ethanol, and hot water extractives content
were ranged from 0.19 to 0.38%, 1.58 to 4.95%, and 0.64 to 1.89% based on oven-dried wood,
respectively. The amounts of ethanol and hot water extract were slightly lower compared to the
values of ethanol-toluene extract of young E. pellita wood from progeny trials, Wonogiri (Fatimah
et al. 2013) and from South Borneo (Lukmandaru et al. 2016) with the values from 3 to 6.4%
(ethanol-toluene extract) and from 0.8 to 3.5% (hot water extract). However, it showed lower
amounts compared to ethanol extract (3.98 to 16.86%) and hot water extract (1.92 to 3.40%) in
a mature tree (Lukmandaru 2018). Further, the levels of the ethanol and hot water extracts were
also smaller than of ethanol-toluene (6.19 - 13.22%) and hot water extract (13.96%) values of
E. pellita from Brazil (Igarza et al. 2006, Oliveira et al. 2010, Andrade et al. 2010).
The variation of the dichloromethane extract was reported in the parallel work (Arisandi
et al. 2019), which the bark part had the highest levels (0.42 ± 0.18%) for radial direction, and in
the top parts (0.33 ± 0.14%) for longitudinal direction. Further, the two-way ANOVA analysis
was performed to obtain the effects of axial and radial direction (Tab. 1). Two-way ANOVA
showed that the radial factor affected (p < 0.01) the ethanol extract amount, as well as interaction
on axial and radial direction affected hot water extract amount (p < 0.05).

Tab. 1: Results from two- way ANOVA for ethanol and hot water extract of E. pellita.
Solvent
Source of variation df
Ethanol Hot water
Axial (A) 1 n.s. ***
Radial (R) 2 ** ***
AxR 2 n.s. *
Error 18
Total 23
df degrees of freedom;
n.s. not significant at 5% level, * P<0.05, **P<0.01, ***P<0.001.

(a) (b)
Fig. 1: (a), (b): The extractive content of ethanol and hot water (% based on oven-dried wood) from
E. pellita (means of four trees) on a radial direction for ethanol and interaction on an axial and radial
direction for hot water with error bar as standard deviation. The same letters on the histograms implied
no significant differences (p <0.05 by Duncan’s test).
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The levels of ethanol extract in the heartwood part were significantly higher than those
of bark and sapwood part (Fig. 1 a-b). Further, the amount of hot water extract in bark at the
top part was significantly found to be the highest. No systematic difference was measured in
the heartwood and sapwood parts for axial direction, except in the bark part (Fig. 1b). In other
eucalypt species, these trends are similar to those described in the previous works with regard to
the ethanol extract in E. globulus (Miranda et al. 2007, Lourenco et al. 2010, Morais and Pereira
2012), E. urograndis hybrid (Gominho et al. 2001), and E. grandis and E. pilularis (Higgins 1984,
Mariana et al. 2005). Furthermore, this present finding showed that the ethanol extract of the
bark part was lower compared to heartwood part. It was a similar trend with ethanol extract of
Juglans regia L. (Hosseini Hashemi 2012). Previously, Kai (1991) reported that heartwood part
is richer in polyphenols and resin acids (diterpenes) than bark part. In the heartwood part (with
lower pH condition), the most of the soluble sugars (xylose, mannose, and arabinose) were derived
from the hydrolyses.
Further, the portion of hot water extract in this experiment was similar to previous works
where the extractive content in the bark was considerably higher compared to other wood parts
(Fengel and Wegener 1989, Sjöström1993). In other eucalypts, the total amount of extractives
content had only a few percents in bole but the amount can be much larger in certain parts of the
tree, e.g., bark, branches, and topwood part (Pereira 1988, Domingues et al. 2010). The content
and composition of extractives can vary in response to age (Pereira 1988), species (Wilkes 1984),
growth rate (Hillis 1971), site (Miranda and Pereira 2002) and wood storage (Gutiérrez et al.
1998). The highest levels of hot water content in the top bark indicated more amounts of sugars,
such as starches (Gominho et al. 2015).

Total phenolic and flavanol contents

The total phenolic contents in the ethanol and hot water extracts ranged from 368.4 to 632.5
and from 199.9 to 429.6 mg GAE/g dried extract, respectively. On the other hand, the values
of flavanols content from the ethanol and hot water extracts were 165.6 to 373.8 and 47.5 to
132.5 mg CE/mg dried extract, respectively. Two-way ANOVA analysis was carried out to
observe the effects of axial and radial direction (Tab. 2).

Tab. 2: Results from two-way ANOVA for ethanol and aqueous extracts for total phenolic and flavanol
contents of E. pellita.
Source of Total phenolics content Total flavanols content
df
variation Ethanol Hot water Ethanol Hot water
Axial (A) 1 n.s. ** n.s. n.s.
Radial (R) 2 ** ** *** ***
AxR 2 n.s. n.s. * **
Error 18
Total 23
df degrees of freedom;
n.s. not significant at 5% level, * P<0.05, **P<0.01, ***P<0.001.

Two way ANOVA showed that the TPC values of ethanol extract were significantly affected
by radial direction, whereas hot water extract was influenced by both axial and radial directions.
On the other hand, the interaction between axial and radial direction significantly affected the
amount of TFC in both ethanol and hot water extracts (Tab. 2).

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Fig. 2: Total phenolic content of ethanol extract Fig. 3: Total phenolic content of hot water extract
based on mg GAE/g dried extract from E. pellita based on mg GAE/g dried extract from E. pellita
(means of four trees) on radial direction with error (means of four trees) on radial direction with error
bar as standard deviation. The same letters on bar as standard deviation. The same letters on
the histograms implied no significant differences the histograms implied no significant differences
(p <0.05 by Duncan’s test). (p <0.05 by Duncan’s test)
.

(a) (b)
Fig. 4: (a), (b): Total flavanols content of ethanol and hot water extract based on mg CE/g dried extract
from E. pellita (means of four trees) interaction on axial and radial direction with error bar as standard
deviation. The same letters on the histograms implied no significant differences (p <0.05 by Duncan’s
test).

By Duncan test, TPC of ethanol extract in the sapwood part was significantly lower than
in the bark and heartwood parts (Fig. 2). Additionally, TPC amounts of hot water extract in the
heartwood part was significantly larger compared to other parts in the radial direction (Fig. 3).
Further, TPC in the top part was significantly higher (347.2 ± 32.4 mg GAE/g dried extract)
than in the bottom part (261.6 ± 12.6 mg GAE/g dried extract) on axial direction (p<0.05).
These findings implied that in the heartwood part are potential sources of phenolic molecules
for antioxidant activities as well as antimicrobial and antifungal activities. Luís et al. (2014)
reported that stump wood has a great potential of phenolic molecules for medicinal purposes.
Gao et al. (2007) reported that sapwood is rich in nutrients such as sucrose and glycosides, unlike
heartwood and outer bark which are typically deficient in these nutrients but are rich in secondary
metabolites such as the flavanols and the other phenolic contents to protect the living tissues
against biological attacks.

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A comparison of the TPC of E. pellita in stem wood and bark with previously published
works in other eucalypt species is interesting. In most cases, TPC in the stem wood and bark
observed here were either similar or below the reported values: in the bark of some Eucalyptus,
such as E. sideroxylon, E. grandis, E. urograndis, and E. maidenii barks, the values were 441,
386, 347, and 204 mg GAE/g ethanol and ethanol–hot water extract, respectively (Santos et
al. 2012, Miranda et al. 2016). Further, Vázquez et al. (2008) and Santos et al. (2011) also
mentioned that TPC values of E. globulus bark of the ethanolic, methanolic, and hot water
extracts were 223, 410 and 155 to 201 mg GAE/g dried extract, respectively. Additionally, these
results are also in agreement with those obtained for E. tereticornis (198 mg GAE/g methanolic
extract) (Puttaswamy et al. 2014), 11 eucalyptus species ranged from 283 to 917 mg GAE/g
of extract (Lima et al. 2017) and in E. camaldulensis, E. globulus, and E. rudis (93, 23, and 3
mg GAE/g of bark), respectively (Conde et al. 1995, Cadahía et al. 1997). Furthermore, the
values of phenolic contents of some Eucalyptus wood, such as E. globules was 262.67 ± 3.06 mg
GAE/g dried ethanolic extract (Luís et al. 2014), E. grandis, E. urograndis, and E. maidenii were
825.47 ± 26.75, 775.59 ± 34.51, 687.89 ± 30.58 mg GAE/g dried extract methanolic / hot water
extract (Santos et al. 2013) and E. camaldulensis, E. globulus, and E. rudis were in the range from
5.22 to 25.63 mg.g-1 (Conde et al. 1995). Compared to other species, TPC of the heartwood,
sapwood, inner bark and outer bark extract of Port Orford Cedar were 136.9, 257.7, 537.5,
489.1 mg GAE/g dried extract (Gao et al. 2007).
With regard to flavanol contents, some studies were published for other eucalyptus
species. Luís et al. (2014) reported the values of flavanols content in the wood and stump bark
varied from 13.93 to 17.00 mg GAE/g dried extract. The content of flavanols in the different
eucalyptus barks were also observed, i.e. E. sideroxylon (395 mg CE/g ethanolic extract) (Miranda
et al. 2016), E. urophylla hybrids (77 to 184 mg CE/g in ethanol extract) (Sartori et al. 2016),
E. tereticornis bark (103 mg tannic acid equivalents / g in hot water extract) (Puttaswamy et al.
2014), and E. camaldulensis, E. globulus, and E. rudis barks (0.13 to 39.21, 3.28 to 7.43, 0.16 to
2.09 mg GAE/dried of bark, respectively) (Cadahía et al. 1997), as well as for 11 other eucalyptus
species in bark (94 mg to 545 mg CE/g dried extract) (Lima et al. 2017). Further, other species
showed that the values TFC trembling aspen bark was 5.2 to 18.3 mg CE/g dried extract (Diouf
et al. 2009) whereas the values in methanol / hot water extract of Maclura tinctoria wood and
bark were 5.1 ± 0.6 and 3.9 ± 0.1 mg CE/dried wood / bark (Lamounier et al. 2012). This study
finding demonstrated the differences between the levels of TPC and TFC on the axial and
radial direction. It should be noted that this study observed the high levels of TPC and TFC,
particularly in the heartwood and bark parts (e.g. Fig. 2 and Fig. 4a-b). Phenolic compounds have
received much attention for their effective antioxidant properties (Lima et al. 2017). Therefore,
identification of those compounds will be necessary in the future work.

Cell wall components

The carbohydrate portion of a cell wall is composed of holocellulose and minor amounts
of other sugar polymers such as pectins and starches. The carbohydrate fraction constitutes
70-75% of wood cell wall. Further, lignin is the most abundant natural non-carbohydrate organic
compound in fibrous materials. The content of holocellulose, α-cellulose, and Klason lignin
ranged from 67.37 to 70.57%, 42.55 to 50.97%, and 28.8 to 32.94%, respectively. Two-way
ANOVA analysis was performed to observe the effects of axial and radial direction (Tab. 3).

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Tab. 3: Results from two-way ANOVA for cell wall components of E. pellita.
Cell wall
Source of variation df
Holocellulose α-cellulose Klason lignin
Axial (A) 1 n.s. n.s. n.s.
Radial (R) 1 n.s. n.s. **
AxR 1 n.s. n.s. n.s.
Error 12
Total 15
df degrees of freedom;
n.s. not significant at 5% level, *P<0.05, **P<0.01.

Tab. 4: Cell wall content on the axial and radial direction of E. pellita.
Radial
Cell wall
Sapwood (%) Heartwood (%)
Holocellulose (b) 70.31 ± 5.44 70.57 ± 3.57
Holocellulose (t) 67.49 ± 3.87 67.37 ± 1.23
α-cellulose (b) 50.97 ± 4.68 48.34 ± 7.09
α-cellulose (t) 42.55 ± 4.52 44.77 ± 5.30
Klason lignin 32.54 ± 1.82 (a) 29.42 ± 1.79 (b)
Note: Average of four trees ± the standard deviation; b = bottom, t = top.

Values followed by the same letter within a column are not significantly different (*P<0.05)
as determined by two-way ANOVA. In the radial position, there was no significant difference in
holocellulose and α-cellulose contents. Difference results were reported by Mariana et al. (2005)
in E. nitens that the heartwood part had lower holocellulose and α-cellulose contents than in
the sapwood part. On the other hand, the amount of Klason lignin content in sapwood part was
significantly larger compared to the heartwood part (Tab. 4). With regard to lignin content, the
value was different compared to a previous work in a mature wood of E. pellita (Lukmandaru
2018). It was observed that the content of Klason lignin in the heartwood part was higher than
that of in the sapwood part. However, similar trends were found in E. nitens, Acacia melanoxylon,
and poplar I-69 (Mariana et al. 2005, Lourenco et al. 2010, Gao et al. 2011).
Compared to other studies of E. pellita wood, the range levels of the holocellulose in
this study was similar to previous studies (Andrade et al. 2010, Oliveria et al. 2010, Fatimah
et al. 2013, Lukmandaru et al. 2016, Lukmandaru 2018). For comparison, in other species,
holocellulose and Klason lignin of 12 eucalyptus species varied from 55.4 to 70.1% and
21.6 to 30.8% (Neiva et al. 2014), as well as α-cellulose, and Klason lignin content in 6 other
eucalyptus species ranged from 46.1 to 48.8% and 28.8 to 31.4%, respectively (Pereira et al. 2013).
Thus, from the point of view of pulp and paper production, the comparatively low lignin content
in the heartwood part would be beneficial regardless of its extractive content.

CONCLUSIONS

The yield of ethanol extract of E. pellita wood was significantly affected by radial direction. It
was found that the levels of ethanol extracts in the heartwood were significantly higher than in the
bark and sapwood. Further, hot water extract was significantly influenced by interaction on axial

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and radial direction. The levels of hot water extract in the bark at the top part were the highest
whereas no significant difference was observed for axial and radial directions with the exception
for the bark part. Further, either in ethanol or hot water extracts, i.e. content the heartwood part
showed a high TPC and TFC levels which indicate a great potential of antioxidative activities. It
was observed that the heartwood part contained the highest levels of both extractive and phenolic
contents. With regard to cell wall components, Klason lignin content of the heartwood part was
significantly lower compared to sapwood part. As the heartwood generally has great proportion,
this condition is an advantage as a raw material for pulp and paper manufacturing.

ACKNOWLEDGEMENTS

This work was supported by JASSO (Japan Student Services Organization) and DPP Grant
2017 (Faculty of Forestry, UGM).

REFERENCES

1. Andrade, M.C.N., Minhoni, M.T.A., Sansigolo, C.A., Zied, D.C., 2010: Chemical
analysis of the wood and bark of different Eucalyptus types before and during the shitake
cultivation. R. Árvore Viçosa MG 34(1): 165-175.
2. Arisandi, R., Takahashi, K., Ashitani, T., Marsoem, S.N., Lukmandaru, G., 2019:
Lipophilic extractives of the stem wood and bark from Eucalyptus pellita F. Muell grown in
Merauke, Indonesia (In submission).
3. Babich, H., Visioli, F., 2003: In vitro cytotoxicity to human cells in culture of some
phenolics from olive oil. Farmers 58: 403-407.
4. Boulekbache-Makhlouf, L., Meudec, E., Mazauric, J.P., Madani, K., Chenier, V., 2013:
Qualitative and semi-quantitative analysis of phenolics in Eucalyptus globulus leaves by high-
performance liquid chromatography coupled with diode array detection and electrospray
ionisation mass spectrometry. Phytochemical Analysis 24: 162-170.
5. Brooker, M.I., Keing, D.A., 2004: Field guide to Eucalyptus (2nd ed.). In Bloomings Book.
Northern Australia, Melbourne.
6. Browning, B.L., 1967: Methods of Wood chemistry Vol. II. A Division of John Wiley and
Sons, Inc. Interscience Publisher. New York.
7. Cadahía, E., Conde, E., de Simon, B.F., García-Vallejo, M.C., 1997: Tannin composition
of Eucalyptus camaldulensis, E. globulus, and E. rudis. Holzforschung 51: 125-129.
8. Conde, E., Cadahía, E., García-Vallejo, M.C., Thomás-Barberán, F., 1995: Low molecular
weight polyphenols in wood and bark of Eucalyptus globulus. Wood and Fiber Science 27(4):
379-383.
9. Diouf, P.N., Stevanovic, T., Cloutier, A., 2009: Antioxidant properties and polyphenol
contents of trembling aspen bark extracts. Wood Science and Technology 43: 457-470.
10. Dombro, D.B., 2010: Eucalyptus pellita: amazonia reforestation’s red mahagony. Colombia:
Planeta verde reforestation S.A.
11. Domingues, R.M.A., Sousa, G.D.A., Freire, C.S.R., Silvestre, A.J.D., Neto, C.P., 2010:
Eucalyptus globulus biomass residues from pulping industry as a source of high value
triterpenic compounds. Industrial Crops and Products 31:65-70.

419
WOOD RESEARCH

12. Fatimah, S., Susanto, M., Lukmandaru, G., 2013: Study of chemical components of
Eucalyptus pellita F. Muell wood of trees from second progeny test in Wonogiri, Central
Java. Jurnal Ilmu Kehutanan 7(1): 57-69 (In Indonesian).
13. Fengel, D., Wegener, G., 1989: Wood: chemistry ultrastructure, reactions. Walter de
Gruyter, Berlin, 613 pp.
14. Gao, H., Shupe, T.F., Eberhardt, T.L., Hse, C.Y., 2007: Antioxidant activity of extracts
from the wood and bark of Port Orford cedar. Journal of Wood Science 53: 147-152.
15. Gao, H., Zhang, L.P., Liu, S.Q., 2011: Comparison of KP pulping properties between
heartwood and sapwood of poplar I-69. Advanced Materials Research 236-238: 1437-1441.
16. Gominho, J., Figueira, J., Rodrigues, J.C., Pereira, H., 2001: Within-tree variation of
heartwood extractives and wood density in the Eucalyptus hybrid urograndis (Eucalyptus
grandis x E. urophylla). Wood and Fiber Science 33(1): 3-8.
17. Gominho, J., Lourenco, A., Miranda, I., Pereira, H., 2015: Radial and axial variation of
heartwood properties and extractives in mature trees of Eucalyptus globulus. BioResources
10(1): 721-731.
18. Gursoy, N., Sarikurkcu, C., Cengiz, M., Solak, M., 2009: Antioxidant activities, metal
contents, total phenolics and flavonoids of seven Morchella species. Food and Chemical
Toxicology 47: 2381-2388.
19. Gutiérrez, A., del Rio, J.C., Francisco, J., Gonzalez, V., 1998: Variation in the composition
of wood extractives from Eucalyptus globulus during seasoning. Journal of Wood Chemistry
and Technology 184(4): 439-446.
20. Harwood, C.E., Alloysius, D., Pomroy, P., Robson, K.W., Haines, M.W., 1997: Early
growth and survival of Eucalyptus pellita provenances in a range of tropical environments,
compared with E. grandis, E. urophylla and Acacia mangium. Common wealth Scientific and
Industrial Research Organization (CSIRO). New Forests 14: 203-129.
21. Harwood, C.E., 1998: Eucalyptus pellita an annotated bibliography. CSIRO Publishing,
Victoria, Australia, 70 pp.
22. Higgins, H.G., 1984: Pulp and Paper. In: Eucalypts for Wood Production (Eds: Hills WE,
Brown AG). Pp 289-312, CSIRO/Academic Press, Australia.
23. Hillis, W.E., 1971: Distribution, properties and formation of some wood extractives. Wood
Science and Technology 5: 272-289.
24. Hosseini Hashemi, S.K., 2012: Comparative chemical analysis of the extractives constituents
in the bark and heartwood of Juglans regia L. from north of Iran. Journal of Advanced
Laboratory Research in Biology 3(2): 98-101.
25. Igarza, U.O., Machado, E.C., Diaz, N.P., Martin, R.G., 2006: Chemical composition of
bark of three species of eucalyptus to three heights of commercial bole: Part 2 Eucalyptus
pellita F. Muell. Rev. Forest. Venez. 50(1): 53-58.
26. Kai, Y., 1991: Chemistry of extractives. In: Wood and cellulosic chemistry (Ed Shiraishi N).
Marcel Dekker Inc. New York.
27. Lamounier, K.C., Cunha, L.C.S., de Morais, S.A.L., de Aquino, F.J.T., Chang, R.,
do Nascimento, E.A., de Souza, M.G.M., Masrtins, C.H.G., Cunha, W.R., 2012:
Chemical analysis and study of phenolics, antioxidant activity, and antibacterial effect of
the wood and bark of Maclura tinctoria L. Evidance-Based Complementary and Alternative
Medicine 2012: 1-7.
28. Lima, L., Miranda, I., Knapic, S., Quilho, T., Pereira, H., 2017: Chemical and anatomical
characterization, and antioxidant properties of barks from 11 Eucalyptus species. European
Journal of Wood and Wood Products 76(2): 783-792.

420
 Vol. 64 (3): 2019

29. Lourenco, A., Gominho, J., Pereira, H., 2010: Pulping and delignification of sapwood and
heartwood from Eucalyptus globulus. Journal of Pulp and Paper Science 36: 3-4.
30. Luís, A., Neiva, D., Pereira, H., Gominho, J., Domingues, F., Duarte, A.P., 2014: Stump
of Eucalyptus globulus as a source of antioxidant and antimicrobial polyphenols. Molecules
19: 16428-16446.
31. Lukmandaru, G., Zumaini, U.F., Soeprijadi, D., Nugroho, W.D., Susanto, M., 2016:
Chemical properties and fiber dimension of Eucalyptus pellita from 2nd generation of
progeny test in Pelaihari, South Borneo, Indonesia. Journal of Korean Wood Science and
Technology 44(4): 571- 588.
32. Lukmandaru, G., 2018: Chemical characteristics of Eucalyptus pellita with heart rot. Wood
Research 63(2): 193-202.
33. Mariana, S., Torres, M., Fernandez, A., Moralez, E., 2005: Effects of Eucalyptus nitens
heartwood in kraft pulping. Tappi Journal 4(2): 8-10.
34. Miranda, I., Pereira, H., 2002: Variation of pulpwood quality with provenances and site in
Eucalyptus globulus. Annals of Forestry Science 59: 283-291.
35. Miranda, I., Gominho, J., Lourenco, A., Pereira, H., 2007: Heartwood, extractives and
pulp yield of three Eucalyptus globulus clones grown in two sites. Appita Journal 60(6):
485- 488.
36. Miranda, I., Lima, L., Quilhó, T., Knapic, S., Pereira, H., 2016: The bark of Eucalyptus
sideroxylon as a source of phenolic extracts with antioxidant properties. Industrial Crops and
Products 82: 81-87.
37. Morais, C.M., Pereira, H., 2012: Variation of extractives content in heartwood and
sapwood of Eucalyptus globulus trees. Wood Science and Technology 46: 709-719.
38. Neiva, D.M., Araujo, S., Laurenco, A., Gominho, J., Pereira, H., 2014: Kraft pulping
and wood chemical composition for 12 Eucalyptus species. Centro de Estudos Florestais.
Instituto Superior de Agronomia. Tapada da Ajuda, Lisboa, Portugal.
39. Oliveira, A.C., Carneiro, A.C.O., Vital, B.R., Almeide, W., Pereira, B.L.C., Cardoso,
M.T., 2010: Quality parameters of Eucalyptus pellita F. Muell wood and charcoal. Scientia
Forestalis 38(87): 431-439.
40. Pereira, H., 1988: Variability in the chemical composition of plantation eucalyptus
(Eucalyptus globulus Labill.). Wood and Fiber and Science 20: 82-90.
41. Pereira, B.L.C., Carneiro, A.D.C.O., Carvalho, A.M.M.L., Colodette, J.L., Oliveira,
A.C., Fontes, M.P.V., 2013: Influence of chemical composition of Eucalyptus wood on
gravimetric yield and charcoal properties. BioResources 8(3): 4574-4592.
42. Pinyopusarerk, K., Gunn, B.V., Williams, E.R., Pryor, L.D., 1992: Comparative
geographical variation in seedling morphology of three closely related Red mahagonies,
Eucalyptus urophylla, E. pellita and E. scies. Australian Journal of Botany 41: 23-34.
43. Puttaswamy, N.Y., Gunashekara, D.R., Ahmed, F., Urooj, A., 2014: Phytochemical
composition and in vitro anti-hyperglycemic potency of Eucalyptus tereticornis bark. Indian
Journal of Nutrition 1:102-107.
44. Rababah, T., Ereifej, K., Esoh, R., Al-u’datt, M., Alrababah, M., Yang, W., 2011:
Antioxidant activities, total phenolics and HPLC analyses of the phenolic compounds of
extracts from common Mediterranean plants. Natural Product Research 25: 596-605.
45. Santos, S.A.O., Freire, C.S., Domingues, M.R.M., Silvestre, A.J.D., Pascoal Neto, C.,
2011: Characterization of phenolic components in polar extracts of Eucalyptus globulus
Labill. bark by high-performance liquid chromatography–mass spectrometry. Journal of
Agricultural and Food Chemistry 59: 9386-9393.

421
WOOD RESEARCH

46. Santos, S., Villaverde, J.J., Freire, C.S.R., Domingues, M.R.M., Pascoal Neto, C.,
Silvestre, A.D., 2012: Phenolic composition and antioxidante activiy of Eucalyptus grandis.
E. urograndis (E. grandis × E. urophylla) and E. maidenii bark extracts. Industrial Crops and
Products 39: 120-127.
47. Santos, S.A.O., Vilela, C., Freire, C.S.R., Pascoal Neto, C., Silvestre, A.J.D., 2013:
Ultra-high performance liquid chromatography coupled to mass spectrometry applied
to the identification of valuable phenolic compounds from Eucalyptus wood. Journal of
Chromatography 938: 65-74.
48. Sartori, C., Mota, G.S., Ferreira, J., Miranda, I., Mori, F.A., Pereira, H., 2016: Chemical
characterization of bark of Eucalyptus urophylla hybrids in view of their valorization in
biorefineries. Holzforschung 70(9): 819-828.
49. Singleton, V.L., Orthofer, R., Lamuela-Raventos, R.M., 1999: Analysis of total phenols and
other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent oxidants
and antioxidants. Academic, San Diego, pp 152-178.
50. Sjöström, E., 1993: Wood Chemistry. Fundamentals and Applications, 2nd edition,
Academic Press. San Diego, USA.
51. Technical Association for the Pulp and Paper Industries, 1992: Acid-insoluble in wood and
pulp. TAPPI Test Method T 222 os - 78, TAPPI Press, Atlanta.
52. Vázquez, G., Fontenla, E., Santos, J., Freire, M.S., González-Álvarez, J., Antorrena, G.,
2008: Antioxidant activity and phenolic content of chestnut (Castanea sativa) shell and
eucalyptus (Eucalyptus globulus) bark extracts. Industrial Crops and Product 28: 279-285.
53. Wilkes, J., 1984: The influence of rate of growth on the density and heartwood extractives
content of eucalypt species. Wood Science and Technology 18: 113-120.

Rizki Arisandi, Sri Nugroho Marsoem, *Ganis Lukmandaru

Universitas Gadjah Mada
Faculty of Forestry
Department of Forest Products Technology
Jl. Agro No.I, Bulaksumur
Yogyakarta 55281
Indonesia +6274 550541
*Corresponding author: glukmandaru@ugm.ac.id

Tatsuya Ashitani, Koetsu Takahashi

Yamagata University
Faculty of Agriculture
1-23 Wakaba-Machi
Tsuruoka, Yamagata 997-855
Japan

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