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Bioresource Technology 99 (2008) 4878–4886

Effect of storage time on the composition and content

of wood extractives in Eucalyptus cultivated in Brazil
Flaviano O. Silvério a,b, Luiz C.A. Barbosa a,*, Célia R.A. Maltha a, Paulo H. Fidêncio c,
Mariluze P. Cruz a, Dorila P. Veloso b, Augusto F. Milanez d
a
Department of Chemistry, Federal University of Viçosa, 36570-000 Viçosa, MG, Brazil
b
Department of Chemistry, Federal University of Minas Gerais, 31270-901 Belo Horizonte, MG, Brazil
c
Federal University of Tocantins, 66, 77402-970 Gurupi, TO, Brazil
d
Company Suzano of Paper and Cellulose, 08613-900 Suzano, SP, Brazil

Received 1 June 2007; received in revised form 14 September 2007; accepted 15 September 2007
Available online 7 November 2007

Abstract

Lipophilic wood extractives commonly referred to as pitch, cause significant problems for the pulp and paper industries. The reduc-
tion of these extractives is an important aspect that concerns industries around the world. In the present work the change in the amount
and chemical composition of lipophilic extractives from Eucalyptus spp. stored for 20, 40, 60, 100, 140 and 180 days after harvesting was
investigated. The results showed a decrease in extractives content with storage time, with the most significant decrease occurring 60 days
after harvesting. In addition, fatty acids and sterols were the main classes of compounds responsible for the significant decrease in extrac-
tive content. Data were analyzed by principal component analysis. PC1 explains approximately 99% of the total variance, and b-sitos-
terol was the major compound responsible for the differentiation. These studies demonstrate that in terms of economical aspects, quality
of the pulp and paper and minimization of pitch formation, the best period of wood storage is 60 days.
 2007 Elsevier Ltd. All rights reserved.

Keywords: Pitch; Fatty acids; Sterols; PCA

1. Introduction The extractive content is considered an important qual-

The search for higher quality products and more com-
petitive prices have increased international competition in
the paper and cellulose sector, consolidating and enlarging
markets for industries of outstanding performance.
Amongst the world’s largest pulp and paper producers,
Brazil has also promoted researches aimed at improving
knowledge on raw materials, processing and final product
on account of the growing economic interest (Almeida
and Silva, 2001; Gonçalves et al., 2001; Caixeta et al.,
2003a, 2003b).
*
Corresponding author. Tel.: +55 031 3899 3068; fax: +55 031 3899
3065.
E-mail address: lcab@ufv.br (L.C.A. Barbosa).
ity parameter for cellulose pulp production (Sjöström and
Alen, 1998; Gullichsen and Paulapuro, 2000; Hillman,
2002; Silvério et al., 2007a, 2007b). Although the amounts
of wood extractives are small, they can negatively influence
the process of pulp production and papermaking, since
they may lead to the formation of pitch deposits (Sjöström
and Alen, 1998; Back and Allen, 2000; Gutiérrez et al.,
1998; Silvestre et al., 1999, 2005; Manji et al., 2005). Pitch
problems are due to the deposition of lipophilic extractives
and other materials possibly derived from wood on the sur-
face of process equipment (Back and Allen, 2000; Gutiérrez
et al., 2001; Gutiérrez and Del Rı́o, 2005). These deposits
agglomerate over time and tend to become released
from the equipment and contaminate the pulp, lowering
the quality of the final product (Gutiérrez et al., 1998,
2001; Del Rı́o et al., 2000; Dorado et al., 2000a, 2000b;

0960-8524/$ - see front matter  2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2007.09.066
 Author's personal copy
F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886
Martinez-Iñigo et al., 2000; Freire et al., 2005). Therefore,
the elimination or reduction of pitch problems has been a
major challenge for companies and researchers.
The wood harvest season has a direct effect on pitch
chemical composition (Olm, 1984; Almeida and Silva,
2001). During the winter, the hydrolysis of wood esters is
considerably reduced and as a consequence the ester frac-
tion of pitch is significantly larger than in the summer
months, when higher temperatures lead to high hydrolysis
rates. High amounts of these esters can increase hydropho-
bicity and viscosity of pitch resulting in the formation of
deposits and inevitably to specks on paper sheets (Olm,
1984).
An alternative used by the industries to reduce pitch for-
mation is to store the wood for long periods of time after
harvesting (Stein, 2003). Thus, triacylglycerols can be
hydrolyzed into their respective fatty acids (Gutiérrez
et al., 2001). Through this process, the amount of esters
of fatty acids affecting pitch deposits decreases during stor-
age and pitch problems are decreased. Wood exposed to
natural conditions for long periods of time can lose its
value due to attacks by insects, fungi and bacteria (Dorado
et al., 2000a, 2000b; Martinez-Iñigo et al., 2000; Gutiérrez
et al., 2001). Such attacks can lead to carbohydrate loss,
mainly cellulose and hemicellulose and result in a lower
pulp yield after processing (Burnes, 2000).
Embodied in a wider project involving the study of pitch
problems in the Brazilian pulp industry, the present work
has investigated the influence of wood storage time, kept
in the field for up to 180 days under natural conditions,
on the content and chemical composition of the lipophilic
extractives. The qualitative and quantitative variation of
the extracts were studied by gas chromatography and mass
spectrometry and the results analyzed by principal compo-
nent analysis (PCA). Physical properties of the pulps and
papers produced from the stored woods were also
monitored.
2. Methods
2.1. Samples
The experiments were conducted under industrial condi-
tions from harvesting to wood chipping, and later the
assays were carried out in laboratory. The studied wood
consisted of a mixture of Eucalyptus grandis (77% w/w)
and E. saligna (23% w/w), approximately 8.5 years old
and 30.5 m high. The studies were carried out with 6.5 m
long logs, stored in piles (6.5 mL · 4.0 mW · 3.0 mH) and
spread out in the log yard. Logs were transported to the
mill at 20, 40, 60, 100, 140 and 180 days after harvest. Each
time the logs arrived at the mill, they were bark-free, then
chopped into small pieces. At the industrial site the wood
chips are classified according to the SCAN-CM 40:94 rec-
ommendations. For laboratory analysis and digester exper-
iments the samples were classified between 2 mm and 5 mm
of thickness. Wood chips were ground to pass a through
4879
1 mm sieve screened in a vibratory sieving apparatus, and
the 40–60 mesh fractions were used for chemical analysis.
2.2. Extraction
Air-dried powdered wood samples (2.00 g) were
extracted with acetone for 6 hours using a Soxhlet appara-
tus following Tappi standard process (T 204 cm-97). The
solvent was removed under reduced pressure in a rotary
evaporator, and the extracts were weighed. All extractions
were carried out in triplicate, and the extraction yields were
expressed as a percentage in relation to the wood’s dry
weight. This procedure was used for samples collected at
20, 40, 60, 100, 140 and 180 days after harvest.
2.2.1. Alkaline hydrolysis
An aliquot of 10 mg of extract was added to a two neck
round-bottomed flask (10 mL), followed by 1.8 mL of
aqueous KOH solution (3 mol L1) and 0.2 mL of metha-
nol. The mixture was refluxed under nitrogen atmosphere
for 1 h. It was subsequently cooled to room temperature,
acidified with aqueous HCl (3 mol L1) to pH  2 and
extracted with dichloromethane (3 · 2 mL). The combined
organic extracts were dried over anhydrous MgSO4, filtered
off and the solvent was completely removed under reduced
pressure in a rotary evaporator (Gutiérrez et al., 1999).
2.2.2. Derivatization
Aliquots of hydrolyzed and non-hydrolyzed extracts
(2.0 mg) were dissolved in pyridine (60 lL) in capped vials
followed by the addition of 100 lL bis(trimethylsilyl)-triflu-
oroacetamide (BSTFA) containing 1% chlorotrimethylsi-
lane (TMSCl). The reaction mixture was heated at 70 C
for 30 min. It was then cooled to room temperature before
GC–MS analysis.
2.2.3. GC–MS analysis
GC–MS analysis were performed on a Shimadzu
PQ5050A GC–MS equipped with an AOC-5000 autoinjec-
tor and a DB-1 J&W capillary column (30 m · 0.25 mm
i.d., 0.25 lm film thickness), using helium as carrier gas
(35 cm/s). The chromatographic conditions were as fol-
lows: injector temperature 290 C; oven initial temperature
80 C (5 min); temperature increase rate 4 C/min; final
temperature 285 C (40 min). The interface temperature
was 290 C and a split ratio of 1:10 was used. The mass
detector operated with ionization for electrons impact
(70 eV) for a scan range of 30–600 a.m.u.
For semi-quantitative analysis, the GC–MS equipment
was calibrated with pure reference compounds, representa-
tive of the major extractives components. Reagents of 97–
99% purity obtained from Sigma-Aldrich (Milwaukee, WI,
USA) were: hexadecanoic acid, hexadecan-1-ol, 16-
hydroxyhexadecanoic acid, 2-hydroxyoctanoic acid, trans-
ferulic acid and b-sitosterol. The calibration was relative
to hexanedioic acid and tetracosane (99% purity obtained
from Sigma-Aldrich, USA), used as internal standards of
 Author's personal copy

4880 F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886

concentration 0.1562 mg mL1, as described by Freire 35 spread out logs piled logs
et al. (2002a). The hexanedioic acid was prepared by oxida- 30

Mass Loss (%)

tion of cyclohexanone with potassium permanganate. This
compound was more than 99% pure as evaluated by GC
analysis. The response factors needed to obtain the correct
quantification of the peak areas were calculated as an aver-
age of the sixteen GC–MS runs with the standard com-
pounds of concentration 0.1562 mg mL1, after silylation
with BSTFA and TMSCl in pyridine as already described
(Freire et al., 2002a, 2002b; Wallis et al., 1999). Com-
pounds were identified as TMS derivatives by comparing
their mass spectra with the GC–MS spectral library (Wiley
330.000, 7th Edition), with data from the literature (Barros,
2003) and when necessary, by injection of standards.
2.3. Principal component analysis (PCA)
PCA is frequently used to reduce the dimensionality of
the data in some chemical problems (Wold et al., 1984;
Ferreira, 2002; Correia and Ferreira, 2007). The objective
of PCA is to compress data within a group of new vari-
ables, which is a linear combination of original variables,
maximizing the description of data variance (Correia and
Ferreira, 2007). The original data matrix (X) of m lines
(objects or samples) and n columns (variables) is decom-
posed in a score matrix, S (m, f), weight matrix, L (n, f),
and error matrix E (m, n): X = SL 0 + E, where f is the
number of significant principal components. The columns
of the score matrix are groups of new orthogonal variables,
the so called principal component (PC). The elements
of the weight matrix are used to indicate the contribution of
the original variables to each PC. The errors correspond to
the variance that is not described.
In order to reduce the number of the most significant
variables in the system, that is, to obtain the principal com-
ponents, PCA was performed assigning the sample data
from the different storage times (objects) and the extractive
contents as variables, which were pre-processed by mean
centering. The principal component was carried out using
the PLS_Toolbox software (version 2.1, Eigenvector
Research, Inc., Wenatchee, Washington, USA).
3. Results and discussion
3.1. Evaluation of wood storage time
25
20
15
10
5
0
15 30 60 90 120
Storage Time (Days)
Fig. 1. Mass loss in Eucalyptus wood stored under natural conditions in
the field over the period of 120 days.
mass loss up to 60 days, and approximately 33.0% of mass
loss at 120 days after wood harvesting. There was no differ-
ence in the mass loss of piled logs as a function of their
position in the pile, top or bottom, over the storage time.
During the drying process, most of the water present in
the wood is removed and therefore the weight per unit of
volume is reduced (Stein, 2003). In the present study the
reduction observed in the wood weight varied from a quar-
ter to a third of the initial weight. This drying process
reduces the transportation costs considerably. However,
lower moisture content can influence wood properties neg-
atively, such as difficulty in debarking (Panshin and De
Zeeuw, 1980; Tsoumis, 1991; Stein, 2003).
According to these results, if only the wood moisture
content is to be taken into consideration, it is recom-
mended to leave the eucalyptus woods in the field (spread
out or piled up) for a maximum period of 60 days.
A greater period is not recommended because after 60 days
the drying rate is relatively slower and as a consequence,
there is an increased risk of wood degradation by microor-
ganisms (Stein, 2003).
3.1.2. Wood lipophilic extractives
Considering the importance of the extractives content
for the classification of wood used for pulp and paper pro-
duction (Back and Allen, 2000), the behavior of the Euca-
lyptus wood extractives was evaluated at 20, 40, 60, 100,
140 and 180 days after harvesting (Fig. 2.)
The extractive percentages up to 40 days of storage
(1.80% w/w) were significantly higher than the value
reported for E. globulus (1.52% w/w), without storage (Gut-
iérrez et al., 1999). However, at 60 days after harvesting, the

3.1.1. Wood drying test

2.5
Before evaluating the extractives content as a function
of storage time, it should be noted that after tree harvest- 2
% of extractives

ing, the moisture content decreased slowly until wood 1.5

moisture reached an equilibrium with air relative humidity, 1
indicated by the results shown in Fig. 1. It was verified that,
0.5
for a storage period up to 15 days, the logs that were spread
0
out showed higher mass loss (16.5%) than the logs piled up
20 40 60 100 140 180
(11.2%), because of evaporation. However, after 60 days of
Storage Time (Days)
storage the results were nearly the same, for both the
spread out and piled logs, with approximately 25.0% of Fig. 2. Variation in the extractives percentage during 180 days of storage.
 Author's personal copy

F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886 4881

extractives content decreased significantly, yielding values
lower than the observed for E. globulus, the most used Euca-
lyptus species for pulp production in the Iberian Peninsula.
These results show that storage time improves wood quality
for pulping process, due to reduction of extractives content,
as they could be associated with pitch formation (Back and
Allen, 2000).
Not only is the total extractives content important, but
knowing their chemical composition is also relevant in
order to find out which compounds are reduced more sig-
nificantly with wood storage, and whether or not these
are involved in the pitch formed during processing. GC–
MS analysis of derivatized fractions, before and after alka-
line hydrolysis, revealed a large number of compounds
present in all fractions (Tables 1 and 2).
Fatty acids, sterols, long chain aliphatic alcohols and
aromatic compounds were the main chemical classes found
in all analyzed extractives, before and after hydrolysis
(Fig. 3, shows data for the hydrolyzed extractives). After
hydrolysis, sterols were found in largest amounts for all
storage times, mainly b-sitosterol and b-sitostanol. The
amount of sterols at 20 days of storage (4540.7 mg kg1)
was much higher than at 180 days after harvesting
(2055.7 mg kg1). At 60 days (2714.3 mg kg1) of storage
the wood had 40.2% less sterols than the amount found
at 20 days of storage.
Many sterols have already been found as components of
several pitch samples (Silvestre et al., 1999; Del Rı́o et al.,
2000) and also in bleached pulps (Wallis et al., 1997, 1999;
Silvestre et al., 1999, 2005; Freire et al., 2006). In the latter
case, the presence of high amounts of b-sitosterol and b-
sitostanol in the wood can affect the characteristic of the
produced pulp, mainly the physicochemical properties of
the cellulose fibers, reducing the quality and the value of
the final product (Back and Allen, 2000).
The same reduction occurred for fatty acids in the stud-
ied period of time. After hydrolysis, wood extracts 20 days
after harvesting had fatty acid concentration
(3439.0 mg kg1) approximately four times higher than at
180 days (920.8 mg kg1); similar results were found also
before hydrolysis (Fig. 3, Table 1). At 60 days after har-
vesting, there was a decrease of approximately 45.1% in
the total fatty acids content, particularly those containing
16–18 carbon atoms (Fig. 4). The qualitative composition
of fatty acids derived from the six storage times did not
show great differences (Table 2).
Hexadecanoic or palmitic (C16:0), octadecanoic or stea-
ric (C18:0), octadec-9-enoic or oleic (C18:1), octadeca-9,12
dienoic or linoleic (C18:2) acids were the most abundant
compounds in all extractives, as already reported for other
eucalyptus wood (Freire et al., 2002a; Cruz et al., 2006;
Silvério et al., 2007a, 2007b). Considerable amounts of
docosanoic (C22:0), tetracosanoic (C24:0) and hexacosa-
noic (C26:0) acids were detected in the samples from the
six storage times. All of these fatty acids were found in
the free and esterified forms. This was confirmed indirectly
by the substantial increase in fatty acid content detected
after the alkaline hydrolysis. Similar results were also

Table 1
Components (mg of compound/kg of dry wood) identified in the acetone extract, before (BH) and after hydrolysis (AH), from wood of Eucalyptus cultived
in the Brazil
Chemical Class Storage time (days)
20 40 60 100 140 180
a b
BH AH BH AH BH AH BH AH BH AH BH AH
Fatty acids
Saturated 727.8 1728.3 465.6 1585.7 177.6 1019.3 182.1 836.2 205.3 817.9 196.7 522.4
Unsaturated 514.6 1382.1 295.1 1061.4 87.4 683.1 134.8 547.2 110.2 554.7 78.7 297.9
Hydroxyacids 55.6 328.6 29.5 266.5 11.6 184.8 13.1 118.6 14.7 142.4 15.0 100.5
Total 1298.0 3439.0 790.2 2913.6 276.6 1887.2 330.0 1502.0 330.2 1515.0 290.4 920.8

Aromatic compounds
Aromatic acids 54.3 260.2 186.9 277.9 143.3 205.6 206 206.9 152.3 165.7 34.7 77.5
Others 2.1 74.9 5.4 33.2 2.1 45.4 9.0 17.8 4.7 36.4 1.3 7.23
Total 56.4 335.1 192.3 311.1 145.4 251.0 215.0 224.7 157.0 202.1 36.0 93.5

Long Chain Aliphatic Alcohols

<C20 31.1 54.27 22.3 50.4 27.5 34.7 22.9 35.4 23.9 28.2 16.2 25.4
>C20 36.8 170.9 66.5 149.2 28.4 92.3 11.8 76.4 11.2 98.8 23.8 64
Total 67.9 225.17 88.8 199.6 55.9 127.0 34.7 111.8 35.1 127.0 40.0 89.4

Sterols 4175.6 4540.7 1434.7 3379.4 1197.6 2714.3 468.6 2901.8 993.0 2467.8 923.6 2055.7

Total 5597.9 8539.9 2506.0 6803.7 1675.5 4979.5 1048.3 4740.3 1515.3 4311.9 1290.0 3159.4
a
BH – before hydrolysis.
b
AH – after hydrolysis.
 Author's personal copy

4882
Table 2
Components (mg of compound/kg of dry wood) identified in the acetone extract, before (BH) and after hydrolysis (AH), from wood of Eucalyptus cultivated in the Brazil
PCA Number Compound Storage time (days)
20 40 60 100 140 180
BH AH BH AH BH AH BH AH BH AH BH AH
1 Octan-1-ol 8.4 13.2 10.1 13.6 7.0 10.2 4.5 9.1 2.1 4.0 1.3 5.4
2 Benzoic acid 4.2 12.6 5.5 2.6 1.7 1.2
3 Octanoic acid 2.3 2.4 8.1 1.1 0.8 0.7

F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886

4 Glycerol 11.7 6.77 7.2 8.2 16.7 1.2 14.8 6.6 19.7 8.5 10.6 3.3
5 Nonanoic acid 2.6 1.6 1.1 1.1 0.8 1.0
6 Decanoic acid 2.5 1.9 1.2 2.2 0.9 1.1
7 2-Phenylpropanoic acid 0.8 7.8 4.6 1.2 0.3 0.1
8 4-Hydroxybenzoic acid 12.2 14.2 5.8 6.4 5.6 1.7
9 Dodecanoic acid 7.8 10.2 5.0 2.7 3.2 1.3
10 4-Hydroxy-3-methoxybenzoic acid 2.6 25.6 4.3 42.1 2.6 33.7 4.5 40.1 4.0 31.7 14.0
11 4-Hydroxy-3,5- 2.1 68.6 5.4 28.5 2.1 42.7 9.0 17.3 4.7 35.2 1.3 15.4
dimethoxybenzaldehyde
12 Azelaic acid 1.9 9.8 1.9 4.1 3.9 2.0
13 3,4,5-Trihydroxybenzoic acid 43.3 3.3 179.6 3.9 138.1 1.8 199.0 3.2 145.4 1.5 29.6 1.2
14 Tetradecanoic acid 9.1 23.1 8.3 17.3 4.5 13.1 2.9 10.8 4.7 9.8 2.2 7.3
15 4-Hydroxy-3,5-dimethoxybenzoic acid 26.3 58.3 28.0 51.5 32.3 10.7
16 cis-Ferulic acid 65.1 41.2 59.0 44.2 38.2 14.7
17 2,5-Dihydroxybenzoic acid 8.4 3.0 2.6 2.5 2.9 5.1
18 Coniferilic alcohol 6.3 4.7 2.7 0.5 1.2 0.6
19 Pentadecanoic acid 8.9 26.7 6.2 20.4 1.1 12.9 3.2 13.9 1.5 11.9 1.9 8.3
20 Pentadec-9-enoic acid 4.4 3.9 3.3 2.1 1.8 1.6
21 Hexadecan-1-ol 4.7 9.9 2.7 5.3 2.0 4.5 1.8 4.2 0.9 2.9 2.1 4.0
22 Hexadec-9-enoic acid 3.8 16.8 4.7 12.6 1.6 11.6 2.7 10.8 2.2 10.4 1.8 9.1
23 Hexadecanoic acid 209.8 426.1 116.7 340.3 46.7 189.4 74.1 186.4 53.1 164.3 37.3 103.2
24 trans-Ferulic acid 122.7 97.8 67.2 57.7 54.4 33.9
25 Heptadecanoic acid 9.6 28.5 6.7 24.3 2.8 16.3 3.9 15.0 3.5 14.4 3.2 10.0
26 Heptadec-9-enoic acid 10.3 6.6 4.9 2.9 1.9 1.3
27 Octadecan-1-ol 6.3 24.4 2.3 23.3 1.8 18.8 1.8 15.5 1.2 12.8 2.2 12.7
28 Octadecanoic acid 77.0 153.0 37.7 159.1 16.9 74.8 19.6 57.5 28.0 74.3 15.9 39.6
29 Octadec-9-enoic acid 379.6 1028.8 202.3 771.4 58.8 488.7 88.9 449.8 75.4 456.1 56.0 209.5
30 Octadeca-9,12-dienoic acid 126.8 314.2 84.2 258.8 23.7 168.9 41.1 75.1 30.8 79.6 19.3 75.2
31 16-Hydroxyhexadecanoic acid 9.9 5.7 15.0 4.6 3.0 0.4 5.8 0.6 2.3 1.6 3.0 0.5
32 Nonadecanoic acid 4.9 12.7 4.0 12.7 1.6 7.2 2.5 7.1 1.4 7.6 0.9 3.2
33 Icosan-1-ol 9.9 15.8 9.8 10.2 10.3 7.0
34 Icos-9-enoic acid 12.0 12.0 9.0 8.6 6.7 2.8
35 Icosanoic acid 28.7 58.1 20.4 57.7 6.6 37.1 10.4 31.4 10.8 31.5 5.4 13.9
36 Henicosanoic acid 21.6 51.9 16.0 47.5 5.4 36.3 7.1 31.7 8.1 31.2 4.5 13.2
37 Docosan-1-ol 79.7 49.1 75.9 20.2 47.7 10.2 35.7 4.7 49.0 14.6 32.2
38 Docosanoic acid 48.3 105.8 33.3 96.3 11.5 72.4 10.8 55.4 15.8 57.1 13.9 34.4
39 Tricosanoic acid 34.1 53.9 27.7 61.4 10.9 36.4 8.1 33.7 13.3 31.6 11.4 20.9
 Author's personal copy

F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886 4883

mg of compound / kg of dry wood

5000 day 20 day 40 day 60 day 100 day 140 day180

3159.4
102.4

103.2

163.9
1850
28.6

31.4

18.8

15.2
41.4
28.1

20.2
5.7
0.9

3.9

1.3

4.8
23
4000

3000

1290.0
788.8
44.0
18.9

10.8
28.6
51.2

74.3
1.7

1.2

7.5

8.6
9.3
2000

1000

4307.6
150.7

147.5

156.8
2261
11.4

36.5

44.8

19.8

20.2
59.0
39.9
30.2

27.7
0
1.9

7.9

1.7

5.7
ST FA AC LCAA

Fig. 3. Major classes of compounds identified in the hydrolyzed extracts

1519.6
867.3

of wood at 20, 40, 60, 100, 140 and 180 days after harvesting (ST: sterols,
33.7
13.6

14.9
42.8

14.8

68.1
1.9

2.5

9.9

4.6

2.9

FA: fatty acids, AC: aromatic compounds, LCAA: long chain aliphatic
alcohols).
4740.3
150.2

151.7

168.0
2663
39.4

28.8

35.2

14.6
55.5
40.8
35.6

26.4
9.0
1.3

6.9

1.8

4.2

mg of compound / kg of dry wood

2000
FA < C15 FA C16-C18 FA > C19
1800
1048.3
401.3
21.4

12.7
30.9

32.8

1600
1.8

3.5

5.5

1.6

1.9
3.6

1400
1200
1000
4979.5
205.3

208.6

163.7
2502
40.1

49.9

18.3

19.6
82.9
52.1
30.3

40.9

800
9.0
1.7

6.2

1.8

7.2

600
400
200
1061.7

1675.5
34.0
15.3

16.1
56.3

10.2

69.4

0
1.5

1.9

6.7

6.7

4.2

BH AH BH AH BH AH BH AH BH AH BH AH

20 40 60 100 140 180

6803.7

Storage Time (Days)

291.1

288.5

128.5

214.8
3070
16.5

74.4
11.1
64.1

55.1

29.9

68.8
39.5
11.4

53.0
2.1

2.8

Fig. 4. Major fatty acids present in the acetone extracts of wood with 20,
40, 60, 100, 140 and 180 days after harvesting, before (BH) and after
alkaline hydrolysis (AH). FA < C15 – fatty acids with less than 16
1191.7

2506.0
125.4
84.6
38.9

10.6
53.9

15.0

11.2
17.7

99.9

carbons; FA C16:C18 – fatty acids with 16–18 carbons and FA > C19 –
2.4

3.9

fatty acids with more than 19 carbons.

8539.9
307.3

102.5
317.5

137.8

321.9
4100
19.8

67.8
15.9

69.4

45.6

78.8
49.4
10.2

65.9
3.2

3.3

found in previous works with E. globulus wood (Freire

et al., 2002a,b, 2006).
The x-hydroxylated fatty acids, such as 19-hydroxynona-
3644.9

5597.9
125.9

195.0

297.1
55.5

40.8
79.1

30.0

15.3
38.6

decanoic, 22-hydroxydocosanoic, 23-hydroxytricosanoic,

6.8

4.9

24-hydroxytetracosanoic and 25-hydroxypentacosanoic

acids were only found after alkaline hydrolysis, while
The peak numbers refer to the PCA’s in Figs. 5 and 6.

16-hydroxyhexadecanoic and 21-hydroxyhenicosanoic

acids were found before and after the hydrolysis. The
19-Hydroxynonadecanoic acid

25-hydroxypentacosanoic acid
24-Hydroxytetracosanoic acid
21-Hydroxyhenicosanoic acid

behavior of these compounds during the wood storage is

2-Hydroxyoctadecanoic acid

22-Hydroxydocosanoic acid

23-Hydroxytricosanoic acid

important since they have been previously detected in

pitch samples (Silvestre et al., 1999; Gutiérrez and Del
Stigmasta-3,5-diene
Pentacosanoic acid
Tetracosanoic acid

Hexacosanoic acid

Rı́o, 2005; Cruz et al., 2006).

Octacosanoic acid
Tetracosan-1-ol

Hexacosan-1-ol

Octacosan-1-ol

Aromatic compounds (Fig. 3, Table 1) represent a small

Campesterol

b-Sitostanol
b-Sitosterol

fraction of the total wood extractives for all storage times

[335.1 mg kg1 (20 days); 311.1 mg kg1 (40 days);
Total

251.0 mg kg1 (60 days); 224.7 mg kg1 (100 days);

202.1 mg kg1 (140 days) and 93.5 mg kg1 (180 days)].
The major compounds identified in the extractives were
3,4,5-trihydroxybenzoic acid, cis- and trans-ferulic acids,
4-hydroxy-3,5-dimethoxybenzoic acid and 4-hydroxy-3-
methoxybenzoic acid. The increase in the amounts
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57

of 4-hydroxy-3,5-dimethoxybenzaldehyde, 4-hydroxy-3-
 Author's personal copy

4884 F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886

methoxybenzoic acid, 4-hydroxy-3,5-dimethoxybenzoic
acid, cis- and trans-ferulic acids after hydrolysis indicate that
they are partially esterified in the extractives (Table 2). Long
chain aliphatic alcohols (free and esterified) represent a small
fraction of the total extractives for the six storage times
(Fig. 3 and Table 1) and might not be important in terms
of pitch deposition during the pulping process.
3.2. PCA
3.2.1. PCA of wood extractives before hydrolysis
As shown in Fig. 5, PC1 explains 99.42% of the total
variance, and b-sitosterol with positive influence on the
PC1 (compound 55, Table 2) was the compound responsi-
ble for the differentiation of wood extractives at 20 days of
storage (BH20) from the other extractives, due to its differ-
ent percentage in relation to the other compounds. How-
ever, 40 days after harvesting (BH40), compound 13
(3,4,5-trihydroxybenzoic acid) with negative influence on
the PC1, and with positive influence on the PC2 (0.42%
of variance), was the most influenced by significant increase
(more than 4 times) in relation to the initial amount (20
days of storage) (Table 2), being differentiated from the
samples containing lower b-sitosterol contents. This could
be explained by the fact that 3,4,5-trihydroxybenzoic acid
is derived from the degradation of lignin. Although micro-
organisms, especially fungus, are known to degrade lignin
during eucalyptus storage (Gutiérrez et al., 2000; Van Beek
et al., 2007), this might not be the case in the present inves-
tigation as the wood was monitored for 180 days and no
visible signs of attack by fungus were observed.
The influence of compound 13 is not observed in the
extractives after hydrolysis, since it is highly polar, due to
the presence of three hydroxyl groups, and consequently
it is retained in the aqueous phase during the alkaline
hydrolysis.
3.2.2. PCA of wood extractives after hydrolysis
In Fig. 6, PC1 (98.47% of total variance) indicates that
compound 55 (b-sitosterol) accounts for the differentiation
of wood extractives at 20 days of storage (AH20) from the
others, with a similar trend observed in PCA of Fig. 5
(before hydrolysis). This can be explained by its significant
amount present at 20 days of storage (4100 mg kg1).
However, at 40 days after harvesting (AH40), compound
29 (octadec-9-enoic acid) had larger influence on the differ-
entiation from the others. At 60 days of wood storage, the
location of the obtained extractives (AH60) in the scores of
Fig. 6 is explained by the presence of almost all com-
pounds. Significantly, the same behavior can be seen at
storage times AH100, AH140 and AH180. Finally, PCA
supplied information based on non-supervised classifica-
tion, clustering samples by similarities (Wold et al., 1984;
Ferreira, 2002; Correia and Ferreira, 2007).
At the industrial level it was observed that the drying
process did not affect significantly the wood chipping, as
attested by the chip classification values obtained. Usually,
typical chip classification values are: oversize = 0.5%
(45 mm, round holes), overthick = 6.0% (8 mm, parallel
cylindrical rods), accept = 90.% (7 mm, round holes),
pin = 2.1% (3 mm, round holes), fines = 1.4% (fines tray).
It is worth noting that the wood used in this experiment
is of low density (430 kg/m3) which facilitates chipping
even at low moisture content.
As the experiments were carried out under industrial
conditions, samples from the pulps and paper produced
with the wood stored for the times previously mentioned
were also collected and their physicochemical properties

0.8
0.8

13 AH40
BH40 0.6
0.6 29
29

0.4 0.4
30
43
BH100 23 47
AH60 28 23
0.2 48
30 0.2 51
PC 2 ( 0.42%)

PC 2 (1.20%)

3743 38
37
44
47
28
38
35 AH140 39
3524
57
52
46
31
39
36
11144 36
2
15
711
54
50
33
27
25
12948
10
8
0 19
50
10
14
25
32
53
22
42
20
52
27
21
41
40
45
34
33
24
26
49
51
57
54
12
18
16
153 56
2
9
8
7
6
5 0 32
40
18
34
14
3
13
41
31
26
19
22
49
45
41
20
21
17
42
6
5
53
17
446 BH20 AH180 16 56
55
BH140
-0.2 -0.2 AH20
BH60

-0.4 -0.4 55

-0.6 BH180 -0.6

AH100

-0.8 -0.8
-0.4 -0.2 0 0.2 0.4 0.6 0.8 1 -1 -0.5 0 0.5 1
PC 1 (99.42%) PC 1 (98.47%)

Fig. 5. Normalized scores (s) and loadings (+) of storage time data as a Fig. 6. Normalized scores (s) and loadings (+) of storage time data as a
function of the chemical composition of extractives before hydrolysis function of the chemical composition of extractives after hydrolysis (AH).
(BH). BH20, BH40, BH60, BH100, BH140 and BH180 correspond to the AH20, AH40, AH60, AH100, AH140 and AH180 correspond to the wood
wood storage times. storage times.
 Author's personal copy
F.O. Silvério et al. / Bioresource Technology 99 (2008) 4878–4886
were evaluated. It was observed that yield for the pulping
process did not vary significantly with the wood storage
time (on average the yield was 56.5%) and the screen reject
was bellow 1% for all samples. The pulp viscosity varied
from 62.8 cP for the pulp obtained from wood stored for
20 days to 70.2 cP for wood stored for 180 days. Although
the observed increase was constant, the results were not
statistically different and the values obtained were within
the expected range generally observed under industrial
conditions.
Analysis carried out on the fibers showed that coarse-
ness (7.35–8.95 mg /100 m), average fiber length (0.90–
0.95 mm), and the number of fibers per gram
(15.75 · 106–17.9 · 106) did not vary significantly when
the wood was stored. The chemical properties of the pulp,
including, solubility in NaOH 5% (average of 10.2), hex-
enuronic acid content (71.9–77.7 mmol kg1), and total lig-
nin content (soluble and Klason lignin, 1.91–2.44) were not
affected by the wood storage.
Tests carried out on the paper produced under industrial
conditions, revealed that the physical properties (drainage,
apparent density, tear index, resistance to air and Klemm
capillary) were not affected by storage time. As these prop-
erties depend mainly on the cellulose characteristics, it can
be considered that the storage period did not affect the cel-
lulose content.
4. Conclusions
The results presented in this work revealed that the
extractives present in eucalyptus wood stored for 180 days
in piled logs or spread out in the log yards decline by
69.6%. The rate of the extractive reduction was greater dur-
ing the first 60 days (47.9%) of storage.
Lipophilic fractions analyzed by GC–MS consisted pri-
marily of fatty acids (saturated, unsaturated and hydroxy
acids) and sterols, in both free and esterified forms. Both
groups of compounds decreased in the extractives during
the storage period. Among the fatty acids, those with
16–18 carbon atoms had the largest decrease with storage,
whereas b-sitosterol was the major compound responsible
for the reduction in the total sterol content of the extrac-
tives. These results indicate that the decrease in these two
classes of compounds is directly related to the decrease in
lipophilic extractive contents. From the results described,
associated with PCA analyses, it is recommended that the
wood should be stored for 60 days before being processed.
Although after this period of time, there was still some
reduction in the extractive lipophilic constituents, storing
the wood for a longer period is not economically viable.
The tests carried out on the pulp and paper properties
obtained from the wood stored after 60 days, have demon-
strated that their qualities did not decrease with storage. In
summary, considering the economical aspects, quality of
the pulp and paper produced and possibility of pitch for-
mation, the best period for storage of the wood in the field
was 60 days.
4885
Acknowledgements
To Conselho Nacional de Desenvolvimento Cientı́fico e
Tecnológico (CNPq) for financial support and research fel-
lowships (LCAB, DPV) and a PhD studentship (FOS);
Fundação de Amparo à Pesquisa do Estado de Minas Ger-
ais (FAPEMIG) for financial support. We thank Fabio C.
Chaves from Plant Biology and Pathology Department at
Rutgers University (USA) for suggestions and corrections
made on the manuscript.
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