Termiticidal activity of Toona sinensis wood vinegar against Coptotermes curvignathus holmgren

Abstract

Termiticidal activity of wood vinegar from Toona sinensis (TS) sawdust has been evaluated against Coptotermes curvignathus. TS wood vinegar was produced at 250-300°C pyrolysis temperature, and a no-choice test was applied for evaluating termiticidal activity with thirty-three active termites. The concentrations of TS wood vinegar in the dried filter paper was prepared to 0% (solvent only), 2%, 4%, 6%, and 8%. The TS wood vinegar showed strong termiticidal activity and that concentrations of wood vinegar significantly influenced mortality of termite. The termite mortality increased when the concentration of wood vinegar increased, at a concentration of 8% all termites dead after 3 days tested. Forty constituents were characteristic on the basis of GC-MS data, and 33 components were identified. The organic compounds in TS wood vinegar were classified into carboxylic acids, phenols, ketones, amides, aldehydes, furans, esters, alcohols, sugar derivative, and ether, and among of this components, acetic acid comprised 57.43%. The contents of organic fraction of TS wood vinegar and largely acetic acid might be responsible for the termiticidal activity. Base on physicochemical properties, TS wood vinegar has a good quality as well.

Full text

Vol. 10 | No. 4 |1088-1093 | October - December | 2017
ISSN: 0974-1496 | e-ISSN: 0976-
0083 | CODEN: RJCABP
http://www.rasayanjournal.com
http://www.rasayanjournal.co.in
Rasayan J. Chem., 10(4), 1088-1093(2017)
http://dx.doi.org/10.7324/RJC.2017.1041866
TERMITICIDAL ACTIVITY OF Toona sinensis WOOD
VINEGAR AGAINST Coptotermes curvignathus Holmgren
Morina Adfa
1,
*, Arif J. Kusnanda
1
, Wendra D. Saputra
1
, Charles Banon
1
,
Mai Efdi
2
, and Mamoru Koketsu
3,4
1
Department of Chemistry, Faculty of Mathematics and Natural Sciences, University of
Bengkulu, Jalan W.R. Supratman, Bengkulu 38371, Indonesia
2
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Andalas
University, 25163 Limau Manis, Padang, Indonesia
3
Department of Chemistry and Biomolecular Sciences, Faculty of Engineering, Gifu
University, 1-1 Yanagido, Gifu 501-1193, Japan
4
Department of Materials Science and Technology, Faculty of Engineering, Gifu University,
1-1 Yanagido, Gifu 501-1193, Japan
*E-mail: morinaadfa@unib.ac.id; morinaadfa@yahoo.com
ABSTRACT
Termiticidal activity of wood vinegar from Toona sinensis (TS) sawdust has been evaluated against Coptotermes
curvignathus. TS wood vinegar was produced at 250-300°C pyrolysis temperature, and a no-choice test was
applied for evaluating termiticidal activity with thirty-three active termites. The concentrations of TS wood
vinegar in the dried filter paper was prepared to 0% (solvent only), 2%, 4%, 6%, and 8%. The TS wood vinegar
showed strong termiticidal activity and that concentrations of wood vinegar significantly influenced mortality of
termite. The termite mortality increased when the concentration of wood vinegar increased, at a concentration of
8% all termites dead after 3 days tested. Forty constituents were characteristic on the basis of GC-MS data, and
33 components were identified. The organic compounds in TS wood vinegar were classified into carboxylic acids,
phenols, ketones, amides, aldehydes, furans, esters, alcohols, sugar derivative, and ether, and among of this
components, acetic acid comprised 57.43%. The contents of organic fraction of TS wood vinegar and largely
acetic acid might be responsible for the termiticidal activity. Base on physicochemical properties, TS wood
vinegar has a good quality as well.
Keywords: Toona sinensis, wood vinegar, termiticidal activity, GC-MS
© RASĀYAN. All rights reserved
INTRODUCTION
Termite is recognized as insect pest in the human institution, agriculture, and forestry crop. Reducing
the use of persistent organic insecticide has led to a search for effective, environmentally friendly
compounds with anti-termite activities. The genus Coptotermes is found in tropical and subtropical
regions of the world and is the most devastating genus pest insects in Asia and Australia
1
. Scientists in
different parts of the world are working for the development and establishment biopesticides or natural
pesticides. Biopesticides are natural plant products that have many activities such as insecticidal
activity
2,3
, suppress the growth of fungal
4
, repellent to pests
5
, a deterrent to feeding
6
, insect growth
regulation, and toxicity to agricultural pests
7
.
Wood vinegar which is also called as pyroligneous acid or liquid smoke is a product by processing of
high-temperature carbonization of wood with absent of oxygen. Wood vinegar contains 10-20% organic
compounds including more than 200 chemical components with mainly acetic acid. It also contains
various kinds of phenol, carbonyl, and alcohol. In pesticide application, wood vinegar has been widely
used as a wood preservative, fungicide, repellent and insecticide, herbicide, and feed-stuff
8
. Yatagai et
al.
9
was reported that wood vinegar produced from mixed chips of Cryptomeria japonica and

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TERMITICIDAL ACTIVITY OF Toona sinensis WOOD VINEGAR Morina Adfa et al.
Pseudotsuga menziesii; Quercus serrata; and Pinus densiflora exhibited high termiticidal activity
against Reticulitermes speratus.
Suren (Toona sinensis) timbers are the most popular used in the furniture and construction in West
Sumatera region caused by durability, beauty, and its color. Therefore sawdust waste can be found every
day at the furniture factories or wood workshops. Previously, we reported termiticidal activity of stem
wood and stem bark methanol extracts of T. sinensis and its isolated compounds with moderate activity
2
.
To continue our study concerning the potential of T. sinensis from Indonesia as biopesticides, in this
study we evaluated the termiticidal activity of T. sinensis wood vinegar against Coptotermes
curvignathus and its chemical components.
EXPERIMENTAL
Materials
Sawdust of Toona sinensis was collected from Kanagarian Sungai Jernih, Kecamatan Gunung Talang,
Kabupaten Solok, West Sumatera-Indonesia and then air dried in Laboratory of Organic Chemistry,
Department of Chemistry, University of Bengkulu. Chemicals (NaOH, oxalic acid anhydrous, and
phenolphthalein) were purchased from Sigma-Aldrich.
Instrumentation
GC-MS data were measured with Agilent Series (Agilent Technologies Inc., GC Seri 6980 and MS Seri
5973). A 60 m capillary column HP-5MS (Agilent 1909S1S-436) with id 0.25 mm and 0.25 µm film
thickness was used and maximal column temperature is 350°C. The carrier gas was helium. Front inlet
mode: split with initial temperature 280°C and split flow 199.6 mL/min. The mass spectrometer was
operated in electron-ionization (EI) mode at 70 eV. The mass spectra were obtained by Acquisition
mode scan of the mass range from 40 to 500.
Procedure
Preparation of Wood Vinegar from Toona sinensis
Wood vinegar was produced from burning sawdust of Suren wood (Toona sinensis) (300 g) in a
pyrolysis reactor at 250-300°C temperatures. This process was repeated three times and the vinegar was
combined. The collected wood vinegar was stored at room temperature and sedimentation for 3 months
to separate tar
10
. The wood vinegar was weighed to calculate the yield, color and odor were observed
as well.
Physicochemical Properties of Wood Vinegar
The pH value of wood vinegar was measured by a pH meter (HANNA HI-98107). Organic acid was
determined by the method of acid-base titration with 0.001 N NaOH and expressed as acetic acid (%).
The density of wood vinegar was measured by the pycnometer
11
.
Analysis of Wood Vinegar by GC-MS
The analysis of chemical components of Toona sinensis (TS) wood vinegar was carried out using GC-
MS. The compounds were identified based on the comparison of their retention time (RT) and mass
spectra of Wiley, NIST library data of the GC-MS system. The percentage of components was
calculated by the GC peak area
Termiticidal Activity of Wood Vinegar
Workers and soldiers of Coptotermes curvignathus Holmgren were collected from trees were attacked
by termites around the University of Bengkulu and identified at School of biological sciences and
technology, Institute Technology Bandung, Indonesia. The colony was maintained at 28°C ± 2 and 80%
± 5 RH in a glass container, until used. The termites were fed wet filter paper during storage in the
laboratory. A no-choice test was employed for evaluating the termiticidal activity of TS wood vinegar
according to the previous method
2
. The concentrations of TS wood vinegar in the dried filter paper was

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TERMITICIDAL ACTIVITY OF Toona sinensis WOOD VINEGAR Morina Adfa et al.
prepared to 0% (solvent only), 2%, 4%, 6% and 8% % (sample mass (mg)/filter paper mass (mg) ×
100%), and was applied to filter papers (Whatman No. 3, diameter 90 mm) after dissolved in 500 µL
methanol. Thirty-three active termites (30 workers and 3 soldiers) from the adult of Coptotermes
curvignathus were added to Petri dishes (90 mm in dia × 20 mm height) and kept in a dark room at
room temperature (28°C ± 2) for 1 week. Filter paper treated with methanol only was used as the control.
The number of dead termites was counted daily and the five replications were performed for each
concentration. The termicidal activity was evaluated from the termite mortality (%) average, and the
LC
50
after three days exposures was calculated with Probit analysis
12
.
Statistical Analysis
Duncan's multiple comparison tests were used to evaluate differences in percent mortality of termites.
Results with P < 0.05 were considered significant. Calculations were performed using statistical
software SPSS 16.0 according to the previous report
13
.
RESULTS AND DISCUSSION
Yield, Physicochemical Properties, and Chemical Components of Toona sinensis Wood Vinegar
The yield of TS wood vinegar was 20.29%, with reddish brown in color, smoky in odor, and transparent.
The total organic acids contain 10.897%, the pH value was 3.25, and density was 1.058 g/cm
3
. Wada
14
has been suggested the criteria for good quality of the wood vinegar such as: pH around 3; specific
gravity around 1.010–1.050 g.mL-1; color (visually) pale yellow, bright brown or reddish brown;
smoky odor; and transparency visually is transparent with no suspended matter. On the other hand,
Mathew and Zakaria
15
have been review physicochemical characteristics of pyroligneous acid produced
from different plant sources (Rubber wood, Birch heartwood, Wallnut tree branches, Moso bamboo,
Walnut shell, Mixed chips of Quercus sp, and Coconut shell) with variation carbonization temperature
where wood vinegar exhibited the pH value 1.8–3.8, and density 1.009–1.09 g/cm
3
. Thereby, the TS
wood vinegar in current study appeared to be of good quality in terms of color, odor, pH, density, and
transparency because of the value as in accordance with the previous report. The quality of wood
vinegar depending on the type of starting materials, pyrolysis time and temperature pyrolysis
16
.
The composition of wood vinegar was characterized by 40 components (Figure-1), and 33 components
were identified. The identified components are listed in Table-1 belong to 10 groups. The GC-MS
indicated that TS wood vinegar were mainly contain carboxylic acids (61.96%, 6 components), phenols
(8.29%, 9 components), ketones (4.75%, 4 components), amides (4.43%, 2 components), aldehydes
(3.58%, 3 components), and a few furans (2.7%, 2 components), esters (2.6%, 2 components), alcohols
(1.98%, 3 components), sugar derivative (1.29%, 1 component), and ether (0.56%, 1 component),
successively. The result showed that organic acids were the major components of TS wood vinegar,
while acetic acid was the largest component (57.43%) among 6 kinds of organic acids. Following
organic acids, phenols were the primary compounds in TS wood vinegar, among them 2,6-
dimethoxyphenol/ syringol (2.41%) was identified as the major phenol compound. Phenols basically
result from the thermal degradation of lignin
17
. Similarly to Akakabe et al.
18
perhaps acetic acid, and
syringol were contributed to the characteristic odor of TS wood vinegar. That result was agreement with
Zhai et al.
10
that reported were organic acids, phenolic compounds, ketones, aldehydes, and benzene
and its derivatives were the major components obtained at the middle temperature (190 to 290°C)
pyrolysis range. Thermal cracking of wood is degradation of hemicellulose, cellulose, and lignin
19
. The
acetic acid is believed to have originated from the acetyl groups in the hemicellulose
20
and was the
largest content of the wood vinegar samples obtained in the current study.
Anti-termite activity of Toona sinensis wood vinegar
We evaluated the termiticidal activity of TS wood vinegar at various concentrations with no choice test.
The mortality of termite after exposure with TS wood vinegar and their statistical analysis was shown
in Table-2. TS wood vinegar exhibited strong termiticidal activity, the termite mortality increased when
the concentration of wood vinegar increased. At a concentration of 2% all termites dead after 7 days

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TERMITICIDAL ACTIVITY OF Toona sinensis WOOD VINEGAR Morina Adfa et al.
tested, after the concentration increased to 4%, 6%, and 8% all termite dead after 6, 5, and 3 days,
successively. The mortality of termite increased significantly after 3 days, with its LC
50
of 2.94%. The
termiticidal activity of TS wood vinegar may due to its components. Acetic acid (57.43%) which contain
more than 50% in TS wood vinegar, propionic acid (3.27%), and syringol (2.41%) might be largely
contributed to termiticidal activity in addition to other minor components. The results are agreement
with Yatagai et al.
9
that acetic acid is the largest content of wood vinegar made from mixed chips of
Cryptomeria japonica and Pseudotsuga menziesii; Quercus serrata; and Pinus densiflora exhibited
high termiticidal activity against R. speratus, and phenol with some substituents revealed higher
termiticidal activity than benzene derivatives, which have no hydroxyl group, an ortho substituent of
phenol plays an important role in termiticidal activity. On the other hand, filtrates from biomass slurry
fuel/wood vinegar from sugi and acacia wood had no important termiticidal activity against C.
formosanus. Though high acetic and lactic acids content of the wood vinegar, the presence of vanillin
in sugi and acacia wood vinegar may have served as an additional food source and promoted termite
attack
20
.
Table-1: Identified chemical components of Toona sinensis wood vinegar
RT (min) Compounds Peak area
(%)
3.879 3,4-Furandiol, tetrahydro, cis 1.04
3.932 Methyl glyoxal 0.94
4.191 Acetic acid 57.43
4.414 Propionic acid 3.27
4.557 1,3-Butanediol 0.59
4.843 Ethanimidic acid, ethyl ester 0.87
4.896 2-Oxo, butanoic acid 0.24
4.965 Succindialdehyde 0.65
5.033 2,4-Dimethyloxetane 0.56
5.197 3-Methylbutanoid acid 0.20
5.346 2,5-Dimethylfuran 1.08
5.399 2-Methyl-propanoic acid/ isobutyric acid 0.25
5.494 2-Methyl-propanal/ Isobutyraldehyde 1.99
5.939 1-Pentanol, 4-amino 0.35
6.029 Butyrolactone 1.73
6.527 Phenol 0.45
6.802 1-Hydroxy-2-pentanone 1.70
7.083 3-Methylcyclopentane-1,2-dione 2.21
7.204 2,3-Dimethyl-2-cyclopenten-1-one 0.17
7.432 2-Methyltetrahydrofuran 1.62
7.686 2-Methoxyphenol/ Guaiacol 1.68
7.808 N,N-Dimetylacetamide 3.89
7.988 4-Ethylcyclohexanone 0.67
8.698 2-Methoxy-4-methylphenol/ Creosol 0.78
8.751 2-Hydroxyphenol/ Catechol 0.67
8.793 Cyclopropanecarboxylic acid 0.57
8.920 1,4:3,6-Dianhydro-,α,-d-glucopyranose 1.29
9.518 4-Ethyl-2-methoxyphenol/ 4-Ethylguaiacol 0.18
10.170 2,6-Dimethoxyphenol/ Syringol 2.41
10.996 3,5-Dimethoxy-4-hydroxytuluen 1.05
11.652 2,6-Dihydroxy-4-methoxyacetophenone 0.72
11.795 4-(2-Hydroxyethyl)-2-methoxyphenol 0.35
11.907 N-Acetylpropanamide 0.54

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TERMITICIDAL ACTIVITY OF Toona sinensis WOOD VINEGAR Morina Adfa et al.
Fig.-1: GC chromatogram of Toona sinensis wood vinegar
Table-2: Termiticidal activity of Toona sinensis wood vinegar against Coptotermes curvignathus
Sample Mortality of termite (%)
a
n
over 1-7 days
1 2 3 4 5 6 7
Control 0
A
0
A
0
A
0
A
0
A
0
A
0
A
Wood vinegar [2%] 8.48
A,B
18.18
A,B,C
29.7
C,D
48.48
E
67.88
F,G
90.91
H,I
100
I
Wood vinegar [4%] 19.39
A,B,C
43.03
D,E
60.61
E,F
78.18
G,H,I
94.55
H,I
100
I
100
I
Wood vinegar [6%] 23.64
B,C
57.58
E,F
83.64
G,H
93.33
H,I
100
I
100
I
100
I
Wood vinegar [8%] 49.09
E
83.64
G,H,I
100
I
100
I
100
I
100
I
100
I
a
Numbers followed by different letters (A–I) on mortality (%) are significantly different at the level of P < 0.05
according to Duncan multiple comparison test, n = 5 using 33 termites per replicate.
CONCLUSION
Our results show that the wood vinegar made from Toona sinensis sawdust exhibited strong termiticidal
activity. The contents of organic components and acetic acid might be responsible for the termiticidal
activity. The T. sinensis wood vinegar might be useful for biopesticide in the future application. Future
investigations of TS wood vinegar will be focused on anti-wood rotting fungi.
2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00
1000000
2000000
3000000
4000000
5000000
6000000
7000000
8000000
9000000
1e+07
1.1e+07
1.2e+07
1.3e+07
1.4e+07
1.5e+07
Time-->
Abundance
TIC: SAMPEL_1.D\data.ms

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TERMITICIDAL ACTIVITY OF Toona sinensis WOOD VINEGAR Morina Adfa et al.
ACKNOWLEDGEMENT
The authors would like to acknowledge the financial support of the Ministry of Research, Technology
and Directorate General of Higher Education the Republic of Indonesia for a research grant
International research collaboration and scientific publication (061/SP2H/LT/DPRM/IV/2017). Thanks
to Mohamad Rafi for his assistance in GC-MS. A.J.K. thank head Department of Chemistry to give
opportunity as a research assistant at Division of Organic Chemistry.
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[RJC-1866/2017]