World Applied Sciences Journal 8 (9): 1056-1059, 2010
© IDOSI Publications, 2010
Laevifonol: A Unique Dimer Oligostilbene from the Stem Bark of Vatica odorata
Wan Zuraida Wan Mohd Zain and
Chemistry Department, Faculty of Applied Sciences,
Universiti Teknologi MARA Pahang, 26400 Jengka, Pahang, Malaysia
Faculty of Forestry, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Abstract: The isolation of laevifonol (dimerstilbene) in Vatica odorata is the second time for its present in
Vatica sp. This compound is a unique oligostilbene formed from a condensation between g-viniferin and
ascorbic acid and was firstly isolated from Shorea laevifonia and recently from Vatica umbonata. The structure
of laevifonol was established on the basis of their spectral data, including UV, IR and NMR spectra and also
in comparison with the previously reported data. Cytotoxic properties was evaluated against murine
leukimia P-388 cells and Artemia salina which resulting not strongly inhibited with IC values of 60.5 µM and
> 796.2 µM, respectively. Antibacterial activity also was screened against two gram positive bacteria (Bacillus
subtilis and Staphylococcus aeureus) and one gram negative bacteria (Escherichia coli). The antibacterial
testing was carried out by using the disc diffusion method. Blanc disc of 6mm diameter was loaded with 1000
µg/ml of the compound applied to the inoculate plate. The compound showed moderate activity against all the
bacteria with inhibition zones of 0.5 mm against E.coli and B.subtilis and 0.1 mm against S. aerues compared
to positive control (Erythromycin 60 µg) with 0.13 mm, 0.03 mm and 0.1 mm each. The present investigation is
apart of our ongoing studies on the oligostilbene of Malaysian Dipterocarpacaeae in which no phytochemical
data was recorded on V. odorata.
Key words: Oligostilbene % Dipterocarpaceae % Vatica odorata % Laevifonol % Biological activity
MATERIALS AND METHODS
Stilbene (3,4’,5-trihydroxystilbene) is a phenolic
General Experimental Procedures: UV spectra were
compound are occuring in the particular families such as
measured with a Varian Conc. 100 instrument. IR spectra
Dipterocarpaceae, Vitaceae, Cyperaceae, Leguminoseae
were determined with a Perkin Elmer FTIR Spectrum One
and Gnetaceae . Vatica is a relatively large genus
spectrometer using KBr pellets. H and
C NMR spectr
belonging to the family Dipterocarpaceae and is
were recorded with a JEOL ECP400 operating at 400 ( H)
distributed mainly in Southest Asia . Genus Vatica,
and 100 (13C) MHz using residual and deuterated solvent
Shorea, Hopea and Vateria have proven to be a rich
peaks as reference standards. Vacuum liquid (VLC) and
source of oligostilbene compounds derived from a
column chromatography were carried out using Merck
stilbene . As a phytoestrogen, oligostilbene has
silica gel 60 GF and silica gel G60 35-70
mesh. For TLC
recently been drawn to attentions because of its various
analysis, precoated silica gel plates (Merck Kieselgel
biological properties that may account for its possible
60 GF254, 0.25 mm) were used.
cardioprotective action, including of smooth muscle cell
proliferation and platelet aggregration . Moreover, this
Plant Materials: Samples of the stem barks of V.odorata
compound shows anti-inflammatory  and anticancer
were collected from Pulau Mata Kail, Belum Forest
activities . Because of its pharmacological functions,
Reserve, Perak, Malaysia. The plant was identified by
oligostilbene is now gaining scientific attention as a
botanist, University Putra Malaysia and a voucher
specimen was deposited in the herbarium (SK 616/03).
Corresponding Author: Dr. Kamaruzaman Jusoff, Faculty of Forestry, Universiti Putra Malaysia, 43400 Serdang, Selangor.
Malaysia. Tel: +609-460 2780
World Appl. Sci. J., 8 (9): 1056-1059, 2010
C NMR (100 MHz CD
): 130.7 (C-1a), 12
(C-2a/C-6a), 114.9 (C-3a/C-5a), 157.6 (C-4a), 93.2 (C-7a),
55.1 (C-8a), 144.8 (C-9a), 106.1 (C-10a/C-14a), 159.9 (C-
11a/C-13a), 101.4 (C-12a), 128.6 (C-1b), 127.1 (C-2b/C-6b),
114.1 (C-3b/C-5b), 157.3 (C-4b), 88.8 (C-7b), 55.1 (C-8b),
131.1 (C-9b), 121.7 (C-10b), 160.0 (C-11b), 95.9 (C-12b),
157.9 (C-13b), 110.0 (C-14b), 171.2 (C-1’), 80.0 (C-2’), 117.7
(C-3’), 88.1 (C-4’), 73.4 (C-5’), 74.6 (C-6’).
HRMS-FAB: m/z [M-H] calcd for C
Cytotoxicity Assay: Laevifonol was tested with brine
shrimp lethality test which carried out according to
Said et al., 1990. Meanwhile, cytotoxic test on compound
against murine leukimia P-388 cells was carried out
according to the method described previously .
Antibacterial Assay: The bacteria used for the tests were
obtained from Institute Medical Research (IMR), Malaysia
which included both gram-positive (B.subtilis and
Fig. 1: Laevifonol
S.aeureus) and gram-negative bacteria (E.coli). A 100 µL
of bacterial suspension was spread on nutrien agar (NA)
Extraction and Isolation: The dried powdered tree bark
plate. Laevifonol (1000 µg/mL) as dissolved in methanol
(0.45 kg) of V. odorata was macerated with acetone
on 6 mm sterile filter paper disc and placed on inculated
(3 x 4L) followed by methanol (3 x 4L) and each
agar. Paper disc with methanol solvent was used as
extract was evaporated under reduced pressure to
negative control meanwhile for positive control
give dark brown residues. The MeOH (75g) extract was
Erythromycin 60 µg was used. The plates were incubated
subjected to fractionation using VLC (silica gel, n-hexane-
at 37°C for 18-24h. After incubation time, zone of
EtOAc = 10:0, 8:2, 6:4, 4:6 and 2:8) into five major fractions
inhibition was measured. This method was referred to
F-J. Fractionation of fraction J (6g) was done by flash
Barry at el. .
column chromatography (silica gel, n-hexane-EtOAc =
10:0, 8:2, 6:4, 4:6 and 2:8 and EtOAc-MeOH = 10:0 to 7:3)
RESULTS AND DISCUSSION
into six subfractions J1-J6. Purification of subfraction J3
Laevifonol was isolated as white crystals, which
by radial chromatography (silica gel, CHCl -MeOH = 1
has a melting point at 300°C and optical rotation
to 1:1) yielded laevifonol (40mg).
["]D =-175(c=0.1; MeOH). The UV spectrum showed the
maximum absorptions at 203, 226, 284 and 284 nm which
(-)-Laevifonol (Figure 1)
suggesting the presence of stilbene skeletal in laevifonol.
["]D:-175°(c 0.1, MeOH).
The IR spectrum showed an absorption band for hydroxyl
Rf: 0.58 (100% EtOAc).
at 3364 cmG , a str
etching vibration for carbon aliphatic at
IR (KBr): 3364, 2913, 1789, 1614, 1516, 1454, 1257, 835 cmG .1
2913 cmG1, a stretching absorption for carbon aromatic at
UV/Vis 8 (MeOH) nm: 203, 226, 278, 284, 298.
1614, 1587, 1516, 1454 cmG ,
1 a stretching absorption for
NMR (400 MHz, CD COCD
tetra-substituted ring at 835 cmG and aryl-O at 1257cm
: 6.76 (2H, m, H-2a/H
6a), 6.74 (2H, m, H-3a/H-5a), 5.03 (1H, br d, J = 7.3 Hz, H-
Beside that, there was a special characteristics for
7a), 3.26 (1H, d, J = 7.4 Hz, H-8a), 5.90 (2H, s, H-10a/H-14a),
adsorption of laevifonol which was an absorption band at
6.15 (1H, t, J = 2.2 Hz, H-12a), 6.96 (2H, d, J = 8.4 Hz, H-
1789cmG for lactone carbonyl.
The H NMR spectrum for laevifonol showed th
2b/H-6b), 6.73 (2H, m, H-3b/H-5b), 5.27 (1H, d, J = 10.6 Hz,
signals was for dimer resveratrol. This data was proved
H-7b), 3.30 (1H, ovlp., H-8b), 6.17 (1H, d, J = 2.2 Hz, H-
by the presence of one multiplet signals at *6.73-6.70
12b), 7.13 (1H, br s, H-14b), 4.41 (1H, br s, H-4’) 4.21 (1H,
integrated as three equivalent revealed at H-2a/H-6a,
m, H-5’), 3.97 (1H, dd, J = 4.4, 10 Hz, H-6’a), 4.05 (1H, dd,
H-3a/5b. Meanwhile one doublet signals at * 6.96 ( J = 8.4
J = 2.2, 10.3 Hz, H-6’b).
Hz, H-2b/H-6b) assigned as two unit 4-hydroxyphenyl.
World Appl. Sci. J., 8 (9): 1056-1059, 2010
Table 1: NMR data of Laevifonol in acetone d6 (400 MH )
*H (multiplicity, J in Hz)
Table 2: Inhibition zone of bacteria treated with Laevifonol, Erythromycin 60 µg and methanol
Inhibition zone (mm)
Erythromycin (60 µg) (Positive control)
Methanol (Negative control)
The presence of 1,3,5-trisubstituted ring group was
* 3.30(1H, H-8b). However two signals at * 3.26 and
assigned by the triplet signals at * 6.15 ( J = 2.2 Hz, H-12a)
* 3.30 for H-8a and H-8b could not count the multiplicity
and multiplet signals at * 5.90 integrated as two
due to it was hidden in the water peak. (* 3.28-* 3.31).
equivalent protons at H-10a and H-14a. The signals for
Laevifonol compound was in same group with
one set aromatic proton with meta signals at * 6.17 (1H,d,
g-viniferin . This type of dimer stilbene has a ring of
J=2.2Hz) and * 7.13 (br s, 1H) each was revealed signals at
2,3-dihydrobenzofurane. However, laevifonol no longer
H-12b and H-14b assigned as 1,2,3,5-tetra-substituted
has this skeletal stilbene as g-viniferin. Vinyl group from
stilbene skeletal has been broken and combined with
signals for aliphatic proton in the
ascorbic acid unit . The presence of ascorbic acid unit
dihydrobenzofurane integrated as dublet at * 5.03
was assigned with integration of four signals for
(br d, J = 7.3 Hz, 1H, H-17a) and * 3.26(H-8a). H NM
aliphatic proton at * 4.41 (br s, 1H, H-4’), * 4.21(m, 1H,H-
spectrum also revealed that that were two signals for
5’, *3.97 (dd, J==4.4Hz, 10.0Hz, H-6a’) and *4.05
aliphatic proton at * 5.27(d, J = 10.6 Hz,1H, H-7b) and at
(dd, J==2.2Hz, 10.3, 1H, H-6b’).
World Appl. Sci. J., 8 (9): 1056-1059, 2010
The C NMR
spectrum gave a 27 signals represented
for 34 carbons, seven of them are equivalent. Seven
equivalent carbon assigned at *128.0 (C-2a/6a), * 114.9
(C-3a/5a), * 55.1(C-8a/8b), * 106.1 (C-10a/14a), * 159.9
(C-11a/13a), * 127.1(c-2b/6b) and * 114.7(C-3b/5b). There
was one signals shifted at downfield (*171.2) revealed a
Symington, C.F., 1974. Foresters’ Manual of
characterictic signal for carbonyl carbon which support IR
Dipterocarps. University Malaya Publication, Kuala
data for lactone carbonyl band absoption. Five signals of
oxyaryl carbon assigned at *157.3-159.9 revealed that
Sotheeswaran, S. and V. Pasupathy, 1993.
laevifonol was a dimer stilbene. Other signals were signals
Distribution of resvertarol oligomers in plants.
for nine aliphatic carbon at *55.2-117.7 and four aromatic
Phytochemistry, 32: 1083-1092.
carbon at *95.9-144.8.
Pace-Asciak, C.R., S. Hahn, E.P. Diamandis, G. Soleas
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Cytotoxicity Activity: A preliminary toxicity test of the
trans-resveratrol and quarcetin block human platelet
laevifonol isolated revealed that it was not strongly
aggregration and icosanoid synthesis: Implication for
inhibited to Artemia salina with IC value of >796.2 µ
protection against coronary heart disease. Clin. Chim.
and cytotoxic evaluation against P-388 cells inidicated IC
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Bertelli, A.A., F. Ferrara, G. Diana, A. Fulgenzi, M.
Corsi, W. Ponti, M.E. Ferrero and A. Bertelli, 1999.
Resveratrol a natural stilbene in grapes and
Antibacterial Activity: As can be seen in Table 2 above,
enhances intraphagocytosis in human
laevifonol showed strongly inhibited against B. subtilis
promonocytes: a co0-factor in anti-inflammatory and
followed by E.coli which compared with positive control
anticancer chemopreventive activity. Intra J. Tissue
as standard. Meanwhile for S. aeureus it showed the same
React, 21: 93-104.
activity with standard. The negative control (methanol)
Lee, M.H., B.Y. Choi, J.K. Kundu, J.K. Shin, H.K. Na
did not inhibit any bacterial growth.
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important chemical characters for the chemotaxonomical
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analysis of Vatica. Moreover, the occurence of
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laevifonol which is a unique oligostilbenoid formed from
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a condensation of g-viniferin and ascorbic acid highlights
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the relationship between Vatica with Shorea.
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L.B. Ghisalbertti and E.L. Latip, 2005. Cytotoxic
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Hopea dryobalanoides. J.Z Naturforsch (in German)
We wish to thank MOHE for the Financial support
of the FRGS grant (Project Code: 011000080002), Mr.
Barry, A.L., M.B. Coyle, C.E.H and R.W. Howkinson,
Shamsul Khamis of Universiti Putra Malaysia for the
1979. Methods of measuring zones of inhibition with
identification of the plant specimen and also Universiti
the Bauer-Kirby disk susceptibility test. J. Clinical
Kebangsaan Malaysia (UKM) for the financial assistance
Microbiol., 10: 885-889.
during the Postgraduate Student Exchange Program at
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L. Makmur and Y.M. Shah, 2004. Oligostilbenoids
Bandung Institute of Technology, Indonesia.
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and Ecol., 32(11): 1051-1053.
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