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ASIAN JOURNAL OF CHEMISTRY
ASIAN JOURNAL OF CHEMISTRY
https://doi.org/10.14233/ajchem.2017.20689
Antidesma ghaesembilla Gaertn. (family: Phyllanthaceae)
is a plant indigenous of Philippines. Locally known as binayuyo,
the young shoots are used as a vegetable and as a spice [1].
The leaves are used as a poultice to treat headaches, scurf,
abdominal swellings and fevers. The stems are emmen-agogue
and the fruit is purgative. The fully ripe fruit can be eaten
raw, cooked or made into jams and jellies [2]. A previous study
reported that the crude methanol extract of A. ghaesembilla
Gaertn. leaves exhibited moderate to strong antioxidant poten-
tial and showed significant hypoglycemic potential [3]. Recently,
the methanolic extract of A. ghaesembilla leaves was reported
to exhibit antithrombotic, cytotoxic and antibacterial activities
[4]. Chemical studies on the leaves of A. ghaesembilla reported
the isolation of a megastigmane, vomifoliol [5] and the flavone
glycosides, vitexin, orientin, isovitexin and homoorientin [6].
We report herein the isolation of β-friedelinol (1), lupeol
(2), squalene (3), polyprenol (4), β-sitosterol (5), long-chain
hydrocarbons (6) and chlorophyll-a (7) from the leaves and
β-sitosterol (5) and triacylglycerols (8) from the fruit of A.
ghaesembilla. To the best of our knowledge this is the first
report on the isolation of compounds 1-8 from A. ghaesembilla.
NMR spectra were recorded on a Varian VNMRS spectro-
meter in CDCl3 at 600 MHz for 1H NMR and 150 MHz for 13C
NMR spectra. Column chromatography was performed with
silica gel 60 (70-230 mesh). Thin layer chromatography was
NOTE
Chemical Constituents of Leaves and Fruit of Antidesma ghaesembilla Gaertn.
RHANNEY L. GONZALES1,2, CHIEN-CHANG SHEN3 and CONSOLACION Y. R AGASA1,4,*
1Chemistry Department, De La Salle University, 2401 Taft Avenue, Manila 1004, Philippines
2Chemistry Department, Central Luzon State University, Munoz, Nueva Ecija 3020, Philippines
3National Research Institute of Chinese Medicine, Ministry of Health and Welfare, 155-1, Li-NongSt., Sec. 2, Taipei 112, Taiwan
4Chemistry Department, De La Salle University Science & Technology Complex, Leandro V. Locsin Campus, Biñan City, Laguna 4024,
Philippines
*Corresponding author: Tel/Fax: +63 2 5360230; E-mail: consolacion.ragasa@dlsu.edu.ph
Received: 6 April 2017; Accepted: 30 June 2017; Published online: 31 August 2017; AJC-18544
Chemical investigation of the dichloromethane extracts of Antidesma ghaesembilla Gaertn. has led to the isolation of β-friedelinol (1),
lupeol (2), squalene (3), polyprenol (4), β-sitosterol (5), long-chain hydrocarbons (6) and chlorophyll-a (7) from the leaves and β-sitosterol
(5) and triacylglycerols (8) from the fruit. The structure of β-friedelinol (1) was elucidated by extensive 1D and 2D NMR spectroscopy,
while those of compounds 2 to 8 were identified by comparison of their NMR data with literature data.
Keywords: Antidesma ghaesembilla, Phyllanthaceae, ββ
ββ
β-Friedelinol, Lupeol, Squalene, Polyprenol, ββ
ββ
β-Sitosterol, Chlorophyll-a.
Asian Journal of Chemistry; Vol. 29, No. 10 (2017), 2333-2334
performed with plastic backed plates coated with silica gel
F254 and the plates were visualized by spraying with vanillin/
H2SO4 solution followed by warming.
Sample collection: Samples of the leaves and fruits of
Antidesma ghaesembilla Gaertn. were collected from Lupao,
Nueva Ecija, Philippines in June 2016. The samples were
authenticated at the Botany Division, Philippine National
Museum.
Isolation procedure: A glass column 12 inches in height
and 0.5 inch internal diameter was used for the chromato-
graphy. The crude extracts were fractionated by silica gel
chromatography using increasing proportions of acetone in
CH2Cl2 at 10 % increment by volume as eluents. Five milliliter
fractions were collected. All fractions were monitored by thin
layer chromatography. Fractions with spots of the same Rf
values were combined and rechromatographed in appropriate
solvent systems until TLC pure isolates were obtained. Final
purifications were conducted using Pasteur pipettes as columns.
One milliliter fractions were collected.
Isolation of chemical constituents from leaves of A. A.
ghaesembilla: The air-dried A. ghaesembilla leaves (323.4 g)
were ground in a blender, soaked in CH2Cl2 for 3 days and
then filtered. The solvent was evaporated under vacuum to afford
a crude extract (2.4 g) which was chromatographed using
increasing proportions of acetone in CH2Cl2 at 10 % increment
by volume. The CH2Cl2 fraction was rechromatographed using
petroleum ether. The less polar fractions were combined and
rechromatographed using petroleum ether to afford long chain
hydrocarbons (6) (8 mg) after washing with petroleum ether.
The more polar fractions were combined and rechromatogra-
phed using petroleum ether to yield squalene (3) (10 mg).The
10 % acetone in CH2Cl2 fraction from the chromatography of
crude extract was rechromatographed (2×) using 1 % EtOAc
in petroleum ether. Final purification was conducted by
rechromatography using 5 % EtOAc in petroleum ether to
afford polyprenol (4) (9 mg). 30 % acetone in CH2Cl2 fraction
from the chromatography of crude extract was rechromato-
graphed (2×) using 5 % EtOAc in petroleum ether to yield β-
friedelinol (1) (12 mg) after washing with petroleum ether.
The 40 % acetone in CH2Cl2 fraction from the chromatography
of the crude extract was rechromatographed by gradient elution
using 5 % EtOAc in petroleum ether, followed by 7.5 % EtOAc
in petroleum ether and finally, 10 % EtOAc in petroleum ether.
The fractions eluted with 5 % EtOAc in petroleum ether were
combined and rechromatographed using 5 % EtOAc in petroleum
ether to provide lupeol (2) (7 mg) after washing with petroleum
ether. The fractions eluted with 7.5 % EtOAc in petroleum
ether were combined and rechromatographed using 10 %
EtOAc in petroleum ether to afford β-sitosterol (5) (15 mg)
after washing with petroleum ether. The fractions eluted with
10 % EtOAc in petroleum ether were combined and rechroma-
tographed using 15 % EtOAc in petroleum ether to yield
chlorophyll-a (7) (3 mg) after washing with petroleum ether,
followed by Et2O.
Isolation of the chemical constituents from fruit of A.
ghaesembilla: The freeze-dried A. ghaesembilla fruit (38.2
g) was ground in a blender, soaked in CH2Cl2 for 3 days and
then filtered. The solvent was evaporated under vacuum to
afford a crude extract (1.60 g) which was chromatographed
using increasing proportions of acetone in CH2Cl2 at 10 % incre-
ment by volume. The CH2Cl2 fraction was rechromatographed
(2×) using 10 % EtOAc in petroleum ether to yield triacylgly-
cerols (8) (71 mg) after washing with petroleum ether. The
30 % acetone in CH2Cl2 fraction was rechromatographed using
10 % EtOAc in petroleum ether. Fractions from this column
were combined and rechromatographed using 12.5 % EtOAc
in petroleum ether to afford β-sitosterol (5) (39 mg) after
washing with petroleum ether.
Silica gel chromatography of dichloromethane extracts
of A. ghaesembilla afforded compounds 1-8. The structure of
β-friedelinol (1) was elucidated by extensive 1D and 2D NMR
spectroscopy and confirmed by comparison of its NMR data
with those reported in the literature for β-friedelinol [7]. The
NMR spectra of lupeol (2) are in accordance with data reported
in the literature for lupeol [8]; squalene (3) for squalene [9];
polyprenol (4) for polyprenol [10]; β-sitosterol (5) for β-
sitosterol [11]; long-chain hydrocarbons (6) for long-chain
hydrocarbons [12]; chlorophyll-a (7) for chlorophyll-a [13]
and triacylglycerols (8) for triacylglycerols [14].
The presence of α-linolenic acid in the triacylglycerols
(8) was deduced from the methyl triplet at δ 0.96 (t, J = 7.8
Hz), the double allylic methylenes at δ 2.78, allylic methylene
protons at δ 2.00 and the olefinic protons at δ 5.34 (m) [15].
Oleic acid was also found in 8 as indicated by the resonances
for the methyl triplet at δ 0.86 (t, J = 6.6 Hz), allylic methylene
protons at δ 2.00 and the olefinic protons at δ 5.34 (m) [15].
ACKNOWLEDGEMENTS
One of the authors, RLG acknowledges a scholarship and
research grant from the Department of Science and Technology
of the Philippines.
REFERENCES
1. Antidesma ghaesembilla (PROSEA Fruits)–Plant Use. Downloaded from
uses.plantnet-project.org/en/Antidesma_ghaesembilla_ (PROSEA_Fruits).
2. Antidesma ghaesembilla-Useful Tropical Plants; Downloaded from
tropical.theferns.info/viewtropical.php?id=Antidesma+ghaesembilla.
3. M.F. Gargantiel and M.C. Ysrael, Int. J. Sci. Technol. Res., 3, 422 (2014).
4. S. Md. A. Kader, M.S.H. Kabir, M. Hasan, Md. S. Uddin, Md . A.A. Ansary,
M.A.A. Noman, F. Zaheed, Md. R. Hossain, M.Z. Habib, Md. I. Hossain,
A. Hasanat and Md. R. Islam, Int. J. Pharm., 6, 45 (2016).
5. S. Maria, F. Islam, N. Qais and C.M. Hasan, Asian J. Chem., 25, 3533
(2013);
https://doi.org/10.14233/ajchem.2013.13872.
6. L.C.V. Cuong, D.T. Trang, N.T. Cuc, N.X. Nhiem, P.H. Yen, H.L.T. Anh,
L.M. Huong, C.V. Minh and P.V. Kiem, Vietnam J. Chem., 53, 94 (2015);
https://doi.org/10.15625/6654.
7. G.F. Sousa, L.P. Duarte, A.F.C. Alcantara, G.D.F. Silva, S.A. Vieira-Filho,
R.R. Silva, D.M. Oliveira and J.A. Takahashi, Molecules, 17, 13439 (2012);
https://doi.org/10.3390/molecules171113439.
8. V.D. Ebajo Jr., C.-C. Shen and C.Y. Ragasa, J. Appl. Pharm. Sci., 5, 33
(2015).
9. C.Y. Ragasa, V.A.S. Ng, M.M. De Los Reyes, E.H. Mandia and C.-C.
Shen, Der Pharm. Lett., 6, 14 (2014).
10. J.A. Rideout, C.Y. Ragasa and H. Ngo, ACGC Chem. Res. Commun.,
16, 40 (2003).
11. C.Y. Ragasa and V.D. Ebajo Jr., J. Appl. Pharm. Sci., 5, 16 (2015).
12. C.Y. Ragasa and K. Lim, Philipp. J. Sci., 134, 63 (2005).
13. C.Y. Ragasa and J. de Jesus, Res. J. Pharm. Biol. Chem. Sci., 5, 701 (2014).
14. V.A.S. Ng, E.M.G. Agoo, C.-C. Shen and C.Y. Ragasa, Int. J. Pharmacog.
Phytochem. Res., 7, 616 (2015).
15. C.Y. Ra gasa, V.A.S. Ng, O.B. Torres, N.S.Y. Sevilla, K.V.M. Uy, M.C.S.
Tan, M.G. Noel and C.-C. Shen, J. Chem. Pharm. Res., 5, 1237 (2013).
2334 Gonzales et al. Asian J. Chem.
