British Journal of Pharmacology and Toxicology 2(5): 270-272, 2011

ISSN: 2044-2467
© Maxwell Scientific Organization, 2011
Submitted: September 08, 2011 Accepted: September 30, 2011 Published: November 25, 2011

Isolation of Steroids from Acetone Extract of Ficus iteophylla

1, 2
I.A. Abdulmalik, 2M.I. Sule, 2A.M. Musa, 3A.H. Yaro, 2M.I. Abdullahi,
1
M.F. Abdulkadir and 1H. Yusuf
1
Department of Applied Science, C.S.T. Kaduna Polytechnic-Nigeria
2
Department of Pharmaceutical and Medicinal Chemistry, Ahmadu Bello University,
Zaria -Nigeria
3
Department of Pharmacology, Faculty of Medicine, Bayero University Kano-Nigeria

Abstract: Two steroids were isolated from the leaves of Ficus iteophylla (Family: moraceae) a plant popularly
used in African traditional medicine to treat variety of illnesses. The leaf part of the plant was investigated
phytochemically using a standard procedure. Schematic fractionation of its ethanol extract by acetone and
methanol and subsequent column chromatography of the acetone fraction over silica gel G (60-120) mesh size
led to the isolation of 3$-cholest-5-ene-3, 23diol (1) and 24 ethyl cholest-5-ene- 3$-ol (2). The structure of
these two compounds was elucidated using 1H-NMR, 13C- NMR and DEPT analysis. To the best of our search
this is the first report on the isolation of these compounds from the leaves of Ficus iteophylla..

Key words: Ficus iteophylla; 1: H- NMR; 13: C- NMR, DEPT; 3$-cholest-5-ene-3; 23diol; 24 ethyl cholest-5-
ene-3$-ol

INTRODUCTION solvent in both cases were removed at reduced pressure to

Ficus iteophylla belongs to family moraceae, The
bark is used to treat dysentery and rheumatic pain
(Burkill, 1997). The root has a wide usage for treating
paralysis, tuberculosis, epilepsies, convulsion, spasm and
pulmonary troubles (Burkill, 1997). The leaf part is
reported to have analgesic, anti-inflammatory activity
(Abdulmalik et al., 2011) and antibacterial activity
(Ahmadu et al., 2006). It is also reported to contain two
furanocoumarines (Ahmadu et al., 2004) and two
flavonoid glycosides (Ahmadu et al., 2006). In
continuation of investigation of bioactive metabolites
from Ficus iteophylla, we report herein the isolation and
identification of steroids from the leaf part of the plant.
MATERIALS AND METHODS
Plant material: The plant samples were collected from
Ahmadu Bello University, Zaria Nigeria in the month of
March, 2006. It was authenticated by comparing with the
existing one by Mallam Musa Muhammad of the
Herbarium section of the Department of Biological
Sciences, Ahmadu Bello University, Zaria, Nigeria.
Extraction procedure: The powdered leaves (850 g) of
the plant was exhaustively extracted with petroleum
ether(60-80ºC) using Soxhlet apparatus, the marc was
dried and extracted with ethanol (96%) in same way. The
give 15 and 50 g of Petroleum Ether (PE) and ethanol
(EE), respectively. The Ethanol Extract (EE) was
successively partitioned with acetone followed by
methanol to give acetone extract coded EEAc (8 g) and
methanol extract coded EEM (16 g).
Column chromatography of acetone extract from
ethanolic extract: The acetone extract from ethanol
extract (EEAc) was dissolved in small quantity of acetone
and was adsorbed on silica gel. It was allowed to dry and
ground into a fine powder. The fine powder was applied
over a well-packed silica gel G (60-120 mesh size)
column. The column was then eluted gradiently with
Hexane: Ethyl acetate mixture, with polarity increased
gradually. Eluents were collected as 30 mL fraction and
the progress of the separation was monitored by thin layer
chromatography, similar fractions were pooled together.
Instruments: The melting point was determined using
Gallemkemp capillary and melting point apparatus and
they were uncorrected. The 600 MHz H-NMR spectra
were recorded in CDCl3 with Teramethyl Silane (TMS) as
internal standard. The 13C NMR and DEPT were recorded
at 400MHz. The DEPT experiments were used to
determine the multiplicities of carbon atoms. Thin layer
chromatography (TLC) was performed on TLC Silica gel
60 F254 pre-coated (Merck). The spots were visualized by
spraying with 10% H2SO4 followed by heating at 100ºC
for 5 min.

Corresponding Author: I.A. Abdulmalik, Department of Applied Science, C.S.T. Kaduna Polytechnic-Nigeria, Tell.: +234-
8069451993
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 Br. J. Pharmacol. Toxicol., 2(5): 270-272, 2011

Table 1: 1H and 13C NMR chemical shift assignments of compounds 1

and 2 and DEPT spectrum of compound 2
1 2
----------------------------------- --------------------------------------------
Carbon 1Ha 13 a
C 1
Ha 13 3
C DEPTb
1 38.9 1.0825 37.2665 CH2
2 37.7 1.5158 31.6687 CH2
3 3.59 79.0 3.5255 71.8239 CH
4 48.8 2.2860 42.3056 CH2
5 158.1 - 140.7623 C
6 5.48 116.8 5.3589 121.7271 CH
7 37.8 1.9951 31.9134 CH2
8 28.8 1.2534 23.0732 CH
9 29.9 1.1831 24.3059 CH
10 - - - C
11 27.9 1.460 21.0852 CH2
12 41.3 1.1600 39.7822 CH2
13 - - - C
14 63.2 0.9814 56.7756 CH
15 35.6 1.5857 29.1625 CH2
16 33.3 1.8573 28.2480 CH2
17 55.5 1.0915 50.1417 CH
18 0.85 18.8 0.6980 11.9817 CH3
19 1.01 17.4 1.0374 11.8602 CH3
Fig. 1: Thin Layer Chromatography for compound 1 and 2) 20 38.8 1.3575 37.2561 CH
21 1.50 27.1 0.9292 19.0351 CH3
22 38.0 1.3229 33.9534 CH2
23 3.15 63.0 1.1718 26.0861 CH2
HO  24 49.3 1.9198 45.8475 CH
25 35.7 1.6587 29.6965 CH
26 1.20 22.6 0.8265 18.7810 CH3
27 1.20 21.3 0.8265 18.7810 CH3
28 1.2288 23.0732 CH2
HO  29 0.8530 19.8174 CH3
a
: Spectra recorded at 600MHz in CDCl3; b: Spectra recorded at 400MHz
in CDCl3
Compound 1
proton signal (*H1.0 - *H1.8) attributed to resonance of
overlapping of methylenes and methines a characteristic
frame work of steroid (Yun-Song et al., 2006). It showed
a multiplet at *H 3.59 and 3.15. The signal at *H 3.59 and
3.15 revealed the presence of two hydroxyl group, these
signal were for proton on carbon a djacent to alcohol.
The signal at *H 3.59 was ascribed to C-3 (XU and Zeng,
2000) and that at *H 3.15 was ascribed to C-23. The C-6
HO   oleifinic proton appeared at *H 5.48. The 1H-NMR
showed vicinal coupling between C-3 methine proton and
Compound 2 C-2 methylene at *H 1.98 and *H 1.85 (López et al.,
2008), the apectrum further displaced signal at *H 0.85 (C-
RESULTS AND DISCUSSION 18) and *H 1.01 (C-19) characteristic of 5-ene-3$-hydroxy
sterols, at *H 1.50 (C-21) and *H 1.20 (C-26 and C-27).
Thin layer Chromatography of Compound 1 and The 13C-NMR showed peaks at *C 11.8, *C 11.7, *C 19.0
Compound 2 (Fig. 1). Chromatographic separation of the corresponding to C-18, C-19 and C-21, respectively. The
acetone fraction over a silica gel G (60-120) mesh size led C-3 and C-23 resonated at *C 79.0, *C 63.2, respectively.
to isolation of two compounds that gave positiv C- 5 resonated at *C 158.1 while C-6 resonated at *C
Salkowski and Liebermann-Burchard test specific for 116.8. Some of the signals are shown in (Table 1). Based
steroids. on above evidence the structure of compound 1 was
Compound 1 was obtained as white crystal, its identified as 3$-cholest-5-ene-3, 23diol.
melting point is 228-230ºC, Rf value is 0.515 Compound 2 was obtained as white crystal, its
(Hexane/Ethylacetate). The structure was established by melting point is 106-108ºC, Rf value is 0.350
1
H-NMR and 13C-NMR at 600MHz in CDCl3. The 1H-N (Hexane/Ethylacetate). The structure was established by
spectrum of the isolated compound showed a series of DEPT, 1H-NMR and 13C-NMR at 400MHz in CDCl3. Its

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1
HNMR revealed the presence of hydroxyl group at *H
3.52 while the oleifinic proton appear at *H 5.35 which
shows that there is double bond between C-5 and C-6.
The spectra further revealed the presence of six methyl
groups *H 0.69. 1.03, 0.92, 0.82, 0.82, and 0.85
corresponding to *C C-18, C-19, C-21 C-26, C-27, and C-
29, repectively. The 13C NMR spectrum showed the
presence of 29 carbon signals in the molecules. The
DEPT spectrum exhibited six methyl, eleven methylene
and nine methine, while the remaining three signals in the
broad band spectrum were due to the quaternary carbon
atom. All the signals are shown in (Table 1). Base on the
evidence above compound 2 was identified as 24-ethyl
cholest-5ene-3$-ol. These two compounds probably could
be responsible for the analgesic and anti-inflammatory
activity already reported.
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