Steroid from the seeds of artabotrys odoratissimus

Article abstract of DOI:10.13005/ojc/310141

A steroid has been isolated from the seeds of Artabotrys odoratissimus (Annonaceae) and its structure has been established as (3β, 22E)-ergosta-5,7,22-trien-3-ol by spectroscopic and chemical studies. By chemical correlations, it has been identified as a known compound ergosterol. This is the first report of isolation of ergosterol from this plant.

Full text of DOI:10.13005/ojc/310141

ISSN: 0970-020 X
CODEN: OJCHEG
015, Vol. 31, No. (1):
Pg. 353-356
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Free Access, Peer Reviewed Research Journal
2
www.orientjchem.org
Steroid from the Seeds of Artabotrys odoratissimus
1
2
SUBEDAR PRASAD and CHITRASEN GUPTA *
1
Department of Chemistry, M.M. T.D. College, Ballia, India.
2
Department of Chemistry, Kutir Post Graduate College, Chakkey, Jaunpur, India.
*
Corresponding author E-mail: chitrasengupta@yahoo.co.in
http://dx.doi.org/10.13005/ojc/310141
(Received: December 16, 2014; Accepted: January 20, 2015)
ABSTRACT
A steroid has been isolated from the seeds of Artabotrys odoratissimus (Annonaceae)
and its structure has been established as (3, 22E)-ergosta-5,7,22-trien-3-ol by spectroscopic
and chemical studies. By chemical correlations, it has been identified as a known compound
ergosterol. This is the first report of isolation of ergosterol from this plant.
Key words: Artabotrys odoratissimus, Annonaceae, Ergosterol.
INTRODUCTION
MATERIALAND METHODS
Isolation procedure
Artabotrys odoratissimus, Wight & Arn.
[
(Syn: A. uncinatus (Lam.) Merr. and Annona
500g seeds of A. odoratissimus were
collected from the campus of Banaras Hindu
University, Varanasi, India, milled and left overnight
in n-hexane to defat. Then filtered and residue was
extracted with n-hexane in a soxhlet extractor for
16 hours. Systematic column chromatographic
fractionation of the n-hexane extract by with mixture
of solvents i.e., n-hexane and benzene (8:2) resulted
in isolation of an ergosteroid, named ergosterol.
hexapetalus (L. f.) Bhandari)] of the family
Annonaceae is a shrub distributed in tropical and
temperate Asia from the southern part of China to
India.The plant, known for its essential oils, is used
for the treatment of malaria and scrofula in traditional
Chinese medicine . Isolation of aliphatic alcohols
and esters , sesquiterpenes , alkaloids ,
anthraquinones and four new triterpenes were
earlier reported from the plant. In previous works,
isolation of flavonol glycosides is also reported
1
2
3
4
5
6
7
Chemical and physical analysis
from the leaves of the plant. In this paper we report
the isolation of an ergosteroid from the n-hexane
extract of defatted seeds of Artabotrys
odoratissimus.
Melting point was taken on a Yazawa hot
1
13
stage microscope. H-NMR and C-NMR spectra
were recorded on 300 MHz or 500 MHz/ 125 MHz
on Bruker 300 or JEOL GSX-500 MHz spectrometer

3
54
PRASAD & GUPTA, Orient. J. Chem., Vol. 31(1), 353-356 (2015)
Further elution of same column yielded
using tetramethyl silane (TMS) as a internal
reference. The chemical shift value are reported in 1.73g of compound as white crystalline solid and
0
delta (d, ppm) units. Silica gel for column recrystalised from methanol, m.p.: 165-168 C, [a] -
D
0
chromatography refers to E. Merck (India) Ltd. (60- 135 (c, 0.1, CHCl ). EIMS : m/z (rel. int.) 396 (100)
20 mesh). Thin layer chromatography plates were (M ), 386 (15), 363 (71), 353 (7), 337 (29), 316 (4),
3
+
1
prepared with silica gel–G of E. Merck (India) Ltd. 301 (5), 283 (9), 271 (25, 256 (50), 253 (22), 213
and spot were visualized in iodine vapour or by 5% (213), 199 (14), 171 (16), 157 (20), 137 (29), 129
o
H SO in ethanol and heating the plates at 110 C (29), 109 (29), 95 (46), 81 (65), 55 (61), 41 (61).
2
4
for 10 minutes. Compound was routinely dried over
P O for 24 hours in vacuum and was tested for
purity by thin layer chromatography. Anhydrous absorption at max 3400 cm for hydroxyl group. It
IR spectrum of compound showed broad
2
5
-
1
sodium sulphate was routinely used for drying the was, therefore, inferred from IR spectrum of
organic solvents.
RESULTSAND DISCUSSION
compound that it bears hydroxyl group in the
molecule.The H-NMR spectrum of compound was
measured in CDCl3 solution at 500 MHz. The
spectrum showed (Table 2) signals for a number of
1
The single spot nature of compound was methyl groups in upfield region, a carbinyl hydrogen
determined by exposure of the TLC plates to iodine signal at d 3.642, 1H, heptet, signals for four olefinic
vapours as well as charring the plates after spraying hydrogens from d 5.199 to 5.60 ppm and a large
0
with 5% ethanolic sulphuric acid at 110 C for 10 number of signals ranging from d 1.2 to 2.5 ppm for
minutes. The compound responded positive to methine and methylinic hydrogens.
Liebermann-Burchard test, when its solution in
chloroform and acetic anhydride was treated gently
A careful analysis of the signals in upfield
with concentrated sulphuric acid; it developed pink region from d 0.62 to 1.05 ppm revealed that a
colouration at the junction of two layers which turned compound possess two tertiary methyl groups at d
to be gray on prolonged keeping, which shows 0.631 and 0.947, 3H singlet, each which is probably
steroidal nature of the compound. The R values of due to 18 and 19–methyl groups of a steroid
f
compound in different solvent systems are tabulated molecule. The four methyl doublets at d 0.824 (J =
in table-1.
6.6Hz); 0.834 (J = 6.6 Hz) 0.918 (J = 6.6 Hz) and
Table 1: R value of compound in different solvent system
f
Solvent system
Ratio
R value
f
Benzene
-
0.14
0.42
0.56
0.60
Benzene : Chloroform
Benzene : Ethyl acetate
Benzene : Methanol
8.5 : 1.5
9 : 1
5 : 5
1
Table 2: H-NMR in CDCl at 500 MHz compound (Ergosterol)
3
S. N.
No. of Carbons
Chemical Shift
Coupling constantJ (Hz)
1.
2.
3.
4.
5.
6.
7.
3 H
3 H
3 H
3 H
3 H
3 H
1 H
0.631 s
0.824 d
0.834 d
0.918 d
0.947 s
-
6.6
6.6
6.6
—
1.037 d
7.2
3.642 heptet

PRASAD & GUPTA, Orient. J. Chem., Vol. 31(1), 353-356 (2015)
355
H
H
H
H
3
H CCOO
HO
Fig. 2: Ergosterol-3-acetate
Fig. 1: Ergosterol
.037 (J = 7.2 Hz) suggests that these are due to
1
signals at d 5.199, 2H, multiplet, d 5.395, and d
5.569, 1H each. The one at d 5.199, 2H multiplet
was consider to be for the hydrogens associated
with the double bond present in the side chain of
the molecule whereas the two signals at 5.395
and d 5.569 were attributed to two conjugated
double bonds which can only be possible in the B
ring of a steroid nucleus and on the basis of the
data presented so for the structure of compound
can be written as:
secondary methyl groups present in the side chain
of an ergostane. Amongst these four secondary
methyl groups one at d 1.037 was attributed to be
due to C-21 methyl group and two secondary methyl
groups at d 0.824 (6.6 Hz) and d 0.834 (6.6 Hz)
were due to terminal isopropyl group of the side
chain present in normal ergostane type of steroid.
The carbinyl hydrogen signal at d 3.642
heptet was obviously considered for the hydrogen
bound to C-3-position and is ± to C -b-hydroxyl
A search of literature has revealed that
3
group. It is a general characteristic of C-3-a-H, which
appears as heptet in nearly all C-3-hydroxylated
steroids. The presence of C-3-hydroxyl is further
supported by acetylation of compound with acetic
anhydride and pyridine at room temperature which
yielded monoacetate of compound, analyzed for
C H O .The IR spectrum, of acetylated compound
this is a known ergosteroid and has earlier reported
8
from: Engleromyces goetzei , Polyporous
9
10
umbellatus , Phycomyces blakesleeanus and
11
Agelas sceptrum .
This is the first report of isolation of
ergosterol from the seeds of Artabotrys
odoratissimus.
30
46
2
showed the presence of ester carboxyl absorption
-
1
at 1736 cm with simultaneous disappearance of
-
1
band for hydroxyl absorption around 3400 cm .This
showed that only oxygen present in compound is
acylable secondary hydroxyl group.
ACKNOWLEDGEMENTS
The authors thank Prof.Yoshinori Fujimoto,
Department of Chemistry and Materials Science,
Tokyo Institute of Technology, Meguro, Tokyo 152-
8551, Japan for providing spectral analysis and
structural information.
The presence of four olefinic protons in
the molecule of compound is evidence by the
REFERENCES
1
2
.
.
Green et al, J. Cancer Chemether, 1972, 3.
Rupprecht et al, Heterocycles, 1986, 24 (3),
Industrial Research, New Delhi, Vol. I. PP.
119 (1952).
1
179.
5.
6.
A Dictionary of the Economic Products of
India vol.1 p 322.
Gupta, C.; Prasad, S.; Sahai, M.; Asai, A; Hara,
N;Fujimoto,Y., Helvetica Chimica Acta , 2010
93 (10) 1925.
3
4
.
.
Mc Cloud, T.G.; Experientia, 1987, 43.
The Wealth of India: raw materials, A
Dictionary of India raw Material and
Industrial Products, Council of Scientific

3
56
PRASAD & GUPTA, Orient. J. Chem., Vol. 31(1), 353-356 (2015)
7.
8.
9.
Singh, A.P.; Sahai, M., Planta Medica, 1996,
2 (2), 192.
Zhan, Z.; Sun, H.; Wu, H.; Yue, J., Acta
Botanica Sinica, 2003, 45 (2), 248.
Lu, W.;Adachi;Kano, K.;Yasuta, A.;Toriizuka,
K.; Ueno, M.; Horikoshi, I., Chem. Pharm.
Bull., 1985, 33 (11), 5083.
Mercer, E.I.; Bartlett, K., Phytochemistry,
1974, 13 (7), 1099.
Hu, J.- F.; Kelly, M.; Hamann, M.T., Steroids,
2002 67 (9), 743.
6
10.
11.