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ISSN: 2349-2759
Research Article
Abstract
1. Introduction
contains them. In fact, the European Foods The powdered leaf material (310g) of
Safety Authority [4] recommends Odontonema strictum (Acanthaceae) were
consuming about 1.5 - 2.4 g/day of extracted with 800 ml of Methanol (MeOH)
phytosterols and/or stanols in order to and DichloroMethane (DCM) for 24 hours.
reduce blood cholesterol. Furthermore, The supernatant was filtered through
FDA has approved the role of foods Whatman N1 filter paper. The extract was
containing phytosterol esters inside a low then left to dry under room temperature.
saturated fat and cholesterol diet in The quantity extracted was 25g.
reducing the risk of heart disease, The crude extract was subjected to the
especially consumption of at least 1.3 qualitative phytochemical analysis (Table
g/day sterols, twice a day [5]. The 1). Phytochemical screening of the extracts
antibacterial activities of stigmasterol and indicated the presence of flavonoids (type
beta sitosterol have been reported in many of flavones), saponins, glycosides, tannins,
reports [6][7][8]. Research has indicated steroids and terpenoids.
that stigmasterol may be useful in Vacuum Liquid chromatography (VLC)
prevention of certain cancers, including was chosen to separate compounds using
ovarian, prostate, breast, and colon silica gel as a stationary phase. 90 g of
cancers. It also possesses potent silica gel for Thin Layer Chromatography
antioxidant, hypoglycemic and thyroid (TLC)(Merck) was mixed with hexane to
inhibiting properties [9]. Corfuff and form slurry and stirred using a stirring
Benedi [10] reported the laxative rod. 25g of the crude extracts was mixed
properties of stigmasterol. - sitosterol is with 1 g of silica gel and the mixture was
used as an antioxidant and an antidiabetic dried in room temperature. A vacuum was
agent [11]. created to allow separation of compounds.
The present study is aimed to extract, Initially, hexane was gradually added into
isolate and characterize by spectroscopic the column to remove fats, waxes and
methods the stigmasterol and - sitosterol some chlorophyll. The polarity was
from Odontonema strictum. increased by addition of EtOAC (0% -100
%). The total volume used was 200 ml. 26
2. Experimental fractions were collected and left to dry at
Collection, Identification and room temperature. Fractions were mixed
preparation of plant materials according to the results obtained from
TLCs. The use of concentrated sulfuric acid
Plant specimens were collected in Lusaka and vanillin as a spraying reagent revealed
(January-February 2014) and identified by the presence of sterols in fractions 7 and 8
Dr. CHUBA and his team of the Department (870 mg).
of Biological Sciences/University of A normal Column Chromatography (CC)
Zambia. Voucher samples were prepared was used to fractionate compounds from
and deposited in the Herbarium of the fractions 7-8 using Hexane- EtOAc- CHCl3:
Department of Biology. The plant samples 5:1:2 as the solvent system. 6 fractions
were shade dried at room temperature were obtained. A white crystal was formed
and powdered into a fine powder in a in one of the eluates. The crystals were
blender. named compound 1.
The compound (1) was subjected to TLC
Extraction, fractionation and isolation using several solvent systems including
Procedures Ethyl acetate: Hexane (1:5), Ethyl acetate:
Hexane: Chloroform (1.5: 2) and it showed
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
to be homogenous compound. The white 3.529 ppm. At 5.197 ppm and at 5.378
crystalline powder (80mg) with melting ppm corresponds to a peak in the form of a
point (134136C) and Rf value 0.55 single in the region of the ethylene protons
(EtAc/Hex: 1/3) was further subjected to suggesting the presence of three protons.
IR, Proton NMR (400MHz), Carbon13 13NMR spectrum of Compound (1) has
NMR (100 MHz) and 2NMR. given signal at 140.943 and 1211.321ppm
Tests for steroid for C5=C6 double bond respectively,
Compound (1) gave a positive test to 71.974 for C3 -hydroxyl group 19.064
Liebermann Buchard and Salkowski and 12.060 for angular methyl carbon
reagents for steroidal nucleus [12][13] atoms for C19 and C18 respectively (table
[14][15]. 1).138.404 ppm for C22 and 129.341ppm
Spectroscopic characterization for C-23. The C5, C6, C22 and C23
Different spectroscopic methods were appeared to be alkene carbons.
used to elucidate the structure of isolated
compound (1), including: IR, 1H NMR and 3. Results and Discussion
13C NMR. The infra red spectrum was From the positive tests for steroids given
recorded on FTIR Perkin Elmer, 1HNMR by compound 1, it is assumed to be a
and 13CNMR spectra were recorded using compound containing steroidal nucleus.
CDCl3 as solvent on Bruker Advance II 400 Compound (1) is white crystalline
NMR spectrometer at the department of substance with melting point 134136C
Chemistry, University of Cape Town. and Rf value 0.55 (EtAc/Hex: 1/3). On
On subjection to IR Spectroscopic analysis, subjection to IR spectroscopic analysis, the
the observed absorption bands are observed absorption bands are
3547.41cm-1 that is characteristic of O-H 3547.41cm that is characteristic of OH
1
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
Table 2: 1H and 13C NMR chemical shift values for compound 1 recorded in CDCl 3 (400 MHz)
a-b.
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Luhata Lokadi Pierre et al, JIPBS, Vol 2 (1), 88-95, 2015
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