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Chemical and Antioxidant Studies of Citrus Macroptera (bengali Name- Shatkara; English Name - Wild Orange; Family-rutaceae)

Sadia A. Chowdhury
Department of Pharmacy, Stamford University, Dhaka-1217.

M. Hossain Sohrab
Analytical Research Division, Bangladesh Council of Scientific and Industrial Research Laboratories (BCSIR), Dhaka-1205,

Bidyut K. Datta
Department of Pharmaceutical Engineering and Technology, University of Dhaka, Dhaka-1000,

Choudhury M. Hasan
Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka-1205, Bangladesh

Abstract/ Executive Summary:

Lupeol (1) and Stigmasterol (2) were isolated from the crude extracts of the stem bark of Citrus macroptera (Family: Rutaceae) for the first time. The n-hexane, dichloromethane and methanol extracts were screened in vitro for antioxidant activity using the 1, 1-diphenyl-2-picrylhydrazyl-hydrate (DPPH) free radical scavenging assay. The hot methanol extract showed potential antioxidant activity (IC50: 178.96 μg/ml) whereas the cold methanol and dichloromethane extracts showed moderate activity (IC50: 242.78 μg/ml and 255.78 μg/ml respectively). Mild antioxidant activity was observed with the n-hexane extract of the stem bark of Citrus macroptera (IC50: 422.94 μg/ml).

Keywords: Citrus macroptera, Rutaceae, Lupeol, Stigmasterol, Antioxidant.

Location: Analytical Research Division, Bangladesh Council of Scientific and Industrial Research Laboratories (BCSIR)

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

In this paper, the isolation and structure elucidation of the Lupeol (1) and Stigmasterol (2) by using spectroscopic techniques and the preliminary antioxidant activities of the organic extractives are being reported.

Methodology:

General experimental procedure: The 1H NMR sprectra was recorded using a Bruker DPX-400 (400 MHz) instrument. For NMR studies deuterated chloroform was used and the δ values for 1H spectra were referred to the residual nondeuterated solvent signals. Plant material: Stem bark of C. macroptera was collected from Asam, India and identified by a taxonomist. Extraction and Isolation: The powdered stem bark (260 g) of C. macroptera was extracted in a Soxhlet apparatus for 7 days with n-hexane, 10 days with dichloromethane and 10 days with methanol. Separately 24 g powdered stem bark was soaked in 250 ml methanol for 7 days for cold extraction. All the extracts were filtered through a cotton plug followed by Whatman filter paper number 1 and then concentrated by using a rotary vacuum evaporator to provide n-hexane (1.28 g), dichloromethane (3.43 g), hot methanol (0.523 g) and cold methanol (2.77 g) extractives. The hot methanol extract (0.523 g) was fractionated by column chromatography over silica gel (Kieselgel 60, mesh 70-230). The column was eluted with petroleum ether, dichloromethane and methanol mixture of increasing polarities to provide 31 fractions. Lupeo (Aratanechemuge, et al, 2004; Haque, 2006) (1) (3.6 mg) was isolated from fractions (11+12) eluted with 72.5-75% dichloromethane in petroleum ether. The Rf value of the compound was determined as 0.53 in chloroform-methanol (97:3) on silica gel PF254 plate and the yield value was 0.0014%. The cold methanol extract (2.77 g) was fractionated by column chromatography over silica gel (Kieselgel 60, mesh 70-230). The column was eluted with n-hexane, ethyl acetate and methanol mixture of increasing polarities to provide 22 fractions. Stigmasterol (Khan, 1991) (2) (2.0 mg) was isolated from fraction-2 eluted with 12.5% ethyl acetate in n-hexane. The Rf value of the compound was determined as 0.33 in toluene-ethyl acetate (95:5) on Silica gel PF254 plate and the yield value was 0.0083%.

Source of Information: Bangladesh J. Sci. Ind. Res. 43(4), 449-454, 2008

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Most of the tannins, flavanoids and phenolic compounds may be responsible for antioxidants properties of many plants (Larson, 1988). In these experiments, antiradical activity may be due to the presence of antioxidant principles in the extracts. The free radical scavenging property may be one of the mechanisms by which this plant is effective in its ethno pharmacological uses against different ailments. Further studies comprising of phytochemical investigations of the used plant and evaluation for antioxidant activity using other methods (e.g. various biochemical assays both in vivo and in vitro) are essential to characterize them as biological antioxidants.

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