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Chemical Investigations of Stem Bark’s Extracts of Aegle Mermelos: (an Initiative Exploration Pro Medicinally Important

Mohammad Mamunur Rahman
Department of Applied Chemistry and Chemical Engineering, University of Dhaka, Bangladesh

Md. Raihan Chowdhury
ACI Pharmaceuticals Limited, Narayanganj-1400, Bangladesh

Partha Saha
ACI Pharmaceuticals Limited, Narayanganj-1400, Bangladesh

Mahedy Hasan Chowdhury
Sunbeams School, Plot# 6-12, Road#13/A, Sector#10, Uttara, Dhaka, Bangladesh

S. M. Estiar Haque
ACI Pharmaceuticals Limited, Narayanganj-1400, Bangladesh

Md. Saddam Nawaz
ACI Pharmaceuticals Limited, Narayanganj-1400, Bangladesh

Abstract/ Executive Summary:

This study describes the chemical investigations of Aegle marmelos (A.marmelos), a plant belonging to the family rutaceae. Although some investigations mainly on leaves and fruits have previously been reported with this plant, but very little investigations on bark of A.marmelos have been reported to date. The stem bark of A. marmelos was extracted with organic solvent and the extracts were fractionated by using standard chromatographic techniques. A total of three compounds were isolated of which two (AM_-1 & AM_-5) have been isolated from different plant, but this is the first report of their occurrences in the bark of A. marmelos. The isolated pure compounds were identified by extensive analyses of their high resolution Nuclear Magnetic Resonance (1H-NMR) spectral data. The powdered stem bark of A. marmelos was extracted with methanol. Chromatographic (vacuum liquid chromatography) fractionation and purification of the crude methanolic extract yielded compound (AM_-1), one coumarin (AM_-5), and one sterol (AM_-2). The purified compound (AM-1) was identified as Lupeol, compound (AM_-5) was identified as 5-Methoxy psoralen and compound (AM_-2) was identified as stigmasterol or 24-ethyl-cholesta-5- 22-dien-3beta-ol.

Keywords: Aegle marmelos, Stem bark, Lupeol, Stigmasterol, 5-Methoxy psoralen, Nuclear magnetic resonance

Location: Department of Applied Chemistry and Chemical Engineering, University of Dhaka

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Plant(s)

Objectives:

To study and describe the chemical investigations of Aegle marmelos (A.marmelos), a plant belonging to the family rutaceae, a medicinal plant.

Methodology:

Chemicals used - The chemicals and reagents used for the purpose were obtained from E Merck (Germany) and BDH (England). All the chemicals and reagents used were of analytical grade. Collection and preparation of plant material - The stem bark of the plant A. marmelos was collected from healthy trees (from Dhaka University Campus) and was collected in fresh condition. It was sun-dried and then dried in an oven at reduced temperature (not more than 50⁰C) to make it suitable for grinding purpose. The coarse powder was then stored in airtight container with marking for identification and kept in cool, dark and dry place for future use. Extraction of the plant material - About 700 g of the powdered material was taken in a clean, round bottomed flask (5 liters) and soaked in 2.5 liter of methanol. The container with its content was sealed by foil and kept for a period of 15 days accompanying occasional shaking and stirring. The whole mixture was then filtered through filter paper and the filtrate was concentrated at 50 0C with a rotary evaporator. The concentrated extract thus obtained is termed as crude extract as well as estimated about 28 g extract was obtained. Fractionation and isolation of compounds - 23 g crude extract was subjected to TLC screening to see the type of compounds present in the extract. The whole portion of the methanolic extract was subjected to Vacuum Liquid Chromatography (VLC) for rapid fractionation. The VLC fractions were screened by TLC to find out interesting fractions. The sample for VLC was prepared by absorbing methanolic extract onto silica gel (Kiesel gel 60, mesh 70-230) allowed to dry and subsequently applied on top of the adsorbent layer. The column was then eluted with n-hexane followed by mixture of n-hexane and ethyl acetate and then ethyl acetate and methanol. The polarity was gradually increased by adding increasing proportions of ethyl acetate and methanol. A total of 25 fractions were collected each in 100 ml beakers. All the VLC fractions were screened by TLC under UV light and by spraying with vanillin-sulfuric acid reagent followed by heating at 110⁰C. Instrumentation for isolation and characterization of compounds - The 1H NMR (400 MHz and 300 MHz) spectra was acquired in CDCl₃ on an Ultra Shield Bruker DPX 400 spectrometer and the chemical shifts are reported in parts per million relative to the residual nondeuterated solvent signals. Checking the homogeneity of the compounds were made by TLC on Kieselgel gel 60 F₂₅₄ pre-coated sheets (E. Merck) and the spots were detected by exposure to UV-lamp at 254 nm. Column chromatography was done on silica gel (60-120 mesh ASTM).

Source of Information: International Journal of Biosciences (IJB); ISSN: 2220-6655 (Print) 2222-5234 (Online); Vol. 2, No. 11, p. 74-80, 2012

Article Link (Online Journal): http://www.innspub.net

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Successive chromatographic separation and purification of the methanolic fraction of bark of A. marmelos yielded a total of three compounds. The structures of these compounds were elucidated as (1) Lupeol (2) stigmasterol 24-ethyl-cholesta-5, 22-dien-3beta-ol and (3) 5-Methoxy psoralen. Medicinal plants are rich sources of bioactive compounds and thus serve as important raw materials for drug production. They constitute a precious natural wealth of a country and contribute a great deal to its health care programmes. Judicious and scientific exploitation of this wealth can significantly improve the general health of the people.

Knowledge Management: Journal