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Chemical Investigation and Microbial Activity of Tinospora Cordifolia Miers

A H Molla
Department of Applied Chemistry and Chemical Engineering, University of Rajshahi, Rajshahi- 6205, Bangladesh

M G Zakaria
Department of Applied Chemistry and Chemical Engineering University of Rajshahi, Rajshahi- 6205, Bangladesh

M T H Molla
Department of Applied Chemistry and Chemical Engineering University of Rajshahi, Rajshahi- 6205, Bangladesh

M T Alam
Department of Applied Chemistry and Chemical Engineering University of Rajshahi, Rajshahi- 6205, Bangladesh

M S Ahsan
Department of Applied Chemistry and Chemical Engineering University of Rajshahi, Rajshahi- 6205, Bangladesh

Abstract/ Executive Summary:

Context: Plants have been used in treating human diseases for thousands of years. Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer, malaria, HIV/AIDS, Alzheimer's, and pain. Objectives: The chemical investigation, antimicrobial activity and toxicity of the active principles isolated from the plant. Materials and Methods: The rectified spirit extract of the fresh stem of the plant Tinospora cordifolia was fractionated using standard chromatographic techniques to afford several fractions. The fraction TC-1 was purified by crystallization and screened. FTIR, 1H NMR, 13C NMR spectral analyses were performed to characterize the compound. The antibacterial and antifungal activities of TC-1 were observed by “Disc diffusion method” against a number of pathogenic bacteria and fungi. Standard antibiotics “Kanamycin” (30μg/disc) and “Fluconasol” (500 μg/disc) were used respectively for comparison. The minimum inhibitory concentration (MIC) of the compound TC-1 was carried out by the “Serial Dilution Technique”. The compound also showed significant activity against the brine shrimp nauplii. Results: The compound TC-1 showed promising antibacterial and antifungal activities against all tested organisms.The minimum inhibitory concentration (MIC) of TC-1 against Bacillus megaterium and Salmonella typhi-A was found to be 128μg/ml in nutrient broth medium. The value of medium lethal concentration, LC50 (9.34 μg/ml) indicated the high toxic effect of the compound TC-1. Conclusion: It may be concluded that the compound TC-1 is an alkaloid having significant activities.

Keywords: Chromatography, Antibacterial activity, Antifungal activity, Toxicity, Minimum inhibitory concentration, Brine shrimp lethality bioassay.

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

Start Date: 00-01-1997

End Date: 00-02-1997

Program Area: Development of Host and Medicinal Plants

Non-commodity: Performance

Objectives:

The chemical investigation, antimicrobial activity and toxicity of the active principles isolated from the plant.

Methodology:

The chemicals used in this study were purchased from BDH Chemical Ltd., England and Merck, Germany and were purified by standard methods prior to use. Infrared (IR) spectrum was recorded with Pye-unicam SP3 -300 Spectrophotometer (KBr), NMR (1H, 500 MHz; 13C, 400 MHz) spectra were obtained in pyrd5 on a JEOL-AX-500 and JEOL-JNM AX 400 spectrometers, respectively. The chemical shifts are given in δ values. Collection of the plant: Fresh stem of Tinospora cordifolia Miers were collected from the adjoining areas of Rajshahi University (Bangladesh) Campus during the month of January-February, 1997. Preparation and extraction of plant materials: The stems of the plant were cut into small pieces and made paste using morter and pestle. It was then extracted with rectified spirit and the solvent was evaporated off under normal pressure and temperature to obtain a gummy mass. Isolation and purification of compounds: The crude rectified spirit extract was subjected to alumina column for fractionation. The sample was successively eluted with n-hexane, n-hexane with increasing portions of petroleum ether, ethyl acetate, and finally with methanol. A number of color bands were observed during the development of the columns. Different fractions were obtained and the fractions were combined on the basis of their preliminary TLC examination. Only the fractions designated as TC-1 were collected and the solvent of the combined fractions was evaporated off under reduced pressure to obtain a crystalline mass which after re-crystallization needle shaped compound, TC-1 having melting point 187ºC was obtained.Source of Test Microorganisms: To determine the antibacterial and antifungal activity of compound TC-1, a number of organisms were collected from “The International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B).” Antibacterial and antifungal screening of TC-1: The antibacterial and antifungal activities of TC-1 were determined using “Disc diffusion method” Bauer et al. (1966) against a number of pathogenic bacteria and fungi. Determination of minimum inhibitory concentration (MIC): The minimum inhibitory concentration (MIC) of the compound TC-1 was determined against gram positive Bacillus megaterium and gram negative Salmonella typhi-A. The test was carried out by the “Serial Dilution Technique” Reiner (1982) using nutrient broth medium. Brine Shrimp Lethality Bioassay: Brine shrimp lethality test is a recent development in the bioassay for the bioactive compounds. Bioactive compounds are almost always toxic in high dose. In this method, sea water was kept in the shrimp tank where constant oxygen supply was carried out and constant temperature (around 37ºC) was maintained. Two days were allowed for the shrimp to hatch to mature nauplii (larva). The hatched shrimps were attracted to the lamp on the other side of the tank which is divided through a perforated dam.

Source of Information: J. bio-sci. 20: 153-160, 2012, ISSN 1023-8654

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The plant Tinospora cordifolia belonging to Menispermaceae family was studied. From the combined fractions A-F, only one compound (TC-1) was isolated and purified. From the different spectral evidences, it has been concluded that the compound TC-1 is an alkaloid. The pure compound TC-1 showed activity against some bacteria and fungi. The minimum inhibitory concentration of this compound was determined. It was also showed significant activity against the brine shrimp nauplii. The plant is being used as folk medicine % of Mortality in the treatment of diabetes, jaundice and skin diseases but unfortunately in this study no such compounds were identified and no such experiment was done for the treatment of those diseases and hence further study on this plant is to be extended.

Knowledge Management: Journal