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Phytochemical and Biological Activity Studies of Tinospora Crispa Stem

Mariam Rahman
Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh

Md. Hasanur Rahman
Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh

Tofail Ahmad Chowdhury*
Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh

Abstract/ Executive Summary:

Three compounds namely, tinosporol A, 8-methoxy palmatine and callecdysterol C were isolated from the methanol extract of the stem of Tinospora crispa. Biological activities of different partitionates of the parent extract was also evaluated. Antimicrobial screening of different partitionates was carried out against sixteen different microorganisms and only nhexane fraction exhibited significant zone of inhibition against Staphylococcus aureus (9 mm), Shigella boydii (9 mm), Shigella dysenteriae (9 mm), Candida albicans (10 mm) and Aspergillus niger (10 mm).The crude methanol extract showed the highest general toxicity with LC50 values of 57.14 µg/mL against brine shrimp lethality bioassay.The total antioxidant capacity of aqueous fraction was found to be 61.61 mg as ascorbic acid equivalentper gram of plant extract.The antioxidant activity of DCM fraction revealed the highest activity having IC50 values 54.74 µg/mL. No significant cytotoxicity was observed on both HeLa and Vero cell lines.

Keywords: Secondary metabolites, Antimicrobial, Lethality bioassay, Antioxidant, Cytotoxicity.

Location: Department of Botany, University of Dhaka

Start Date:

End Date:

Program Area: Resource Development and Management

Commodity: Medicinal Plants

Objectives:

The paper reports the isolation of three known compounds and biological activities of different extracts of the stems of T. crispa of the variety from Bangladesh.

Methodology:

Collection of sample The stems of the plant were collected from Sylhet, Bangladesh and taxonomic identification was made by the renowned plant taxonomist Professor Dr. Mohammad Zashim Uddin, Department of Botany, University of Dhaka. The collected stems were dried and crushed into powder.

Phytochemical screening Phytochemical screening was carried out using standard procedure14 for identifying the chemical constituents. The presence of phytochemicals such as tannins, phlabotannins, saponins, flavonoids, steroids, terpenoids and alkaloids were detected.

Extraction and Isolation The dried powder (500 g) of T. crispa stems were extracted with methanol and partitioned with n-hexane and fractionation of methanol extract was done with dichloromethane (DCM) and ethyl acetate (EtOAc). The extracts were separately concentrated to dry mass using rotary vacuum evaporator at 40°C under reduced pressure. DCM extract (3.25 g) and EtOAc extract (4.65g) were subjected to column chromatography and 15 fractions (P-1 to P-15) were obtained from the TLC study. The fraction P-5 (~40 mg) showed single spot with tailing and was subjected to preparative thin layer chromatography (PTLC) and a white crystalline compound was obtained which was labeled as 3 (~4.5mg). The fraction P7 appeared to contain two spots and was subjected to sub column. One of the fraction was found with single spot under TLC and yielded white powdered compound after washing with solvent (1~5mg). The fraction P-9 gave with two spots on TLC and was subjected to sub-column, and finally a single yellow crystalline compound was obtained 2(~4.5mg).

Biological activity screening Freshly prepared stem powder was extracted successively with n-hexane (HEX), dichloromethane (DCM), ethyl acetate (EA) and methanol (ME) at room temperature. All the extracts were evaporated to dryness and used for antimicrobial activity screening, brine shrimp lethality bioassay, cytotoxicity assay on cancer cell lines and determination of total antioxidant capacity using their individual standard procedures.

Cytotoxicity assay was examined against HeLa cell line (a human cervical carcinoma cell) and Vero cell line (kidney epithelial cells extracted from an African green monkey) in Center for Advanced Research (CARS), University of Dhaka.

Source of Information: Dhaka Univ. J. Sci. 68(2): 167- 170, 2020 (July)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Tinsporol A, 8-Methoxy palmatine and callecdysterol C were isolated from the stems of T. crispa using different chromatographic separation techniques. The isolated compounds were identified by extensive analysis of their high resolution 1H-NMR (400 MHz), 13C-NMR (100 MHz) spectroscopic data. Different types of tests were carried out to evaluate the biological activities of the stems of T. crispa. The HEX extract revealed a significant antibacterial activity against gram negative bacteria, C. albicans and antifungal activity against A. niger. ME extract demonstrated highest lethality with LC50 value of 57.13 μg/mL. Different extracts of T. crispa stem showed moderate antioxidant activity.

Knowledge Management: Journal