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Antimicrobial, Cytotoxicity, Free Radical Scavenging Activities and Compounds Isolated from the Roots of Hemidesmus Indicus (linn.) R. Br.

A. Saha
Institute of National Analytical Research and Service (INARS), BCSIR Laboratories Dhaka, Dr. Qudrat-i-Khuda Road, Dhanmondi, Dhaka-1205, Bangladesh, and Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh.

J. Tasnim
Institute of National Analytical Research and Service (INARS), BCSIR Laboratories Dhaka, Dr. Qudrat-i-Khuda Road, Dhanmondi, Dhaka-1205, Bangladesh, and Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh.

S. Ahmed
Institute of National Analytical Research and Service (INARS), BCSIR Laboratories Dhaka, Dr. Qudrat-i-Khuda Road, Dhanmondi, Dhaka-1205, Bangladesh.

T. Muslim
Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh.

M. A. Rahman
Department of Chemistry, University of Dhaka, Dhaka-1000, Bangladesh.

N. Sultana
Principal Scientific Officer
Institute of National Analytical Research and Service (INARS), BCSIR Laboratories Dhaka, Dr. Qudrat-i-Khuda Road, Dhanmondi, Dhaka-1205, Bangladesh.

Abstract/ Executive Summary:

Hemidesmus indicus Linn. R. Br. is a twining shrub traditionally used for the treatment of various diseases and ailments in the rural area of Bangladesh. Different extracts (methanol, hexane, ethyl acetate, n-butanol and water) from the roots of this shrub tested for their antimicrobial, cytotoxicity and free radical scavenging activities. All extracts showed moderate free radical scavenging activity and SC₅₀ values ranging from 24.98µg/mL to 93.14µg/mL, and sufficient potency for their brine shrimp lethality activity and IC₅₀ values from 1.32µg/mL to 8.67µg/mL. The n-hexane extract showed significant antimicrobial activity against all tested organisms, with a zone inhibition ranging from 10 mm to 27 mm at the concentration of 400µg/disc. Bioassay guided fraction of the extracts has led to isolation of two triterpeniods, 3β-O-acetyl β-amyrin, lupeol 3- β -acetate and a phenolic compound, 2-hydroxy-4-methoxy benzaldehyde. The structures of the compounds were elucidated on the basis of spectroscopic methods as well as comparison with available data in the literature

Keywords: Hemidesmus indicus, Antimicrobial activity, Free radical scavenging activity, Cytotoxicity, Triterpeniods, 2-hydroxy-4-methoxy benzaldehyde.

Location:

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To investigate the antimicrobial, cytotoxicity and free radical scavenging activities of different extracts of the roots of H. indicus and
To isolate and identify of compounds from this species through bioactivity guided fraction of its extract.

Methodology:

The roots of Hemidesmus indicus L. R.Br. were collected from Modhupur, Tangail, Bangladesh in December 2012. The plant was identified and confirmed by Prof. Dr. Md. Abul Hassan, Department of Botany, University of Dhaka. The fresh roots were taken into laboratory and cut into small pieces and were air dried. The roots were ground to powder by a Cyclotec grinding machine. The powders were stored in air tight bottle and these were used throughout the investigation.Melting points were measured on Stuart Scientific SMP3 melting point apparatus and were uncorrected. The IR data for all compounds were obtained from dissolved sample in methanol solvent using a Shimadzu FT-IR 8400S spectrometer. UV spectra and absorbance were performed with a Perkin Elmer Lambda 25 UV-visible spectrophotometer. H (400 MHz) and C (100.60 MHz) NMR spectra were recorded on a Bruker DPX- 400 (400 MHz) instrument, with chemical shift data reported in ppm relative to the solvent used. General laboratory solvents were distilled from glass before use. Column chromatography (CC) and Vacuum liquid chromatography (VLC) were performed using Merck silica gel (0.063–0.2 mm) and silica ge 60H (15μm), respectively. Silica gel 60 F₂₅₄ coated on aluminum plates for thin layer chromatography (TLC) was supplied by Merck. The reagents used in the present work were of analytical grade (Merck and BDH). Vitamin C was purchased from Sigma Aldrich Chem Co.
The dried, ground root (600g) was extracted with methanol at room temperature for three days. The extract was filtered and evaporated to dryness using a rotary evaporator under reduced pressure. The free radical scavenging activity of the roots of H. indicus were assayed spectrophotometrically and these were carried out as previously described method 11. The ascorbic acid was used as a positive control. Each treatment was replicated thrice. Antimicrobial activities of all extracts (methanol, n-hexane, ethyl acetate and n-butanol) of the roots of H. indicus were carried out by the disc diffusion methods 12, 13. Five bacterial species which include two Gram-positive and three Gram-negative bacterial strains and two fungus Aspergillus niger and Aspergillus flavus were taken for the test. The bacteria were Bacillus cereus (ATCC-10876), Staphylococcus aureus (ATCC-9144), Escherichia coli (ATCC-25922), Pseudomonas aeruginosa (ATCC-27853), and Salmonella typhi (ATCC-6539). Each organism was maintained on nutrient agar slant. The samples were dissolved separately in chloroform and applied to sterile filter paper disc at a concentration of 400μg/disc. Kanamycin disc (30μg/disc) was used at standard in antibacterial and Ketoconazole disc (30μg/disc) was used at standard in antifungal study. The sample disc, standard disc and control discs were placed gently on the previously marked zones in the agar plates pre-inoculated with test bacteria. The plates were then kept in the refrigerator at 4⁰C for about 24 hours upside down to allow sufficient diffusion of the materials from discs to the surrounding agar medium. The plates were then inverted and kept in an incubator at 37⁰C for 24hours. The antimicrobial potency of the test agents was measured by their activity to prevent the growth of the microorganisms surrounding the discs which gives clear zone of inhibition expressed in mm. The experiment was carried out in triplicate and the mean values were taken. The zones of inhibition were calculated as mean ±S.D. (n=3). Brine Shrimp Lethality Bioassay of all the extracts (methanol, nbutanol, n-hexane and ethyl acetate) were carried out against Artemia sarina in a 1-day in vivo assay. For the experiment 4mg of each of the extracts was dissolved in dimethyl sulfoxide (DMSO) in vials to get stock solutions. Solutions of varying concentrations such as 400, 200, 100, 50, 25, 12.50, 6.25, 3.125, 1.563, 0.781µg/ml were obtained by serial dilution technique. The median lethal concentration LC₅₀ of the test samples after 24 hours was obtained by a plot of percentage of the shrimps killed against the logarithm of the sample concentration. Here vincristine sulphate was used as a standard.

Source of Information: International Journal of Pharmaceutical Sciences and Research, 2015; Vol. 6(1): 117-122. E-ISSN: 0975-8232; P-ISSN: 2320-5148

Article Link (Online Journal): http://ijpsr.com/?action=download_pdf&postid=13756

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It has been reported that 2-hydroxy-4-methoxybenzaldehyde had significant antimicrobial and antioxidant activities which could potentially be developed as an antimicrobial and antioxidant agent in future. The present study had further supported the traditional uses of this plant as antimicrobial and antioxidant agents.

Knowledge Management: Journal