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In Vitro Comparative Analysis of Antibacterial Activity of Different Fractions of Corchorus Capsularis and Corchorus Olitorius Leaves Extracts

Riad Raihan Abir
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

Mafruha Marjia
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

Nadira Naznin Rakhi
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh, 2Department of Biotechnology & Genetic Engineering, Bangabandhu Sheikh Mujibur Rahman Science & Technology University, Gopalganj, Bangladesh.

Otun Saha
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

M Anwar Hossain
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

M Mizanur Rahaman*
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

Abstract/ Executive Summary:

High prevalence of antibiotic resistance is necessitating the investigation of novel antimicrobials from natural herbs and plant. So, this present study investigated two of indigenously cultivated jute plants, Corchorus capsularis (white jute)and Corchorus olitorius (tossa jute) for their antibacterial activity. Lipophilic extracts of leaves were prepared and fractionated by column chromatography resulting in 6 fractions of the extract of C. olitorius leaves (At, Bt, Ct, Dt, Et and Ft) and 11 fractions in case of white jute leaves (Aw, Bw, Cw, Dw, Ew, Fw, Gw, Hw, Iw, Jw and Kw). Each fraction of both of the leaves extracts were used to tested by agar well diffusion assay against Staphylococcus aureus and Escherichia coli along with a control organism, E. coli DH5±. While the fractions of C. olitorius leaves showed higher antibacterial activity against S. aureus, fractions of the extract of C. capsularis leaves were more effective against E. coli. The At fraction of C. olitorius extract showed the highest inhibition zone of 19 ± 2.80 mm against S. aureus and Dw fraction of C. capsularis extract had the highest inhibition zone of 15 ± 2.3 mm against E. coli (p <0.05). The extract of C. olitorius leaves showed comparatively higher antibacterial effect than that of C. capsularis leaves. Considering the promising finding regarding the antibacterial effectiveness, these fractions of the leave extract should be analyzed further to isolate the exact bioactive component to develop the lead component of new generation antibacterial drugs.

Keywords: Corchorus, Antibacterial activity, Gram-positive, Gram-negative bacteria, Leave extracts.

Location: Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Jute

Objectives:

This study targeted to investigate both the species of Corchorus to compares their antibacterial activity against both Gram-positive and Gram-negative bacteria. Phase separation by chromatography techniques was used to imply about the bioactive molecule of these leave extracts.

Methodology:

Sample preparation: Leaves of both white jute and tossa jute were collected and processed in a same manner. At first the collected leaves were washed with distilled water and dried under constant heat at 30° C for 1 week. The shade dried leaves were powdered using a mechanical grinder and blender which converted them into powder form. Extraction of bioactive compounds: The 70% n-hexane was used as the solvent for extraction of bioactive compounds. 30 g of each powdered samples were soaked into 150 mL of each of the solvents and kept at room temperature for 2 weeks. After 2 weeks the soaked samples were filtered using cotton wool filtration and Whatman Filter Paper Grade 1 filters to filtrate out the extracted samples. The filtered samples were then passed into rotary evaporator to dry out the existing solvents from the samples to isolate the extract for further experiment. Fractionation of leaf extracts: Both thin-layer chromatography and silica gel column chromatography were applied to fractionate the leaf extracts as previously described. In both chromatography techniques, silica gel was used as stationary phase and the mixture of chloroform and methanol was used as the mobile phase. Thin layer chromatography (TLC) of leaf extracts was firstly done to determine the total fractions from each extract. TLC plates were exposed under short and long length wavelength UV light to detect the separated components. Rf (retention factor) for each component separated by the chromatography technique was calculated by the following equation: Rf = (distance traveled by the compound)/(distance traveled by solvent front). On the other hand, silica gel column chromatography was done using two different preparations of chloroform (C) and methanol (M), C:M = 99:1 and C:M = 95:5 as solvents for C. capsularis and C:M = 99:1 for C. olitorius leaves extract. Column chromatography was done to collect and isolate each fraction from each of jute leaves extracts. After collecting each fraction (1-3 mL), these fractions were again subject to thin layer chromatography for confirming the separation of the components. The alike or same fractions were combined on the basis of TLC experiments 12. Antibiotic susceptibility test: Fractions separated and confirmed by chromatography techniques were evaporated by rotatory evaporator for excluding the solvent from the fractions. Later this dried preparation of both white jute leaves extract and tossa jute leaves extract were used to test for antimicrobial activity by agar well diffusion method 13-14. For this test, two test isolates including a single Gram-positive isolate S. aureus and a single Gram-negative isolate, E. coli strain DH69 (NCBI Accession No.:MN620472) were taken from Microbial genetics and Bioinformatics Laboratory (MGBL) at University of Dhaka, Bangladesh. E. coli DH5± was used as the control organism. Statistical analyses: All the measurements were carried out in duplicate. The mean values and standard deviations were calculated and the data were expressed as mean ± SD. The data analysis and statistical analysis were done using SPSS software15. Differences were considered significant at the p <0.05 level.

Source of Information: Bangladesh J Microbiol, Volume 36, Number 2, December 2019, pp 69-73

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The composition of the solvent along with the purity of the component by the extraction process may be responsible for better antibacterial activity. Even though the exact biochemical nature of the bioactive compounds has not been identified in this study, the comparative analysis of antibacterial effects of different fraction indicates the nature of polarity of the active compounds. So, this study will help to isolate and characterize the exact component of the antibacterial potential for developing an attractive treatment alternative.

Knowledge Management: Journal