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Antimicrobial Activities and Gc-ms Chemical Profiling of Methanol Extracts of Different Parts of Morus Alba (tut) in Bangladesh

Md. Sabbir Hasan
Natural Products Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Mahci Al Bashera
Natural Products Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Shyama Prosad Moulick
Natural Products Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Fatema Tuz Jubyda
Applied Botany Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Md. Jahidul Islam
Fruits and Food Processing and Preservation Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Mst. Sarmina Yeasmin
Oils, Fats and Waxes Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Md. Badrul Islam
Natural Products Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi Laboratories, Rajshahi-6206, Bangladesh

Abstract/ Executive Summary:

Morus alba has potential therapeutic value in different medicinal parts. We investigated the chemical differences presence in the methanolic extract (ME) of leaves, stem and root M. alba using GC-MS technique. GC-MS-based phytochemical profiling revealed the presence of potent bioactive components in methanolic extracts of different parts (leaves, stem and root) of M. alba. Further, GC-MS study confirmed the occurrence of twenty to thirty bioactive constituents ranging from 0.06 to 32% based on their peak area. Among different methanolic extract of M. alba maximum number of phytocompounds were observed in leaves extract with glycerin (32.77%) as the predominant compound and lowest in root bark (SBP) extract which is dominated by hexadecanoic acid methyl ester (30.55%) respectively. Furthermore, all the study extracts were also screened for their antimicrobial activity and only stem bark extract (SBP) were found to active against both gram-positive and gram-negative bacteria but all are inactive against fungal strain.

Keywords: Morus alba, GC-MS study, antimicrobial activity, anti-fungal activity, Methanollic extracts.

Location: Rajshahi, Bangladesh.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To evaluate the chemical composition of methanol extracts of leaves, stem, and root bark of M. alba by GC-MS as well as the in-vitro antimicrobial activities.

Methodology:

Chemicals- Methanol was purchased from Sigma Chemical Co. USA. All other chemicals and other solvents were of analytical grade. All chemicals were used without further purification. Plant materials- Fresh plants with roots, stems, and leaves of M. alba were collected from Rajshahi, Bangladesh. Stem and root barks were peeled and washed along with leaves using distilled water. The raw materials were then grounded and dried separately at 50°C for at least 24hrs to obtain leaf powder (LP), stem barks powder (SBP) and root barks powder (RBP). A fine powder was obtained by further grounding and then stored separately in glass bottles at room temperature. Preparation of methanol extracts of LP, SBP, and RBP- The dried fine powder of LP (50g) was extracted with methanol by sonication. The extract was filtered through Whatman No. 1 filter paper in a Buchner funnel. The filtrate was evaporated to dryness under reduced pressure in a rotatory vacuum evaporator. The remaining residues were successively extracted with methanol under the same conditions. The dried extracts were kept in dark at 4°C until further analysis. Methanol extracts of SBP and RBP were also obtained by following the same procedure. GC-MS analysis- GC Ms analysis of the respective plant extract was carried out by GCMS-QP₂₀₂₀. The chromatography method was carried out by a capillary column that is characterized by 30m in length and 0.25 mm in inner diameter. The column is prepared with 5% diphenyl and 95% dimethyl poly-siloxane. The injection temperature was set at 220°C. Initially, the oven temperature was set at 80°C and it remained isothermal for 2 minutes, then programmed to 150°C at the rate of 5°C/min and finally held at 280°C. The ion source temperature was 280°C. Helium was used as a carrier gas with a flow rate of 1.72ml/min. 4 micro-liter of the sample was injected in the split-less mode in the ratio of 1:100. For GC-MS detection, the ionizing energy gained by the detector was 45 m/z. A scan interval of 0.30 seconds. The total GC running time was 50 minutes. Interpretation of mass spectrum of GC-MS was done by using the database of National Institute Standard and Technology (NIST). The mass spectrum of an unknown component was compared with the spectrum of the known component stored in the NIST library 2008 and 2014 editions. Antimicrobial activity- Antimicrobial susceptibility pattern of the study compounds was measured in vitro by employing the modified Kirby-Bayer method (Bayer et al. 1966). It is frequently used to determine the drug sensitivity of microorganisms isolated from the infectious process and to interpret their disease potential. This method allows for the rapid determination of the efficacy of a drug by measuring the diameter of the zone of inhibition that results from the diffusion of the agent into the medium surrounding the disc. Commercially available antimicrobial discs (ciprofloxacine) were used as a positive control for bacteria whereas amphotericin B was used as a control for fungus. A single colony of the respective culture of the organisms was inoculated in Mueller-Hinton broth (for bacterial culture) and PDB (for fungal culture) to match the equivalent turbidity standard to that of McFarland 0.5 Standard. A sterile cotton swab was dipped into the suspension of these cultures and the medium of choice was Mueller-Hinton agar for bacteria and PDA for fungus, with a pH of 7.3, which was poured into plates to a uniform depth of 5mm. The swab (inoculums) was then heavily inoculated over the entire surface of the plate to obtain a confluent growth of the organism. Antibiotic control discs and the discs impregnated with study compounds (1, 2, and 3) were applied aseptically to the surface of the inoculated plates and at an appropriate special arrangement with the help of sterile forceps. The plates were then inverted and incubated at 37⁰C for 24h for bacterial growth and 28⁰C for 48h for fungal growth.

Source of Information: International Journal of Biosciences; Vol. 18, No. 5, p. 156-166, 202

Article Link (Online Journal): http://dx.doi.org/10.12692/ijb/18.5.156-166

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

GCMS analysis and antimicrobial study of the methanolic extract of Morus alba revealed that the stem bark extract contains a vast array of bioactive constituents that may contribute to various pharmacological bioactivity. Although various compounds including free fatty acid ester, sterol, terpenoids are identified in the three samples, only stem bark extract showed potential antibacterial activity. The identified compounds in the respective plant extract were reported earlier to have antioxidant, antimicrobial, antifungal, antiproliferative, hemolytic activity and so on. The inhibition of bacterial growth by the extract in our study supports the property of the identified compounds present in other plants. We can conclude that Morus alba is a potential and promising natural resource in the Indian subcontinent containing bioactive constituents that support the traditional use of the plant in the field of ethno pharmacology. The work is in progress to ascertain its biological efficacy so that we can pave a way to remark our medicinal plants as source of therapeutically valued metabolites against various diseases.

Knowledge Management: Journal