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In Vitro Antioxidant Studies Uncover Potential in Leaf and Bark Extracts of Artocarpus Chaplasha Roxb

MD. MIZANUR RAHMAN
Department of Botany, Jahangirnagar University, Savar, Dhaka,

MAHBUBUL KABIR HIMEL
Department of Botany, Jahangirnagar University, Savar, Dhaka,

SANZIDA MUBASSARA3 ,
Department of Botany, Jahangirnagar University, Savar, Dhaka

MD. FOKHRUL ISLAM
Assistant Professor, Department of Pharmacy, Jahangirnagar University, Savar, Dhaka, Bangladesh

APARNA SHIL
Department of Botany, Jahangirnagar University, Savar, Dhaka

Abstract/ Executive Summary:

The study was conducted at the Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh during the period of 2011 to 2013. Traditional medicine plays a significant role in healthcare across the world and especially in regions of Asia. Many of these natural remedies are plant-based and have led to modern drug discovery in the 19th century and hence the surge in research into the bioactivity of plant products. Leaf and bark samples of Artocarpus chaplasha were extracted with methanol and subjected to screening as a potential source of antioxidants and antibacterial activity. Phenolic acid and flavonoid content was evaluated by Folin- Ciocalteu reagent and aluminium chloride colorimetry respectively. 2, 2- diphenyl-1-picrylhydrazyl (DPPH) was used to assess free radical scavenging activity, whilst a method developed from antibiotic susceptibility testing was conducted to investigate antibacterial activity. Brine Shrimp lethality assay was performed to determine cytotoxicity. The bark extract contained higher levels of both polyphenols and free radical scavenging activity with a calculated IC50 of 26.21 μg/mL. Both leaf and bark extracts displayed moderate antibacterial activity, but were induced cytotoxicity. The results indicated that extracts from A. chaplasha do exhibit antioxidant properties; however in their current form they are not suitable as natural remedies due to potentially harmful effects. However, profiling of polyphenols found in A. chaplasha could lead to the discovery of novel antioxidant and/or antibacterial compounds.

Keywords: Artocarpus chaplasha, Antioxidant, DPPH, IC50, Cytotoxicity and Antimicrobial.

Location: Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh

Start Date: 00-00-2011

End Date: 00-00-2013

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

s, cytotoxicity of the leaf and bark extracts in other vehicles need to be tested using other mammalian cells to understand their potential medicinal importance. Research in this area will strengthen our understanding of the antioxidant potential of the considered plant materials and its medicinal value, as well as will indicate further implementation of the findings on exploring medicinally essential plants.

Methodology:

The study was conducted at the Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh between 2011 and 2013.Artocarpus chaplasha Roxb plant material was collected from Jahangirnagar University campus and identified by relevant authority of Bangladesh National Herbarium, Dhaka, Bangladesh (plant accession number - DACB37589). Folin-Ciocalteu reagent was purchased from Merck Millipore, Germany. 2, 2-diphenyl-1-picrylhydrazyl (DPPH), Gallic acid, quercetin and ascorbic acid were obtained from Sigma-Aldrich (MO, USA). Identified bacterial strains were collected from the Microbiology and Pathology Laboratories of Department of Botany, Jahangirnagar University, Savar, Dhaka, Bangladesh. Leaves and young green bark of Artocarpus Chaplasha were collected and the specimens were thinly sliced and initially sun-dried before being transferred to an oven at 70°C. Samples were ground to a fine powder and transferred to Soxhlet apparatus where they were extracted for 8 hours using 80% methanol. Total Phenolic compound assay: The phenolic content of the samples was determined using FolinCiocalteu Reagent (FCR) and the methodology adapted from Singleton et al.(1999). Folin-Ciocalteu reagent was diluted in distilled water (10%, v/v) to create a working solution. 0.5 ml of plant extract diluted in distilled water(10%, w/v) was mixed with 5 ml Folin-Ciocalteu working solution and 4 ml aqueous sodium carbonate (1M). Following incubation at room temperature for 15 minutes, the absorbance was measured at 765 nm. Gallic acid was used to produce a standard curve using preparations between 0-250 mg/L in methanol: distilled water (1:1, v:v). Regression analysis of the standard curve was performed; results were calculated and expressed as Gallic acid equivalent (GAE) in mg/g of dry mass of the plant extract.

Flavonoid content: Flavonoid content of the plant extracts was determined by an aluminium chloride colorimetric method developed by Chang et al. (2002). The plant extract was diluted in methanol (10%, v/v) and 0.5 ml was mixed with 1.5 ml methanol (99.8%), 100 µl aqueous aluminium chloride, 100 µl potassium acetate (1M) and 2.8 ml distilled water. Following incubation at room temperature for 30 minutes, the absorbance was measured at 415 nm. A standard curve was produced using concentrations between 12.5-100 µg/mL of quercetin in methanol. The equation of the regression line was used to calculate the result, which were expressed as quercetin equivalent (QE) in mg/g of dry mass of plant extract. Total antioxidant capacity: A method developed by Prieto et al. (1999) was used to measure antioxidant activity in the samples. A reaction mixture was created (0.6M sulphuric acid, 28mM sodium phosphate and 4mM ammonium molybdate) and 3 mL was added to 0.3 mL of plant extract. The solution was incubated for 90 minutes at 95°C, following which the solutions were left to cool to room temperature before absorbance was measured at 695 nm. Ascorbic acid was used to produce a standard curve, and the equation for the line of regression was used to calculate the results, expressed as ascorbic acid equivalent (AAE) in mg/g of dry mass of plant extract. DPPH free radical scavenging activity: Free radical scavenging activity was determined using a method established by Brand-Williams et al. (1995). Plant extract (200 µl) was mixed with 2 mL DPPH solution (0.5 mM). Absorbance was measured at 517 nm after 30 minutes of incubation at room temperature. Percentage of DPPH scavenging was calculated from [(A0-A1)/ A0] x100, where A0 is the value produced by the control (DPPH solution) and A1 is the value produced by the sample (Ahmed et al., 2012). Free radical scavenging activity curves were produced and IC50 values calculated. Antibacterial sensitivity test: Traditional disc diffusion antibacterial sensitivity test was performed to assess the activity of the extracts. The plant extract was diluted in methanol (5 mg/ml) and applied to blank antimicrobial susceptibility disks in volumes 120 µl, and air dried. This created disks containing 600 µg of extract.100 µl of bacterial suspension was distributed evenly over nutrient agar using the spread plate method. Extract-loaded and control disks (Ciprofloxacin, 5 µg/disk) were placed on the agar, and plates incubated upside-down at 4°C for 24 hours to enable impregnation. Plates were inverted and incubated for a further 24 hours at 37°C, following which zones of inhibition were measured.

Source of Information: Int. J. Sustain. Agril. Tech. 15(5): 01-08, May 2019

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Achievement/Findings:

Leaf and bark extracts of Artocarpus chaplasha exhibit antioxidant and antibacterial activity in vitro, and should be investigated for novel active polyphenols. There is potential to develop extracts of A. chaplasha to produce natural remedies, but in their current form they are not advised for use due to concerns of toxicity.

Knowledge Management: Journal