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Preliminary Phyto-chemical Screening and Comparative Antioxidant Potential of Alstonia Scholaris Leaves Extracts

Md. Tohidur Rahman
Department of Chemistry, University of Rajshahi-6205, Bangladesh

Md. Asadul Islam
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Md. Abdul Jalil
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Nazim Uddin Ahmed
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Md. Abdurrahim
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Md. Mahmudul Hassan Mondol
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Ali Ahsan Muzahid
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

Md. Badrul Islam
Drugs and Toxins Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR), Rajshahi-6206, Bangladesh

ABM Hamidul Haque
Department of Chemistry, University of Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

Yet to now natural products, or its’ derivatives are in service to humankind as the best source of molecules to control the pest and disease problems of human beings. The present work investigated total phenolic content, total flavonoid content, reducing power capacity, total antioxidant capacity, DPPH free radial scavenging activity of four different extracts of the plant A. scholaris which belongs to the family Apocynaceae. Total phenolic content (21.92± 0.13 mg GAE/g) and total flavonoid content (16.61± 0.06 mg CatE/g) were found to be highest in Dia-ion resin adsorbed fraction, and the same fraction showed the highest total antioxidant activity with absorbance 1.049±0.014 at 100 mg/mL as well as highest DPPH radical scavenging activity with IC50 value 24.90 mg/mL. The Iron reducing power of the different extractives and standard exhibited the following order: Ascorbic acid> DRAF > CLF > EAF > PEF and the total antioxidant activity of different extractives and standard exhibited the following order: Ascorbic acid> DRAF > PEF > EAF > CLF.

Keywords: Alstonia scholaris, Extracts, Phytochemical screenning, Total phenolic, Total flavanoids.

Location: BCSIR, Rajshahi

Start Date:

End Date:

Program Area: Physical and Mechanical Properties

Commodity: Medicinal Plants

Objectives:

To study the antioxidant efficacy of different extracts of medicinal plant Alstonia scholaris grown in Bangladesh based on phyto-chemical screening and in vitro antioxidant tests.

Methodology:

All the reagents and chemicals used for the presence work were purchased from Thomas Baker (Mumbai, India), Bdh (England), Fluka (Switzerland) and E. Merck (Germany) supplied by local vendors. Commercial alcohol (rectified spirit) and absolute alcohol were available from Carew and Company, Darsana, Chuadanga. The solvents used mainly in this work were benzene, acetone, tetrahydrofuran (THF), ethyl acetate, chloroform, n-hexane, petroleum ether, methanol, absolute alcohol, toluene etc. The solvents were dried and distilled when necessary. Collection of plant materials- The Alstonia scholaris plant leaves were collected from the cultivated adjacent areas of BCSIR, Rajshahi. The collected materials were washed thoroughly in water, chopped, air dried for a week at 35-40^oC and pulverized in electric grinder. Dried ground leaves of Alstonia scholaris were exhaustively extracted with ethanol in Soxhlet apparatus. The resulting juicy extract was filtered through Whatman paper No.1 and concentrated under reduced pressure at 45°C using the Buchi Rotavapor R-200 to obtain a crude residue (23.5%).The process was repeated several time for maximum yield. Then water triturate part was collected from crude extract. The water triturate fraction was passed through a previously well packed dia-ion resin column which was selectivity collect only the phenolic group containing compounds. Then the materials, which were bound in resin column, collected by passing methanol solvent. Then Petroleum ether, Ethyl acetate and Chloroform solvents were passing through the residue respectively. Finally Petroleum ether, Ethyl acetate and Chloroform triturate were collected. Preliminary phyto-chemical analysis- Preliminary phytochemical screening of crude ethanol extract and its different fractions was done using various published qualitative chemical tests procedures for several classes of natural products (Sofowara, 1993 & Harborne, 1973). Estimation of total phenolic- Total phenolic content of different extractives of A. scholaris were determined employing the method as described by Singleton et al. (1965) which involves Folin-Ciocalteu reagent as oxidizing agent and gallic acid as standard. The absorbance of the solution was measured at 760 nm using a UV spectrophotometer (ThermoSpectronic 20, USA) against blank. Estimation of total flavanoids- Total flavonoid content of different extractives of A. scholaris leaves were determined by aluminium chloride colorimetric method using catechin as reference compound (Zhishen et al., 1999). A volume of 125µL of extract is added to 75 μL of a 5% NaNO₂ solution. The mixture was allowed to stand for 6 min, then 150 μL of aluminium trichloride (10%) was added and incubated for 5 min, followed by the addition of 750 µL of NaOH (1M). The final volume of the solution was adjusted to 2500 μL with distilled water. After 15 min of incubation the mixture turned to pink and the absorbance was measured at 510 nm. The total flavonoids content was expressed as mg cat.E/g DM. Determination of reducing power capacity- The reducing power of methanol extract and different fractions of A. scholaris leaves were evaluated by the method of Oyaizu, (1986) using potassium ferricyanide [K3Fe (CN) 6] (1%) solution. In this assay, the yellow color of the test solution changes to various shades of green and blue depending on the reducing power of antioxidant samples. The iron reducing power of different extracts of A. scholaris leaves were estimated by measuring the formulation of Perl’s Prussium blue at 700 nm. Determination of total antioxidant capacity- Total antioxidant activity of different extracts of A. scholaris leaves were determined by the method of Prieto et al., (1999) with some modifications. The phosphomolybdenum method usually detects antioxidants such as ascorbic acid, some phenolics, α- tocopherol and carotinoids. This method was based on the reduction of Mo (VI) to Mo (V) by the antioxidant compound. 3 ml of reaction mixture containing 0.6 M sulfuric acid, 28 mM sodium phosphate and 1 % ammonium molybdate was added into the experimental samples and incubated at 95^oC for 90 minutes to complete the reaction and measured the absorbance of the solution at 695 nm. Determination of free radical scavenging activity- DPPH free radical scavenging activity of the ethanolic extract and its different fractions were carried following the method described by Braca et al., (2001). Briefly, 0.1 ml of extract with various concentrations was added to 3 ml of a 0.004% methanol solution of DPPH. Absorbance at 517 nm was determined after 30 min, and the percentage inhibition activity was calculated from [(A₀– A₁)/A₀]×100, where A₀ is the absorbance of the control and A₁ is the absorbance of the extract/ standard. IC₅₀ value was calculated from the equation of line obtained by plotting a graph of concentration (µg/ml) versus % inhibition.

Source of Information: International Journal of Bio-sciences, Vol. 15, No. 4, p. 161-171, 2019

Article Link (Online Journal): http://dx.doi.org/10.12692/ijb/15.4.161-171

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The four different extractives of Alstonia scholaris showed the presence of phyto-constituents from moderate to high level in Dia-ion resin absorbed fraction, whereas minute to high in other extracts. The results of the present study also revealed that the different fractions of ethanolic extracts of Alstonia scholaris leaves exhibits considerable total antioxidant, iron reducing and free radical scavenging capacity. This result can be strong scientific evidence to use this plant as a useful source of both biological and pharmacological references. Yet, further studies are necessary to clarify a mechanistic way how the plant contributes in these properties.

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