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Comparison and Evaluation of Total Phenolic, Flavanoid Content and Antioxidant Activity of Crude Methanol and Ethyl Acetate Extracts of Carica Papaya Leaves

Tahira Foyzun
Department of Pharmacy, Southeast University, Dhaka, Bangladesh

Koly Aktar
Department of Pharmacy, Southeast University, Dhaka, Bangladesh

Abstract/ Executive Summary:

The present study describes the evaluation and comparison of antioxidant activity of the methanolic and ethyl acetate leaf extract (CME and CEE) of Carica papaya. The antioxidant potential was evaluated in terms of total phenolic and flavonoid content, DPPH radical scavenging potential, iron reducing power assay and total antioxidant capacity by standard procedures. Total phenolic and flavonoid contents were estimated as gallic acid and catechin equivalents respectively. Maximum phenolic (42.96 GAE/mg of plant extract) & flavonoid (39.56 CE/mg of plant extract) contents was found in CME. Both CME and CEE showed the potent antioxidant activity with IC50 5.96 µg/ml & 6.79 µg/ ml respectively which are very close to standard ascorbic acid (IC50 2.16 µg/ml). The CME showed potent total antioxidant activity and iron reducing power assay than that of CEE. The results suggested that both the extracts of the leaves of C. papaya possess potent antioxidant activities and can be a good source of natural antioxidant.

Keywords: Papaya. Phenolic content, Flavanoid content, Total antioxidant capacity, DPPH

Location: Narsingdi, Bangladesh

Start Date: 00-03-2016

End Date:

Program Area: Quality and Nutrition

Commodity: Papaya

Objectives:

This study is therefore designed to evaluate the total phenolic content, total flavanoid content and antioxidant activity in terms of DPPH radical scavenging assay, total antioxidant determination and reducing power capacity assessment of C. papaya leaves.

Methodology:

Green leaves of C. papaya were collected from the local area of Narsingdi, Bangladesh during the month March 2016. The plant was identified by an expert taxonomist. 2.2 Preparation of Plant Sample Having collected, the leaves of C. papaya were washed thoroughly in tap water and shade dried for several days with occasional sun drying. These were then dried in an oven for 24 hours at considerably low temperature (not more than 45°c) for better grinding. Dry samples of leaves were ground into fine powder in a grinding mill. The coarse powder was then stored in an air tight container and kept in cool and dry place for further use. 2.3 Extraction and Solvent Evaporation The powdered plant materials were extracted by cold extraction process and taken in an amber colored reagent bottle and soaked in to 500 ml of methanol and ethyl acetate separately. The bottles with their contents were sealed and kept for 7 days with occasional shaking and stirring. The whole mixtures were filtered through cotton and Whatman no.1 filter paper and were concentrated with a rotary evaporator under pressure at 50°c temperature to afford crude extract known as crude methanolic extract (CME) and crude ethyl acetate extract (CEE) of C. papaya leaves. 2.4 Determination of Total Phenolics Total phenolic content of methanolic and ethyl acetate extracts of leaves of C. papaya were determined employing the method as described by Singleton et al. 1965 involving Folin-Ciocalteu reagent as oxidizing agent and gallic acid as standard. Firstly, 0.5 ml of plant extract or standard of different concentration solution was taken in a test tube and 2.5 ml of Folin – ciocalteu (diluted 10 times with water) reagent solution was added into the test tube. Then 2.5 ml of sodium carbonate (7.5%) solution was added and incubated for 20 minutes at 25°c to complete the reaction. Then the absorbance of the solution was measured at 760 nm using a spectrophotometer against blank. 2.5 Determination of Total Flavonoid Total flavonoid content was determined by following the procedure of Dewanto et al. 2002. Catechin was used as standard and the flavonoid content of the extracts were expressed as mg of catechin equivalent/gm of dried extract. Firstly, 1ml of extract was placed in a volumetric flask, then 5ml of distilled water added followed by 0.3ml of 5% NaNO2. After 5 minutes, 0.6 ml of 10% AlCl3 was added and volume made up with distilled water. The solution was mixed and absorbance was measured at 510 nm. 2.6. Determination of antioxidant activity 2.6.1. DPPH (1, 1-diphenyl-2-picrylhydrazyl) Radical Scavenging Assay The antioxidant activity of both methanol and ethyl acetate extracts were determined in terms of hydrogen donating ability, using the DPPH method with a minor modification [30- 31] . Firstly, 2 ml of methanol solution of plant extract or standard at different concentration was taken in a test tube. Then 3 ml of methanol solution of DPPH was added into the test tube. The test tube was incubated at room temperature for 30 minutes in dark place to complete the reaction. Then the absorbance of the solution was measured at 517 nm using a spectrophotometer against blank. A typical blank solution contained all reagents except plant extract or standard solution. The following equation was used to determine the percentage of the radical scavenging activity of each extract. DPPH radical scavenging activity (%) = [(Abscontrol − Abssample)]/(Abscontrol)] × 100 Where, Abscontrol = absorbance of DPPH radical + methanol; Abssample = absorbance of DPPH radical + sample extract /standard. The concentrations of sample required to scavenge 50% of the DPPH free radical (IC50) was determined from the curve of percent inhibitions plotted against the respective concentration.

Source of Information: Journal of Pharmacognosy and Phytochemistry 2017; 6(2): 117-124

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, the study demonstrates that the crude methanol and ethyl acetate leaves extracts of C. papaya are different in their antioxidant effects. The results indicate that the methanolic extract possess the highest antioxidant activity than that of ethyl acetate extract. However, further pharmacological & toxicity studies are necessary to confirm this suggestion. Phytochemicals analysis should be carried out to characterize the compounds in C. papaya that act as antioxidant agents. This work suggested that C. papaya may be considered as a useful source of human health, as an antioxidant agent.

Knowledge Management: Journal