Purification and Identification of the Active Principles in Ethanol Extract Fractions of Phyllanthus Amarus Leaves

Abstract

All organisms have their own antioxidant defense system that protects them against free radical damage by enzymes such as superoxide dismutase and catalase or compounds such as ascorbic acid, tocopherol and glutathione. However, our body system sometimes cannot work efficiently to mop up these free radicals thus, leading to unavoidable damages caused by oxidation. As a result, consumption of additional antioxidants through herbs such as Phyllanthus amarus, foods or supplements (phytochemicals) are beneficial to the body system, defending it against these harmful ROS. This research is aimed at ascertaining the presence of various antioxidants in ethanolic extract fractions of Phyllanthus amarus leaves. Phyllanthus amarus leaves were collected from Federal University Wukari premise, Taraba State, Nigeria and dried. The dried leaves were crushed into a fine powder manually using mortar and pestle. The ethanolic extract of Phyllanthus amarus leaves was placed under fan to evaporate the ethanol before subjecting it to further assays. The results obtained from this study revealed that for beta carotene inhibition assay, fraction 4b (445.9459%) with the solvent combination Chloroform:ethyl acetate (50:50) has the highest bleaching inhibition capacity while fraction 4a (81.08109%) has the lowest bleaching inhibition capacity. Fraction 2a (56.9565 mg/mL) has the highest phenolic content while fraction 6b (7.608669 mg/mL) has the lowest phenolic content. The assay for total flavonoid content showed that fraction 3a (145.71428 mg/mL) has the highest flavonoid content while fraction 8a (121.42857 mg/mL) has the lowest flavonoid content. The results for anti-lipid peroxidation increased with increasing extract concentration. Fraction 3b (55.732 mg/mL) with the solvent combination (chloroform 100:00) has the highest antioxidant capacity while fraction 6a (2.775 mg/mL) with the solvent combination of ethyl acetate: ethanol (50:50) has the lowest antioxidant activity. The correlation between total antioxidants capacity and total Phenolic content and that of antioxidant and flavonoid was found to be weak negative (r 2 =-2.058, r 2 =-0.024 respectively), while the correlation between phenolics and Betacarotene and that of the total antioxidant capacity and total flavonoid content was found to be weak positive (r 2 =0.245 and r 2 =0.313 respectively). The results of the analysis revealed that Phyllanthus amarus leaves contain significant nutritional components that are good for human health, such as flavonoids which is a subclass of the phytochemical, polyphenols. It is reasonable to conclude that fractions 2a and 2b has the highest antioxidant activities which have good potentials for drug development, especially against diseases of oxidative stress origin.