Penentuan Profil Bioautografi Antioksidan Fraksi n-Heksan dan Etil Asetat dari Fraksinasi Bertingkat Daun Pugun Tano (Picria Fel-Terrae Lour.)

Abstract

Background: Free radicals are highly reactive molecules that can damage body cells, proteins and fats. Therefore, the presence of antioxidants is very important to neutralize free radicals. Antioxidants can be obtained from natural sources, one of which is pugun tano leaves. Thin layer chromatography (TLC) bioautography test was conducted to identify active compounds that have activity as antioxidants. Objective: To determine the n-hexane fraction and ethyl acetate fraction of pugun tano leaves can counteract DPPH free radicals using TLC bioautography method. Methods: The research include plant identification, sample collection, sample processing, extract preparation and fractionation, characterization of simplisia and extracts, phytochemical screening of simplisia, extracts, and fractions of n-hexane and ethyl acetate, antioxidant activity test with thin layer chromatogaphy (TLC) bioautography using 3 types of mobile phases namely chloroform acetone: formic acid (10:2:1), mixture of chloroform: methanol: water (64:50:10), mixture of ethyl astetate: methanol: water (8: 1: 1), campuran chloroform: ethyl astetate: n-butanol: water (5: 2: 2: 1) and mixture of n-hexane: ethyl acetate (8:2). Results: Characterization of pugun tano leaf simplisia obtained water content 7.98%, water soluble juice content 20.8%, ethanol soluble juice content 9.23%, total ash content 1.87%, and acid insoluble ash content 0.3%. Characterization of ethanol extract of pugun tano leaves obtained a moisture content of 9.28%, total ash content of 0.13%, and acid insoluble ash content of 0.09%. Phytochemical screening of simplisia and pugun tano leaf extract contains flavonoids, glycosides, saponins, steroids and tannins, in the n-hexane fraction contains flavonoids and steroids while in the ethyl acetate fraction contains flavonoids, saponins and steroids. The results of the antioxidant activity test by TLC bioautography obtained effective stain separation obtained in the ethyl acetate fraction using the mobile phase chloroform: acetone: formic acid (10: 2: 1) which is most effective as an antioxidant. Conclusion: The compound group that has antioxidant activity is the flavonoid compound group in the ethyl acetate fraction of pugun tano leaves.