Activity-Guided Isolation of Antioxidant Compounds from Andrographis stenophylla Leaf

The antioxidant potency of various extracts of Andrographis stenophylla leaf was evaluated in vitro using ferric thiocyanate method. Reductive ability and free radical scavenging activity of the extracts were also investigated. Amounts of phenolic compounds in each of the extracts were determined using Folin-Ciocalteau reagent and compared to observe the correlation between antioxidant activities and total phenolic content. Methanol extract exhibited maximum antioxidant activity and was found to contain 2% of total phenolic compounds. Methanol extract was subjected to column chromatographic separation over silica gel G using ethyl acetate: formic acid: acetic acid: water. Fractions thus obtained were screened for their antioxidant activity. Among the eleven fractions screened, fraction C was more active than the standard butylated hydroxyanisole. Fraction C on further fractionation with n-butanol: acetic acid: water afforded two flavanoids namely acacetine and isosakuranetine. Fraction A was also shown to possess good antioxidant activity which was developed using TLC and indicated the presence of a terpenoid, Andrographolide. The structures of the isolated compounds were confirmed by UV, IR, MS, 1H and 13C NMR spectral data. This is the first report wherein Andrographolide, Acacetine and Isosakuranetine are isolated from Andrographis stenophylla leaf