Simultaneous RP-HPLC quantification of four phenolics in Elephantopus scaber L. And their in vitro pharmacological validation

Abstract

Elephantopus scaber (L.) is a widely used traditional medicinal plant, grows in hotter parts of the Indian subcontinent, Tropical Africa and East Asia. The present study was aimed RP-HPLC quantification of phenolics and its pharmacological validation in Elephantopus scaber (L.). The chromatographic separation was obtained using RP-C18 column, using mobile phase acetic acid: water (1.0: 99.0 V/V) as solvent-A and acetonitrile as solvent-B and 0.6ml/min flow rate was used for proper separation. HPLC method was developed and validated according to ICH guidelines. Four important phenolic compounds i.e chlorogenic acid, ferulic acid, gallic acid and protocateuchine were identified and quantified with the help of developed RP-HPLC method. Result reveals the presence of chlorogenic acid (3.481mg/g), ferulic acid (0.842 mg/g), gallic acid (0.668 mg/g) and protocateuchic acid (0.086 mg/g). Total phenolic and flavonoid content in the methanolic extract was found to be 16.24±1.61 mg/g GAE and 12.87±0.043 mg/g QE. In vitro antioxidant and antidiabetic assays were performed as per standard protocols. The IC50 value for the in vitro DPPH method was found to be (0.167±1.21 mg/ml), in DNS assay (0.522±0.04 mg/ml) and (0.364±0.03 mg/ml) for the starch-Iodine method in methanolic extract of E. scaber. This implies that Elephantopus scaber which has been generally considered as weeds and exploited for beneficial purposes not only traditionally but also would be used in medicinally based on their biochemical active compounds. Hence, the study aids in authentication of E. scaber by identification of marker compounds with validated traditional claims.