Quantification of the secondary metabolites by HPTLC, analgesic and antipyretic activity evaluation of Ficus racemosa L. leaves

Abstract

There have been no reports of simultaneous quantification of kaempferol and quercetin in Ficus racemosa L. Objective of the present work included to perform extraction using petroleum ether (PEE) and ethanol (EE), to perform preliminary phytochemical test, quantitative estimation of phytochemicals and simultaneous estimation of kaempferol and quercetin in Ficus racemosa L. using high performance thin layer chromatography (HPTLC) method and to assess the analgesic and antipyretic activity of the extracts. PEE and EE obtained from cold maceration technique. Extracts were subjected to qualitative test and estimated quantitatively. Extracts were applied on silica gel G 60 F254 plate; solvent-Toluene: Chloroform: Acetone 4.5:4.5:1 (v/v) and scanned at 254 nm. The extracts were also subjected to analgesic activity (hot plate method and tail immersion method) and antipyretic (Brewer’s yeast induced pyrexia model and Lipopolysaccharide induced pyrexia mode) activity. Findings of preliminary phytochemical test and quantitative estimation of phytochemicals confirms the presence of flavonoids in both the extracts (PEE and EE). The amount of flavonoids in PEE and EE were found to be 24.58 ± 0.60 mg/100 g and 21.01 ± 0.58 mg/100 g respectively. HPTLC system resulted well resolved bands for quercetin (Rf 0.38) and kaempferol (Rf 0.55). The validated HPTLC method was found suitable for kaempferol and quercetin quantification in Ficus racemosa L. Hot plate reaction time response was improved in a dose-dependent manner in a group of rats treated with PEE suggesting its analgesic potential. In view of analgesic potential by tail immersion method, PEE (400 mg/kg b.w.) showed no significant difference in tail flick reaction time as shown by standard treated animal (Aspirin 100 mg/kg b.w.). The findings of antipyretic activity suggested that PEE (400 mg/kg b.w.) have no significant difference in treating pyrexia when compared with Paracetamol (100 mg/kg). PEE (400 mg/kg b.w.) was found to be most potent. It may be due to the presence of comparatively high amount of flavonoids in PEE confirmed by the preliminary phytochemical test, quantitative estimation of phytochemicals and HPTLC quantification the PEE was found to be more effective when compared to EE.