Quantification of Lupeol as Secondary Metabolite by HPTLC Technique and Assessment of Antimicrobial Potential of Ethyl Acetate Extract of Betula alnoides Bark

Abstract

The present work includes extraction of Betula alnoides bark using ethyl acetate as a solvent, preliminary phytochemical test, quantification of phytochemicals and quantification of lupeol in Betula alnoides by High Performance Thin Layer Chromatography (HPTLC) instrument along with the assessment of the antimicrobial potential of Ethyl Acetate Extract (EAE). The marc obtained after defatting of the coarsely powdered crude drug in Petroleum ether (60-80) was extracted using ethyl acetate. Afterward, preliminary phytochemical tests were done. For High Performance Thin Layer Chromatography (HPTLC), the solvent used was n-hexane: ethyl acetate (8:2 v/v) and scanning was performed at wavelength 254 nm. EAE was screened for antimicrobial potential. The extraction yield was 3.45% w/w. The result of the phytochemical analysis confirmed the presence of some important phytochemicals in EAE. A clear and resolved peak of lupeol was observed at Rf 0.61. The developed method was validated as per ICH guidelines. The concentration (%) of the marker compound (lupeol) was found to be 0.0168. Disk diffusion method using Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus subtilis as bacterial strains and Candida albicans, Aspergillus flavus and Epidermophyton floccosum as fungal strains against ciprofloxacin (for antibacterial activity) and fluconazole (for antifungal activity) as standard drugs was employed. The finding suggested that EAE possess significant antibacterial and antifungal activity when comparison was made with standard drugs. The proposed elucidated mechanism behind this action may be due to the presence of triterpenoids in EAE.