Quantification of Gallic Acid, Rutin and Quercetin in Hydro-Ethanolic Extract of Holarrhena Antidysenterica using High Performance Thin Layer Chromatography (HPTLC)

Holarrhena antidysenterica possesses antidiarrhoeal and antibacterial activity. The present study was focused to detect bioactive compounds by high-performance liquid chromatography (HPLC). HPLC is a tool to separate mixture of compounds in analytical chemistry (identify, quantify, purify) but cannot be used to establish biological activity. HPLC was performed to separate mixture of compounds with varying molecular sizes like gallic acid, quercetin and rutin in hydro-ethanolic extracts of seeds of Holarrhena antidysenterica. These markers were detected, quantified and compared with standards at 200 -450 nm wavelength using mobile phase Toulene: Ethyl Acetate: Formic Acid: Methanol (3:3:0.3:0.2) with CAMAG Scanner III system. The hydro-ethanolic seeds extracts of Holarrhena antidysenterica showed peak Rf values which were coinciding with standard Rf value of gallic acid, quercetin and rutin. Comparable Rf values were found for gallic acid, quercetin and rutin in sample and reference standard. This study confirmed the presence of gallic acid, quercetin and rutin in Holarrhena antidysenterica seeds hydro-ethanolic extract might be important for preventing diarrhoea. In conclusion, the hydro-ethanolic extract of seeds of Holarrhena antidysenterica could be a potential source of antidiarrhoeal, antibacterial, astringent and anti-inflammatory drugs.