Journal of Chromatography & Separation Techniques
Open Access

Abstract

Determination of Quercetin a Biomarker in Hepatoprotective Polyherbal Formulation through High Performance Thin Layer Chromatography

Arti Gupta, Navin R Sheth, Sonia Pandey and Jitendra Singh Yadav

Background: Quercetin was determined in bioactive fractions of Ocimum gratissimum, Butea monosperma, Bauhinia variegate and polyherbal hepatoprotective formulation by HPTLC method. Methods: Polyherbal hepatoprotective formulation was developed by using five bioactive fractionated extracts of three plants namely Butea monosperma, Bauhinia variegate and O. gratissimum. All three plants contain quercetin. Chromatographic separation was performed on aluminium foil plates coated with 200 μm silica gel 60F254 Linear ascending development with toluene:ethyl acetate:formic acid, 5:4:0.1 (v/v/v) was performed at room temperature (25 ± 2°C) in a twin-trough glass chamber saturated with mobile phase vapor. Compact bands (Rf=0.38) were obtained for quercetin. Spectro densitometric scanning was performed in fluorescence mode at 380 nm. The method was validated for precision, recovery, specificity, detection and quantification limits. Results: Linear regression analysis of the calibration plots showed a good linear relationship (R²=0.9843 ± 0.0001) between peak area and concentration in the range 0.5-2.5 μg/band, respectively. The limits of detection and quantification were 0.089 and 0.26 μ/band. The recovery of the method was 97.33-99.11%. Conclusion: The above method was a rapid and cost effective quality-control tool for routine analysis of quercetin in herbal extracts and in pharmaceutical dosage form.