Method Development And Validation For Simultaneous Estimation Of Quercetin And Rutin In Bark Of Saraca Asoca And Herbal Formulation By HPTLC
Journal Title: Indo American Journal of Pharmaceutical Research - Year 2013, Vol 3, Issue 12
Abstract
A simple, accurate and precise High-performance thin layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of Quercetin and Rutin in the methanolic extract of bark of Saraca asoca and standard herbal formulation - Ashokarishta. Fresh bark of Saraca asoca was obtained from Zandu Foundation, Gujarat and a comparative study was performed for presence of both the standards - Quercetin and Rutin with the standard formulation of Ashokarishta. The HPTLC of these bio-flavonoids was performed on F254 TLC plates with Toluene: Ethyl acetate: Methanol: Formic acid (8:8:3:1) as mobile phase. Densitometric determination of bio-flavonoids was performed at λ= 240 nm in reflectance/absorbance mode. The linear regression analysis data for the calibration plots showed a good linear relationship in the concentration range of 100-1000 ng/band with r2= 0.993 and 0.99 for Quercetin and Rutin, respectively. The average recovery for Quercetin was 98.593%and for Rutin was 97.66%. The proposed HPTLC method provides good resolution of Quercetin and Rutin from other constituents present in methanolic extract of Bark of Saraca asoca and can be used for quantification of Quercetin and Rutin present in the extract and standard herbal formulation. Method was found to be specific, precise, accurate, sensitive, selective, robust and rugged for determination of bio-flavonoids.
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