A simple, accurate and precise high-performance thin-layer chromatography (HPTLC) method was developed and validated for the simultaneous determination of quercetin and kaempferol in the methanolic extract of leaves of Centella asiatica.Leaves of two different chemotypes of Centella asiatica were obtained from two different regions viz. Maharashtra and Gujarat state, and a comparative study was performed for the presence of both the standards — quercetin and kaempferol.The HPTLC of flavonoids was performed on F254 TLC plates with toluene-ethyl acetate-chloroform-formic acid (6:4:4:1, ν/ν) as a mobile phase. Densitometric determination of 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 = 0.9764 and 0.9823 for quercetin and kaempferol, respectively. The accuracy of the method was confirmed by conducting recovery studies at three different levels using the standard addition method. The average recovery for quercetin was 98.60% and for kaempferol was 90.08%. The proposed HPTLC method provides good resolution of quercetin and kaempferol from other constituents present in methanolic extract of leaves of Centella asiatica and can be used for quantification of quercetin and kaempferol present in the extract. Statistical analysis of the data showed that the method is sensitive and selective for determination of flavonoids.
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