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Ashwini Surendra Shinde*


A high performance thin layer chromatography (HP-TLC) method was developed for identification, quantification and validation of lupeol from methanolic and hydro-methanolic extract of Elephantopus scaber L. TLC Si gel 60 F254 precoated plate was used as a stationary phase. Toluene: Ethyl acetate: Formic acid (12:7:1 v/v/v) was used as a mobile phase for better separation and resolution. Quantification was performed by densitometric scanning at 540 nm after derivatizating with anisaldehyde sulphuric acid reagent (ASR). Compact spot at Rf 0.67 was obtained corresponding to standard lupeol in the plant sample. According to the ICH guidelines, this method was validated in terms of specificity, precision and accuracy assay. Linearity range for lupeol was 100 – 800 ng/band with correlation of coefficient (r2) = 0.99319. The LOD and LOQ were found to be 9.34 and 28.31 ng/band, respectively. The lupeol content in methanol and hydromethanol extract were found to be 386.0 and 301.3 μg/mg, respectively. The HP-TLC method was found to be rapid, simple, specific, precise, accurate and convenient method for screening of active principles present in the plant extracts. This method can also be used for routine analysis of quality control of herbal formulations containing E. scaber.

Keywords: Elephantopus scaber L., HP-TLC, Lupeol, Method development, Quantification, Validation.

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