Quantification of piperine in different varieties of Piper nigrum by a validated high-performance thin-layer chromatography-densitometry method

Piperine is the most important alkaloid molecule present in Piper nigrum. In this study, a simple, rapid, quantitative high-performance thin-layer chromatography (HPTLC) technique proposed by International Organization for Standardization (ISO) was optimized and validated for the quantitative estimation of piperine in P. nigrum. Densitometric analysis of piperine was carried out in the absorbance mode at 254 nm. The method gave spot at R-F = 0.63, corresponding to piperine in different samples. The limit of detection and limit of quantification per spot were confirmed with the mobile phase toluene-ethyl acetate (3:2, V/V). Linear regression analysis data for the calibration plot for piperine showed a good linear relationship with a correlation coefficient (r(2)) of 0.99077 in the concentration range of 200-1000 ng per spot. The method was validated for sensitivity, linearity, accuracy, precision and specificity as per the International Council for Harmonisation guidelines. This simple, precise and accurate HPTLC method provides a novel approach for the routine analysis of black pepper for quality control and standardization purposes, which may be further useful for industrial perspectives.