Estimation of crotepoxide in the fruits of Piper attenuatum buch.-ham. using high-performance thin layer chromatography and high- performance liquid chromatography

Background: Crotepoxide is a potent molecule for the development of anticancer pharmacodynamics therefore; estimation process for such novel compound is highly desired for future research work in the field of natural products. Introduction: Piper attenuatum is an important species used as a drug in Ayurvedic system of medicine. Pipoxide chlorohydrin, galbelgin, 8-hentriacontanol, several aristolactams, and 4,5-dioxaporphines are the main chemical constituents reported from this plant. Method: Methods based on HPTLC and HPLC with PDA detector for rapid quantitative estimation of crotepoxide in the methanolic extract of Piper attenuatum fruits has been described. Result: The linearity for crotepoxide was found to be in the concentration range of 0.2 to 1.2 μg/spot with coefficient of determination 0.9988 for HPTLC and 1 to 32 μg/mL with coefficient of determination 0.9990 for HPLC with respect to peak area. Limit of detection (LOD) and limit of quantification (LOQ) were found to be 60 ng/spot and 198 ng/spot, for HPTLC and 15 ng/mL and 50 ng/mL for HPLC, respectively. Repeatability studies have shown 0.14% RSD and SE 2.06 for HPTLC and 0.65% RSD for HPLC. Recovery values of 96.06 to 103.02% for HPTLC and 98.05 to 100.09% for HPLC indicate excellent accuracy of the methods. Conclusion: HPTLC and HPLC analyses of the fruit extract had shown percent content of crotepoxide 2.5 and 2.43, respectively, which appears to be the highest from any plant source. The developed methods are accurate, precise, cost-effective, and can be successfully applied for the estimation of crotepoxide.