Quantification of Piperine by TLC Densitometer Method and Forced Degradation Study in a Classical Ayurvedic Formulation-Trikatu Churna

Developed a thin layer chromatography (TLC) method for the quantification of piperine in Trikatu Churna formulation it is a major and important ingredient in the formulation. TLC methods for the determination of piperine in the Trikatu churna along with its raw materials have been developed, as per the ICH guideline. The developed method has been validated, experimented with parameters like linearity, accuracy, the limit of detection, the limit of quantification, inter-day and intra-day assay precision, repeatability of measurement, and repeatability of a sample application. Performed a degradation study in different conditions. Prepared a calibration curve in the concentration range (100-800 ng/spot) with correlation coefficients r2 (0.997) with Rf value 0.46 ± 0.03. The Limit of detection (LOD) and Limit of quantification (LOQ) value has 100 ng and 329 ng, respectively. The maximum degradation found in the bench top (98%) follow with an acidic condition (86%), almost similar in the range of condition basic, oxidation and wet (75-77%), and a minimum in dry heat condition (49%). The freeze-thaw stability study, the accelerated stability study, the real-time stability study result of these conditions almost is the same range (92-99%).