Validation of an LCMS method for stability evaluations of piperine in Murchita utpalashatpala ghrita

Background: Utpalashatphala ghrita (USG) is a traditional ayurvedic medicine containing piperine as its main chemical constituent responsible for the activity. Murchita is an ayurvedic process to detoxify and purify the lipids (ghee, oil). It involves heating ghee with specific ayurvedic ingredients for a specified duration and temperature. It is essential to understand the stability of piperine during and after the murchita process. Objective: The objective of this study was to develop and validate an LCMS method to assess the stability of piperine after the murchita process. Method: LCMS system with electrospray ionization and ion-trap as the analyser was used to estimate the piperine content. Analysis was carried out in the multiple reaction monitoring (MRM) mode. Chromatographic mobile phase composed of a mixture of 55 % formic acid (0.1 %) in Milli-Q water (pH 3) and 45 % methanol. The LC-MS method was validated as per ICH Q2 (R2) guidelines. Stability of ghrita was assessed at various stability conditions. Result: The developed LCMS method was accurate, precise and linear over a range of 0.5–32 μg/mL. LOQ of the method was 125 ng/mL and the run time was 10 min. Utpalashatphala ghrita (USG) and Murchita- Utpalashatphala ghrita (M-USG) were prepared as specified under Bhaishajya Ratnavali. Our study revealed that the murchita process degraded piperine by 26 %. However, the stability of piperine was found to be better in the murchita ghrita (M -USG), compared to the unprocessed USG. Conclusion: Study effectively demonstrated that the murchita process enhanced the long-term stability of piperine.