Objective: To develop simple, selective, precise and stability indicating high-performance thin-layer chromatographic method of analysis of piperine and validation as per ICH guidelines.
Methods: The method employed TLC plates precoated with silica gel 60F-254 as the stationary phase. Ideal solvent system was developed after several trials. The method was validated as per ICH guidelines for linearity, accuracy, precision, specificity and limits of detection and quantification. Standard piperine was subjected to different ICH prescribed stress conditions (acid and base hydrolysis, oxidation, dry and wet heat degradation and photo-degradation) and establishment of stability indicating HPTLC assay.
Results: The solvent system consists of toluene: ethyl acetate: diethyl ether (12:6:1) was found to give well separated spots of piperine at Rf 0.26. The linear regression analysis data for the calibration plots showed good linear relationship with r 0.997 respects to peak height and peak area, respectively, in the concentration range 200-700 ng per spot. The limits of detection and quantification were 5.7 ng and 17.27 ng per spot, respectively. The degraded product peaks were well resolved from the standard piperine with significant difference in their Rf values.
Conclusion: The stability studies indicate that standard piperine was susceptible to alkali hydrolysis, acid hydrolysis, oxidative stress degradation, photolytic degradation and dry heat degradation. The developed novel HPTLC method could effectively separate piperine from its degradation products thus it can be employed as a stability indicating one.
Chanchal Garg, Sachin Thakur, Saurabh Satija, Meenu Mehta, Munish Garg. Stability indicating HPTLC studies of piperine through method development and analysis. International Journal of Pharmaceutical Science and Research, Volume 1, Issue 7, 2016, Pages 24-29