Pippli Churna (PC) is a most popular Ayurvedic formulations among the Ayurvedic medicines, Piperine is one of the major constituent of Pippli Churna. The process of development of HPLC fingerprints for Pippli Churna extract is introduced in detail. The three laboratory batches and three marketed batches were taken in this study to estimate the % of piperine in this indigenous formulation. The selection of a suitable chromatographic system, the screening for important parameters, and gradient optimization to method validation, and an integrated and universal HPLC fingerprint approach was performed. This improves the separation quality of the fingerprint. The detection wavelength of piperine was 343 nm. Thethree laboratory batches and three marketed batches were taken in this study to estimate the % of piperine in this indigenous formulation. The results of the method validation, based on the relative standard deviation of relative retention times and relative peak areas, were acceptable. Calibration curves showed good linearregression (R2 __ampersandsigngt; 0.999) within test range. The LODs and the LOQs for the piperine were 0.063 mg/ml and 0.071 mg/ml. This strategy is used for the estimation of piperine in Pippli Churna(PC) formulation and identifies and assessedits quality. Results of statistical analysis show present HPLC method for determination of Piperine is simple, precise, accurate and suitable for routine analysis of Piperine in PC. The developed fingerprints can be used as a standard and Piperine can be used as a possible marker compound for fingerprinting of PC.