Development and validated stability indicating rp-hplc method for quantitative determination of ascorbic acid and piperine in dhathryaristam

Dhathryaristam is an ayurvedic medicine described in Charaka samhita chikitsasthana. The preparation consists of Phyllanthus emblica (amlaki, Indian goose berry) and Piper longam (Indian long pepper). The aim of this research was to determine a finger print methods for ascorbic acid and piperine using simple mobile phase((buffer: acetonitrile) in raw material as well as formulation. The efficient separation of chemical marker was carried out using reverse phase C18 eluted with gradient mobile phase. The separation occurred at 4.1min ascorbic acid and 5.6min piperine. Method validation was demonstrated by system suitability, accuracy, precision, repeatability and recovery. The calibration curve was found to be within the tested ranges. The recoveries were between 98–102%. The photo degradation studies exhibit the formation of various degraded product. The quantification of ascorbic acid in raw material was 1.917μg/ml in Phyllanthus emblica and marketed formulation D-I and D-II are 1.877μg/ml and 1.871μg/ml respectively. The quantification of piperine in raw material was found to be 10.106μg/ml in Piper longam in two marketed formulation D-I and D-II are 10.845 μg/ml and 10.106μg/ml respectively. The developed and validated method can be effectively applied to the quantitative determination of herbal formulation containing ascorbic acid and piperine.