A Validated HPLC Method for the Quantification of Hypericin in Hypericum perforatum

Abstract

A simple, sensitive and specific RP-HPLC method has been developed for the quantification of hypericin in Hypericum perforatum. The method involved evaluation of hypericin after resolving it by RP-HPLC with UV detector at 590 nm using acetonitrile:methanol: 10 mM ammonium acetate (pH 5.0) in the ratio 54:36:10 (v/v/v) as mobile phase. The method was validated as per the ICH guidelines for linearity, precision (inter-day, intra-day and inter-system), robustness, accuracy, limit of detection and limit of quantification. The calibration was linear with correlation coefficient of 0.998 over a concentration range of 10-80 μg mL^-1 for hypericin. The proposed method for the quantification of hypericin was found to be simple, precise, specific, sensitive and accurate and the method can be used for the routine analysis of hypericin for quality control of raw material of H. perforatum, several Unani and Ayurvedic formulations containing it as an ingredient.