A novel RP-HPLC method for the determination of bharangin in Ghantu bharangi crude extracts.

An accurate, simple, reproducible and sensitive RP-HPLC method for the determination of bharangin has been developed and validated. The separation of bharangin and 2-nitroaniline (internal standard) was achieved on Supelcosil LC-18 (3micro, 150 x 4.6 mm i.d.) column using UV detection at 388 nm. The mobile phase was consisting of methanol and 0.01 M KH(2)PO(4) buffer (pH 3.0, adjusted with ortho-phosphoric acid) (75:25, % v/v). The linear range of detection for bharangin was found to be 10-50 ng/ml. Intra-and inter-days assay relative standard deviations were less than 3.21. The method has been successfully applied to the determination of bharangin in various crude extracts. The method has been shown to be linear, reproducible, specific, and rugged.