Simultaneous Determination of Strychnine and Brucine in Herbal Formulation by RP-HPLC

A simple, precise and accurate reverse phase HPLC method has been established for simultaneous quantitative determination of strychnine and brucine in herbal formulation (tablets). The HPLC method was performed on a Phenomenex C-18 ODS column (250 mm×4.6 mm ID, 5 μm), with isocratic elution using a mixture of acetonitrile: phosphate buffer in the ratio of 15:85 v/v at flow rate of 1 mL/min with UV detection at 264 nm. Strychnine and brucine were well resolved on the stationary phase, and the retention times were 6.263 minutes for strychnine and 4.987 minutes for brucine. The calibration curves were found to be linear (Correlation coefficient, r=0.9997 for strychnine and 0.9996 for brucine) in the concentration range 20-120 μg/mL for strychnine and brucine. The herbal preparation was quantified by external standard calibration method. The assay was validated for the parameters like accuracy, precision, robustness and system suitability parameters. The proposed method can be useful in the routine analysis for the determination of strychnine and brucine in herbal formulation.