APPLICATION OF RP-HPLC METHOD FOR THE DETERMINATION OF BOLDINE IN ALPHONSEA SCLEROCARPA THWAITES

Abstract

Determination of Boldine in Alphonsea sclerocarpa Thwaites was followed through by Reverse Phase High Performance Liquid Chromatography (RP- HPLC). Separation and determination of Boldine in chloroform, bark & leaf extracts of Alphonsea sclerocarpa was accomplished on an inertsil ODS(150mmX 4.6mm, 5µm i.d.) column system employing isocratic elution mode with a mobile phase consisting of Acetonitrile and 0.1% Octane sulphonic acid buffer (70:30) (v/v) at a constant flow rate of 1ml/min. Calibration graph of Boldine in the concentration range of 84-420µg.ml^-1 was obtained by using the linear relationship between concentration and peak area measured at a wavelength of 302nm using UV detector. The entire chromatographic run was carried out at ambient temperature. The developed RP-HPLC method was successfully validated as per ICH guidelines (ICH, Q2 (R1)) The retention time of boldine was found to be 2.1 min. Percentage recovery was found to be 100.82, 100.04 &100.44 at 50, 100, 150 % levels respectively. The %RSD for intermediate precision and repeatability was found to be 0.9 and 0.8 respectively. Limit of Detection and Limit of Quantitation was found to be 5.22 and 15.82 µg.ml^-1. The method developed was validated and successfully applied for the quantitative analysis of Boldine in Chloroform-Bark and leaf extracts of Alphonsea sclerocarpa Thwaites.