Abstract

SEPARATION, IDENTIFICATION AND QUANTIFICATION OF LAWSONE AND METABOLITES BY CHROMATOGRAPHIC METHODS

Dr. Gandhimathi Muruganathan*, Bharathi Mandala, Thengungal Kochupapy Ravi

Abstract

Two simple, precise and accurate chromatographic methods were developed and validated for the simultaneous estimation of Lawsone and its metabolites. In RP-HPLC the separation was carried out by C18 column by using mobile phase (0.06%v/v trifluoro acetic acid buffer): methanol: acetonitrile (60:30:10 v/v/v) with pH 3.7 and flow rate of 0.8ml/min in isocratic mode. The PDA detection wavelength was 235nm and 20 μl of sample was injected. For HPTLC the solvent system comprised of toluene: ethyl acetate: ethanol: formic acid in the ratio of 8:2:1:1 drop % v/v/v/v detection was done at 235nm. The Rf value of lawsone, salicylic acid and catechol were found to be 0.37, 0.33 and 0.49 respectively. Linearity was found to be 10-160 μg/ml for Lawsone, 5-80μg/ml for salicylic acid, 40-640 μg/ml for Catechol in HPLC; 20-320 μg/ml for Lawsone, 10-160μg/ml for salicylic acid, 80-1280 μg/ml for Catechol in HPTLC. The above methods were validated according to ICH guidelines and successfully employed for estimation of lawsone and its metabolites.

Keywords: Lawsone, salicylic acid, catechol, RP-HPLC and HPTLC.