CHEMOMETRICS ASSISTED RP-HPLC METHOD DEVELOPMENT FOR THE SEPARATION OF SECOND GENERATION TYROSINE KINASE INHIBITORS IN BULK DRUGS AND PHARMACEUTICAL FORMULATIONS

Abstract

The present study describes a newly developed, optimized and validated isocratic RP-HPLC method for the separation of two tyrosine kinase inhibitors (Dasatinib-DST and Nilotinib-NLT) with Methyl paraben-MPB as internal standard (IS), in bulk and pharmaceutical formulations with the aid of Chemometrics, multi criteria decision making (MCDM) approach. The separation was achieved by using Phenomenex Enable C18 column (15×4.6 mm id, 5$\mu$m particle size) and PDA-UV-detection at 277nm.The range of independent variables used for the optimization were MeOH: 60-70%, pH: 2-2.5 and flow rate:0.3-0.8ml/min. The influence of these independent variables on the output responses: capacity factor of the first peak (k1), resolution (Rs) and separation ($\alpha $) of the second peak and retention time (tR3) were evaluated. Using this strategy, mathematical model was defined, and response surface were derived for the separation. The coefficients of determination R² were more than 0.9258 for all the models. The four responses were simultaneously optimized by using Derringer's desirability functions and MCDM approach. Optimum conditions chosen for assay were MeOH, 0.01mM KH2PO4 (pH 2.5±0.5) adjusted with diluted orthophosphoric acid solution (68.03:31.97v/v) and flow rate of 0.8 mL/min. Peak area ratio of the analyte and internal standard was used for the quantification of pharmaceutical formulation samples. Total chromatographic analysis time per sample was approximately 9.0 min with DST, MPB (IS) and NLT eluting with retention times of 2.7, 3.2, and 6.0 minutes respectively. The optimized assay condition was validated as per ICH guidelines and applied for the quantitative analysis Sprycel-DST tablet and Tasigna -NLT capsule.