Chemometrics Assisted RP- HPLC Method for the Simultaneous Determination of Levocetirizine, Ambroxol and Montelukast in Pharmaceutical Formulation

This paper deals with multiple response simultaneous optimization using the Derringer’s desirability function for the development of a reversed-phase HPLC methods for the simultaneous determination of Ambroxol(AMB) and Montelukast(MLS) with Levocetirizine (LCT) in commercial pharmaceutical preparations with Probenecid(PRO) as Internal standard. The ranges of the independent variables used for the optimization were MeCN: 30-40%, buffer conc.: 10-20 mM and flow rate: 0.8-1.2 ml/min. The influence of these independent variables on the output responses: capacity factor of the first peak (k1), resolutions (Rs2,3), and Retention time (tR4) were evaluated. Using this strategy, mathematical model were defined and response surface were derived for the separation. The coefficient of determination R2 was more than 0.8972 for all the models. The three responses were simultaneously optimized by using Derringer’s desirability functions. Optimum conditions chosen for assay were MeCN, MeOH, 11.39 mM K2HPO4 (pH 7.0 ) solution (40:30:30 v/v/v) and flow rate 0.81 ml/min. Total chromatographic analysis time per sample was approximately 4.56 min with PRO(IS), AMB, LCT and MLS eluting with retention times of 2.44,3.44,4.11 and 4.56 minutes respectively. The LODs were 0.20, 0.16, 0.23 and 0.34 ng /mL and the LOQs were 0.56, 0.47, 0.58 and 1.12 ng/mL for PRO (IS), AMB, LCT and MLS respectively. The optimized assay condition was validated as per the ICH guidelines and applied for the quantitative analysis of Airitis plus capsules, and Montair-LC