RP-HPLC Method development and validation for simultaneous estimation of montelukast and ebastine in pharmaceutical dosage form

Abstract

Combination therapy of Montelukast (MNK) and Ebastine (EBA) provide anti-asthmatic effector the maintain treatment of asthma and relieve symptoms of allergies. The objective of this study is development of a new simple, accurate, sensitive, and reproducible RP-HPLC method for simultaneous estimation of MNK and EBA in pharmaceutical formulation (tablet) using Ofloxacin (OFL) as an internal standard and validate the same as per ICH guidelines. The chromatogram separation was achieved on Qualisil-5 BDS C8 column (250 mm × 4.6 mm, 5µm) column with mobile phase acetonitrile: water (pH 2.8 with TFA) in the composition of 84:16 v/v at a flow rate of 1 mL/min using PDA detector at 254 nm at ambient column temperature, keeping the injection volume 20 μL. The retention time of OFL, MNK, and EBA was observed to be 2.107 min, 2.517 min, and 3.819 min, respectively. All the criteria for the validation (linearity, accuracy, precision, and robustness) were observed to be within the acceptation range. The calibration plots were obtained between 5-60 µg/mL for MNK and 5-60 µg/mL for EBA with r2 values of 0.999 in each case. The recovery of MNK and EBA was found to be 98.99% and 99.40%, respectively with a % RSD of <2. This RP-HPLC method was found to be rapid, specific, precise, and accurate and can be used for the routine analysis of MNK and EBA in bulk as well as in tablet dosage form. The separation was complete with a shorter analysis time along with well good resolved peak.