Rapid RP-HPLC Method for Simultaneous estimation of Umeclidinium and Vilanterol in human plasma

Abstract

A simple, precise, accurate method was developed for the estimation of Umeclidinium andVilanterol in human plasma using Empagliflozin as internal standard by RP-HPLC (Reverse phase-High performance Liquid Chromatographic) technique. Chromatographic conditions used are stationary phase KromasilC18 (250 x 4.6 mm, 5m)coulmn, mobile phase of 0.01N di sodium hydrogen phosphate (pH: 3.0): Acetonitrile in the ratio of 65:35(v/v) and flow rate was maintained at 1.0ml/min, detection wave length was 215nm, column temperature was set at 30oC and diluent was mobile phase. Retention time of Umeclidinium and Vilanterol and were found to be 2.471min and 3.303min. %CV of the Umeclidinium and Vilanterol were found to be 0.33% and 0.43%. %Recovery was obtained as 99.34% and 99.029%. The linearity concentration is in the range of 1.25-50µg/mL of Umeclidinium, 0.25-10.0µg/mL of Vilanterol respectively. The lower limits of quantification were 1.25µg/mL of Umeclidiniumand 0.25µg/mL of Vilanterol which reach the level of both drugs possibly found in human plasma. Further, the reported method was validated as per the ICH guidelines and found to be well within the acceptable range.