Method Development and Validation for the Determination of Fluticasone and Salmeterol in API and Combined Tablet Dosage Form by RP-HPLC

Abstract

Analytical Method Development and Validation forSalmeterol and Fluticasone in bulk and Combined Dosage Form by RP-HPLC, New method was established for simultaneous estimation ofSalmeterol and Fluticasone by RP-HPLC method. The chromatographic conditions were successfully developed for the separation ofSalmeterol and Fluticasone by using Phenomenex Luna C18 (4.6mm×250mm, 5µm) particle size, flow rate was 1.0 ml/min, mobile phase ratio was (40:60 v/v) Acetonitrile: TEA buffer pH-4.2 (pH was adjusted with orthophosphoric acid), detection wavelength was 220nm. The instrument used was WATERS Alliance 2695 separation module, Software: Empower 2, 996 PDA detectors. The retention times were found to be 2.246mins and 5.461mins respectively. The % purity ofSalmeterol and Fluticasone was found to be 101.27% and 99.76% respectively. The system suitability parameters forSalmeterol and Fluticasone such as theoretical plates and tailing factor were found to be 5387, 0.97 and 5398 and 1.26, the resolution was found to be 2.97. The linearity study nSalmeterol and Fluticasone was found in concentration range of 30µg-70µg and 60µg-140µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 100.14% and 100.56%, %RSD for repeatability was 0.1 and 0.5, % RSD for intermediate precision was 0.1 and 0.1 respectively. The precision study was precise, robust, and repeatable. LOD value was 0.56 and 1.2, and LOQ value was 1.7 and 3.6 respectively. Hence the suggested RP-HPLC method can be used for routine analysis ofSalmeterol and Fluticasone in API and Pharmaceutical dosage form