Analytical Method Development and Validation of Stability Indicating RP-HPLC Method for Estimation of Dolutegravir in Tablet Dosage Form Using QbD Approach

Abstract

A Simple, accurate, specific and rugged reverse phase liquid chromatographic method was developed and validated for the estimation of dolutegravir in tablet dosage form using QbD approach. In the HPLC method, analysis of the drug was carried out on the X bridge C18,150 mm x 4.6 mm, 5 μm column using a mixture of Acetonitrile : phosphate buffer (pH 6.5) in the ratio of 70:30 v/v as the mobile phase at the flow rate 1.2 mL/min. Column oven temperature was 35°C and the column eluent was detected by using UV detector at detection wavelength 258 nm. The retention time of dolutegravir was found to be 8.5 and linearity of the method was found to be between 50-150 μg/mL. Quality by Design approach was applied to reduce the development trials for optimization of analytical method. Miscellaneous Response Surface Methodology was applied in Design Expert software. Column Temperature and Acetonitrile and buffer ratio in mobile phase were two critical parameters observed. The response was on number of Plate counts and Retention time of Dolutegravir Sodium. Other graphs such as Contour plot and 3D surface were obtained from software, have been useful to understand the response of factors on analytical method. Chromatographic peak purity results indicated the absence of co-eluting peaks with the main peak of Dolutegravir sodium The method was validated as per ICH guideline and applied successfully for the estimation of dolutegravir from tablet formulation.