DEVELOPMENT AND VALIDATION OF RP-HPLC CHROMATOGRAPHIC METHOD FOR THE ESTIMATION OF DOLUTEGRAVIR IN BULK AND TABLET DOSAGE FORM

Kolhe Mahesh Hari*, Phalke Shubhangi, Kakad Sudarshan Balasaheb and Dukre Tushar Pradip

ABSTRACT

A simple, sensitive, rapid and selective isocratic reversed phase High Performance Liquid Chromatographic (HPLC) method has been developed for Dolutegravir from bulk drug using a mobile phase consisting mixture of Methanol: Buffer KH2PO4 (80:20v/v) (pH 3) Composition of buffer: (0.272gm in 200ml HPLC water and pH adjusted to 3 using ortho Phosphoric acid) at the flow rate of 0.8 mL/min. A Cosmosil C18 (250 cm x 4.6 mm, 5 μm) column was used as stationary phase. The retention time of Dolutegravir found to be 5.698 min. The eluent were detected at 258 nm. Linearity was observed in the concentration range of 10-50 ppm for Dolutegravir. Percent recoveries obtained for Dolutegravir were 97%. The correlation coefficient for Dolutegravir was found to be 0.998. After performing analysis by different analysts, it was found that the RP-HPLC method for the determination of Dolutegravir was found to be Rugged. Percent RSD for robustness was well within the acceptable USP limits, ensuring that the proposed method was robust. The LOD were 0.060 μg/ml Dolutegravir,. For Dolutegravir, the LOQ were found to be 0.184 μg/ml. This demonstrated that the developed RP-HPLC method was simple, linear, precise, accurate, robust, and Rugged, could be conveniently adopted for the routine quality control analysis of Dolutegravir, from its pharmaceutical formulations and bulk drug.

Keywords: Dolutegravir (DTG), HPLC Method, Mobile phase.