Development and validation of high performance liquid chromatographic method for the determination of Dolutegravir in human plasma

A rapid, specific and accurate high performance liquid chromatographic method for the determination of dolutegravir in human plasma using hydrochlorothiazide as internal standard was developed and validated by UV detection. The extraction process involved a liquid-liquid extraction using methyl-t-butyl ether. Both dolutegravir and the internal standard were eluted under isocratic mode using a 150 X 4.6 mm i.d, 5 μm Phenomenex ODS 2 C18 column. The mobile phase composed a mixture of 30:70 % v/v 20 mM Sodium acetate buffer (pH 4.0) and Methanol at a flow rate of 1.0 mL/minute. The wavelength of detection is 254 nm. The injection volume is 20 μL. The runtime of the method is 6 minutes with retention times 2.08 minutes and 4.16 for Hydrochlorothiazide and Dolutegravir respectively. The method showed good linearity in the range of 101.90 to 7004.49 ng/mL. The recovery of Dolutegravir is 59.21 % with a co-efficient of variation 3.72 % and recovery of internal standard was 60.61 % with a co-efficient of variation 3.33 %