Development and Validation of Dolutegravir SodiumIn Human Spiked Plasma by Using HPLC Method

A rapid and simple high performance liquid chromatography (HPLC) method was developed and validated for the measurement of dolutegravir sodium from tampered human plasma using a UV detector. A mobile phase comprising Acetonitrile: Phosphate Buffer pH3 (50:50%, v/v) at a flow rate of 1 ml/min was used to develop and optimise chromatographic conditions on the RESTEK C18 Column (250 × 4.6 mm, 5 um). Internal standard pioglitazone and dolutegravir sodium were extracted from human plasma by protein precipitation using the organic solvent acetonitrile (ACN). Dolutegravir sodium was successfully extracted from plasma components and internal standards. The detection was performed at 258 nm. For the calibration curve, which was linear in the 0.2–8 µg/mL range, heteroscedasticity was minimised using weighted least square regression with a weighing factor (1/x). The November 2022 ICH M10 guidelines for bioanalytical method validation's standard curve requirements were met by this process. At all three tested levels (LQC, MQC, and HQC), the percentage RSD and %RE were less than 15 and between ± 15, respectively, indicating the statistical accuracy and precision of the approach. The results of recovery studies show that extraction is effective.The results of the stability evaluation indicate that the dolutegravir sodium concentrations in the stability samples varied between 85 and 115% of the nominal concentration with percentage RSD. below fifteen. The method is in compliance with ICH M10 guidelines and can be applied to dolutegravir sodium bioavailability and bioequivalence tests.