Analytical Method Development and Validation for Rilpivirine and Dolutegravir in Combine Dosage Forms by RP- HPLC

Pakir Pranaya*
Guru Nanak Institute of Pharmacy, Ibrahimpatnam, Rangareddy, Telangana, India

 A B S T R A C T
The chromatographic conditions were successfully developed for the separation of Rilpivirine and Dolutegravir by using ACE C18 column (4.6×150mm) 5µ, flow rate was 1.2 ml/min, mobile phase ratio was (70:30 v/v) methanol: Phosphate buffer pH 3 (pH was adjusted with orthophosphoricacid), detection wavelength was 240nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2690, photo diode array detector 996, Empower-software version-2. The retention times were found to be 2.344 mins and 3.284 mins. The % purity of Rilpivirine and Dolutegravir was found to be 101.27% and 99.97% respectively.  The  system  suitability  parameters  for  Rilpivirine  and  Dolutegravir  such  as theoretical plates and tailing factor were found to be 4668, 1.3 and 6089 and 1.2, the resolution was found to be 6.0. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study n Rilpivirine and Dolutegravir was found in concentration range of 50µ g-250µ g and 5µ g-50µ g and correlation coefficient (r²) was found to be 0.999 and 0.999, % recovery was found to be 99.56% and 99.48%, %RSD for repeatability was 0.2 and 0.2, % RSD for intermediate precision was 0.2 and 0.1respectively. The precision study was precise, robust, and repeatable. LOD value was 3.17 and 5.68, and LOQ value was 0.0172 and 0.2125 respectively. Hence  the  suggested RP-HPLC  method  can  be  used  for  routine  analysis  of Rilpivirine and Dolutegravir in API and Pharmaceutical dosage form.
Keywords: ACE C18 column, Rilpivirine and Dolutegravir, RP-HPLC