Development and Validation of RP-HPLC Stability Indicating Method for Simultaneous Estimation of Dolutegravir and Lamivudine in Bulk and Pharmaceutical Dosage Form

Aims: Dolutegravir (DVR) and Lamivudine (LMN) is anti-viral drug combination used in treatment of HIV-I infection. FDA approved dosage regime for DVR and LMN is 50mg and 300mg respectively. The aim of present research work is to develop and validate a reverse phase high performance liquid chromatography (RP-HPLC) method for simultaneous estimation of DVR and LMN in bulk and pharmaceutical dosage forms. Further the stability indicating nature of method has been evaluated.

Methodology: A chromatographic separation was achieved on Hypersil BDS C18, 250 × 4.6 mm 3.5 μm particle size, column as stationary phase and mobile phase composed of Phosphate Buffer pH 3.0: Acetonitrile (60:40%V/V) with flow rate of 1.5mL/min with 20µL injection volume. The analytes were estimated at 232nm using PDA detector. The DVR and LMN solutions were exposed to various forced degradation stress conditions to evaluate the stability behavior of the product. The method was also validated as per ICH Guideline (Q2R).

Results: The retention time for DVR and LMN was found 3.94 and 2.62 min, respectively. The developed method was found linear within concentration range of 27.5 to 82.5 μg/ml (50-150%) for DVR and 167.5 to 502.5 μg/ml (50-150%) for LMN. The% recovery (Accuracy) was found between 99.50%-101.23% and 100.09%-101.51% for DVR and LMN respectively in range of 50-150% for both drugs. The%RSD for the accuracy at all levels was less than 2.0%. LOD and LOQ, for DVR were found to 0.669 and 2.028 μg/mL, respectively, and for LMN 0.102 and 0.308 μg/mL, respectively.

Conclusion: The developed RP-HPLC method indicates no interference from the excipients and degradants peaks. All the degradant peaks were having been efficiently resolved through the use of the evolved analytical method with changed retention times. The results obtained were statistically analyzed and meet the acceptance criteria as specified in ICH Q2R1 guideline. Hence, developed method can be successfully applied for the analysis of estimation of DVR and LMN in bulk as well as pharmaceutical dosage form.