HIGH-THROUGHPUT UPLC METHOD FOR DETERMINATION OF BEDAQUILINE IN PLASMA USING PROTEIN PRECIPITATION

Abstract

Background: Bedaquiline, a diarylquinoline inhibitor of ATP synthase is an important drug in the treatment of multidrug resistant tuberculosis (MDR TB) but due to its lipophilic property and low plasma levels, sensitive measurement of this drug is difficult. Method: To achieve the first UPLC assay for bedaquiline in human plasma, the method was developed and is now completely validated based on ICH guidelines. The separation was carried out on a Thermo Hypurity C18 column using acetonitrile–ammonium acetate (60:40, v/v) as the mobile phase with bedaquiline d6 as the internal standard. Results and Discussion: The method achieved excellent linearity with r2 of 0.999, over the range of 25-225 µg/mL, accuracy of 94-110% and precision of < 7%CV. Different types of plasma (lipemic, hemolyzed etc.) all had a recovery of around 70%. Freeze–thaw, bench-top exposure and storage of processed samples did not cause a loss of stability. Using a simple precipitation clean up and UPLC. Using the Lean Six Sigma (DMAIC) concept, the workflow was methodically optimized for efficiency, reproducibility and compliance to regulations. Conclusion: This assay appears to be an alternative and the procedure provides a quick, selective and reproducible method for pharmacokinetic study and therapeutic drug monitoring where TB patients are infected with MDR TB than previous HPLC methods.