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Bioanalysis of bedaquiline in human plasma by liquid chromatography-tandem mass spectrometry: Application to
Viritha Bezawada1,2, Padma Mogili2, Srinivasa Rao Polagani3
1Aurora Degree and PG College, Hyderabad, Telangana, India.
Introduction:
A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for estimation of bedaquiline (BDQ) in human plasma using the deuterated analogue of the analyte, bedaquiline-d6 (BDQ-d6) as the internal standard.
Methods:
The plasma sample of 50 µL was extracted by liquid-liquid extraction using methyl tertiary butyl ether (MTBE). The separation was achieved on Zodiac C18 (50 x 4.6 mm, 5 µm) column with a mobile phase consisting of methanol and 5 mM ammonium formate in 0.1 % formic acid (w/v) (90:10, v/v) at a flow rate of 1.0 mL/min. Protonated analyte and internal standard were detected on a triple quadrupole mass spectrometer using multiple reaction monitoring (MRM) mode.
Results:
The linearity of the method was established in the concentration range of 5---1800 ng/mL with correlation coefficient, r2 ≥ 0.99. All the validated parameters were found well within the limits.
Discussion:
The method was applied for the first time to evaluate the pharmacokinetic parameters after single oral dose of BDQ 100 mg under fed conditions in healthy human volunteers, and the results were further authenticated by incurred sample reanalysis.
