Abstract:

Objective To establish an high performance liquid chromatography-mass spectrometry/mass spectrometry (HPLC-MS/MS) method for the determination of bedaquiline in human plasma and to evaluate the application of the method.Methods The protein of plasma samples were precipitated with acetonitrile. HPLC-MS/MS was performed using an Agilent ZORBAX SB-C18 column (2.1mm×100mm, 3.5μm) as the analytical column. The mobile phase consisted of methanol-5mmol/L ammonium formate (containing 0.1% formic acid solution) (85∶15, v/v). Electrospray ionization source (ESI) was applied and operated in the positive multiple reaction monitoring (MRM) mode. One analysis was completed within 3min.Results Chromatograms showed no endogenous interfering peaks with blank samples. The mass-to-charge ratio (m/z) of the parental ion and the quantitative ion of bedaquiline was 555.2→58.3. The linear calibration curve was obtained over the concentration on range of 0.1-5.0μg/ml. The inter- and intra-day precision (RSD) were less than 10%. The accuracy were 97.8%-107.0%. 48 plasm samples of 12 tuberculosis patients were detected using the established method. When the dosage was 400mg daily, the peak concentration of bedaquiline was (3.17±1.14)μg/ml, and the trough concentration was (1.12±0.64)μg/ml; when the dosage was 200mg per time, 3 times a week, the peak concentration of bedaquiline was (2.01±0.87)μg/ml, and the trough concentration was (0.65±0.30)μg/ml. The concentrations of bedaquiline in the 48 samples were in the range of calibration curve (0.1-5.0μg/ml).Conclusion A HPLC-MS/MS method was established for the quick analysis and determination of bedaquiline (a new anti-tuberculosis drug) in plasma samples of tuberculosis patients.

Key words: Chromatography, high pressure liquid, Tandem mass spectrometry, Pharmacokinetics, Drug monitoring, Bedaquiline