| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1212088 | Journal of Chromatography B | 2015 | 5 Pages |
•A method was validated to assay ribavirin, sofosbuvir and its metabolite in plasma.•The plasma sample was prepared by a protein precipitation.•The method could also be used for clinical pharmacokinetic study.
A rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) method for the determination of ribavirin, sofosbuvir and its metabolite GS-331007 in rat plasma was established. The analytes and the internal standard (midazolam) were separated on an Acquity UPLC BEH C18 chromatography column (2.1 mm × 50 mm, 1.7 μm) using gradient elution with a mobile phase of acetonitrile and 0.1% formic acid in water at a flow rate of 0.4 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z 245.1 → 113.1 for ribavirin, m/z 530.3 → 243.1 for sofosbuvir, m/z 261.5 → 113.1 for GS-331007 and m/z 326.2 → 291.1 for midazolam (IS) using a positive electrospray ionization interface. The method was validated over a concentration range of 5–1000 ng/mL for ribavirin, 10–2000 ng/mL for sofosbuvir and 10–2000 ng/mL for GS-331007. Total time for each chromatograph was 3.0 min. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD) <10.0% and the accuracy values ranged from −10.6% to 11.6%. The method was successfully applied to a pharmacokinetic study of ribavirin, sofosbuvir and GS-331007 in rats.
