title: Evaluation of a rapid method for the simultaneous quantification of ribavirin, sofosbuvir and its metabolite in rat plasma by UPLC-MS/MS.
abstract: 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 (2.1mm×50mm, 1.7μm) using gradient elution with a mobile phase of acetonitrile and 0.1% formic acid in water at a flow rate of 0.4mL/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 5-1000ng/mL for ribavirin, 10-2000ng/mL for sofosbuvir and 10-2000ng/mL for GS-331007. Total time for each chromatograph was 3.0min. 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.
authors.affiliation: Department of Hepatopathy, Ningbo No.2 Hospital, Ningbo 315000, China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Shi
authors.affiliation: Department of Hepatopathy, Ningbo No.2 Hospital, Ningbo 315000, China.
authors.forename: Dedong
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Digestive Department, Ningbo No.2 Hospital, Ningbo 315000, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Lou
authors.affiliation: Pharmaceutical Preparation Section, Ningbo No.2 Hospital, Ningbo 315000, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhu
authors.affiliation: Department of Hepatopathy, Ningbo No.2 Hospital, Ningbo 315000, China. Electronic address: 21466738@qq.com.
authors.forename: Ai-rong
authors.initials: AR
authors.lastname: Hu
authors.affiliation: Department of Paediatrics, Ningbo Women and Children's Hospital, Ningbo 315000, China. Electronic address: 109315536@qq.com.
authors.forename: Dongmei
authors.initials: D
authors.lastname: Gan
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000097:blood* / Q000493:pharmacokinetics; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D057230:Limit of Detection; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012254:Ribavirin / Q000097:blood* / Q000493:pharmacokinetics; D000069474:Sofosbuvir / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000998:Antiviral Agents; D012254:Ribavirin; D000069474:Sofosbuvir
keywords: GS-331007; Pharmacokinetics; Plasma; Ribavirin; Sofosbuvir; UPLC–MS/MS