title: Simultaneous determination of ledipasvir, sofosbuvir and its metabolite in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study.
abstract: In this work, a rapid and sensitive ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the determination of ledipasvir, sofosbuvir and its metabolite GS-331007 in rat plasma was developed. The analytes and the internal standard (diazepam) 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 889.8→130.1 for ledipasvir, m/z 530.3→243.1 for sofosbuvir, m/z 261.5→113.1 for GS-331007 and m/z 285.2→193.1 for diazepam (IS) using a positive electrospray ionization interface. The method was validated over a concentration range of 2-500ng/mL for ledipasvir, 10-2000ng/mL for sofosbuvir and 10-2000ng/mL for GS-331007. Total time for each chromatography 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)<10.2% and the accuracy values ranged from -9.8% to 11.2%. The method was successfully applied to a pharmacokinetic study of ledipasvir, sofosbuvir and GS-331007 in rats.
authors.affiliation: The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China.
authors.forename: Chenwei
authors.initials: C
authors.lastname: Pan
authors.affiliation: The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
authors.forename: Yongping
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Wenzhou People's Hospital, Wenzhou 325027, China.
authors.forename: Weilai
authors.initials: W
authors.lastname: Chen
authors.affiliation: The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China.
authors.forename: Guangyao
authors.initials: G
authors.lastname: Zhou
authors.affiliation: The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China.
authors.forename: Lingxiang
authors.initials: L
authors.lastname: Jin
authors.affiliation: The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China. Electronic address: linwei196061@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Lin
authors.affiliation: The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China. Electronic address: wzzhenzhenpan@126.com.
authors.forename: Zhenzhen
authors.initials: Z
authors.lastname: Pan
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000097:blood* / Q000493:pharmacokinetics; D001562:Benzimidazoles / Q000097:blood* / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D005449:Fluorenes / Q000097:blood* / Q000493:pharmacokinetics; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D000069474:Sofosbuvir / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry
chemical_list: D000998:Antiviral Agents; D001562:Benzimidazoles; D005449:Fluorenes; C586541:ledipasvir; D000069474:Sofosbuvir
keywords: GS-331007; Ledipasvir; Pharmacokinetics; Plasma; Sofosbuvir; UPLC–MS/MS