title: A validated LC-MS/MS method for the simultaneous determination of BMS-791325, a hepatitis C virus NS5B RNA polymerase inhibitor, and its metabolite in plasma.
abstract: BMS-791325 is a hepatitis C virus (HCV) non-structural protein 5B (NS5B) RNA polymerase inhibitor that is being developed for the treatment of HCV infection. A rugged and accurate LC-MS/MS method was developed and validated for the quantitation of BMS-791325 and its metabolite, BMS-794712, in rat and dog plasma. This method utilized stable-isotope labeled [D6]-BMS-791325 and [13CD3]-BMS-794712 as internal standards. The samples were extracted using liquid-liquid extraction with n-butyl-chloride. Chromatographic separation was achieved with gradient elution on a Waters Atlantis dC18 analytical column (2.1mm×50mm, 3.0μm). Analytes and their stable isotope labeled internal standards were detected by positive ion electrospray tandem mass spectrometry. The standard curves, which ranged from 5.00 to 2000ng/mL for BMS-791325 and from 1.00 to 400ng/mL for BMS-794712, were fitted to a 1/x2weighted linear regression model. For both species, the intra-assay precision was within ±4.3% CV, the inter-assay precision was within ±6.2% CV, and the assay accuracy was within ±10.8% of the nominal values for BMS-791325 and BMS-794712. The validated method was successfully applied to support pre-clinical toxicokinetic studies.
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Long
authors.initials: L
authors.lastname: Yuan
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA. Electronic address: hao.jiang@bms.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Naiyu
authors.initials: N
authors.lastname: Zheng
authors.affiliation: Bioanalytical & Discovery Analytical Sciences, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Yuan-Qing
authors.initials: YQ
authors.lastname: Xia
authors.affiliation: Bioanalytical & Discovery Analytical Sciences, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ouyang
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Jianing
authors.initials: J
authors.lastname: Zeng
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Billy
authors.initials: B
authors.lastname: Akinsanya
authors.affiliation: Drug Safety Evaluation, Bristol-Myers Squibb, New Brunswick, NJ 08903, USA.
authors.forename: Jack L
authors.initials: JL
authors.lastname: Valentine
authors.affiliation: Drug Safety Evaluation, Bristol-Myers Squibb, Mt. Vernon, IN 47620, USA.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Moehlenkamp
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Yuzhong
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Aubry
authors.affiliation: Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
keywords: BMS-791325; HCV; LC–MS/MS; Metabolite; Quantitation