title: A rugged and accurate liquid chromatography-tandem mass spectrometry method for the determination of asunaprevir, an NS3 protease inhibitor, in plasma.
abstract: Asunaprevir (BMS-650032) is a potent hepatitis C virus (HCV) non-structural protein protease inhibitor currently in Phase III clinical trials for the treatment of HCV infection. A rugged and accurate LC-MS/MS method was developed and validated for the quantitation of asunaprevir in rat, dog, monkey, rabbit and mouse plasma. A systematic method screening and optimization strategy was applied to achieve optimized mass spectrometry, chromatography, and sample extraction conditions. The validated method utilized stable-isotope labeled D9-asunaprevir as the internal standard. The samples were extracted by liquid-liquid extraction using 10% ethyl acetate in hexane. Chromatographic separation was achieved with gradient elution on a Waters Atlantis dC18 analytical column. Analyte and its internal standard were detected by positive ion electrospray tandem mass spectrometry. The standard curve, which ranged from 5.00 to 2000ng/mL for asunaprevir, was fitted to a 1/x(2) weighted linear regression model. The intra-assay precision was within ±3.6% CV, inter-assay precision was within ±4.0% CV, and the assay accuracy was within ±8.1% of the nominal values in all the species. The method was successfully applied to support multiple pre-clinical toxicokinetic studies in different species.
authors: Analytical and Bioanalytical Development, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, United States.
authors: Long
authors: L
authors: Yuan
authors: Hao
authors: H
authors: Jiang
authors: Zheng
authors: Z
authors: Ouyang
authors: Yuan-Qing
authors: YQ
authors: Xia
authors: Jianing
authors: J
authors: Zeng
authors: Qianping
authors: Q
authors: Peng
authors: Robert W
authors: RW
authors: Lange
authors: Yuzhong
authors: Y
authors: Deng
authors: Mark E
authors: ME
authors: Arnold
authors: Anne-Françoise
authors: AF
authors: Aubry
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods; D004285:Dogs; D000882:Haplorhini; D016174:Hepacivirus / Q000201:enzymology*; D006526:Hepatitis C / Q000188:drug therapy; D007546:Isoquinolines / Q000097:blood*; D051379:Mice; D011480:Protease Inhibitors / Q000097:blood*; D011817:Rabbits; D051381:Rats; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000998:Antiviral Agents; D007546:Isoquinolines; D011480:Protease Inhibitors; D013449:Sulfonamides; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C571889:asunaprevir