title: Validation of an electrospray ionisation LC-MS/MS method for quantitative analysis of telaprevir and its R-diastereomer.
abstract: A sensitive high-performance reverse phase liquid chromatography-positive ion electrospray tandem mass spectrometry method was developed and validated for the quantification of telaprevir and its inactive R-diastereomer (VRT-127394) in human plasma. The analytes and the internal standard (telaprevir-d11) were extracted from plasma by liquid-liquid extraction using tert-Butyl methyl ether (TBME). Chromatographic separation was achieved on a reversed-phase Accucore C18 column with a gradient programme consisting of water:ammonia (25%), 100:0.01 (v/v) (mobile phase A) and ACN:MeOH:ammonia (25%), 15:85:0.01 (v/v/v) (mobile phase B). The MS acquisition was performed with selective reaction monitoring mode using the respective [M+H](+) ions, m/z 680.59→322.42 for telaprevir and VRT-127394, and 691.15→110.13 for telaprevir-d11. The assay exhibited a linear dynamic range of 5-5000ng/mL for telaprevir and VRT-127394. Acceptable precision (%RSD<6.5%) and accuracy (94-108%) were obtained for concentrations over the range of the standard curve. A procedure was established to stabilise the plasma to prevent ex vivo interconversion of the isomers.
authors.affiliation: Department of Molecular & Clinical Pharmacology, University of Liverpool, Liverpool, UK.
authors.forename: Sujan Dilly
authors.initials: SD
authors.lastname: Penchala
authors.forename: John
authors.initials: J
authors.lastname: Tjia
authors.forename: Omar
authors.initials: O
authors.lastname: El Sherif
authors.forename: David J
authors.initials: DJ
authors.lastname: Back
authors.forename: Saye H
authors.initials: SH
authors.lastname: Khoo
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Else
mesh_terms: D000998:Antiviral Agents / Q000097:blood* / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D016174:Hepacivirus / Q000201:enzymology*; D006526:Hepatitis C / Q000188:drug therapy; D006801:Humans; D057230:Limit of Detection; D009842:Oligopeptides / Q000097:blood* / Q000737:chemistry; D011480:Protease Inhibitors / Q000097:blood* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000998:Antiviral Agents; D009842:Oligopeptides; D011480:Protease Inhibitors; C486464:telaprevir
keywords: Ex vivo interconversion; Hepatitis C; LC–MS/MS; Method validation; Telaprevir; VRT-127394