title: Novel rapid liquid chromatography tandem masspectrometry method for vemurafenib and metabolites in human plasma, including metabolite concentrations at steady state.
abstract: A novel, rapid and sensitive liquid chromatography tandem-mass spectrometry method for quantification of vemurafenib in human plasma, that also for the first time allows for metabolite semi-quantification, was developed and validated to support clinical trials and therapeutic drug monitoring. Vemurafenib was analysed by precipitation with methanol followed by a 1.9 min isocratic liquid chromatography tandem masspectrometry analysis using an Acquity BEH C18 column with methanol and formic acid using isotope labelled internal standards. Analytes were detected in multireaction monitoring mode on a Xevo TQ. Semi-quantification of vemurafenib metabolites was performed using the same analytical system and sample preparation with gradient elution. The vemurafenib method was successfully validated in the range 0.5-100 μg/mL according to international guidelines. The metabolite method was partially validated owing to the lack of commercially available reference materials. For the first time concentration levels at steady state for melanoma patients treated with vemurafenib is presented. The low abundance of vemurafenib metabolites suggests that they lack clinical significance. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, SE, 581 85, Linköping, Sweden.
authors.forename: Svante
authors.initials: S
authors.lastname: Vikingsson
authors.affiliation: Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, SE, 581 85, Linköping, Sweden.|Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, SE, 587 58, Linköping, Sweden.
authors.forename: Malin
authors.initials: M
authors.lastname: Strömqvist
authors.affiliation: Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, SE, 581 85, Linköping, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Svedberg
authors.affiliation: Department of Oncology-Pathology, Karolinska Institutet, Karolinska University Hospital Solna, SE, 171 64, Stockholm, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Hansson
authors.affiliation: Department of Oncology-Pathology, Karolinska Institutet, Karolinska University Hospital Solna, SE, 171 64, Stockholm, Sweden.
authors.forename: Veronica
authors.initials: V
authors.lastname: Höiom
authors.affiliation: Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, SE, 581 85, Linköping, Sweden.|Department of Forensic Genetics and Forensic Toxicology, National Board of Forensic Medicine, SE, 587 58, Linköping, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Gréen
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D004791:Enzyme Inhibitors / Q000097:blood*; D006801:Humans; D007211:Indoles / Q000097:blood*; D057230:Limit of Detection; D015203:Reproducibility of Results; D013449:Sulfonamides / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000077484:Vemurafenib
chemical_list: D004791:Enzyme Inhibitors; D007211:Indoles; D013449:Sulfonamides; D000077484:Vemurafenib
keywords: BRAFV600E; LC-MS/MS; melanoma; metabolites; validation