title: Development and validation of a rapid and simple LC-MS/MS method for quantification of vemurafenib in human plasma: application to a human pharmacokinetic study.
abstract: BACKGROUND
  Vemurafenib (Zelboraf) is a new tyrosine kinase inhibitor that selectively targets activated BRAF V600E gene and is indicated for the treatment of advanced BRAF mutation-positive melanoma. We developed a simple method for vemurafenib quantification using liquid chromatography-tandem mass spectrometry. A stability study of vemurafenib in human plasma was also performed.
  
  
  METHODS
  (13)C(6)-vemurafenib was used as the internal standard. A single-step protein precipitation was used for plasma sample preparation. Chromatography was performed on an Acquity UPLC system (Waters) with chromatographic separation by the use of an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7-mm particle size; Waters). Quantification was performed using the monitoring of multiple reactions of following transitions: m/z 488.2 → 381.0 for vemurafenib and m/z 494.2 → 387.0 for internal standard.
  
  
  RESULTS
  This method was linear over the range from 1.0 to 100.0 mcg/mL. The lower limit of quantification was 0.1 mcg/mL for vemurafenib in plasma. Vemurafenib remained stable for 1 month at all levels tested, when stored indifferently at room temperature (20 °C), at +4 °C, or at -20 °C. This method was used successfully to perform a plasma pharmacokinetic study of vemurafenib in a patient after oral administration at a steady state.
  
  
  CONCLUSIONS
  This liquid chromatography-tandem mass spectrometry method for vemurafenib quantification in human plasma is simple, rapid, specific, sensitive, accurate, precise, and reliable.
authors: Departments of *Pharmacology and CIC-1421, and †Pharmacy, Pitié-Salpêtrière Hospital, AP-HP; ‡Department of Pharmacology and UMR ICAN 1166, Faculty of Medicine, Sorbonne University, UPMC Univ Paris 06; and §INSERM, CIC-1421, and UMR ICAN 1166, Paris, France.
authors: Kevin
authors: K
authors: Bihan
authors: Chloé
authors: C
authors: Sauzay
authors: Lauriane
authors: L
authors: Goldwirt
authors: Fanny
authors: F
authors: Charbonnier-Beaupel
authors: Jean-Sebastien
authors: JS
authors: Hulot
authors: Christian
authors: C
authors: Funck-Brentano
authors: Noël
authors: N
authors: Zahr
mesh_terms: D000970:Antineoplastic Agents / Q000097:blood* / Q000493:pharmacokinetics*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D007211:Indoles / Q000097:blood* / Q000493:pharmacokinetics*; D047428:Protein Kinase Inhibitors / Q000097:blood* / Q000494:pharmacology*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D013449:Sulfonamides / Q000097:blood* / Q000493:pharmacokinetics*; D053719:Tandem Mass Spectrometry; D000077484:Vemurafenib
chemical_list: D000970:Antineoplastic Agents; D007202:Indicators and Reagents; D007211:Indoles; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D000077484:Vemurafenib