title: A validated liquid chromatography tandem mass spectrometry method for quantification of erlotinib, OSI-420 and didesmethyl erlotinib and semi-quantification of erlotinib metabolites in human plasma.
abstract: A liquid chromatography tandem mass spectrometry method was developed and validated for quantification of erlotinib and its metabolites in human plasma. The method is suitable for therapeutic drug monitoring and pharmacokinetic studies. The substances were extracted using protein precipitation, separated on a BEH XBridge C18 column (100 ×2.1 mm, 1.7 μm) by gradient elution at 0.7 mL/min of acetonitrile and 5 mM ammonium acetate. The concentration was determined using a Waters Xevo triple quadrupole mass spectrometer in a multi reaction monitoring mode. The total run time was 7 min. Deuterated erlotinib and OSI-597 were used as internal standard for erlotinib and its metabolites, respectively. Erlotinib, OSI-420 and didesmethyl erlotinib were quantified in the concentration range 25-5000 ng/mL, 0.5-500 ng/mL and 0.15-10 ng/mL, respectively. Precision and accuracy was <14% except for OSI-420 at LLOQ (17%). Extraction recovery was above 89%, 99% and 89% for erlotinib, OSI-420 and didesmethyl erlotinib, respectively. The human liver microsomes generated 14 metabolites, three of them not previously reported. Twelve metabolites were measured semi-quantitatively and validated with respect to selectivity, precision and stability.
authors.affiliation: Clinical Pharmacology, Division of Drug Research, Department of Medical and Health Sciences, Linköping University, SE-581 85 Linköping, Sweden. Electronic address: anna.svedberg@liu.se.
authors.forename: Anna
authors.initials: A
authors.lastname: Svedberg
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; School of Biotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, SE-171 21 Solna, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Gréen
authors.affiliation: Department of Pulmonary Medicine, University Hospital, SE-581 85 Linköping, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Vikström
authors.affiliation: School of Biotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, SE-171 21 Solna, Sweden.
authors.forename: Joakim
authors.initials: J
authors.lastname: Lundeberg
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
mesh_terms: D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D016903:Drug Monitoring / Q000379:methods; D000069347:Erlotinib Hydrochloride; D006801:Humans; D008862:Microsomes, Liver / Q000378:metabolism; D010949:Plasma / Q000737:chemistry*; D011799:Quinazolines / Q000097:blood* / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: C487471:OSI-420; D011799:Quinazolines; D000069347:Erlotinib Hydrochloride
keywords: EGFR; Human liver microsomes; LC–MS/MS; Non-small cell lung cancer; Tyrosine kinase inhibitor