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 (100×2.1mm, 1.7μm) by gradient elution at 0.7mL/min of acetonitrile and 5mM 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 7min. 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 25–5000ng/mL, 0.5–500ng/mL and 0.15–10ng/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.
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