Liquid chromatography-tandem mass spectrometry method development and validation for the determination of erlotinib in human plasma and its application in pharmacokinetic study

A simple, rapid, specific and precise liquid chromatography–tandem mass spectrophotometric (LC-MS/MS) method was developed and validated for quantification of Erlotinib, in human plasma. Erlotinib d6 was used as internal standard, added to plasma sample prior to extraction using acetonitrile as a precipitating agent. Chromatographic separation was achieved on Phenomenex Luna C18 column (50 mm × 1 mm, 5 μm) with acteonitrile: 10 mM ammonium formate buffer (80: 20, v/v) as an isocratic mobile phase with a flow rate of 0.5 mL/min. Quantitation was performed by transition of 394.0 → 278.0 (m/z) for Erlotinib and 400.0 → 278.0 (m/z) for Erlotinib d6. The lower limit of quantitation was 10 ng/mL with a 100 mL plasma sample. The concentrations of nine working standards showed linearity between 10 and 5000 ng/mL (r2 ≥ 0.9992). Chromatographic separation was achieved within 3.50 min. The average extraction recoveries of 3 quality control concentrations were 94.25% for Erlotinib and 93.55% for Erlotinib d6. The coefficient of variation was ≤15% for intraand inter-batch assays. The developed method was successfully applied to the determination of Erlotinib pharmacokinetics after oral administration.