Development and validation of an HPLC-MS/MS method to simultaneously quantify brigatinib, lorlatinib, pralsetinib and selpercatinib in human K2-EDTA plasma.

A liquid chromatography-tandem mass spectrometry method was developed and validated to quantify the small molecule inhibitors (SMIs) brigatinib, lorlatinib, pralsetinib and selpercatinib, which are used in patients with oncogenic driven non-small cell lung cancer. Chromatographic separation was performed on a HyPURITY® C18 analytical column with a gradient elution using ammonium acetate in water and in methanol, both acidified with formic acid 0,1%. Detection and quantification were performed by a triple quad mass spectrometer with an electrospray ionization interface. The assay was validated over a linear range of 50 - 2,500 ng/mL for brigatinib, 25 - 1,000 ng/mL for lorlatinib, 100 - 10,000 ng/mL for pralsetinib, and 50 - 5,000 ng/mL for selpercatinib. All four SMIs were stable for at least seven days at cooling conditions (2 - 8 °C), and at least 24 hours at room temperature (15 - 25 °C) in K2-EDTA plasma. At freezing conditions (-20°C), all SMIs were stable for at least 30 days, except for the lowest quality control (QC LOW ) of pralsetinib. The QC LOW of pralsetinib was stable for at least 7 days at -20°C. This method provides an efficient and simple way to quantify four SMIs with a single assay in clinical practice.