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Simple determination of plasma ibrutinib concentration using high-performance liquid chromatography.

Ibrutinib is an oral inhibitor of Bruton tyrosine kinase, which is one of the key drugs used for the treatment of chronic lymphocytic leukemia and mantle cell lymphoma. In this study, we aimed to develop a simple method for determining plasma ibrutinib concentration. The analysis required extraction of a 200 μL plasma sample and precipitation of proteins using solid-phase extraction. Ibrutinib and nilotinib, which was used as an internal standard, were separated using high-performance liquid chromatography (HPLC) using a mobile phase of acetonitrile-0.5% monopotassium phosphate (KH2 PO4 , pH 3.0; 52:48, v/v) on a Capcell Pack C18 MG II (250 × 4.6 mm) monitored at 260 nm, at a flow rate of 1.0 mL/min. The calibration curve was linear at the plasma concentration range of 10-500 ng/mL with a coefficient of determination (r2 ) of 0.9999. The coefficients of intra-day and inter-day validation were 4.0-6.6 and 2.6-7.7%, respectively. The assay accuracy was -4.4-8.6%, and the recovery was >84%. This HPLC method coupled with ultraviolet (UV) detection for determining ibrutinib plasma concentration has several advantages such as simplicity and applicability to routine therapeutic drug monitoring at hospital laboratories.