Determination of Three Tyrosine Kinase Inhibitors and One Active Metabolite by an Identical and Validated Ultra-performance Liquid Chromatography-DAD Method in Human Plasma

Tyrosine kinase inhibitors (TKIs) are a class of targeted drugs with antiangiogenic and antitumor activities. Due to inter-individual metabolic variability, an accurate therapeutic drug monitoring (TDM) represents a key element for the patient treatment. Here a fast and easily accessible method for the quantification of 3 TKIs (with one active metabolite) in human plasma after extraction is described. Sample pre-treatments were performed by solid phase extraction (Oasis (R) MCX mu Elution technology). Chromatographic separation was performed on a Waters Acquity UPLC (R) system with diode array detection (DAD) using a gradient of ammonium formate-acetonitrile on BEH C18 2.1x50 mm column. The analytical methods were validated by using the accuracy profiles approach (beta-expectation set at 95%). The methods were successfully validated for sunitinib (10 - 250 ng/mL), N-desethyl sunitinib (15 - 250 ng/mL), axitinib (15 - 250 ng/mL) and pazopanib (20 - 200 mu g/mL). The first concentration levels validated were considered as limit of quantification (LOQ). The validated method will be used in a clinical research study to determine TKI plasma levels and in this way help physicians to optimize the posology in order to achieve the best therapeutic response for their patients.