Development and Validation of a Simple Method for Simultaneously Measuring the Concentrations of BCR-ABL and Bruton Tyrosine Kinase Inhibitors in Dried Blood Spot (DBS): A Pilot Study to Obtain Candidate Conversion Equations for Predicting Plasma Concentration Based on DBS Concentration

摘要

Background:Dried blood spots (DBSs) are promising candidates for therapeutic drug monitoring. In this study, a simple method for the simultaneous measurement of tyrosine kinase inhibitors (TKIs), including bosutinib, dasatinib, ibrutinib, imatinib, nilotinib, and ponatinib, using DBS was developed and validated. The prediction of the plasma concentration of TKIs based on the TKI concentrations in the DBS was assessed using the developed measurement method.Methods:DBS was prepared using venous blood on Whatman 903 cards. One whole DBS sample containing the equivalent of 40 mu L of blood was used for the analysis. The analytical method was validated according to the relevant guidelines. For clinical validation, 96 clinical samples were analyzed. The regression equation was derived from a weighted Deming regression analysis, and correction factors for calculating the estimated plasma concentrations (EPCs) of the analytes from their concentrations in the DBS and the predictive performance of EPC were evaluated using 2 conversion equations.Results:This method was successfully validated. Hematocrit had no significant effect on the method's accuracy or precision. Ibrutinib was stable in the DBS for up to 8 weeks at room temperature, whereas all BCR-ABL TKIs were stable for 12 weeks. All BCR-ABL TKIs exhibited similar predictive performance for EPCs using both calculation methods. Good agreement between EPCs and the measured plasma concentrations of bosutinib, imatinib, and ponatinib was observed with both conversion equations. However, Bland-Altman analysis showed that blood sampling time affected the EPC accuracy for dasatinib and nilotinib.Conclusions:A simple method for the simultaneous determination of BCR-ABL and Bruton TKI concentrations in DBS was developed and validated. Owing to the small clinical sample size, further clinical validation is needed to determine the predictive performance of EPCs for the 6 TKIs.