LAUSR

The hematocrit-bias still remains one of the most discussed issues when it comes to dried blood spot (DBS) analysis. Therefore, many attempts to cope with this issue have been made, among which the development of novel sampling tools such as the Capitainer-B (further referred to as MF (microfluidic)-DBS) devices. These are designed to allow a straightforward absorption of a fixed volume (13.5 μL) of blood by a preperforated paper disc, which can be analyzed afterward. The aim of this study was to evaluate the potential of these devices to nullify the hematocrit-based area bias and to investigate whether the amount of blood applied has an influence on the device performance. An LC-MS/MS method for the quantification of caffeine and paraxanthine in MF-DBS was fully validated, meeting all preset acceptance criteria. In a next step, using a set of 133 authentic, venous patient samples with a hematocrit range of 18.8-55.0, concentrations of both compounds in MF-DBS were compared to those in corresponding partial-punch pipetted DBS (PI-DBS) and liquid blood samples. When compared to blood as a reference, the concentrations obtained in MF-DBS were not affected by a bias in function of the evaluated hematocrit, in contrast to those obtained from partial-punch PI-DBS. Furthermore, analysis of samples resulting from spiking different volumes of whole blood at different hematocrit levels, revealed that the amount of blood applied at the device inlet has no influence on the performance of the devices. Therefore, it can be concluded from this study, being the first in which the impact of the hematocrit and the applied volume is evaluated by analyzing authentic, venous patient samples, that MF-DBS devices effectively assist in eliminating the hematocrit-based area bias, independently from the applied blood volume.