Utilization of volumetric absorptive microsampling and dried plasma spot for quantification of anti-fungal triazole agents in pediatric patients by using liquid chromatography-tandem mass spectrometry

Background: Recently, increasing attention has been paid to the use of microsampling techniques for therapeutic drug monitoring (TDM) in neonatal and pediatric populations. Volumetric Absorptive Microsampling (VAMS) has been introduced in the market under the name Mitra® (Neoteryx). These devices consist of porous absorbent tips that allow collection of fixed blood volumes (10-30 µL) to overcome the DBS-related hematocrit effect. Here, the authors analyzed the concentrations of triazole agents (voriconazole, posaconazole, and isavuconazole) in VAMS and dried plasma spot (DPS) samples.

Methods: Fifty whole blood samples were obtained from pediatric patients subjected to systemic anti-fungal therapy. VAMS were collected by dipping the tip into whole blood before centrifugation for plasma recovery. Then, 30 µL of plasma was carefully spotted on filter paper to obtain DPS. Anti-fungal concentrations were measured using a validated LC-MS/MS kit (MassTox® Antimycotic Drugs/EXTENDED) provided by Chromsystems (Chromsystems Instruments & Chemicals). Drug concentrations in VAMS and DPS samples were compared to those in fresh plasma using Passing-Bablok and Bland-Altman tests.

Results: Plasma concentrations of voriconazole, posaconazole, and isavuconazole were positively and significantly correlated with those obtained in VAMS and DPS samples (Spearman r range, 0.82-0.94, p < 0.001). Data were further analyzed using the Bland-Altman test, which showed a % mean difference compared to fresh plasma of -15.06-10.98 (range). The stability of both VAMS and DPS was ensured for at least 14 d at room temperature.

Conclusions: These results demonstrate that VAMS and DPS can be used for the TDM of anti-fungal agents. Owing to their stability, both sampling devices can be easily stored and shipped, without the need for refrigeration, to TDM laboratories that facilitate remote TDM applications. Finally, VAMS could be particularly suitable for pediatric and neonatal patients because they allow the collection of a few microliters of blood, thus improving ethical and compliance limitations.