Capillary dried blood microsampling is not suited for dihydropyrimidine dehydrogenase (DPD) phenotyping

Dihydropyrimidine dehydrogenase (DPD) phenotyping through uracil and dihydrouracil determination is a well-established approach to identify (partial) DPD deficiencies prior to fluoropyrimidine chemotherapy. However, preanalytical stability has challenged this test for years. This study therefore investigated whether dried blood spots (DBS) can improve preanalytical stability.

Uracil, dihydrouracil, and uridine were determined in 6 mm DBS sub-punch extracts by liquid chromatography-tandem mass spectrometry. Paired venous and capillary DBS were collected from 15 healthy volunteers across three days to evaluate venous-capillary DBS differences. The impact of blood spotting, drying and preanalytical stability for up to two weeks was assessed using venous DBS of the same volunteers.

Uracil was elevated in all capillary DBS, with a median of 219 % relative to venous DBS. In addition, the variation between capillary DBS replicates was 29 %, opposed to only 7 % in venous DBS. For dihydrouracil, a small bias of -7 % was observed, while uridine showed no difference, with similar inter-spot variation in venous and capillary DBS. Generation and drying of DBS had statistically significant yet minor effects on all analytes. Venous DBS enhanced preanalytical stability, yielding median uracil levels of 105 % and 107 % after 1 and 2 weeks at room temperature, and no differences for dihydrouracil or uridine relative to overnight dried DBS.

Capillary DBS are unsuitable for uracil determination in DPD phenotyping owing to poor agreement with venous DBS and substantial variability. Venous DBS, however, demonstrate superior preanalytical stability over liquid samples and may provide a practical solution for managing preanalytical variables.