Hematocrit, blood volume, and surface area of dried blood spots – a quantitative model

The use of the dried blood spot (DBS) sampling technique has extended the scope of clinical research, particularly in children. The effects of different hematocrit levels (25–55%) and different blood volumes (7.5–30 μL) on the surface area of the blood spots were investigated using ImageJ® software. Variation in hematocrit levels between patients and inaccuracies in blood volumes applied to Guthrie cards can have a marked effect on analyte concentrations measured in DBS samples. The current study presents a validated model that links blood volume and hematocrit to the surface area of the blood spot. The final model showed that both factors affect the blood spot surface area, however, the positive effect of blood volume is higher than the negative effect of hematocrit. The measurement of surface area could be added as an additional quality control step in clinical studies that have adopted fixed volume DBS sampling for the quantification of the analytes. This approach can be used in estimating the hematocrit if this is not known for a patient or estimating the volume in spots that are visually different in size from the norm, i.e. technical error. The current study presents a validated model that links blood volume and HCT to the surface area of DBS. The final model showed that both factors affect the blood spot surface area, however, the positive effect of blood volume is higher than the negative effect of HCT. This approach can be used in estimating HCT if this is not known for a patient or estimating volume in spots which are visually different in size from the norm.