Elution of normal and pathological human red blood cells (RBCs) was performed by gravitational field-flow fractionation (GFFF). The reproducibility of the retention factor was lower than 10% and elution at high and low flow-rates confirmed the existence of "lifting forces". No direct correlation between size and retention was observed for normal RBCs in the absence of density information. Elution of pathological human RBCs, known to be modified in shape, density and rigidity, was performed. The elution parameters confirmed that the retention mechanism of RBCs is at least density dependent but that other factors can be involved, such as shape or deformity. Moreover, peak profile description parameters (standard deviation and asymmetry) can be qualitatively related to some biophysical parameters. Numerous elution characteristics can be linked to cell properties described in the literature and although GFFF appeared to have limited capabilities in terms of size analysis it appeared to be a versatile tool for studying cell biophysical characteristics.