Spontaneous and aspirated blood samples were collected from the welling points of surface efferent arterioles in the rat kidney. The amount of tubular reabsorbate in each sample was calculated from measurements of the inulin concentration in the sample and systemic arterial plasma. Both spontaneous and aspirated samples contained substantial amounts of tubular reabsorbate (4% +/- 1% and 11% +/- 1%, respectively). Accordingly, the single nephron filtration fraction values derived from measurements of albumin concentrations alone were significantly lower than those calculated considering the addition of the amount of tubular reabsorbate in each sample (0.26 +/- 0.01 vs. 0.30 +/- 0.01 and 0.25 +/- 0.01 vs. 0.34 +/- 0.01 for the spontaneous and aspirated samples, respectively). Increasing the rate of blood collection from the welling point by aspiration was associated with a significant decrease in the hydraulic pressure in the same vessel. The efferent arteriolar blood samples were therefore probably contaminated with tubular reabsorbate as a result of retrograde flow from the peritubular capillaries. It was confirmed that single nephron filtration fractions derived from hematocrit measurements were significantly higher than those based on albumin concentration measurements, supporting that the hematocrit of afferent arteriolar blood of the superficial nephrons is higher than that of systemic arterial blood.