Two experiments were conducted to assess the endogenous fraction of purine derivative (PD) excretion, urinary recovery, and intestinal digestibility of purines in Nellore heifers. For both experiments, 8 Nellore heifers fitted with ruminal and abomasal cannulas were allocated to two 4 × 4 Latin squares. The diets were based on corn silage and concentrate (60 and 40% DM basis, respectively); feces and urine samples were obtained by total collection, and abomasal DM flow was estimated using indigestible NDF as an internal marker. In Exp. I, 4 of the 8 heifers (BW 258 ± 20 kg) were also fitted with ileal cannula. The planned treatments were 4 different DMI: 1.2, 1.6, 2.0, and 2.4% of BW (DM basis). The endogenous losses and purine recovery as urinary PD were estimated using linear regression between daily urinary PD excretion (Y) and daily abomasal flow of purine bases (X), expressed in millimoles per kilogram of BW(0.75). In Exp. II, the same 8 Nellore heifers (BW of 296 ± 15 kg) were fed at 1.37% BW (DM basis). The treatments were the infusion of purines (RNA from torula yeast, type VI, Sigma) into the abomasum in increasing amounts (0, 33, 66, and 100 mmol/d). All statistical analyses were performed using the PROC MIXED procedure in SAS. In Exp. I, the DMI range was 1.16 to 1.84% of BW and did not affect (P > 0.05) the apparent RNA digestibility in the small intestine, which had a mean of 75.6%, and a true digestibility of 93.0%. The mean ratio of the N-RNA to the total-N in the ruminal bacteria was 0.137. The daily urinary PD excretion (Y, mmol/kg of BW(0.75)) was a function of RNA flow in the abomasum (X, mmol/kg of BW(0.75)): Y = 0.860X + 0.460, where 0.860 and 0.460 were the PD recovery of purines and the endogenous fraction (in mmol/kg of BW(0.75)), respectively. In Exp. II, the daily urinary PD excretion was a function of RNA flow in the abomasum: Y = 0.741X + 0.301, where 0.741 and 0.301 were the recovery of PD in urine of infused purines and the endogenous losses (in mmol/kg of BW(0.75)), respectively. In conclusion, our data suggest that in Nellore heifers the respective values of endogenous PD excretion (mmol/kg of BW(0.75)), urinary recovery of the purines absorbed in the abomasum, and true digestibility of RNA in the small intestine were 0.30, 0.80, and 0.93.