Symmetric dimethylarginine (SDMA) is an early marker for renal lesions in mammals. The objectives of this study were 1) to establish a reference interval of SDMA in healthy brook trout, Salvelinus fontinalis (Mitchill, 1814), and 2) to assess its sensibility as a marker of renal pathology. Plasma SDMA was quantified by liquid chromatography–mass spectrometry in 25 adult brook trout, including 20 fish displaying no renal histologic lesions, and five fish displaying chronic nonactive microscopic granulomas. The fish size (P = 0.30) and weight (P = 0.12) were not statistically different among groups, nor were SDMA values (P = 0.22). However, brook trout without microscopic renal lesions tended to have lower SDMA values (no lesions: mean = 24.9 µg/dL; lesions: mean = 31.4 µg/dL). The reference interval (90% confidence interval [90% CI]) for SDMA concentration in brook trout ranged between 10.0 µg/dL (90% CI: 5.4–14.7) and 39.8 µg/dL (90% CI: 34.8–43.9). These values were higher than those previously reported in other vertebrate species. Further research is needed to evaluate the use of SDMA as a marker of renal function in fishes.