We investigated the prevalence of exercise-associated hyponatremia (EAH) in 145 male ultra-marathoners at the '100-km ultra-run' in Biel, Switzerland. Changes in body mass, urinary specific gravity, haemoglobin, haematocrit, plasma [Na(+)], and plasma volume were determined. Seven runners (4.8%) developed asymptomatic EAH. Body mass, haematocrit and haemoglobin decreased, plasma [Na(+)] remained unchanged and plasma volume increased. Δ body mass correlated with both post race plasma [Na(+)] and Δ plasma [Na(+)]. Δ plasma volume was associated with post race plasma [Na(+)]. The athletes consumed 0.65 (0.30) L/h; fluid intake correlated significantly and negatively (r = -0.50, p < 0.0001) to race time. Fluid intake was neither associated with post race plasma [Na(+)] nor with Δ plasma [Na(+)], but was related to Δ body mass. To conclude, the prevalence of EAH was low at ~5% in these male 100 km ultra-marathoners. EAH was asymptomatic and would not have been detected without the measurement of plasma [Na(+)].