Abstract In this study, creatinine-based equations to evaluate glomerular filtration rate (eGFR) were proposed to more accurately assess kidney function, and cystatin C, a parameter not dependent on muscular mass, was introduced to improve GFR calculation in professional cyclists during a long-lasting race. Nine cyclists participating in the 2011 Giro d'Italia were recruited. Blood and anthropometrical data were collected the day before (T-1) the race, on the 12th day (T 12) and on the 22nd day (T 22) of the race. Haemoglobin and haematocrit were registered. Haemodilution was observed at T 12, whilst stabilization was evident at T 22. Creatinine, cystatin C concentrations and eGFR values were not modified during the observed period; only GFR evaluated with the Cockcroft-Gault (CG) formula and expressed as ml/min/1.73 m(2) significantly decreased (p < 0.05) at T 22 in comparison with T-1, probably as a consequence of weight decrease. Cystatin C levels were in the reference range, while creatinine concentrations were lower. The lowest eGFR values were observed with CG normalized and the Modification of Diet in Renal Disease (MDRD) formulas. A good correlation was observed between the MDRD and the Chronic Kidney Disease-Epidemiology Collaboration (CKD-EPI) equations and between CG normalized and both CKD-EPI and MDRD formulas. The worst correlation was registered between CKD-EPI creatinine and cystatin C and all the other equations. In conclusion, adaptive mechanisms of renal function allow athletes to maintain stable creatinine, cystatin C and eGFR values during a long-lasting race. The use of GFR equations to evaluate general health status of sportsmen should be recommended with caution, considering also weight modification during competition.