The study aimed to assess the effect of a single bout of incremental exercise on irisin and BDNF plasma concentrations as related to erythrocyte purine nucleotides concentration at rest and after exercise. Master endurance master athletes (training experience 38 ± 6 years) and a group of untrained participants completed a single bout of progressive incremental exercise test until exhaustion. The dual-energy x-ray absorptiometry and blood collection were performed. Blood was taken twice at rest and 10 min after exercise. Concentrations of ATP, ADP, and AMP were assessed in the erythrocytes. Hypoxanthine and uric acid were determined in plasma using the high-performance liquid chromatography. Plasma concentrations of irisin and BDNF were assessed through the immunoenzymatic method. The ATP level, ATP/ADP ratio and AEC value were significantly higher in the athletic group. A significantly higher concentration of BDNF was it also noted in the trained group that correlated with the erythrocyte energy status at rest. The single session of exercise induced a significant increase in ATP erythrocyte levels in both groups. Both exerkines significantly correlated at rest with red blood cell adenine nucleotides and degradation products (BDNF positively and irisin negatively). The blood concentration of BDNF and irisin, in response to exercise, was not significantly different between groups. Obtained data revealed a higher erythrocyte energy status and lower purine degradation products concentration in master athletes. Also resting plasma exerkines differed substantially between groups. In conclusion, longterm training resulted in exercise adaptation reflected by a higher erythrocyte energy status, lower purine degradation products concentration and modified concentration of