The aim of this study was to examine the impacts on blood lactate concentration, measured heart rate and assessment of perceived exertion during split sessions of equal relative load, as also their relationship to the specific sport practised. Nineteen regional-level athletes (nine middle and long-distance runners (cyclic group) and ten field-sport team players (acyclic group)) performed four high-intensity interval training (HIIT) sessions with work-interval durations of 10 s, 50 s, 90 s and 130 s. The sessions were carried out at their usual training sites with a separation of at least 48 hours. Blood lactate concentration was measured at rest and 3 min after the completion of each protocol. Heart rate was measured continuously during all sessions with a sampling rate of 1 s, and rating of perceived exertion (RPE) was requested at the end of the trial. The results showed an increase in blood lactate concentration, peak heart rate and rating of perceived exertion during long protocols as compared with short ones. No differences were observed in dependent variables between cyclic and acyclic groups. Significant but moderate correlations were observed between post-exercise blood lactate concentration, peak heart rate and RPE.