We tested the hypothesis that the respiratory compensation point can be accurately determined in healthy participants during incremental cycling exercise using non-invasive near-infrared spectroscopy-derived measures of deoxygenated hemoglobin (deoxyHb).Validation study.118 healthy men (average age 47±19 yrs, range 20–79 yrs) performed an incremental cycling test to exhaustion. Breath-by-breath pulmonary oxygen uptake (V˙O2) and other ventilatory and gas exchange variables were measured and used to determine respiratory compensation point. Vastus lateralis deoxyHb was monitored using a frequency domain multi-distance system near-infrared spectroscopy device and deoxyHb data were modeled with a piece-wise double-linear function from which the deoxyHb deflection point (deoxyHbDP) was determined. The absolute (Lmin−1) and relative (% maximal V˙O2 [V˙O2max]) V˙O2 values associated with the respiratory compensation point and deoxyHbDP were determined for each individual.DeoxyHb increased as a function of exercise intensity up to a point (deoxyHbDP) after which the signal displayed a “near-plateau”. The deoxyHbDP corresponded to a V˙O2 of 2.25±0.69Lmin−1 (74±12% V˙O2max) which was not significantly different from the V˙O2 at respiratory compensation point (2.28±0.70Lmin−1 and 74±10% V˙O2max, p<0.05). Both indices were highly correlated (r2=0.86) and Bland Altman analyses confirmed a non-significant bias for V˙O2 (−0.024Lmin−1) concomitant with a small imprecision of 0.26Lmin−1.During incremental cycling exercise, the V˙O2 associated with the onset of a plateau in near-infrared spectroscopy-derived deoxyHb occurs in coincidence with the V˙O2 at respiratory compensation point suggesting that respiratory compensation point can be accurately estimated, non-invasively, using near-infrared spectroscopy-derived deoxyHb in alternative to the use of ventilatory-based techniques.