The Q-LAC analyzer, a portable device employing near-infrared spectroscopy (NIRS), was designed to measure muscle oxygen saturation (SmO2) during physical exercise. This study aimed to assess the reliability of the Q-LAC analyzer in determining SmO2 levels during incremental cycling exercise. Thirteen physically active males (age 28.1 ± 5.3 y; height 181.2 ± 5.7 cm; body mass 79.9 ± 11.1 kg; BMI 24.2 ± 2.4 kg/m2) participated in this study. A submaximal incremental exercise test (SIET) on an electromagnetic cycle ergometer with a seven-day interval was performed twice. SmO2 levels in the vastus lateralis (VL) muscle of the dominant leg were simultaneously recorded using the Q-LAC device during both tests. The study calculated the intraclass correlation coefficient (ICC), the coefficient of variation (CV), the standard error of measurement (SEM), the smallest worthwhile change (SWC), and mean detectable change (MDC). A within-within-subjects ANOVA revealed no statistically significant effects for session (F1,12 = 0.97, p = 0.34, η2 = 0.07) and the interaction between session and workload (F4,48 = 0.19, p = 0.94, η2 = 0.02). ICC values ranged from 0.72 to 0.91. Furthermore, the analysis of CV, SEM, and SWC indicated that SmO2 measurements obtained with the Q-LAC device exhibit good reliability but marginal sensitivity. In conclusion, the portable Q-LAC analyzer provides consistent measurements of muscle oxygen saturation during low-to-moderate-intensity exercise on a cycle ergometer.