This study investigated changes in muscle oxidative metabolism and microvascular responsiveness induced by glucose ingestion in the upper and lower limbs using near-infrared spectroscopy (NIRS). Fourteen individuals (aged 27 ± 1.4 years) underwent 5 vascular occlusion tests (VOT) (pre-intervention (Pre), 30 min, 60 min, 90 min, and 120 min after glucose challenge). NIRS-derived oxygen saturation (StO₂) was measured on the forearm and leg muscle at each VOT. Muscle oxidative metabolism was determined by the StO₂downslope during cuff inflation (deoxygenation slope); microvascular responsiveness was estimated by the StO₂upslope (reperfusion slope) following cuff deflation. There was a significant increase in arm (p < 0.05; 1-β = 0.860) and leg (p < 0.05; 1-β = 1.000) oxidative metabolism activity as represented by the faster deoxygenation slope at 60, 90, and 120 min (0.08 ± 0.03, 0.08 ± 0.03, 0.08 ± 0.02%·s^–1, respectively) (leg) and at 90 min (0.16 ± 0.08%·s⁻¹) (arm) observed after glucose ingestion when compared with their respective Pre values (leg = 0.06 ± 0.02; arm = 0.11 ± 0.04%·s⁻¹). There was a significant increase in arm (p < 0.05; 1-β = 0.880) and leg (p < 0.05; 1-β = 0.983) reperfusion slope at 60 min (arm = 3.63 ± 2.1%·s⁻¹; leg = 1.56 ± 0.6%·s⁻¹), 90 min (arm = 3.91 ± 2.1%·s⁻¹; leg = 1.60 ± 0.6%·s⁻¹), and 120 min (arm = 3.91 ± 1.6%·s⁻¹; leg = 1.54 ± 0.6%·s⁻¹) when compared with their Pre values (arm = 2.79 ± 1.7%·s⁻¹; leg = 1.26 ± 0.5%·s⁻¹). Our findings showed that NIRS–VOT technique is capable of detecting postprandial changes in muscle oxidative metabolism activity and microvascular reactivity in the upper and lower limb.Novelty NIRS-VOT is a promising noninvasive clinical approach that may help in the early, limb-specific detection of impairments in glucose oxidation and microvascular function.