Non-invasive evaluation of peripheral perfusion may be useful in many contexts including peri-operative monitoring. We validated a novel non-invasive spectroscopy technique to assess peripheral perfusion. This method, which is based on the measurement of tissue saturation variations after an ischemic period, was compared to strain gauge plethysmography and radionuclide plethysmography. The technique uses near-infrared spectroscopy (NIRS) to determine the rate of change of forearm tissue saturation during reactive hyperemia. In a prospective crossover study, 25 subjects were simultaneously evaluated with NIRS and strain gauge plethysmography. Six baseline flow measurements were performed to assess the reproducibility of each method. Twenty-seven serial measurements were then made to evaluate flow variation during forearm reactive hyperemia. Strain gauge and NIRS methods showed excellent reproducibility with intra-class correlation coefficients of 0.96 and 0.93, respectively. The NIRS technique appears well suited for the non-invasive evaluation of limb perfusion.