Alterations to cerebral blood flow (CBF) have been implicated in diverse neurological conditions. Near-infrared spectroscopy (NIRS)-measured regional cerebral tissue oxygen saturation ([Formula: see text]) provides an estimate of oxygenation of interrogated cerebral volume useful in identifying variations in oxygen supply to cerebral tissue and in monitoring cerebrovascular function. [Formula: see text]-inhalation-based hypercapnic breathing challenges were used to simulate CBF dysregulation, utilizing NIRS to monitor the CBF autoregulatory response. A breathing circuit was designed to administer [Formula: see text]-compressed air mixtures and assess CBF regulatory responses to hypercapnia in 26 healthy young adults. One to three hypercapnic challenges of 5 or 10 min duration were delivered to each subject while continuously monitoring [Formula: see text], partial pressure of end tidal [Formula: see text] ([Formula: see text]), and vital signs. Change in [Formula: see text] ([Formula: see text]) during [Formula: see text] inhalation positively correlated to [Formula: see text] ([Formula: see text]). Grouping subjects into three exercise factor levels (h/week), (1) 0, (2) [Formula: see text] and [Formula: see text], and (3) [Formula: see text] showed significantly greater [Formula: see text] responses to [Formula: see text] challenges for level 3 subjects but similar [Formula: see text] responses for the three groups. Exercising greater than 10 h/week may produce a higher resting cerebrovascular reactivity (CVR) to [Formula: see text] inhalation. Establishing baseline values of [Formula: see text] and CVR to [Formula: see text] may aid in early detection of CBF changes.