Hypoxic physiological states may occur during anesthetic events of snakes but accurate monitoring of oxygenation is challenging. Oxygenation levels of nine Louisiana pine snakes (Pituophis ruthveni) were assessed using transcutaneous regional oxygen saturation (rSO₂) at the level of the liver (rSO₂^Liver) and at the halfway point of the body (rSO₂^Half ). Reflectance pulse oximetry measured SpO₂, with a sensor overlying the heart. Values were compared with the venous partial pressure of oxygen (PvO₂). Measurements were taken during four phases, simulating an anesthetic event: phase 1, breathing room air; phase 2, while supplied with supplemental oxygen via face mask; phase 3, during ketamine and dexmedetomidine sedation; and phase 4, after receiving atipamezole. There were no significant changes in any oxygenation parameters between concurrent phases, but respiratory rate significantly decreased (P = 0.02) between phases 1 and 2. Strong positive associations were found between both rSO₂^Liver and rSO₂^Half when compared with PvO₂ irrespective of phase (r = 0.72, r = 0.63 respectively), but not with SpO₂ (r = 0.3). Strength of correlation varied with each phase but was uniformly strongest for rSO₂^Liver. The measurement of rSO₂ appears superior compared with traditional pulse oximetry for assessing oxygenation levels of snakes.