BACKGROUND: Most portable bi-level positive airway pressure devices are not equipped with air-oxygen blenders for precisely regulating oxygen concentrations, and supplemental oxygen must be added to increase the F IO 2 . Very few studies have investigated the factors that affect F IO 2 , and their conclusions have been inconsistent. We investigated in vitro noninvasive ventilation (NIV) parameters and their effects on F IO 2 , particularly the effect of the oxygen injection site. METHODS: NIV was simulated with a test lung and manikin setup. F IO 2 was measured with 4 oxygen injection sites (mask, in front of the exhalation valve, at the humidifier outlet, and proximal to the ventilator), with 3 exhalation valve types, with 2 oxygen flows, and with 4 combinations of inspiratory and expiratory pressure. RESULTS: Oxygen flow, inspiratory and expiratory pressure, and exhalation valve type all affected F IO 2 . For a given oxygen flow, the oxygen injection site was the most important factor that affected F IO 2 . The oxygen injection site that was closest to the patient (on the mask) had the higher F IO 2 (P < .001). CONCLUSIONS: The oxygen injection site had the greatest effect on F IO 2 during NIV.