BACKGROUND: During O 2 therapy at low flow in patients who breathe spontaneously, the fraction of delivered O 2 (F DO 2 ) is unknown. In recent years, F DO 2 prediction formulas have been proposed. However, they do not take into account the effect of inspiratory flow (V I ) on the F DO 2 . The aim of this study was to validate a new F DO 2 prediction formula, which takes into account the V I and compares it with other F DO 2 prediction formulas. METHODS: During a bench study, spontaneous breathing was generated with a mechanical test lung connected to a mechanical ventilator set to volume control mode. O 2 flow from a wall-mounted tube was delivered through a heat-and-moisture exchanger filter. A flow sensor recorded each breath of the V I in ambient temperature and barometric pressure conditions. Three parameters [O 2 flow at 2, 3, 4, 5, 6 L/min; minute ventilation at 5, 10, 15, 20 L/min; and ratio of the inspiratory time (T I ) to the total breathing cycle time (T tot ) (T I /T tot ) of 0.33 (T I /T tot value) and 0.50 (T I /T tot value)] were modified to generate many ventilatory patterns. An O 2 analyzer continuously examined the F DO 2 . RESULTS: When the O 2 flow and/or T I /T tot increased, the F DO 2 increased. When the minute ventilation increased, the F DO 2 decreased. The results of the Bland-Altman method for the F DO 2 , calculated by using our mathematical model and the measured F DO 2 , showed that the mean ؎ SD bias value was equal to 1.49 ؎ 0.84%, and the limits of agreement ranged from ؊0.17% to 3.14%. The intraclass correlation coefficients were 0.991 for T I /T tot ؍ 0.33 and 0.994 for T I /T tot ؍ 0.50, and the coefficient of variation was 2.1% for T I /T tot ؍ 0.33 and 1.3% for T I /T tot ؍ 0.50. The results of the Bland-Altman method for the F DO 2 calculated by using the Shapiro formula and the F DO 2 measured on the bench indicated that the bias value was 0.075 ؎ 8.66% and the limits of agreement ranged from ؊16.89% to 17.04%. For the Vincent formula, the bias value was 3.08% ؎ 8.56% and the limits of agreement ranged from ؊13.69% to 19.84%. CONCLUSIONS: The V I has a major impact on F DO 2 during O 2 therapy at low flow. F DO 2 comparisons between frequently used prediction formulas and F DO 2 measured on the bench indicated greater differences. Uncritical use of these formulas should be used cautiously to predict F DO 2 . In this study, our prediction formula indicated a good accuracy for predicting F DO 2 during supplemental oxygenation through a heat-and-moisture exchanger in patients who breathe spontaneously.