Endotracheal tube (ETT) suction is the most frequently performed invasive procedure in ventilated newborn infants and is associated with adverse effects related to negative tracheal pressure. We aimed to measure suction catheter gas flow and intratracheal pressure during ETT suction of a test lung and develop a mathematical model to predict tracheal pressure from catheter and ETT dimensions and applied pressure. Tracheal pressure and catheter flow were recorded during suction of ETT sizes 2.5-4.0 mm connected to a test lung with catheters 5-8 French Gauge and applied pressures of 80-200 mm Hg. The fraction of applied pressure transmitted to the trachea was calculated for each combination, and data fitted to three nonlinear models for analysis. Tracheal pressure was directly proportional to applied pressure (r = 0.82-0.99), and catheter flow fitted a turbulent flow model (R = 0.85-0.96). With each ETT, increasing catheter size resulted in greater catheter flow (p < 0.0001) and thus lower intratracheal pressure (p < 0.0001). The fraction of applied pressure transmitted to the trachea was accurately modeled using ETT and catheter dimensions (R = 0.98-0.99). Negative tracheal pressure during in vitro ETT suction is directly proportional to applied pressure. This relationship is determined by ETT and catheter dimensions.