Objectives <p>To determine the effect of prone positioning on the duration of respiratory support in neonates receiving non-invasive mode of ventilation. </p> Methods <p>This open label randomized controlled trial was conducted in the NICU of a tertiary care hospital. Preterm neonates who fulfilled the eligibility criteria for enrolment were included within 6&#xa0;h of starting non-invasive respiratory support. Neonates in intervention group were given respiratory support in the prone position. After every 90&#xa0;min of prone positioning, the position was changed to side lying/supine position for 30&#xa0;min in the intervention group. The comparator group were given respiratory support in the supine position with 2 hourly changes in position (supine-side lying-prone). The primary outcome was duration of non-invasive ventilation.</p> Results <p>Duration of non-invasive ventilation did not reduce in the intervention group. {[h; 80 (36 to 127) vs. 93.5 (42 to 142)], median difference (h) -8, 95% CI: -30 to 13, <i>p</i> = 0.47} Neonates in the prone group had significantly lower peak FiO₂ requirements [30 (30–30) vs. 35 (30–50), <i>p</i> = 0.01] and frequency of apnea [episodes per day; 0.4 (0.2–1) vs. 1 (0.4–1.6), <i>p</i> = 0.04].</p> Conclusions <p>Prone positioning did not reduce the duration of non-invasive respiratory support in preterm neonates.</p> Clinical Trial Registration: <p>CTRI/2021/10/037183</p>