Abstract Aim: Off-label drug use in neonates remains a global problem. The retrospective, multicenter study was to study the rates and reasons of off-label drug use among representative NICUs in Henan, China. Methods: Prescriptions were randomized sampled from eight NICUs. Patient characteristics and diseases were collected. Classifications of off-label drug use including age, frequency, dose, indication and route of administration were analyzed. Results: 1770 neonates sampled from eight NICUs were evaluable in the study, with a mean body weight 2.98 kg (SD = 0.80, range 0.78-6.0), 92.03% of whom were exposed to ≥ 1 off-label prescription. 5813 prescriptions with 113 drugs were registered, 84.62% of which were off label. 63.03% of the off-label use was in relation to age. Conclusion: Off-label drug use among neonates remains fairly prevalent in Henan, China. Initiative and effective implementation is urgently required to improve this situation.