Introduction: Preterm infants and sick neonates treated in neonatal intensive care units may undergo numerous pain- ful interventions. Despite rapidly growing knowledge about consequences of untreated pain, pain management of neonates is far from ideal. Objective: To determine the frequency and nature of painful procedures and corresponding analgesic therapies in neonates treated in a neonatal intensive care unit of a university teaching hospital in Hungary. Methods: A prospective observational study was performed between September and December 2019. We collected data of all painful procedures, pharmacological and non -pharmacological analgesic therapy performed on neonates during the first 14 days of hospitalization. For data collection, we used a questionnaire designed for this purpose, which was completed in real time by the medical staff. Results: 143 children were enrolled. 43 types of painful interventions were performed, a total of 13,314 times, of which 12,953 were the first, 361 multiple attempts. Each neonate was subjected to a mean of 93.1 interventions in the first 2 weeks of hospitalization, representing an average of 8.2 painful procedures per day per child. Pain relief was performed a total of 4190 times, in 31.5% of the interventions. Of this, 55.5% were continuous pharmacological, 40.7% non -pharmacological, 2.5% occasional drug, and 1.3% combination therapy. Ventilated neonates and preterm infants with shorter gestational age and lower birth weight had the most painful procedures.