Newborns are exposed to pain during heel blood collection. The experience of pain in newborns can lead to both physiological and behavioral changes. The aim of this study was examine the effect of music (classical music and lullaby) on pain during the heel blood collection in premature infants. This single blind randomized controlled study was conducted between October, 2022 and April, 2023 in the Neonatal Intensive Care Unit at a university hospital. A total of 60 premature infants were randomly allocated to three groups: the Control Group, the Classical Music Group, and the Lullaby Group. Pain, oxygenation saturation levels, heart rate and respiratory rate were measured before, during, after the heel blood collection procedure. There were no statistically significant differences in the demographic data of the infants in the classical music, lullaby, and control groups. After heel blood collection procedure, pain levels, heart rate and respiratory rate of premature infants in the lullaby groups measurement were lower than the classical and control group. No significant difference was found in terms of oxygen saturation between the groups. In premature infants, in the lullaby group listening to during heel blood collection was more effective in reducing pain compared to the control group and classical music. This indicates lullaby can potentially be an effective method in alleviating pain during heel blood collection.