Purpose This study compared the pain-relieving effects of human milk, sucrose, and distilled water during examinations for retinopathy of prematurity. Methods Forty-five preterm infants were randomly assigned to receive a pacifier dipped in one of three solutions: human milk (n=14), 24% sucrose (n=15), or distilled water (n=16), 2 minutes before an eye examination. Their pain score, pulse rate, and oxygen saturation were measured at three time points: 5 minutes before the examination, 30 seconds after speculum introduction, and 2 minutes after the examination. Results The infants’ mean gestational age and weight at birth were 33.1±2.1 weeks and 1,842±470 g, respectively. There were no between-group differences in pain relief during the eye examination. The pain score significantly increased both during (p