Abstract: Objective: To analyze the effect of white noise interference method in relieving puncture pain of premature infants. Methods: Totally 78 premature infants were recruited in this study, 38 infants in experimental group and 40 infants in control group. White noise interference method was used in experimental group, and posture comfort method was implemented in control group. The Premature Infant Pain Profile (PIPP) and cry incidence after puncture were used to evaluate the effect. Results: There were significant differences in heart rate, oxygen saturation and PIPP scores between experimental group and control group (P<0.05). The cry incidence was higher in control group than that in experimental group (80% vs 65.8%, P<0.05). Conclusion: The white noise interference method could be useful in relieving puncture pain of premature infants.