Purpose: Newborns routinely experience pain associated with invasive procedures such as blood sampling, venipuncture, heelstick, or venous cannulation. This study was done to provide data for a nursing intervention to alleviate newborn pain clinically by investigating the effect of local anesthetic cream during venipuncture. Methods: Participants were 70 newborns hospitalized in the nursery. Informed Consent was obtained from parents of the newborns. Venipuncture for regular blood sampling was carried out for a test on 2 groups; the experimental, placebo group. The neonatal infant pain scale (NIPS), and duration of crying were measured to assess pain reaction. All neonatal behaviors were recorded on videotape. Results: There were significant differences in pain behavior during venipuncture (t=-4.752, p<.001), immediately after sampling (t=-5.591, p<.001), 3 minutes after puncture (t=-2.469, p=.017), and in duration of crying (t=-3.005, p=.004). Conclusion: Results show that local administration of EMLA cream before venipuncture causes a reduction in neonatal pain response, indicating that the EMLA cream has the effect of pain relief.