Purpose: The aims of this study were to test the efficacy of vapocoolant spray to decrease the symptoms associated with pain in newborns undergoing heel stick and intramuscular injection and compare the pain relief effect of oral glucose. Methods: Randomized, controlled study including sixty newborns undergoing heel stick and intramuscular injection. Group 1 was heelsticked, Group 2 was intramuscular injected, Group A did not recieve any treatment, Group B recieved 30% glucose solution orally, Group C was applied vapocoolant spray symptoms and signs associated with pain at heel stick and intramuscular injection were measured with the premature Infant Pain Profile (PIPP) scale. Results: There was no significant difference in the PIPP score between intramuscular injected group control and heel stick group control (P=0.07). The mean PIPP score of Group 1A (control) 10.6+/-2.4, Group 1B 5.5+/-2.0, Group 1C 5.2+/-1.8. The mean PIPP score 1B and 1C were significantly lower than control (1B P<0.001, 1C P<0.001). The mean PIPP score of Group 2A (control) 12.5+/-1.4, Group 2B 7.0+/-1.7, Group 2C 6.4+/-1.6. The mean PIPP score 2B and 2C were significantly lower than control (2B P<0.001, 2C P<0.001). Conclusion: The antinociceptive effect of vapocoolant sparay is as effective as 30% oral glucose solution for pain control. So this study support the use of vapocoolant spray for reducing pain during painful procedure in the neonatal intensive care units.