Background: Evidence demonstrates that controlling pain in the preterm infants during the neonatal periods is improving physiological, behavioral and hormonal outcomes. Eye shield and massage may play an important role as a non-pharmacological pain management during venipuncture. Aim: Is to investigate effect of eye shield and massage on preterm infants' pain response during venipuncture. Patients and Methods:Time seriesquasi experimental research design was carried out on a randomized sample of 100 newborn infants attending the Neonatal Intensive Care Unit (NICU) of El Manial University Hospital (Kasr Al Ainy), (30control group,30 eye shield group and 30 massage group). Neonatal assessment tool and Preterm Infant Pain Profile (PIPP) were utilized for data collection. Results: There was a significant mean difference between control, eye shield and massage groups regarding PIPP pain assessment scores in three time frames (before venipuncture T 0 , during T 1 and after T 2) at P˂ 0.00 but there wasn't a significant mean differences after 5 min (T 3). Massage group had the fewest mean PIPP score during four time frames (3.50±0.97 before venipuncture, 8.16±1.91 during venipuncture, 4.30±4.30 after venipuncture and 3.22±0.81 after 5 minutes of venipuncture). Conclusion: ANOVA indicated that massage and eye shield groups showed a significant reduction in preterm infant's pain response than control group during venipuncture but massage was more effective than eye shield. Recommendations: Further studies needed to evaluate the effect of massage in combination with eye shield on preterm infant pain response.