The administration of vaccination, being an invasive procedure, can induce considerable pain in preterm infants. This study was conducted to evaluate the effect of grasp reflex stimulation on pain and crying time during hepatitis B vaccine administration in infants. This parallel randomized controlled trial was conducted between November 1, 2022, and April 1, 2023, at the Neonatal Intensive Care Unit of a public hospital in Turkey. The study included preterm infants whose parents provided both verbal and written consent. The control group (n =31) underwent a routine vaccination procedure, while the experimental group (n =28) underwent grasp reflex stimulation in addition to the routine vaccination procedure. Data were collected through the infant information form, stopwatch, and Premature Infant Pain Profile Scale-Revised (PIPP-R) form. The mean PIPP-R score during the procedure was 11.67±2.05 in the experimental group and 15.51±1.36 in the control group (P ≤.001). After the procedure, the mean PIPP-R score was 10.89±2.06 in the experimental group and 14.67±1.55 in the control group (P ≤.001). The mean duration of crying was 19.57±6.17 in the experimental group and 27.12±6.19 in the control group (P ≤.001). The mean PIPP-R and crying time of the control group were higher. Grasp reflex stimulation applied to preterm infants during vaccine administration decreased pain and crying time during and after the procedure. Grasp reflex stimulation can be applied as a care approach for procedures that may cause pain in infants and may help calm infants as part of pain management in nursing. The results of this study provide new information to the literature regarding the impact of grasp reflex stimulation on pain and the duration of crying induced by the hepatitis B vaccine in preterm infants. To our knowledge, this is the first study to confirm the effectiveness and feasibility of grasping reflex stimulation in hepatitis B vaccine administration in preterm infants. This study may contribute to future evidence-based studies. Grasping reflex stimulation can be tried as a parent-provided intervention in younger or older infants or in those who are critically ill.