Background: Myocardial work (MCW), estimated using non-invasive pressure-strain loops, is a novel approach for assessing left ventricular function. MCW offers potential advantages over left ventricular ejection fraction and global longitudinal strain (GLS). This study aimed to establish a reference range for MCW in newborns. Methods and Results:Overall, 113 healthy newborns (mean [±SD] birth weight 3,120±378 g) were included in the study. After entering peripheral systolic and diastolic blood pressure (BP) values, global constructive work (GCW), global work index (GWI), global work efficiency (GWE), and global wasted work (GWW) were calculated using EchoPAC software (version 204). Considering a mean [±SD] GLS of −16.3±2.8%, the mean [±SD] GWI, GCW, GWW, and GWE was 698.7±147.9 mmHg%, 1,008.5±200.1 mmHg%, 58.1±28.1 mmHg%, and 93.1±2.9%, respectively. All MCW parameters in newborns were lower than those previously reported in children and adults. However, GWI and GCW were more closely correlated with BP and GLS, as in adulthood and children. Estimation of the correlation between MCW and the frame rate index revealed no significant correlations among MCW parameters.Conclusions: Cardiac function in newborns was evaluated by assessing MCW. With the establishment of reference ranges and normative MCW data for newborns, routine clinical use and rotational mechanics are likely to become increasingly common. Future studies are needed to determine whether MCW is useful in screening for cardiac illness among newborns.