<b><i>Background:</i></b> Hepcidin, an iron-regulatory hormone, plays a key role in preventing iron overload. Few studies have investigated the regulation of hepcidin in low-birth-weight (LBW) infants who are vulnerable to iron imbalance. <b><i>Objectives:</i></b> To identify perinatal factors associated with serum hepcidin levels in LBW infants. <b><i>Methods:</i></b> Ninety-two LBW infants with a median gestational age (GA) of 32.6 weeks and birth weight of 1,587 g were prospectively enrolled. Serum hepcidin-25 (Hep25) levels were measured from umbilical cord blood using liquid chromatography-tandem mass spectrometry. The relationship between Hep25 levels and prematurity or other possible hepcidin-regulatory factors was evaluated. <b><i>Results:</i></b> The median Hep25 level was 7.3 ng/mL (interquartile range: 2.85-16.38). log(Hep25) correlated with birth weight (<i>r</i> = 0.229, <i>p</i> = 0.028), log(interleukin-6 [IL-6]) (<i>r</i> = 0.408, <i>p</i> < 0.001), log(erythropoietin) (<i>r</i> = -0.302, <i>p</i> = 0.004), transferrin saturation (<i>r</i> = 0.29, <i>p</i> = 0.005), soluble transferrin receptor (<i>r</i> = -0.500, <i>p</i> < 0.001), and log(ferritin) (<i>r</i> = 0.696, <i>p</i> < 0.001). Serum iron and hemoglobin levels did not correlate with log(Hep25). Hep25 levels were higher among infants with chorioamnionitis and infants born vaginally and lower among infants born to mothers with pregnancy-induced hypertension than among infants without the respective characteristics. Stepwise multiple linear regression analysis confirmed the significant association of log(Hep25) with GA, log(IL-6), log(erythropoietin), and soluble transferrin receptor. <b><i>Conclusions:</i></b> Among LBW infants, GA, IL-6, erythropoietin, and soluble transferrin receptor were associated with Hep25 levels. Therefore, prematurity, inflammation, hypoxia, and erythropoietic activity may be important perinatal factors that affect hepcidin levels.