Although hepcidin, a recently discovered peptide hormone, is considered a major regulator of iron metabolism and the anemia of chronic inflammation, its role in the anemia of pregnancy has not been characterized. Our objective was to characterize the role of hepcidin in the anemia of pregnancy. We examined the relationships between urinary hepcidin, iron status indicators, hemoglobin, erythropoietin, alpha-1 acid glycoprotein, and C-reactive protein in a cross-sectional study conducted among 149 pregnant rural Bangladeshi women in biospecimens obtained during home visits. Urinary hepcidin was measured using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Urinary hepcidin, as log (intensity per mmol/L creatinine), was correlated with log ferritin (r= 0.33, p< 0.001), the transferrin receptor index (r=− 0.22, p= 0.007), and log alpha-1 acid glycoprotein (r= 0.20, p= 0.01), but not hemoglobin (r= 0.07, p= 0.40), log transferrin receptor (r=− 0.07, p= 0.41), log erythropoietin (r=− 0.01, p= 0.88) or log C-reactive protein (r= 0.06, p= 0.48). The strength of the relationship between hepcidin and ferritin was maintained in a multiple linear regression analyses after enhancing the sample with women selected for low iron stores (n= 41). Among pregnant women in a community-based study in rural Bangladesh, urinary hepcidin levels were related to iron status and AGP but not hemoglobin, erythropoietin, or C-reactive protein.