Hepatic hepcidin-25 production is stimulated by systemic inflammation, and it interferes with the body's utilization of iron, leading to anemia. A 1-year prospective study was conducted to elucidate an association of serum hepcidin-25 concentration with mortality in anemic patients with non-Hodgkin lymphoma (NHL). Serum hepcidin-25 levels were measured in 50 NHL patients using liquid chromatography-tandem mass spectrometry. The patients were stratified into a high- and a low-hepcidin-25 group according to the median of serum hepcidin-25 concentrations. Factors associated with hemoglobin (Hb) were determined by multivariate regression analysis, incorporating serum hepcidin-25 and inflammatory markers including ferritin and interleukin-6 (IL-6) as covariates. The association between serum hepcidin-25 and mortality was analyzed using both the Kaplan-Meier method and a multivariate proportional hazards regression model. The median of serum hepcidin-25 concentrations was 49.8 (0.6-269) ng/mL, a level approximately nine times greater than the reference value for healthy individuals. Hb level was significantly lower in the high than in the low-hepcidin-25 group. Serum hepcidin-25 was extracted as the significant factor associated with Hb, but neither ferritin nor IL-6 was. The cumulative mortality was significantly greater in the high than in the low-hepcidin-25 group (56.0 vs. 24.0 %; P = 0.0222). The mortality risk for the presence of high hepcidin-25 was four times greater (hazard ratio [95 % confidence interval]: 3.66 [1.12-16.4]). In conclusion, serum hepcidin-25 levels are elevated in anemic NHL patients, and in this study, the group with higher hepcidin-25 levels manifested advanced anemia and poor survival.