Heart valve calcification is an important predictor for all-cause and cardiovascular mortality in hemodialysis patients. Recently, serum β2-microglobulin has been associated with cardiovascular disease in the non-hemodialysis population, but the relationship between serum β2-microglobulin and valve calcification remains unknown. In this cross-sectional study, we recorded the patients' clinical parameters, including serum β2-microglobulin, and related these parameters to the number of calcified valves detected by echocardiography. The patients included 80 males and 35 females (age 67 ± 10 years; duration on hemodialysis 96 ± 67 months). Calcification of the aortic and mitral valves was observed in 89 (77.4%) and 59 patients (51.3%), respectively. Fifty-one patients (44.3%) showed calcification of both valves. In univariate analysis, age (r = 0.301, P = 0.001), serum albumin (r = −0.219, P = 0.01), calcium (r = 0.205, P = 0.02), high sensitivity C-reactive protein (r = 0.209, P = 0.02), and β2-microglobulin (r = 0.206, P = 0.02) significantly correlated with the number of calcified valves. Stepwise multiple regression analysis showed that age (β = 0.389, P < 0.001) and calcium (β = 0.223, P = 0.01) were independent determinants for valve calcification (r2 = 0.195). In addition, carotid intima media thickness was significantly higher in patients with valve calcification compared with those without valve calcification. Our results suggested the impacts of calcium metabolism and malnutrition-inflammation complex syndrome on valve calcification. In addition, serum β2-microglobulin may be another potential marker of cardiovascular complications in patients on hemodialysis.