Objective: Serum magnesium (Mg2+) levels are associated with insulin resistance, hypertension, lipid abnormalities, and inflammation. However, limited studies have indicated the relationship between Mg2+ and multiple system indexes. The purpose of this study was to investigate the association between Mg2+ and allostatic load (AL) in hemodialysis patients.Methods: A cross-sectional survey was conducted on hemodialysis patients from different centers in Anhui Province, China, between January and December 2020. A total of 3,025 hemodialysis patients were recruited. Their clinical data were measured before hemodialysis. Information was collected by an online self-reported questionnaire and medical record. Serum Mg2+ was divided into three groups by tertiles. A score of AL greater than or equal to 3 was defined as high AL. A binary logistic regression model was applied to examine the relationship between serum Mg2+ and AL.Results: A total of 1,222 patients undergoing hemodialysis were included, 60% of whom were males (733/1,222). The mean (standard deviation) age of patients was 55.90 (12.75). The median level of serum Mg2+ was 1.22 mmol/L. The rate of high AL levels was 23.4%. Serum Mg2+ was negatively correlated with body mass index, fasting blood glucose (Glu), and C-reactive protein and positively correlated with high-density lipoprotein, low-density lipoprotein, total cholesterol, diastolic blood pressure (DBP), and serum phosphorus. After adjusting for gender, anxiety, diabetes, family residence, lipid-lowering agents, antihypertensive medications, albumin, and Glu, the binary logistic regression model showed that patients with lower levels of serum Mg2+ were more likely have high AL (OR for the T1 group of serum Mg2+:1.945, 95% CI: 1.365–2.773, and OR for the T2 group of serum Mg2+:1.556, 95% CI: 1.099–2.201).Conclusion: Our data support the hypothesis that higher serum Mg2+ concentrations may contribute to lower health risk in hemodialysis populations. Further randomized controlled trials and cohort studies are warranted to verify whether Mg2+ supplementation could be part of routine examinations in hemodialysis populations.