Vitamin D metabolism is dependent on magnesium (Mg) as a cofactor; therefore, poor Mg status may alter the relationship between vitamin D metabolite serum 25-hydroxyvitamin D (s25OHD) and serum parathyroid hormone (sPTH). We hypothesized that low dietary Mg intake may alter sPTH response to s25OHD in a population with excess body weight, thereby leading to a worsening of cardiometabolic health. To explore this hypothesis, we conducted a cross-sectional study on adults who were either overweight or obese (owt/ob). Dietary Mg intake was measured using a Mg food frequency questionnaire (MgFFQ). Body composition information was measured using Dual Energy X-ray Absorptiometry (DXA). Blood samples were obtained for all biochemical analyses. A total of 57 participants, 22 to 65 years of age, with a body mass index between 25 to 45 kg/m were divided into 3 groups, according to dietary Mg intake percentiles (Low Mg Group = <33 percentile, Medium Mg Group = 33 to 66 percentile, High Mg Group = >66 percentile). Higher s25OHD was negatively associated with lower sPTH in the High Mg Intake group (r = -0.472, P = .041), but not in other groups. A positive relationship between s25OHD and serum high-molecular weight adiponectin concentrations was observed in the High Mg Group (r = 0.532, r = 0.022), but not in other groups. Serum Interleukin-6 concentrations were negatively associated with s25OHD (r = -0.316, P = .017) for the entire study group. Based on these results, our study demonstrated that a low dietary Mg intake may alter PTH response to 25OHD.