Magnesium (Mg) status is currently assessed by various biochemical biomarkers, most of which, however, have some limitations. We developed an in vitro blood load test as a new Mg biomarker using a Mg stable isotope. This test is based on the hypothesis that cellular Mg uptake is increased in Mg deficiency. For this purpose, Wister male rats were fed either Mg-deficient or Mg-adequate diets for 1 mo and blood was sampled and incubated with the (25)Mg isotope (10 mg/L) for 2 h at 37 degrees C. Erythrocytes, lymphocytes and platelets were isolated and (25)Mg concentrations were determined by inductively coupled plasma/mass spectrometry. The feasibility of this approach was then tested on human blood. (25)Mg enrichments in erythrocytes, lymphocytes and platelets from Mg-deficient rats were greater than those from controls. (25)Mg enrichment was low in human erythrocytes (3%) compared with rat erythrocytes (38%), whereas high (25)Mg enrichments were obtained in human lymphocytes and platelets, suggesting that lymphocytes and platelets may be more appropriate cells than erythrocytes for examining Mg status in humans with this approach. More studies are required to validate the utilization of this test as a Mg status biomarker in humans.