To observe the effects of fructose-1,6-diphosphate (FDP) on serum levels of cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB), as well as the concentration of calcium in cardiomyocytes (Myo[Ca(2+)]) and activity of sarcoplosnic Ca(2+)-ATPase (SRCa(2+)-ATPase) in Adriamycin (ADR)-treated rats. Rats were intraperitoneally injected with ADR (2.5 mg/kg every other day for 6 times) and then with different dosages of FDP (every other day for twenty-one times). Bi-antibodies sandwich Enzyme linked immune absorption assay (ELISA) was performed to detect serum level of cTnI. CK-MB was detected by monoclonal antibody, Myo[Ca(2+)] was detected by fluorescent spectrophotometry and the activity of SRCa(2+)-ATPase was detected by inorganic phosphate method. FDP (300, 600, 1200 mg/kg) significantly reduced the serum levels of cTnI and CK-MB, while at the same time decreased calcium concentration and increased SRCa(2+)-ATPase activity in cardiomyocytes of ADR-treated rats (P<0.01). FDP might alleviate the cardiotoxic effects induced by ADR through decreasing calcium level as well as increasing SRCa(2+)-ATPase activity in cardiomyocytes.