We have devised an enzyme immunoassay for the determination of human myoglobin. This assay provides superb reproducibility and a recovery rate without the influence of any anti-coagulants and hemolysis. The mean blood myoglobin level in healthy humans, determined by this assay, was 35.5 ± 23.9 ng/ml and the urine myoglobin level was found to be distributed over a broad range under 373.5 ng/ml. In cases of trauma sustained by living bodies, the blood myoglobin level was found to increase in accordance to the severity of trauma. Further, the myoglobin level in the blood began its increase during the early stage of the injury, and greater increases were shown in the elderly. In autopsy specimens, the ratio of the cardiac blood myoglobin level to the cerebrospinal fluid myoglobin level had a negative correlation with the postmortem passage of time and, as in the case of living bodies, the myoglobin level in the blood was considered a useful indicator of the severity of damage. However, since myoglobin level was found to vary, depending on the sites from where the sample blood was taken, the evaluation of the data should be done carefully. Finally, the simultaneous determination of the creatine phosphokinase (CPK) activity in autopsied blood samples would be of help for estimating the pestmortem passage of time and the progress of putrefaction.