A differential study between antemortem bleeding and a postmortem infiltration of hemoglobin

In a differential study to distinguish antemortem bleeding from a postmortem infiltration of hemoglobin, glycophorins, a component of the erythrocytic membrane, were extracted from two experimental skin tissue models, i.e., skin samples taken on autopsy into which healthy human blood was injected, and skin samples undergoing a postmortem infiltration of hemoglobin; this extraction accomplished by utilizing an anti-glycophorin serum over set periods of time, after which differences between the two models were then evaluated. In all of the bleeding samples from day 0 to day 9, and in 40% of the 12-day-old bleeding samples, glycophorins were clearly detected qualitatively by counterimmunoelectrophoresis and by double-immunodiffusion. In the remaining 12-, 15-, and 18-day-old bleeding samples, glycophorins were only faintly detected by counterimmunoelectrophoresis. Additionally, in a quantitative study by rocket-immunoelectrophoresis, the mean glycophorin value of 0-day-old bleeding samples was 118.5 +/- 5.6 micrograms per 0.5 g of tissue. Further, the recovery of glycophorins from the bleeding samples was satisfactory for up to 9 days, though after 12 days there was a rapid glycophorin decrease. In 15- and 18-day-old bleeding samples, the mean glycophorin values were 19.3 +/- 1.0 and 17.8 +/- 1.0 micrograms per 5 g of tissue, or about 16% and 15% of the 0-day values, respectively. In contrast, no glycophorins were detected in any of the hemoglobin-infiltrated skin samples or our control samples at any time interval. These results suggest that a differential diagnosis between antemortem cutaneous bleeding and postmortem hemoglobin infiltration into the tissue is achievable within 9 to 12 postmortem days.