Identification of fetal hemoglobin and simultaneous estimation of bloodstain age by high-performance liquid chromatography.

A method using reverse-phase high-performance liquid chromatography (HPLC) for the identification of fetal hemoglobin (Hb F) and the simultaneous estimation of bloodstain age is described. Umbilical cord and neonatal bloodstains can be differentiated from adult stains by the presence of gamma-globin chains which are characteristic of Hb F. With this method, cord and neonatal blood could be distinguished from adult blood in stains up to 32 weeks old. The age of the stain was estimated from the ratio of the peak area of the alpha-globin chain to that of heme on the same chromatogram. The ratio decreased gradually with an increase in the age of the stain up to 20 weeks old. Studies performed at each time period revealed no significant difference in the ratios of cord and neonatal bloodstains or in the ratios of cord and adult stains. The regression equation calculated from the ratios (y) and the ages of stains in weeks (x) expressed logarithmically is y = 2.5758-0.2497 In (x) and the coefficient of correlation is -0.7491 (n = 252, P less than 0.001). The present method, having the advantages of simplicity, speed and sensitivity, should be of great value to forensic science.