Identification of fetal hemoglobin in blood stains by high performance liquid chromatography

Abstract

A new method for the identification of fetal hemoglobin (Hb F) in blood stains by reverse-phase high performance liquid chromatography is described. Differentiation between fetal and adult blood stains is based on the existence of gamma-chain peaks which are characteristics of Hb F. Very few gamma chains appeared on chromatograms of all the adult blood stains examined. The level of Hb F could be determined by measuring the total of chromatogram gamma-globin chain areas, and expressing it as a percentage of total Hb. Levels in six cord blood stains on filter paper ranged from 81.1% to 91.3% and remained constant for at least 12 weeks. This method is of great value for its simplicity, sensitivity and speed, and most importantly for its reliability in the field of forensic medicine.

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

“…Samples numbered 1-6 were from cord blood and those numbered 7-11, from peripheral blood of neonates less than 3 weeks old. Data from cord samples up to 12 weeks old have been reported in a previous paper [13]. The chromatographic profiles illustrate that globin chain denaturation proceeds gradually in bloodstains but little change is noted in Hb F levels for up to 20 weeks.…”

“…The percentage ratio of Hb F to total Hb was determined by measuring relative peak areas of non-aglobin chains on the chromatograms. Other technical details have been described elsewhere [13].…”

“…Identification of fetal and/or neonatal bloodstains is also important, especially in cases of infanticide, criminal abortion or concealed delivery. We have previously described a method for the identification of fetal hemoglobin (Hb F) in bloodstains by high-performance liquid chromatography (HPLC) and discussed its usefulness [13]. In the present paper, we describe a one-step HPLC method which can simultaneously identify Hb F in bloodstains and be used to estimate bloodstain age.…”

Species identification of blood and bloodstains by high-performance liquid chromatography

“…Human blood and blood of other primates and various other species (see Table 1) were collected in EDTA or heparin, with the exception of rainbow trout and sardine blood, which was used neat. The methods used for preparing hemolysates, bloodstains on filter paper and the extracts from these bloodstains have been described in a previous paper [7]. Mixed bloodstain samples were made by dropping equal volumes of human blood and blood of other species (pig, cow, dog, cat and chicken) onto filter paper.…”

“…The usefulness of HPLC for the identification of human fetal hemoglobin in bloodstains has recently been described [7] and in this paper, species identification of blood and bloodstains using reverse-phase HPLC is described.…”