Capillary electrophoresis system

Hemoglobin separation and quantification of hemoglobin types were performed using the CAPILLARYS 2 FLEX PIERCING equipped with 8 capillaries according to the manufacturer’s instructions. Briefly, the blood sample was added to a lysis solution, and an aliquot was introduced using aspiration at the anodic end into the capillary tube prior to application of 9,800 V. Detection of hemoglobin was performed via measurement of the absorbance at 415 nm at the cathodic end of the capillary tube. The hemoglobin A₂ fraction was adjusted at the end of the analysis to appear in zone 9 in the middle of the window. The relative quantity (percent of total hemoglobin) and presumptive identification of the hemoglobin types (located in various zones from zone 1 to 15) were recorded from the resulting electropherogram. The positions of different hemoglobin types were identified in the pertinent zones. The mean (±SD) hemoglobin A₂ proportions in the normal and genetically proven beta thalassemia heterozygotes were 2.8±0.3% and 5.8±0.7%, respectively, and the proportion of hemoglobin E heterozygotes was 24.0±3%.^12-14