Hemoglobin fractionation by hplc

Introduction: High performance liquid chromatography (HPLC) is a new technique which has demonstrated advantages in terms of accuracy and precision over conventional procedures employed in newborn and adult hemoglobinopathy screening programs for the identification of Hb variants. It has prompted the need to reassess our knowledge of hemoglobin fractions reference ranges as it relates to HPLC HB quantitation. Materials and Methods: Fifty microliter aliquots of fresh whole blood samples collected in ethylenediamine tetraacetic acid (EDTA) containing vacutainer tubes from normal newborn and adult individuals being screened at the were treated with a hemolyzate reagent, centrifuged at 12,000 rpm at 4°C for 15 minutes and passed through a 0.25 to 0.45 diameter syringe filter unit into a 1.5 ml tube. Results: In this study, the HPLC hemoglobin reference ranges derived from 100 normal adults (ages 18-50) and infants of upto 300 days of age were studied. And HPLC were chosen to establish the hemoglobin reference ranges.. The percentage values are as follows: Hb A mean of 92.3 percent; HbA1 mean 3.6 percent; Hb F mean of 3.2 percent; Hb A2 mean of 2.1 percent, respectively.
Conclusion: Application of the HPLC ranges to confront other abnormalities will prove most useful during blood screening processes. The HPLC hemoglobin ranges for normal adult and newborns have been established.