Determinazione delle frazioni dell’emoglobina alla nascita mediante cromatografia ad alta risoluzione ed elettroforesi capillare: risultati preliminari sulla quantificazione dell’HbA

AUTORI

ABSTRACT

Determination of hemoglobin fractions at birth using HPLC and capillary electrophoresis: preliminary results on the HbA quantification

Introduction: the hemoglobin fractions that can be normally detected at birth are HbF and HbA. Early assessment of hemoglobin profile is required because of clinical symptoms or to confirm prenatal diagnosis. Recently, the need to investigate the hemoglobin status in the cord blood for possible storage in dedicated Cord Blood Banks emerged. The aim of this study is to compare High Performance Liquid Chromatography (HPLC) and Capillary Electrophoresis (CE) methods for the determination of HbA at birth.
Methods: the qualitative-quantitative determination of hemoglobin fractions from 242 newborns was performed by HPLC (Variant II system, Bio-Rad Laboratories) and CE (Capillarys, Sebia).
Results: there is an increasing trend of the percentage of HbA as the gestational weeks at birth increase with both HPLC and CE; two significantly different macro groups can be identified (37-39 weeks versus 40-41 weeks) with relative reference intervals that are wide and method dependent. The HPLC method HbA results are significantly lower than those obtained with the CE method with an average difference of 5.7%.
Discussion: the Cord Blood Bank requires verification at birth of the absence of globin defects to store the cord blood units. According to national recommendations, globin defects are certainly excluded when the HbA percentage is ≥22% regardless of the method used. Discordance between methods represents a non-trivial issue in this area and would possibly lead to method-dependent reference intervals. A molecular characterization of the samples showing a HbA percentage <22% could then be suggested, in order to provide robust cut-offs, eventually differentiated by gestational weeks, for carriers of β thalassemia.

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