The Contribution of High Performance Liquid Chromatography to the Incidental Discovery of Hemoglobin Variants during Glycated Hemoglobin Assay

Abstract

HbA1c is assayed in the biochemistry laboratory using a high pressure liquid chromatography (HPLC) technique in ion exchange, and given the correlation between its value and the risk of complication, it is crucial to know it, advantages and it limits. This work highlights the contribution of HPLC in the detection of qualitative abnormalities of hemoglobin during the determination of HbA1c using the ADAMS HA-8180V ARKRAY® device. This descriptive prospective study, carried out at the Biochemistry-Toxicology department of the Mohammed V Military Hospital in Rabat, focused on all samples for which routine HbA1c testing is prescribed during the past year, external samples and from all departments. For each abnormality noted on the chromatogram, a complete study of hemoglobin was performed using other electrophoretic technique at alkaline and acid pH on the Capillarys 2 Flex Piercing® Sebia® and the Hydrasys 2 Scan Focusing® Sebia®. Almost 0.6% of the variants were detected during the HbA1c assay out of 12,944 blood samples.Only 0.04% were not identified by the machine. After confirmation, variant C was predominantly present (n = 30), followed by S (n = 28) then O-Arab (n = 8) and l 'Hb D (n =2). The HPLC technique, for measurement of HbA1c, represents a reliable screening tool for the most common variants of Hb, with however limitations that encourage vigilance in the analysis and interpretation of the results.