Evaluation of a new semi-automated high-performance liquid chromatography method for glycosylated haemoglobins

The authors report data on the evaluation of a new ion-exchange high performance liquid chromatographic system (HLC-723GHb Glycopack-Dyamat), recently introduced by Bio-Rad Laboratories (Segrate, Milano, Italy) to measure the glycosylated haemoglobins in blood samples. The system operates on haemolysates, which have to be performed separately, by diluting 5 μl of blood with 1 ml of haemolysing reagent. Each analysis takes about 8 min and the results are printed-out, together with a chromatogram. Precision and accuracy (comparison with the minicolumn ion-exchange method) were found to be excellent. The analysis if not affected by the presence of the so-called 'labile fractions', as demonstrated by experiments of incubation with glucose or saline solutions at 37° C. Some haemolysates containing abnormal haemoglobins (HbS, HbE, HbJ Paris and Hb Lepore) were also analysed. The effects of the different mutants on the glycated haemoglobins assay are discussed. The HbF content is also measured by this system, but the accuracy, concerning this particular determination, at the lower concentrations (HbF <2%) is not satisfactory, and needs further improvement.