A RAPID ANION-EXCHANGE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY METHOD FOR THE MEASUREMENT OF HBA(2) IN WHOLE-BLOOD

We developed a binary gradient high-performance liquid chromatography (HPLC) method for measuring HbA(2) in whole blood samples using a Phar macia Mono Q column (1 mt) and measurement at 415 nm. The assay requir es a simple lysis and centrifugation step before injection onto the co lumn. We found good agreement of results between the HPLC method and t he Helena column chromatography method. The within batch precision was 2.6% and between batch precision was 4.6%. We found that using 30 mM Tris buffers (pH 7.8) with a sodium chloride gradient resulted in shor t analysis times and good chromatographic separation of HbA(2), HbS an d HbA. We conclude that this is a robust assay for the diagnosis of be ta-thalassaemia.