A High-performance Liquid Chromatography Method for Hemoglobin A1c

Abstract

Hemoglobin'A1c (HbA1c) is a glycosylated derivative of hemoglobin and is one of a family of derivatives whose concentrations are elevated in patients with diabetes mellltus. Published methods for the measurement of HbA1c are relatively tedious and require modest amounts of blood. A high-performance liquid Chromatographie (HPLC) method for the determination of HbA1c is presented. The method is rapid (20 minutes), precise (coefficient of variation of 5-10 per cent), uses small amounts of sample (3μI.), and can be automated. A sample-preparation technique using filtration was developed that shortened and simplified preparation of venous blood and allowed use of capillary samples. HbA1c was measured by this method in three age-stratified groups of controls and a group of insulin-requiring juvenile diabetics. There was clear separation of HbA1c values between all normals (5.9 ± 1.3, 5.6 ± 0.7, 7.1 ± 0.9 per cent) and the diabetics (12.1 ± 2.4 per cent). Use of this method can facilitate large-scale clinical investigations and permit biochemical investigations of the metabolism and formation of hemoglobin A1c where small sample sizes are necessary.