Changes in glycated haemoglobin levels in diabetic rats measured with an automatic affinity HPLC

Abstract

1. The level of glycated haemoglobin (GHb) in diabetic rats was measured using a newly developed automatic high-performance liquid chromatography (HPLC) with a boronate affinity column that requires only 2.5 min per sample for analysis. 2. Levels of GHb were 2.7% in normal 7-week-old Sprague-Dawley rats. These levels increased gradually following the abrupt induction of hyperglycaemia by intravenous injection of streptozotocin (STZ), reaching a maximal level of 10.1% after 6 weeks. 3. Glycosylated haemoglobin (HbA1) levels measured by cation-exchange chromatography were also increased by STZ treatment, although HbA1 values were lower than GHb measured by affinity column HPLC. 4. In Wistar fatty rats, GHb levels declined gradually over 5 weeks following the administration of pioglitazone (0.75 or 2.25 mg/kg per day) as a food admixture, which reduced plasma glucose (PG) levels to normal levels within 1 week. Glycated haemoglobin levels after 5 weeks treatment with pioglitazone correlated better with the area under the curve for PG over the entire 5 week treatment period than with the PG level at the end of treatment. 5. In addition, GHb determined by affinity column HPLC correlated well with HbA1 measured by cation-exchange chromatography, although the GHb value was higher than the HbA1 value. 6. Glycated haemoglobin levels in db/db and KKAy mice were higher than those in control normoglycaemic animals and were also higher than HbA1 values measured by the cation-exchange method, although the two values did show good correlation. 7. These results indicate that the newly developed affinity column HPLC system is useful for evaluating total GHb levels in rats as an index of antidiabetic treatment.