Hemoglobin glycation is due to the prolonged exposure to hyperglycemia resulting from defects in
insulin secretion. Ketoacidosis is another complication associated with insulin deficiency in diabetes mellitus.
Furthermore, it was recently demonstrated that glycation occurs in vitro with bicarbonyl compounds derived
from fatty acid oxidation. To study the relationship between the glycation of hemoglobin and diabetic
ketoacidosis, we tested the in vitro formation of HbA1 by acetone and $-hydroxibutirate. Incubations were C
carried out with four groups of hemoglobin samples: glucose (Group 1), acetone (Group 2), $-hydroxibutirate
(Group 3) and glucose plus acetone (Group 4). Concentrations of HbA1 were detected by turbidimetric C
inhibition immunoassay using specific HbA1 antibodies and by fluorescence testing using four pairs of filters: C
excitation 320/emission 460; excitation 355/emission 460; excitation 320/emission 538 and excitation 485/emission
538. HbA1 was produced in hemoglobin samples incubated with acetone and $-hydroxibutirate alone, C
suggesting that HbA1 can be non-enzymatically formed in vitro from ketone bodies in the absence of reducing C
sugars, providing evidence that ketone bodies contribute to the development of complications of type 1
diabetes. In addition, results from the comparison of the antiglycating agents showed that glycylglycine
potently inhibits HbA1 formation. A significant unexpected observation emerged when ketone bodies related C
samples increased the fluorescent intensity even more so than the classic glycated samples caused by glucose.
The chemical structures of these unknown fluorescent compounds remain to be investigated.