A rapid semi-automated method for determining glycosylation in whole blood

A rapid, semi-automated method for determining hexoses which are bound to components of whole blood; e.g., hemoglobin, is described. The method is an improvement on similar lengthy manual methods (1,2) in which 5-hydroxymethylfurfural is produced and measured. Heating of diluted blood samples at 15 lb./sq.in. above atmospheric pressure in a pressure cooker for 20 minutes is shown to give the same results as those obtained with other methods in which 2-5 hours heating at atmospheric pressure is specified (1,2,3). The trichloracetic acid precipitation step needed in the manual methods is avoided by continuous flow dialysis on a Technicom Auto Analyzer II system. Various experimental factors have been studied and comparisons made with a commonly-used ion-exchange resin column technique. Two different methods of calibration, one using pure 5-hydroxymethylfurfural standards, and the other using patients' samples with pre-determined, known values as standards, are presented.