| 摘要 |
An ion-exchange system and method is provided for isolating glycosylated hemoglobin (Hb A1) from other hemoglobins in human blood together with a quantitative determination of glycosylated hemoglobin. Separation of the glycosylated hemoglobin is accomplished by the ion-exchange system with little temperature dependence in the range of from about 15 DEG -37 DEG C. and without rigid control of pH and ionic strength. The ion-exchange system lowers the pH of the human blood to about pH=6.9 through the use of a zwitterionic buffer. The ion-exchange system also contains a cation-exchange resin. The preferred composition of the ion-exchange system contains about 0.05 molar 3-(N-morpholino)propanesulfonic acid as the buffer and carboxymethyl dextran as the ion-exchange resin present in an amount of from about 30 milliequivalents to about 50 milliequivalents of binding capacity per liter thereof. According to the method provided, a lysed preparation of human blood is added to the ion-exchange system and mixed, causing the non-glycosylated hemoglobin to bind to the ion-exchange resin and leaving the glycosylated hemoglobin free in solution. The solution containing glycosylated hemoglobin is separated from the ion-exchange resin by filtration. The fractional amount of glycosylated hemoglobin present in the human blood is determined by comparing the absorbance of the glycosylated hemoglobin fraction with the absorbance of a diluted sample of the lysed human blood. The use of a reference material prepared from human blood and containing a known amount of glycosylated hemoglobin facilitates the determination of the unknown concentrations in human blood.
|
