Chemistry: analytical and immunological testing – Hemoglobin – myoglobin – or occult blood – Glycosylated hemoglobin
Patent
1981-03-16
1983-10-04
Turk, Arnold
Chemistry: analytical and immunological testing
Hemoglobin, myoglobin, or occult blood
Glycosylated hemoglobin
106197C, 210692, 210927, 252184, 260112B, 436178, 521 28, G01N 3372
Patent
active
044079613
ABSTRACT:
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.degree.-37.degree. 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.
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Sankar, Analytical Chemistry, vol. 50, No. 13, Nov. 1978, pp. 1922-1924.
Gunter Jr. Charles D.
Turk Arnold
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