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Clinical Chemistry 26: 466-472, 1980;
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Clinical Chemistry, Vol 26, 466-472, Copyright © 1980 by American Association for Clinical Chemistry

Accuracy, precision, and stability in measurement of hemoglobin A1C by "high-performance" cation-exchange chromatography

RS Schifreen, JM Hickingbotham and GN Bowers Jr

We present a "high-performance" liquid-chromatographic method for determination of HbA1c with use of a carboxylate cation-exchange column, ethanolic mobile phases, and a hemolysate-storage reagent that stabilizes a sample for up to five weeks at 4 degrees C. Temperature control and the addition of ethanol to the phosphate step-gradient increase precision by stabilizing peak shape and column activity. Lengthy equilibration of the column between samples is not necessary, because the pKa of the cation-exchange resin is decreased from 6.10 in aqueous media to 5.59 in ethanol/water (5/95 by vol) at 25 degrees C. We identified interfering heme compounds, distinct from HbA1d or HbA1e, that elute with the glycosylated hemoglobins and developed a method of correcting for their presence. The day-to-day coefficient of variation was 3.5%, and the reference interval for a mixed group of 20 healthy adults between the ages of 20 and 56 was 3.2--5.2% HbA1c.


The following articles in journals at HighWire Press have cited this article:


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 Clin. Chem.Home page
K. Zurbriggen, M. Schmugge, M. Schmid, S. Durka, P. Kleinert, T. Kuster, C. W. Heizmann, and H. Troxler
Analysis of Minor Hemoglobins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry
Clin. Chem., June 1, 2005; 51(6): 989 - 996.
[Abstract] [Full Text] [PDF]

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 ScienceHome page
F. Regnier
High-performance liquid chromatography of biopolymers
Science, October 21, 1983; 222(4621): 245 - 252.
[Abstract] [PDF]


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Copyright © 1980 by the American Association for Clinical Chemistry.