Effects of hemoglobin C, D, E and S traits on measurements of hemoglobin A1c by twelve methods

Clin Chim Acta. 2016 Apr 1:455:80-3. doi: 10.1016/j.cca.2016.01.031. Epub 2016 Jan 29.


Background: Hemoglobin C, D Punjab, E or S trait can interfere with hemoglobin A1c (HbA1c) results. We assessed whether they affect results obtained with 12 current assay methods.

Methods: Hemoglobin AA (HbAA), HbAC, HbAD Punjab, HbAE and HbAS samples were analyzed on one enzymatic, nine ion-exchange HPLC and two Capillary Electrophoresis methods. Trinity ultra(2) boronate affinity HPLC was the comparative method. An overall test of coincidence of least-squared linear regression lines was performed to determine if HbA1c results were statistically significantly different from those of HbAA samples. Clinically significant interference was defined as >7% difference from HbAA at 6 or 9% HbA1c compared to ultra(2) using Deming regression.

Results: All methods showed statistically significant effects for one or more variants. Clinically significant effects were observed for the Tosoh G8 variant mode and GX (all variants), GX V1.22 (all but HbAE) and G11 variant mode (HbAC). All other methods (Abbott Architect c Enzymatic, Bio-Rad D-100, Variant II NU and Variant II Turbo 2.0, Menarini HA-8180T thalassemia mode and HA-8180V variant mode, Sebia Capillarys 2 and Capillarys 3) showed no clinically significant differences.

Conclusions: Several methods showed clinically significant interference with HbA1c results from one or more variants which could adversely affect patient care.

Keywords: Capillary Electrophoresis; Diabetes; HPLC; Hemoglobin A(1c); Hemoglobin variants; Immunoassay.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Chromatography, Affinity / methods*
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, Ion Exchange / methods*
  • Electrophoresis, Capillary / methods*
  • Glycated Hemoglobin / analysis*
  • Hemoglobins / chemistry*
  • Humans


  • Glycated Hemoglobin A
  • Hemoglobins
  • hemoglobin A1c protein, human