New HPLC instrument performance evaluation in HbA1c determination and comparison with capillary electrophoresis

Scand J Clin Lab Invest. 2018 Sep;78(5):393-397. doi: 10.1080/00365513.2018.1487072. Epub 2018 Jul 11.

Abstract

Glycated hemoglobin (HbA1c) measurement provides the most important medium to long-term marker of time-averaged glycemic status. Its relationship to clinical outcome in diabetes has been convincingly demonstrated for both type 1 and type 2 diabetes. The main HbA1c measurement methods for clinical routine are as follows: ion-exchange chromatography; affinity chromatography, capillary electrophoresis, immunoassay and enzymatic methods. In this study, we evaluated the analytical performances of a new HPLC instrument (Tosoh HLC-723 G11 in the VAR mode) in HbA1c analysis and compared it with a capillary electrophoresis instrument (Sebia Capillarys 2 Flex Piercing). HbA1c analysis was performed in parallel by both methods for 250 samples randomly chosen from healthy and diabetic subjects at 'Tor Vergata' University Hospital of Rome. Tosoh HLC-723 G11 showed good reproducibility for 10 days both in quality controls and in samples analyzed (%CV < 2%). We found good linearity for HbA1c values ranging from 15 mmol/mol (3.5%) to 178 mmol/mol (18.5%), with a correlation coefficient R2 = 1. In a comparison between Tosoh HLC-723 G11 and Capillarys 2FP a good correlation (r = 0.99) was found; however, Tosoh HLC-723 G11 showed higher values in the low range of HbA1c and lower in the high range (Tosoh HLC-723 G11 = 4.3043 + 0.913 Capillarys 2FP; p < 0.001). Tosoh HLC-723 G11 showed good repeatability, reproducibility, accuracy and automated simplicity, and it seemed suitable for routine use in clinical chemistry laboratories.

Keywords: Diabetes; analytical performance; capillary electrophoresis; glycated hemoglobin; ion-exchange HPLC.

Publication types

  • Comparative Study

MeSH terms

  • Biomarkers / blood
  • Chromatography, High Pressure Liquid / standards*
  • Diabetes Mellitus, Type 1 / blood
  • Diabetes Mellitus, Type 1 / diagnosis
  • Diabetes Mellitus, Type 2 / blood
  • Diabetes Mellitus, Type 2 / diagnosis
  • Electrophoresis, Capillary / standards*
  • Glycated Hemoglobin / analysis*
  • Glycated Hemoglobin / metabolism
  • Humans
  • Hyperglycemia / blood
  • Hyperglycemia / diagnosis
  • Reproducibility of Results

Substances

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