Modified HPLC-ESI-MS Method for Glycated Hemoglobin Quantification Based on the IFCC Reference Measurement Procedure and Its Application for Quantitative Analyses in Clinical Laboratories of China

J Clin Lab Anal. 2016 Sep;30(5):457-63. doi: 10.1002/jcla.21879. Epub 2015 Oct 29.

Abstract

Background: The level of glycated hemoglobin (HbA1c ) has been recognized as an important indicator of long-term glycemic control. However, the HbA1c measurement is not currently included as a diagnostic determinant in China. Current study aims to assess a candidate modified International Federation of Clinical Chemistry reference method for the forthcoming standardization of HbA1c measurements in China.

Methods: The HbA1c concentration was measured using a modified high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method. The modified method replaces the propylcyanide column with a C18 reversed-phase column, which has a lower cost and is more commonly used in China, and uses 0.1% (26.5 mmol/l) formic acid instead of trifluoroacetic acid. Moreover, in order to minimize matrix interference and reduce the running time, a solid-phase extraction was employed. The discrepancies between HbA1c measurements using conventional methods and the HPLC-ESI-MS method were clarified in clinical samples from healthy people and diabetic patients. Corresponding samples were distributed to 89 hospitals in Beijing for external quality assessment.

Results: The linearity, reliability, and accuracy of the modified HPLC-ESI-MS method with a shortened running time of 6 min were successfully validated. Out of 89 hospitals evaluated, the relative biases of HbA1c concentrations were < 8% for 74 hospitals and < 5% for 60 hospitals. Compared with other conventional methods, HbA1c concentrations determined by HPLC methods were similar to the values obtained from the current HPLC-ESI-MS method.

Conclusion: The HPLC-ESI-MS method represents an improvement over existing methods and provides a simple, stable, and rapid HbA1c measurement with strong signal intensities and reduced ion suppression.

Keywords: diabetes; external quality assessment; glycated hemoglobin; high-performance liquid chromatography; mass spectrometry.

Publication types

  • Multicenter Study

MeSH terms

  • China
  • Chromatography, High Pressure Liquid / methods*
  • Clinical Laboratory Services*
  • Diabetes Mellitus / blood*
  • Female
  • Glycated Hemoglobin / analysis
  • Hospitals
  • Humans
  • Linear Models
  • Male
  • Reference Values
  • Spectrometry, Mass, Electrospray Ionization / methods*

Substances

  • Glycated Hemoglobin A