Modified HPLC-electrospray ionization/mass spectrometry method for HbA1c based on IFCC reference measurement procedure

Clinical Chemistry
Patricia KaiserHans Reinauer

Abstract

Monitoring of hemoglobin A(1c) (HbA(1c)) is important in the management of diabetes. The IFCC reference measurement procedure for HbA(1c) is based on the ratio of glycated to nonglycated N-terminal hexapeptides of the beta-chains of hemoglobin after digestion with Glu-C endoproteinase. We developed a modification of the original reference measurement procedure with HPLC-electrospray ionization/mass spectrometry (ESI/MS). We performed chromatographic separation of the hexapeptides using a C12 reversed-phase column and a binary gradient system consisting of a mixture of H(2)O/acetonitrile/formic acid. Using this method, we obtained higher signal intensities and improved system stability compared with the reference measurement procedure. In the range of 3% to 14% HbA(1c), intralaboratory CVs were 0.71% to 1.86%. Deviations from IFCC target values were -0.87 to 1.00 relative %. These values fulfill acceptability criteria for HbA(1c) determination set by the IFCC Working Group on HbA(1c) Standardization. This procedure for the determination of HbA(1c) improves the existing reference measurement procedure.

Citations

Mar 31, 2015·Clinical Biochemistry·Kyunghoon LeeJunghan Song
Jan 21, 2017·Clinical Chemistry and Laboratory Medicine : CCLM·Yeo-Min YunJunghan Song
Jun 5, 2013·Clinical Chemistry and Laboratory Medicine : CCLM·Federica Braga, Mauro Panteghini
Oct 5, 2019·Analytical and Bioanalytical Chemistry·Terumichi NakagawaMitsuru Murata
Dec 14, 2018·Clinical Chemistry and Laboratory Medicine : CCLM·Hong LiuBih-Yann Ng
May 16, 2019·Clinical Chemistry and Laboratory Medicine : CCLM·Vincent DelatourPhilippe Gillery
Oct 15, 2014·Annals of Clinical Biochemistry·Joanne E AdawayLaura J Owen

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