Ratio of the Ion Peak Intensity of Glycated to Non-glycated Hexapeptides from the N-Terminal of Hemoglobin β-Chain Measured by LC-ESI Mass Spectrometry (TMS特集号)

概要

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Liquid chromatography-electrospray ionization mass spectrometry (LC-ESIMS) was used to measure the ratio of the ion peak intensity of the glycated and non-glycated hexapeptides from the amino terminal of hemoglobin (Hb) β-chain to establish an accurate method of measuring glycated hemoglobin (HbA1c). Chemically synthesized glycated and non-glycated hexapeptides were mixed in various ratios, and the mixtures were analyzed by on-line reverse phase LC connected to ESIMS. We found that the ratio of the peak intensity calculated by the equation, 0.5 × peak area of doubly charged ion + 1 × peak area of singly charged ion for each peptide was the most reproducible. The slope of the resulting curves from the equation was nearly equal to 1. This calculation should improve the accuracy of the reference method for HbA1c, proposed by Kobold et al., based on LC/ESIMS analysis of the specific N-terminal residues of the Hb β-chains, which are released by enzymatic cleavage of the intact Hb molecule with endoproteinase Glu-C. The measurement of HbA1c both by the MS method proposed in the present paper and by a conventional HPLC method revealed that the values obtained by HPLC were ca. 20% higher than those obtained by MS.
日本質量分析学会の論文
1999-12-01