Assessment of the effect of hemoglobin variants on routine HbA1c measurements by electrospray ionization mass spectrometry

Abstract

We applied electrospray ionization mass spectrometry (ESI-MS) to identify hemoglobin (Hb) variants, and to assess the effect of the variants on routine measurements of a glycated Hb, HbA1c. Over the past 8 years, we have diagnosed 83 cases, including 42 kinds of variant Hb, using MS as the main technology. Of these variants, 3 were new, and 9 were the first cases in Japan. Some abnormal Hbs cause diseases, and most cause erroneous values of HbA1c measured by various methods. ESI-MS was also successfully used for the accurate determination of glycated Hb. We and other groups proposed methods to examine glycated Hb by ESI-MS using enzyme-digested peptides, or intact globin without enzyme digestion. Using the peptide method, we clarified the extent of discrepancies among the HbA1c values measured by conventional methods and accurate values for samples containing various Hb variants identified by the MS method. We applied the globin method to measure the ratio of the glycated component of a variant chain and that of a normal chain obtained from the same erythrocytes. Although the glycation degree on most variant chains was similar to that on normal chains obtained from the same erythrocytes, the content of the glycated component of a particular variant beta-chain was approximately three times as much as that of the normal beta-chain. Abnormal Hbs cause erroneous values for HbA1c to various extents with commercial measurement methods, and MS may offer an unrivaled strategy to correct these errors.