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Hemoglobin
international journal for hemoglobin research
Volume 33, 2009 - Issue 1
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Hb Belleville [β10(A7)Ala→Thr] Affects The Determination of Hb A1c by Routine Cation Exchange High Performance Liquid Chromatography

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Pages 45-50 | Received 19 May 2008, Accepted 07 Aug 2008, Published online: 07 Jul 2009
 

Abstract

When analyzed by routine cation exchange high performance liquid chromatography (HPLC), the Hb A1c peak of a Caucasian diabetic patient from Yorkshire, UK, appeared to be an incompletely resolved doublet. One component (5.5%) eluted at the normal time for Hb A1c, whereas the other component (6.6%) eluted slightly later. The HPLC trace was otherwise normal. Analysis of the diabetic patient's blood and a tryptic digest thereof by electrospray ionization mass spectrometry (ESI-MS) identified the Hb Belleville trait. To relate Hb A1c determined by HPLC to α and β chain glycation determined by ESI-MS, several normal blood samples (5–14% Hb A1c) were analyzed by both techniques. The Hb A1c levels derived from the α and β chain glycation levels of the diabetic patient (12.9 and 12.4%, respectively) agreed with the sum of the two peaks (12.1%) in the HPLC trace. Similarly, Hb Belleville was detected and identified in another Caucasian, also from Yorkshire, with normal Hb A1c.

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