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Updated: Jan 27, 2026

Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Published on: January 18, 2020
Hb Belleville [beta 10(A7)Ala-->Thr] affects the determination of Hb A1c by routine cation exchange high performance
Jonathan P Williams1, Hannah Jackson, Brian N Green
1Department of Chemistry, University of Warwick, Coventry, Warwickshire, UK. J.P.Williams@warwick.ac.uk
When analyzed by routine cation exchange high performance liquid chromatography (HPLC), the Hb A(1c) 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 A(1c), 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 A(1c) determined by HPLC to alpha and beta chain glycation determined by ESI-MS, several normal blood samples (5-14% Hb A(1c)) were analyzed by both techniques. The Hb A(1c) levels derived from the alpha and beta 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 A(1c).
When analyzed by routine cation exchange high performance liquid chromatography (HPLC), the Hb A(1c) 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 A(1c), 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 A(1c) determined by HPLC to alpha and beta chain glycation determined by ESI-MS, several normal blood samples (5-14% Hb A(1c)) were analyzed by both techniques. The Hb A(1c) levels derived from the alpha and beta 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 A(1c).
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