Effect of HbE and HbH on HbA1C level by ionic exchange HPLC comparing to immunoturbidimetry.
BACKGROUND
The hemoglobin (Hb)A1C level is widely used to monitor diabetes mellitus patients. The N-terminal amino acid valine of its beta chain is glycated. The assay of HbA1C is based on differences in the charge, chemical and structural properties of the protein.


METHODS
There are fully automated instruments available in clinical chemistry laboratory to assay HbA1C level. The effect of hemoglobinopathies was studied between an ionic exchange high-pressure liquid chromatography (HPLC) (Bio-Rad Laboratories, USA) and immunoturbidimetry (BM/Hitachi 912 with Roche HBA1CII, Germany-Japan) assay. The influence of high level HbF relative to the HbA1C level by ionic exchange HPLC is known. The effect of HbE and HbH to the HbA1C level by ionic exchange HPLC comparing to immunoturbimetry was examined. The evaluation was performed on 34 normal controls (A2A), 17 beta thalassemia traits (A2 upward arrow A), 36 HbE heterozygotes (EA), 37 HbE homozygotes (EE), 36 beta thalassemia/HbE (EF/EFA), 11 EABart's diseases (EABart's), 34 Hb H diseases (A2/CSAH) and 13 cord blood samples (FA).


CONCLUSIONS
Hemoglobinopathies can impact on the assay of HbA1C level such as HbE and HbH to ionic exchange HPLC. Although not studied as yet, this effect may influence the other methods such as affinity chromatography.
Pravatmuang
P
P
Department of Medicine, Siriraj Hospital, Mahidol University, 2 Prannok, 10700, Bangkok, Thailand. sippm@mahidol.ac.th
Sae-Ngow
B
B
Whanpuch
T
T
Leowattana
W
W
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006442:Glycated Hemoglobin / Q000032:analysis*; D006446:Hemoglobin E / Q000737:chemistry*; D006447:Hemoglobin H / Q000737:chemistry*; D006801:Humans; D009391:Nephelometry and Turbidimetry / Q000379:methods*
D006442:Glycated Hemoglobin; D006446:Hemoglobin E; D006447:Hemoglobin H