Determination of haemoglobin A(1c) by liquid chromatography using a new cation-exchange column.
The use of a new cation-exchange column, ProPac SCX-10, for the determination of haemoglobin A(1c) (HbA(1c)) by high-performance liquid chromatography is described. After optimization of the analytical method for the separation of the various isoforms of haemoglobin with the ProPac SCX-10 column, the method was applied to the determination of HbA(1c) in blood from 59 volunteers. Three of the 59 had previously been diagnosed as diabetics. Interference studies for carbamylation, acetylation and pre-HbA(1c) were carried out via "in-vitro" experiments. No interference due to carbamylation was observed at the urea values normally found in uremic patients undergoing dialysis. No interference from pre-HbA(1c) was detected either. The method is able to separate haemoglobin A (alpha(2)beta(2)), haemoglobin S (haemoglobin from sickle cell anaemia patients) and haemoglobin A(2) (alpha(2)delta(2)) without interference. The method of Hampel was applied to detect outliers. A value of 3.29+/-0.44% (2sigma) for HbA(1c) was obtained in the analysis of 56 blood samples from non-diabetics. This average value is lower than that reported by most of the methods currently used in routine analyses.
Beatriz de la Calle Guntiñas
Maria
M
European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Retieseweg, 2440 Geel, Belgium. beatriz.de-la-calle@irmm.jrc.be
Wissiack
René
R
Bordin
Guy
G
Rodríguez
Adela Rosa
AR
D002411:Cation Exchange Resins; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006442:Glycated Hemoglobin / Q000032:analysis*; D006801:Humans
D002411:Cation Exchange Resins; D006442:Glycated Hemoglobin