基于国际临床化学和检验医学联合会（IFCC）参考测量程序的糖化血红蛋白（HbA1c）改良高效液相色谱-电喷雾电离/质谱法
Modified HPLC-electrospray ionization/mass spectrometry method for HbA1c based on IFCC reference measurement procedure.
作者信息
Kaiser Patricia, Akerboom Theodorus, Molnar Petra, Reinauer Hans
机构信息
Reference Measurement Laboratory, Instand e.V., Düsseldorf, Germany.
出版信息
Clin Chem. 2008 Jun;54(6):1018-22. doi: 10.1373/clinchem.2007.100875. Epub 2008 Apr 10.
BACKGROUND
Monitoring of hemoglobin A(1c) (HbA(1c)) is important in the management of diabetes. The IFCC reference measurement procedure for HbA(1c) is based on the ratio of glycated to nonglycated N-terminal hexapeptides of the beta-chains of hemoglobin after digestion with Glu-C endoproteinase. We developed a modification of the original reference measurement procedure with HPLC-electrospray ionization/mass spectrometry (ESI/MS).
METHOD
We performed chromatographic separation of the hexapeptides using a C12 reversed-phase column and a binary gradient system consisting of a mixture of H(2)O/acetonitrile/formic acid.
RESULTS
Using this method, we obtained higher signal intensities and improved system stability compared with the reference measurement procedure. In the range of 3% to 14% HbA(1c), intralaboratory CVs were 0.71% to 1.86%. Deviations from IFCC target values were -0.87 to 1.00 relative %. These values fulfill acceptability criteria for HbA(1c) determination set by the IFCC Working Group on HbA(1c) Standardization.
CONCLUSIONS
This procedure for the determination of HbA(1c) improves the existing reference measurement procedure.
背景
糖化血红蛋白A1c（HbA1c）监测在糖尿病管理中至关重要。IFCC的HbA1c参考测量程序基于血红蛋白β链经Glu-C内蛋白酶消化后糖化与非糖化N端六肽的比例。我们开发了一种采用高效液相色谱-电喷雾电离/质谱（ESI/MS）对原始参考测量程序的改进方法。
方法
我们使用C12反相柱和由水/乙腈/甲酸混合物组成的二元梯度系统对六肽进行色谱分离。
结果
与参考测量程序相比，使用该方法我们获得了更高的信号强度并提高了系统稳定性。在HbA1c为3%至14%的范围内，实验室内变异系数为0.71%至1.86%。与IFCC目标值的偏差为相对-0.87至1.00%。这些值符合IFCC HbA1c标准化工作组设定的HbA1c测定可接受标准。
结论
这种HbA1c测定程序改进了现有的参考测量程序。
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