Long-term evaluation of electrospray ionization mass spectrometric analysis of glycated hemoglobin.
Long-term evaluation of electrospray ionization mass spectrometric analysis of glycated hemoglobin.
复制标题
糖化血红蛋白电喷雾电离质谱分析的长期评估。
作者：
N. B. Roberts;A. Amara;M. A. Morris;B. Green
背景
电喷雾电离质谱（ESIMS）已成功地应用于血红蛋白（Hb）的变体和葡萄糖加合物（质量差为162 Da）的存在下，单独的血红蛋白α和β链的鉴定。为了确定ESIMS作为糖化血红蛋白（HbA 1c）定量的常规和/或参考方法的潜力，我们在常规实验室环境中进行了为期4个月的详细评估。
方法
我们优化了一种适用于HbA 1c常规定量分析的ESIMS方法。我们在4个月内确定了可靠性和再现性，并评估了自动进样的潜力。然后，我们将来自糖尿病患者的1022个血液样本的值与常规的基于HPLC的离子交换程序（HA-8140; Menarini）进行比较。
结果
通过ESIMS测量HbA 1c的结果可在3分钟内获得。手动和自动进样的分析不精密度（CV）为1.6-5.0%。在m/z 980-1400范围内收集的数据证实了α链相对于β链的较低糖化（0.66：1）。每个珠蛋白链仅观察到一个糖化。总的糖化血红蛋白（即，在1022个血液样品上通过ESIMS（x）测量的α-和β-链糖化的平均值低于通过HPLC（y）测量的α-和β-链糖化的平均值：y = 1.0432x + 0.4815。然而，ESIMS测量的β链糖化比离子交换HPLC测量的值高20%，并且显示出与离子交换糖尿病控制和并发症试验（DCCT）校正和英国国家外部质量评估计划DCCT平均返回值的密切一致性，特别是在5-10% HbA 1c时。
结论
ESIMS提供了HbA 1c的精确测量，特别是β链的糖化。该方法具有耐用性，可作为证实HbA 1c测量和/或校准的程序。
BACKGROUND
Electrospray ionization mass spectrometry (ESIMS) has been successfully applied to the identification of hemoglobin (Hb) variants and the presence of glucose adducts (mass difference of 162 Da) on the separate Hb alpha and beta chains. To establish the potential of ESIMS as a routine and/or a reference method for the quantification of glycohemoglobin (HbA1c), we carried out a detailed evaluation over a 4-month period in a routine laboratory environment.
METHODS
We optimized a procedure using ESIMS suitable for the routine quantitative analysis of HbA1c. We determined reliability and reproducibility over 4 months and assessed the potential for automated sample injection. We then compared values of 1022 blood samples from diabetic patients with a routine HPLC-based ion-exchange procedure (HA-8140; Menarini).
RESULTS
Results of HbA1c measurement by ESIMS were available within 3 min. The analytical imprecision (CV) was 1.6-5.0% for both manual and automated injections. Data collection over the m/z 980-1400 range confirmed lower glycation of the alpha chain relative to the beta chain (0.66:1). Only one glycation was observed per globin chain. The overall glycohemoglobin (i.e., the average of alpha- and beta-chain glycations) measured by ESIMS (x) on 1022 blood samples was lower than by HPLC (y): y = 1.0432x + 0.4815. However, the ss-chain glycation measured by ESIMS was up to 20% higher than the value measured by ion-exchange HPLC and showed a close conformity, particularly at 5-10% HbA1c, with the ion-exchange Diabetes Control and Complications Trial (DCCT)-corrected and the United Kingdom National External Quality Assessment Scheme DCCT mean return values.
CONCLUSIONS
ESIMS provides a precise measurement of HbA1c and, in particular, glycation of the beta chain. The method is robust and could be proposed as a procedure to substantiate HbA1c measurement and/or calibration.
通过组合 HPLC/电喷雾质谱法加快罕见变异血红蛋白的表征。
影响因子：
1
作者：
Witkowska,HE;Bitsch,F;Shackleton,CH
通讯作者：
Shackleton,CH