Song Zhixin, Xie Baoyuan, Ma Huaian, Zhang Rui, Li Pengfei, Liu Lihong, Yue Yuhong, Zhang Jianping, Tong Qing, Wang Qingtao
Beijing Fangshan District Liangxiang Hospital, Beijing, China.
Eye Hospital, China Academy of Chinese Medical Science, Beijing, China.
J Clin Lab Anal. 2016 Sep;30(5):457-63. doi: 10.1002/jcla.21879. Epub 2015 Oct 29.
The level of glycated hemoglobin (HbA1c ) has been recognized as an important indicator of long-term glycemic control. However, the HbA1c measurement is not currently included as a diagnostic determinant in China. Current study aims to assess a candidate modified International Federation of Clinical Chemistry reference method for the forthcoming standardization of HbA1c measurements in China.
The HbA1c concentration was measured using a modified high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method. The modified method replaces the propylcyanide column with a C18 reversed-phase column, which has a lower cost and is more commonly used in China, and uses 0.1% (26.5 mmol/l) formic acid instead of trifluoroacetic acid. Moreover, in order to minimize matrix interference and reduce the running time, a solid-phase extraction was employed. The discrepancies between HbA1c measurements using conventional methods and the HPLC-ESI-MS method were clarified in clinical samples from healthy people and diabetic patients. Corresponding samples were distributed to 89 hospitals in Beijing for external quality assessment.
The linearity, reliability, and accuracy of the modified HPLC-ESI-MS method with a shortened running time of 6 min were successfully validated. Out of 89 hospitals evaluated, the relative biases of HbA1c concentrations were < 8% for 74 hospitals and < 5% for 60 hospitals. Compared with other conventional methods, HbA1c concentrations determined by HPLC methods were similar to the values obtained from the current HPLC-ESI-MS method.
The HPLC-ESI-MS method represents an improvement over existing methods and provides a simple, stable, and rapid HbA1c measurement with strong signal intensities and reduced ion suppression.
糖化血红蛋白（HbA1c）水平已被公认为长期血糖控制的重要指标。然而，目前在中国，HbA1c检测尚未被纳入诊断决定因素。当前研究旨在评估一种候选的改良国际临床化学联合会参考方法，用于中国即将开展的HbA1c检测标准化工作。
采用改良的高效液相色谱 - 电喷雾电离 - 质谱（HPLC - ESI - MS）法测量HbA1c浓度。改良方法用C18反相柱替代丙基氰柱，C18反相柱成本更低且在中国更常用，同时使用0.1%（26.5 mmol/L）的甲酸代替三氟乙酸。此外，为了最大限度减少基质干扰并缩短运行时间，采用了固相萃取。在健康人和糖尿病患者的临床样本中明确了使用传统方法与HPLC - ESI - MS法测量HbA1c之间的差异。相应样本分发给北京的89家医院进行外部质量评估。
成功验证了运行时间缩短至6分钟的改良HPLC - ESI - MS法的线性、可靠性和准确性。在评估的89家医院中，74家医院HbA1c浓度的相对偏差<8%，60家医院<5%。与其他传统方法相比，HPLC法测定的HbA1c浓度与当前HPLC - ESI - MS法获得的值相似。
HPLC - ESI - MS法是对现有方法的改进，提供了一种简单、稳定且快速的HbA1c测量方法，具有强信号强度和减少的离子抑制。
