Guo Chao, Zhang Qianchun, Bao Linchun, Li Hongliang, Zheng Qingfeng, Guo Zhenpeng, Chen Yi
Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Anal Methods. 2020 Jul 2;12(25):3277-3284. doi: 10.1039/d0ay00384k.
As a fast, high-performance and cost-effective separation technique, capillary electrophoresis (CE) is applicable to the screening and diagnosis of diseases such as thalassemia. However, it is often not preferred due to its unrepeatable and/or irreproducible migration times. Herein, we propose a stable version of CE that uses migration charge density instead of the migration time to plot the electropherogram. The peak position is now independent of the applied voltage or current and the capillary geometry and is also insensitive to temperature. Its applicability was demonstrated in the quantitative analysis of human hemoglobin. On a laboratory-built device, with a running buffer simply consisting of 3.0 M acetic acid and 0.1% (w/v) hydroxyethyl cellulose, it allows a direct injection of whole blood samples and all the concerned globin chains, α, β, Aγ and Gγ can be well separated in 15 minutes. The resolution of α/β, β/Aγ, and Aγ/Gγ reached 4.4, 3.1 and 5.3, respectively. The intra- and inter-day precisions for the peak position based on the migration charge densities were below 0.6%. Its diagnostic applicability was validated in the analysis of several real blood samples from newborns, children and adults, and its capacity was demonstrated to screen and define the type of thalassemia.
作为一种快速、高性能且经济高效的分离技术，毛细管电泳（CE）适用于地中海贫血等疾病的筛查和诊断。然而，由于其迁移时间不可重复和/或不可再现，它往往不被优先选用。在此，我们提出了一种稳定版的毛细管电泳技术，该技术使用迁移电荷密度而非迁移时间来绘制电泳图。现在，峰位置与施加的电压或电流以及毛细管几何形状无关，并且对温度也不敏感。其适用性在人血红蛋白的定量分析中得到了证明。在一台实验室自制的设备上，运行缓冲液仅由3.0 M乙酸和0.1%（w/v）羟乙基纤维素组成，它允许直接进样全血样本，并且所有相关的珠蛋白链，α、β、Aγ和Gγ在15分钟内可以得到很好的分离。α/β、β/Aγ和Aγ/Gγ的分辨率分别达到4.4、3.1和5.3。基于迁移电荷密度的峰位置的日内和日间精密度均低于0.6%。其诊断适用性在对来自新生儿、儿童和成人的几份实际血样的分析中得到了验证，并且展示了其筛查和确定地中海贫血类型的能力。
