Determination of human blood glucose levels using microchip electrophoresis
Determination of human blood glucose levels using microchip electrophoresis
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DOI：
10.1002/elps.200600795
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发表时间：
2007-08-01
期刊：
影响因子：
2.9
通讯作者：
Baba, Yoshinobu
中科院分区：
文献类型：
--
作者：
Maeda, Eilki;Kataoka, Masatoshi;Baba, Yoshinobu
使用带有塑料芯片的微芯片电泳开发了一种高性能的人体血糖水平监测系统。将还原胺化作为葡萄糖标记与荧光 2-氨基吖啶酮 (AMAC) 和葡萄糖-硼酸盐复合物形成相结合，实现了即使在血液样本等复杂基质中也能高度选择性地检测葡萄糖。在 AMAC 标记血浆的电泳图上观察到的单峰迁移时间与葡萄糖标准溶液的迁移时间非常相似。用己糖激酶或葡萄糖激酶处理血浆进行葡萄糖磷酸化，导致峰从大约 145 秒移动到 70 秒，分别对应于葡萄糖和葡萄糖-6-磷酸。双对数图显示葡萄糖浓度和荧光强度在1-300μM葡萄糖范围内呈线性关系（r（2）= 0.9963；p < 0.01），检测限为0.92μM。此外，将根据三名受试者的标准曲线估算的血糖浓度与使用葡萄糖脱氢酶的常规比色分析获得的结果进行比较。根据简单线性回归分析，方法之间观察到良好的相关性（P < 0.05）。该测定的重现性约为 6.3-9.1% (RSD)，日内和日间重现性分别为 1.6-8.4 和 5.2-7.2%。该系统使我们能够以高灵敏度和准确度测定血糖，并将适用于临床诊断。
A high-performance monitoring system for human blood glucose levels was developed using microchip electrophoresis with a plastic chip. The combination of reductive arnination as glucose labeling with fluorescent 2-aminoacridone (AMAC) and glucose-borate complex formation realized the highly selective detection of glucose even in a complex matrix such as a blood sample. The migration time of a single peak, observed on an electropherogram of AMAC-labeled plasma, closely resembled that of glucose standard solution. The treatment of plasma with hexokinase or glucokinase for glucose phosphorylation resulted in a peak shift from approximately 145 to 70 s, corresponding to glucose and glucose-6-phosphate, respectively. A double-logarithm plot revealed a linear relationship between glucose concentration and fluorescence intensity in the range of 1-300 mu M of glucose (r(2) = 0.9963; p < 0.01), and the detection limit was 0.92 mu M. Furthermore, blood glucose concentrations estimated from the standard curves of three subjects were compared with results obtained by conventional colorimetric analysis using glucose dehydrogenase. Good correlation was observed between methods according to simple linear regression analysis (P < 0.05). The reproducibility of the assay was about 6.3-9.1% (RSD) and the within-days and between-days reproducibility were 1.6-8.4 and 5.2-7.2%, respectively. This system enables us to determine blood glucose with high sensitivity and accuracy, and will be applicable to clinical diagnosis.