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Determination of human blood glucose levels using microchip electrophoresis
Author(s) -
Maeda Eiki,
Kataoka Masatoshi,
Hino Mami,
Kajimoto Kazuaki,
Kaji Noritada,
Tokeshi Manabu,
Kido Junichi,
Shinohara Yasuo,
Baba Yoshinobu
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600795
Subject(s) - chromatography , hexokinase , detection limit , chemistry , reproducibility , capillary electrophoresis , glucokinase , linear range , linear relationship , electropherogram , analytical chemistry (journal) , biochemistry , enzyme , glycolysis , statistics , mathematics
A high‐performance monitoring system for human blood glucose levels was developed using microchip electrophoresis with a plastic chip. The combination of reductive amination 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 μM of glucose ( r 2 = 0.9963; p <0.01), and the detection limit was 0.92 μM. 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.