Tzeng Huey-Fen, Chen Jin-Yi, Huang Shih-Wei, Wang Yu-Jing, Yang Chung-Shi
Department of Applied Chemistry, National Chi Nan University, Puli, Nantou, Taiwan.
Electrophoresis. 2007 Apr;28(8):1221-8. doi: 10.1002/elps.200600683.
A simple and rapid MEKC method was developed for the simultaneous determination of myo-inositol, scyllo-inositol, and glucose. Prior to electrophoretic separation, the nonfluorescent inositols and glucose were derivatized by N-methylisatoic anhydride at 25 degrees C for 10 min so that they could be detected by a fluorescence detector during separation. The good separation with high efficiency by MEKC was achieved in 13 min with a glycine buffer containing SDS and PEG 4000. Several parameters affecting the separation were studied, including the pH of BGE, the concentrations of glycine, SDS, and PEG 4000, and the applied voltage. Using glycerol as an internal standard, the linear ranges of the method for myo-inositol, scyllo-inositol, and glucose were 0.03-10, 0.01-5, and 0.05-20 mM; the concentration LODs of myo-inositol, scyllo-inositol, and glucose were 0.020, 0.0078, and 0.026 mM, respectively. The method was applied to analyze extracellular myo-inositol and glucose in the microdialysates from rat brain cortex of ischemia animal model and intracellular myo-inositol and scyllo-inositol in the rat brain extract.
建立了一种简单快速的毛细管电泳法，用于同时测定肌醇、 scyllo - 肌醇和葡萄糖。在电泳分离之前，非荧光性的肌醇和葡萄糖在25℃下用N - 甲基异邻苯二甲酸酐衍生化10分钟，以便在分离过程中能用荧光检测器进行检测。使用含有十二烷基硫酸钠（SDS）和聚乙二醇4000（PEG 4000）的甘氨酸缓冲液，在13分钟内通过毛细管电泳实现了高效的良好分离。研究了影响分离的几个参数，包括背景电解质（BGE）的pH值、甘氨酸、SDS和PEG 4000的浓度以及施加电压。以甘油作为内标，该方法测定肌醇、scyllo - 肌醇和葡萄糖的线性范围分别为0.03 - 10、0.01 - 5和0.05 - 20 mM；肌醇、scyllo - 肌醇和葡萄糖的浓度检测限分别为0.020、0.0078和0.026 mM。该方法应用于分析缺血动物模型大鼠脑皮质微透析液中的细胞外肌醇和葡萄糖，以及大鼠脑提取物中的细胞内肌醇和scyllo - 肌醇。
