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采用无接触电导检测的毛细管电泳法对细胞培养后营养培养基以及人脂肪组织微透析液中葡萄糖和乳酸水平的控制。

The Control of Glucose and Lactate Levels in Nutrient Medium After Cell Incubation and in Microdialysates of Human Adipose Tissue by Capillary Electrophoresis with Contactless Conductivity Detection.

作者信息

Tůma Petr

机构信息

Department of Hygiene, Third Faculty of Medicine, Charles University, Prague, Czech Republic.

出版信息

Methods Mol Biol. 2019;1972:95-108. doi: 10.1007/978-1-4939-9213-3_7.

Abstract

Two methods of capillary electrophoresis with contactless conductivity detection have been developed for monitoring the levels of glucose and lactate in clinical samples. The separations are performed in uncoated fused silica capillaries with inner diameter 10 or 20 μm, total length 31.5 cm, length to detector 18 cm, using an Agilent electrophoretic instrument with an integrated contactless conductivity detector. Glucose is determined in optimized background electrolyte, 50 mM NaOH with pH 12.6 and 2-deoxyglucose is used as an internal standard; the determination of lactate is performed in 40 mM CHES/NaOH with pH 9.4 and lithium cations as an internal standard. Both substances are determined in minimal volumes of (1) nutrient media after cell incubation, and (2) microdialysates of human adipose tissue; after dilution and filtration as the only treatment of the sample. The migration time of glucose is 2.5 min and that of lactate is 1.5 min with detection limits at the micromolar concentration level. The developed techniques are suitable for sequential monitoring of glucose and lactate over time during metabolic experiments.

摘要

已开发出两种采用非接触式电导检测的毛细管电泳方法,用于监测临床样本中葡萄糖和乳酸的水平。分离在未涂层的熔融石英毛细管中进行,内径为10或20μm,总长31.5cm,到检测器的长度为18cm,使用配备集成非接触式电导检测器的安捷伦电泳仪。葡萄糖在优化的背景电解质中测定,即pH值为12.6的50mM NaOH,并使用2-脱氧葡萄糖作为内标;乳酸的测定在pH值为9.4的40mM CHES/NaOH中进行,并使用锂阳离子作为内标。两种物质均在最小体积的(1)细胞培养后的营养培养基和(2)人脂肪组织的微透析液中测定;样品仅经过稀释和过滤处理。葡萄糖的迁移时间为2.5分钟,乳酸的迁移时间为1.5分钟,检测限为微摩尔浓度水平。所开发的技术适用于在代谢实验期间随时间顺序监测葡萄糖和乳酸。

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