On-Chip Derivatization for Determining Glutathione in Single Cell Using Microchip Electrophoresis and Laser Induced Fluorescence Detection

Abstract

Abstract:

A simple method for the detection of glutathione(GSH) in single human erythrocyte was developed by using on-chip labeling by adding the 2,3-naphthalene-dicarboxaldehyde(NDA) simply in microchip electrophoresis buffer,with functional integration of cell sampling,single cell loading docking,lysing,dynamic labeling,electrophoresis separation and LIF detection.After lysing,the reaction between the released GSH and NDA included in electrophoresis buffer can be completed within a migrating distance of 0.5 cm in separation channel during electrophoresis.The average separation efficiency for GSH was 2.4×10⁶ plates/m) which was not significantly different from off-chip derivatization.The GSH migration time on the on-chip mode and off-chip method were 116 s and 110 s,respectively.The present method can minimize the dilution of the contents of single cell during derivatization,simplify the cell treatment and save the sample and reagent.A half life of 5 days of GSH in human erythrocyte was found by using this method.

Key words: On-chip derivatizatian, Microfluidic chip, Single cell, Glutathione

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LING Yun-Yang, YIN Xue-Feng, FANG Zhao-Lun. On-Chip Derivatization for Determining Glutathione in Single Cell Using Microchip Electrophoresis and Laser Induced Fluorescence Detection[J]. Chem. J. Chinese Universities, 2004, 25(S1): 167.

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On-Chip Derivatization for Determining Glutathione in Single Cell Using Microchip Electrophoresis and Laser Induced Fluorescence Detection

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