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Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser‐induced fluorescence detection
Author(s) -
Zhang Hua,
Jin Wenrui
Publication year - 2004
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.200305642
Subject(s) - derivatization , capillary electrophoresis , chromatography , fluorescein isothiocyanate , amino acid , chemistry , analyte , electroporation , fluorescein , fluorescence , laser induced fluorescence , reagent , lysis , electrophoresis , biochemistry , mass spectrometry , organic chemistry , physics , quantum mechanics , gene
A method for monitoring amino acids in single erythrocytes is described. For intracellular derivatization, reagent fluorescein isothiocyanate (FITC) was introduced into living cells by electroporation. For an 8 μm erythrocyte, the analytes were diluted by a factor of only 1.6. After completion of the derivatization reaction, a single cell was injected into the separation capillary tip and lysed there. The derivatized amino acids were separated by capillary electrophoresis, followed by laser‐induced fluorescence detection. Nine amino acids were quantitatively determined, with amounts of amino acids ranging from 3.8–32 amol/single cell.