Assay of glutathione in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection

Citation
Wr. Jin et al., Assay of glutathione in individual mouse peritoneal macrophages by capillary zone electrophoresis with electrochemical detection, ANALYT BIOC, 285(2), 2000, pp. 255-259
Citations number
25
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
ANALYTICAL BIOCHEMISTRY
ISSN journal
0003-2697 → ACNP
Volume
285
Issue
2
Year of publication
2000
Pages
255 - 259
Database
ISI
SICI code
0003-2697(20001015)285:2<255:AOGIIM>2.0.ZU;2-2
Abstract
Glutathione (GSH) in individual mouse peritoneal macrophages was determined by capillary zone electrophoresis with electrochemical end-column amperome tric detection at a gold/mercury amalgam microelectrode. A capillary of 20 mum inner diameter was suitable for determination of GSH in an individual m acrophage with a good signal-to-noise ratio. Individual macrophages could b e drawn into the capillary with the aid of a inverted microscope. Lysing ce lls was studied in different buffer solutions. 0.01 mol/liter NaOH was sele cted to lyse macrophages. In this method, the usual calibration curve of GS H could not be used for the quantification of GSH in individual macrophages . It was found that standard GSH injected after analyzing each cell could b e served as external standard. The whole cell injection and the lack of nec essity of a derivatization reaction lead to more accurate and precise resul ts. The average amount of GSH in an individual mouse peritoneal macrophage is 5.8 fmol, which is consistent with the literature value, (C) 2000 Academ ic Press.