Assay of Glutathione in Individual Mouse Peritoneal Macrophages by Capillary Zone Electrophoresis with Electrochemical Detection

✍ Scribed by Wenrui Jin; Qian Dong; Xiaoying Ye; Daiqing Yu

• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 57 KB
• Volume: 285
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4749

✦ Synopsis

Glutathione (GSH) in individual mouse peritoneal macrophages was determined by capillary zone electrophoresis with electrochemical end-column amperometric detection at a gold/mercury amalgam microelectrode. A capillary of 20 m inner diameter was suitable for determination of GSH in an individual macrophage with a good signal-to-noise ratio. Individual macrophages could be drawn into the capillary with the aid of a inverted microscope. Lysing cells was studied in different buffer solutions. 0.01 mol/liter NaOH was selected to lyse macrophages. In this method, the usual calibration curve of GSH could not be used for the quantification of GSH in individual macrophages. It was found that standard GSH injected after analyzing each cell could be served as external standard. The whole cell injection and the lack of necessity of a derivatization reaction lead to more accurate and precise results. The average amount of GSH in an individual mouse peritoneal macrophage is 5.8 fmol, which is consistent with the literature value.