This HPLC assay with o-phthalaldehyde precolumn derivatization is used to measure the total, oxidized, and protein-bound forms of glutathione in human blood, plasma, and rat tissue. Total glutathione (i.e., sum of reduced, oxidized, and protein-bound fractions) was determined after reduction with dithiothreitol and protein precipitation with perchloric acid (PCA). A preliminary selective blockage of free sulfhydryl groups with N- ethylmaleimide was necessary to evaluate the different oxidized forms. The assay showed high sensitivity (0.999). Samples, after PCA acidification, were stable at room temperature and 4°C for 3 days, and at -20°C and -80°C for >1 month. The method (involving automated derivatization) not only is very rapid and simple but also allows immediate processing of many different biological samples.
|Number of pages||7|
|Publication status||Published - 1995|
- chromatography, reversed-phase
- sample handling
ASJC Scopus subject areas
- Clinical Biochemistry