Specific thiol determination by micellar electrokinetic chromatography and on-column detection reaction with 2,2'-dipyridyldisulfide.
A new method for specific determination of glutathione using micellar electrokinetic chromatography and on-column reaction with 2,2'-dipyridyldisulfide is described. 2,2'-Dipyridyldisulfide and a sample of glutathione are injected consecutively into the capillary as two discrete plugs separated with a short plug of background electrolyte. Due to the differences in the mobilities of the 2,2'-dipyridyldisulfide and glutathione, on-column mixing and reaction occur. Glutathione is in this reaction quantitatively transformed into a mixed disulfide concomitantly with formation of an equimolar amount of the 2-thiopyridone which is further separated by micellar electrokinetic chromatography and determined spectrophotometrically at 343 nm. The concentration of glutathione is thus estimated indirectly from the result of 2-thiopyridone determination.
Glatz
Z
Z
Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic. glatz@chemi.muni.cz
Maslanová
H
H
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives / Q000737:chemistry*; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D004220:Disulfides / Q000737:chemistry*; D005978:Glutathione / Q000097:blood*; D006801:Humans; D013056:Spectrophotometry, Ultraviolet
D004220:Disulfides; C001105:2,2'-dipyridyl disulfide; D015082:2,2'-Dipyridyl; D005978:Glutathione