Specific Determination of Glutathione using Micellar Elektrokinetic Chromatography and On-Column Reaction with 2,2-dipyridyldisulphide

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Authors

GLATZ Zdeněk MAŠLAŇOVÁ Hana

Year of publication 1999
Type Article in Proceedings
Conference Book of Abstract, 1st International Symposium on Separations in the Biosciences, Amsterdam, Netherlands
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords Micellar Elektrokinetic Chromatography; On-Column Derivatization; Thiols; Glutathione
Description The on-column reaction of glutathione with the reactive disulphide 2,2-dipyridyldisulphide was used for the specific detection of glutathione during micellar electrokinetic chromatography: In this set-up the front end of the capillary is used as the reaction chamber. 2,2-dipyridyldisulphide and a sample of glutathione are injected consecutively in the capillary as two discrete plugs separated with short plug of background electrolyte. Due to the differences in the mobilities of the 2,2-dipyridyldisulphide and glutathione on column mixing and reaction occur. This reaction is rapid and quantitative, because thiol form is stabilised by thiol-thione tautomerism. This fact also causes a large shift of the absorption maxima toward longer wavelength. Glutathione is thus quantitatively transformed into a mixed disulphide concomitantly with formation of equimolar amounts of the 2-thiopyridone that is further separated by micellar electrokinetic chromatography and determined spectrophotometrically at 343 nm.
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