Specific thiol determination by micellar electrokinetic chromatography and on-column detection reaction with 2,2-dipyridyldisulphide
| Authors | |
|---|---|
| Year of publication | 2000 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography A |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | thiol; on-column derivatization; MEKC |
| Description | A new method for specific determination of glutathione using micellar electrokinetic chromatography and on-column reaction with 2,2 -dipyridyldisulphide is described. 2,2-dipyridyldisulphide 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-dipyridyldisulphide and glutathione, on-column mixing and reaction occur. Glutathione is in this reaction quantitatively transformed into a mixed disulphide concomitantly with formation of 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. |
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