Specific thiol determination by micellar electrokinetic chromatography and on-column detection reaction with 2,2′-dipyridyldisulfide

• Published in: Journal of Chromatography A 2000-10, Vol.895 (1), p.179-187
• Main Authors: Glatz, Zdeněk, Mašlaňová, Hana
• Format: Article
• Language: English
• Subjects: 2,2'-Dipyridyl - analogs & derivatives; 2,2'-Dipyridyl - chemistry; 2,2′-Dipyridyldisulfide; Analysis; Analytical chemistry; Biological and medical sciences; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, Micellar Electrokinetic Capillary - methods; Disulfides - chemistry; Exact sciences and technology; General pharmacology; Glutathione; Glutathione - blood; Humans; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Miscellaneous. Technology; Other chromatographic methods; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Pharmacology. Drug treatments; Spectrophotometry, Ultraviolet; Sulfur compounds; Thiols
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(00)00475-1

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.