Specific determination of cysteine in human urine by capillary micellar electrokinetic chromatography

• Published in: Journal of separation science 2003-06, Vol.26 (8), p.734-738
• Main Authors: Ševčíková, Petra, Glatz, Zdeněk
• Format: Article
• Language: English
• Subjects: 2,2′‐Dipyridyl disulfide; 2′-Dipyridyl disulfide; Biological and medical sciences; Capillary micellar electrokinetic chromatography; Cysteine; Investigative techniques, diagnostic techniques (general aspects); Medical sciences; Miscellaneous. Technology; On-column detection; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Thiol
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200301372

This paper describes a method for the analysis of cysteine in human urine using capillary micellar electrokinetic chromatography and on‐column reaction with 2,2′‐dipyridyl disulfide. In this reaction cysteine is quantitatively transformed into a mixed disulfide concomitantly with formation of an equimolar amount of 2‐thiopyridone that is further separated by capillary micellar electrokinetic chromatography and determined spectrophotometrically at 343 nm. The concentration of cysteine is thus estimated indirectly from the result of 2‐thiopyridone determination. The linear detection range for concentration versus peak area for the assay is from 0.05 to 5 mM (correlation coefficient 0.989) with a detection limit of 2.5 μM and a limit of quantitation of 8.5 μM. The inter‐day reproducibility of the peak area was 2.18% and the inter‐day reproducibility of the migration time 0.51%. The method is relatively rapid, simple, and can be easily automated. Moreover, its detection limit covers the concentration range at which cysteine is present in biological samples such as human urine.