Monitoring of dithiothreitol clearance by means of micellar electrokinetic chromatography
| Authors | |
|---|---|
| Year of publication | 2002 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography B |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | DTT; MEKC; on-column; |
| Description | A new method for specific determination of dithiothreitol (DTT) using micellar electrokinetic chromatography and on-column reaction with reactive disulfide - 2,2-dipyridyldisulfide is described. DTT is in this reaction quantitatively transformed into a mixed disulfide concomitantly with formation of equimolar amount of the 2-thiopyridone that is further separated by micellar electrokinetic chromatography and determined spectrophotometrically at 343 nm. The concentration of DTT 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 2.5 mM (correlation coefficient 0.993) with a detection limit of 2.5 mM. The inter-day reproducibility of the peak area was 1.35 % and the inter-day reproducibility of the migration time 0.56 %. The method can be applied for DTT monitoring both in chemical and biological systems. |
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