Determination of homocysteine in human plasma by mekc and in-capillary detection reaction with 2,2-dipyridyldisulfide
| Authors | |
|---|---|
| Year of publication | 2003 |
| Type | Article in Periodical |
| Magazine / Source | Journal of Chromatography A |
| MU Faculty or unit | |
| Citation | |
| Field | Biochemistry |
| Keywords | Capillary electrophoresis; on-column derivatization; homocysteine |
| Description | In this paper, we present a new method for homocysteine quantitation in human plasma based on in-capillary reaction of homocysteine with 2,2-dipyridyldisulfide. Homocysteine is in this so called thiol-exchange reaction quantitatively transformed in mixed disulfide concomitantly with formation of equimolar amount of 2-thiopyridone that is further separated by MEKC and determined specifically at 343 nm. The concentration of homocysteine is thus estimated indirectly from the result of 2-thiopyridone determination. The linear detection range for concentration versus peak area for the assay was from 0.03 to 3 mM (correlation coefficient 0.994) with a detection limit of 6 mM and a limit of quantitation 20 mM. The inter-day reproducibility of the peak area and the migration time were 1.37 % and 0.05 %, respectively. The method is simple, relatively rapid and can be easily automated. Moreover the common CE apparatus with UV detector can be used to distinguish between normal and pathological hyperhomocysteinemia plasma samples. |
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