Determination of homocysteine in human plasma by mekc and in-capillary detection reaction with 2,2-dipyridyldisulfide

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Publikace nespadá pod Pedagogickou fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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ŠEVČÍKOVÁ Petra GLATZ Zdeněk TOMANDL Josef

Rok publikování 2003
Druh Článek v odborném periodiku
Časopis / Zdroj Journal of Chromatography A
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Obor Biochemie
Klíčová slova Capillary electrophoresis; on-column derivatization; homocysteine
Popis 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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