Specific thiol determination by on-column derivatization during MEKC

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Authors

ŠEVČÍKOVÁ Petra GLATZ Zdeněk

Year of publication 2002
Type Article in Proceedings
Conference Book of abstracts of 6th Meeting of biochemists and molecular biologists
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords thiol; MEKC
Description The latest development in derivatization in capillary electrophoresis is on-column mode. In this set-up a derivatization reaction is performed during the electrophoretic separation. In principle, the front end of the capillary is used as the reaction chamber. The analyte and the reagent are injected separately and are mixed because of their different electrophoretic mobilities. Various on-column reactions have been demonstrated so far however only for derivatization of the amino group containing compounds. In all these applications the simplified derivatization scheme is the same. The detector nonresponding analytes A, B and C are converted into the derivatives AX, BX and CX that have enhanced detector response properties, in the chemical reaction with the derivatization reagent X : A + B + C + .... + X Ž AX + BX + CX + .... In this study a new type of on-column reaction detection is reported. The analytes A, B, and C react with the reagent XX forming the derivatives AX, BX and CX, and a reaction by-product X. In contrast to the methods mentioned above, this highly absorbing by-product X is used for the detection : A + B + C + .... + XX Ž AX + BX + CX + .... n X Considering that the by-product X is identical for all analytes, the separation of the analytes has to be performed before the reaction. As an example of the utility of this concept the thiol-disulfide exchange reaction of 2,2-dipyridyldisulphide DPDS was used for detection of thiol RSH.
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