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Coupled column chromatography for separation and determination of enantiomers of phenylcarbamic acid derivatives in serum.

Columns packed with vancomycin coupled to an achiral C18 column and beta-cyclodextrin were used for the separation and the determination of enantiomers of alkoxysubstituted esters of phenylcarbamic acid in blood serum. The method involves off-line SPE, the separation of the racemate on a reversed-phase stationary phase, and the separation of the enantiomers on a chiral stationary phase. The limit of detection was 1.0 microg/ml for the vancomycin column and 10.0 microg/ml for the beta-cyclodextrin column in standard solution. In vitro degradation studies of enantiomers demonstrated a difference in the concentation of the enantiomers after the treatment. It was found that the rate constants of R(-)- and S(+)-forms of enantiomers are not significantly different.