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Enantioselective determination of (R)- and (S)-sotalol in human plasma by on-line coupling of a restricted-access material precolumn to a cellobiohydrolase I-based chiral stationary phase.

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Department of Pharmaceutical Chemistry, Albert-Ludwigs-University, Albertstr. 25, D-79104 Freiburg im Breisgau, Germany.


A liquid chromatographic column-switching method for the enantioselective determination of (RS)-sotalol in plasma was developed and validated. The method is based on the on-line coupling of a precolumn filled with the restricted access material LiChrospher ADS to a cellobiohydrolase I-based chiral stationary phase (CSP). The plasma samples were injected onto the precolumn using a mobile phase containing 1% methanol in 10 mM phosphate buffer at pH 7.4 for 10 min for the removal of matrix components. The analytes were transferred to the CSP for their enantiomeric separation by backflushing the precolumn with 15% 2-propanol in 10 mM phosphate buffer (pH 7.0) including 0.05 mM EDTA. The quantitative determination of the sotalol enantiomers was possible upon addition of the internal standard (S)-atenolol. The method was validated showing a good linearity in the concentration range from 25 to 1000 microg l(-1) for each enantiomer. The average values of the intra- and inter-day variability were 1.17% and 3.42%, respectively, for (R)-sotalol and 1.24% and 1.99%, respectively, for (S)-sotalol. The applicability of the method to real world samples has been proven by means of two pharmacokinetic studies. They revealed that the pharmacokinetic properties of the sotalol enantiomers do not differ significantly neither for healthy young volunteers after single dose application nor for elder patients in the steady state.

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