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Rapid chiral high-performance liquid chromatographic assay for salmeterol and alpha-hydroxysalmeterol. Application to
M Zhang1, J P Fawcett, J P Shaw
1School of Pharmacy, University of Otago, Dunedin, New Zealand.
Abstract:
A rapid and sensitive chiral high-performance liquid chromatographic (HPLC) assay for the simultaneous determination of salmeterol and its principal human metabolite alpha-hydroxysalmeterol is described. The two pairs of enantiomers were resolved on a chiral-cellobiohydrolase column and detected by electrochemical detection at +700 mV. Standard curves were linear over the concentration range 0.1 to 4.0 microM for alpha-hydroxysalmeterol enantiomers and 2.5 to 40.0 microM for salmeterol enantiomers. Intra- and inter-day coefficients of variation were < 10%. The method was applied to a study of human hepatic metabolism in vitro which showed that microsomal metabolism of salmeterol to alpha-hydroxysalmeterol is not stereoselective.