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Chiral reversed phase high-performance liquid chromatography for determining propranolol enantiomers in transgenic Chinese hamster CHL cell lines expressing human cytochrome P450
J. Biochem. Biophys. Methods 54(2002)369-376,-0001,():
An enantioselective assay for S-(-)-and R-(+)-propranolol in transgenic Chinese hamster CHL cell lines, expressing human cytochrome P450 (CYP), was developed. The method involves extraction of propranolol from the S9 incubates, using S-(+)-propafenone as internal standard, chiral derivatization with 2,3,4,6-tetra-O-h-D-glucopranosyl isothiocyanate and quantitation by reversed phase high-performance liquid chromatography system with UV detection (k=220nm). A baseline separation of propranolol enantiomers was achieved on a 5-Am reverse-phase ODS column, with a mixture of methanol/water/glacial acetic acid (67:33:0.05, v/v) as mobile phase. The assay is linear from 5 to 500 AM for each enantiomer. The analytical method affords average recoveries of 99.2% and 98.8% for S-(-)-and R-(+)-propranolol, respectively. The limit of quantitation for the method is 5 AM for both S-(-)-and R-(+)-propranolol. The reproducibility of the assay is satisfactory (RSD<10%). The method allowed study of the depletion of S-(-)-and R-(+)-propranolol in transgenic Chinese hamster CHL cell lines expressing CYP3A4, CYP2C18 and CYP2C9.
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