Stereoselective determination of propranolol enantiomer in transgenic cell lines expressing human cytochrome P450.
- Author:
Xiao-feng YAN
1
;
Quan ZHOU
;
Tong-wei YAO
;
Su ZENG
;
Hang-Ping YAO
;
Ying-Nian YU
Author Information
- Publication Type:Journal Article
- MeSH: Chromatography, High Pressure Liquid; Cytochrome P-450 Enzyme System; genetics; physiology; Humans; Propranolol; analysis; metabolism; Reproducibility of Results; Stereoisomerism; Transgenes
- From: Journal of Zhejiang University. Medical sciences 2003;32(2):116-120
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a chiro chromatography for studying the stereoselective metabolism of propranolol (PL) in S(9) incubates prepared from transgenic cell lines expressing human cytochrome P450.
METHODSThe concentration of each enantiomer in S(9) incubates was determined through precolumn derivatization with GITC, followed by RP-HPLC assay using S-(+)-propafenone as internal standard.
RESULTSBaseline separations among the diastereomers of S(-)-P, internal standard and R(+)-PL were achieved on Shimpack CLC C(18)ODS column, with UV detection and methanol:water:glacial acetic acid (67/33/0.05,v/v/v) as mobile phase. The assay was simple, accurate, precise and specific. The linear range was from 5 to 500 micromol/L for each enantiomer. The limit of quantitation (LOQ) for the method was 5 micromol/L for the S(-)-and R(+)-PL, respectively (n=5, RSD<10%). The analytical method afforded average recoveries of 98.7 and 98.1% for S(-)- and R(+)-PL, respectively. The reproducibility of the assay was good (RSD<10%). The time-dependent studies showed that PL had the stereoselectivity of S-(-)-isomer in metabolism via CYP2C18 and the stereoselectivity of R-(+)-isomer in metabolism via CYP2C9.
CONCLUSIONThe method allows to study of stereoselective metabolism of PL in vitro.