Subject Area物理化学
Determination of paeonol O-demethylation activities in vitro by ultra-performance liquid chromatography with MS detection (UPLC-MS)
Liu HX(刘慧鑫); Yang L(杨凌); Yang; L
Conference Name3rd asian pacific regional meeting of the international society for the study of xenobiotics
Conference Date2009-5-10
Conference Place泰国
Funding Organization国际药物代谢学会
AbstractO-demethylation of paeonol has been implicated as an important mechanism of paeonol elimination. To characterize paeonol metabolism by O-demethylation, a rapid and specific ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS) method was developed for the qualitative and quantitative determination of paeonol O-demethylation activity in human liver microsomes (HLMs). HLMs incubation system with paeonol resulted in the formation of a single product peak. Following the purification, this peak was confirmed as O-demethylated metabolite of paeonol (2,4-Dihydroxyacetophenone, DHA) by MS and NMR. The method was validated for the determination of DHA with respect to specificity, linearity, detection limit, recovery, stability, precision and accuracy. The chromatographic separation was achieved on a UPLC BEH C18 column (50 mm × 2.1 mm i.d., 1.7 μm), with phase of acetonitrile-water (ratio 30:70). Selective ion reaction (SIR) monitor was specific for paeonol, DHA and I.S.. The method was linear over the concentration range 0.5-100 μM for DHA in spiked HLMs. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. DHA were stable at 4°C for at least 72 h in spiked liver microsomes samples. The method was successfully used to determine the kinetics of DHA activities toward paeonol in liver microsomes from different speices.
WOS IDWOS:000269483300123
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Document Type会议论文
Corresponding AuthorYang; L
Recommended Citation
GB/T 7714
Liu HX,Yang L,Yang,et al. Determination of paeonol O-demethylation activities in vitro by ultra-performance liquid chromatography with MS detection (UPLC-MS)[C],2009:63/1.
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