OBJECTIVE A simple assay method for paeonol in mice serum by reversed-phase high performance liquid
chromatography (RP-HPLC)
was established.METHODS Serum sample was extracted one time with chloroform for RP-HPLC analysis. The RP-HPLC assay was conducted using Lichrospher C 18 column( 4.6 mm×250 mm, 5 μm) with methanol-acetonitrile-water(30∶40∶30 ) as mobile phase at a flow rate of 0.8 mL·min -1 . 20 μL of sample solution were injected into the C 18 column where the temperature was 25℃.The
detection wavelength was 274 nm.RESULTS The limit of detection for paeonol was 0.01 mg·L -1 in serum sample.The linearity range for paeonol was obtained from 0.02 mg·L -1 to 12.80 mg·L -1 . The linear regression equation was Y= 2712.1836 X- 5.7137 ,r= 0.9999 ,n=5. The recoveries of paeonol were ( 101.2 ± 1.2 )%. The average recovery of paeonol in samples was ( 79.7 ± 3.0 )%. The relative standard deviations of the intra-day and inter-day were less than 10%(n=5).CONCLUSION The method is sensitive, simple, rapid, and accurate, which may be used for blood concentration monitoring and pharmacokinetics study for paeonol.