Determination of Lehmannine in Mice Plasma and Tissue by RP-HPLC

OBJECTIVE:To establish a method for the determination of the concentration of lehmannine in mice plasma and tissues. METHODS: RP-HPLC assay was carried out using a VP-ODS C18 column at 40 ℃. The mobile phase consisted of acetonitrile-0.02 mol·L-1 ammonium acetate buffer-triethylamine (30∶70∶0.04, V/V/V) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 210 nm. 20 mice were intragastrically administered with lehmannine (40 mg· kg-1), then the samples were taken from plasma and tissues at different time for the determination of the concentration of lehmannine. RESULTS: The linear ranges of lehmannine in plasma and tissue were 0.175～7.00 μg·mL-1 and 0.088～3.50 μg·g-1, respectively (r0.999 4 for both). The recoveries of both were higher than 88.77%, and the intra-day RSD and inter-day RSD were less than 8.82 % and 11.07%, respectively. After ig administration of lehmannine, there was an extensive distribution of lehmannine in mice, which were detected in plasma, heart, kidney, liver and brain. The concentration of lehmannine was significantly higher in plasma than in tissue. CONCLUSIONS: The developed method was simple, reliable and accurate. It could be applied to study pharmacokinetics and distribution of lehmannine in tissues.