RP-HPLC法测定小鼠血浆和组织中拉马宁碱的浓度

目的:建立测定拉马宁碱在小鼠血浆和组织中浓度的方法。方法:采用反相高效液相色谱(RP-HPLC)法,色谱柱为VP-ODS C18,流动相为乙腈-0.02mol·L-1醋酸铵缓冲液-三乙胺(30∶70∶0.04),流速为1.0mL·min-1,检测波长为210nm,柱温为40℃。20只小鼠灌胃给予拉马宁碱(40mg·kg-1),不同时间点取样测定血浆和组织中的药物浓度。结果:拉马宁碱在血浆和组织中检测深度的线性范围分别为0.175～7.00μg·mL-1和0.088～3.50μg·g-1(r均>0.9994),回收率均>89.77%,日内、日间RSD值分别小于8.82%、11.07%。拉马宁碱在小鼠体内分布较广,在血浆、心脏、肾脏、肝脏和大脑中均能检测到,且在血浆中的药物浓度明显高于组织中。结论:本方法简单、可靠、准确度高,可用于拉马宁碱的药动学和组织分布研究。

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--7.00ug·mL^-1 and 0.088--3.50ug·mL^-1, respectively （r〉0.999 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 pharmacokineties arid distribution of lehmannine in tissues.