RP-HPLC法测定人肝脏微粒体中异丙酚的药物浓度

RP-HPLC determination of propofol concentration in human liver microsomes

目的:建立高效液相色谱法测定人肝脏微粒体中异丙酚的药物浓度,为进一步研究异丙酚的药物代谢提供新方法,并对异丙酚的体外代谢进行初步研究。方法:异丙酚与人肝脏微粒体共孵育后用乙腈提取,采用 HP1100色谱系统,以甲醇-乙腈-双蒸水(10:50:40)为流动相,流速1.2 mL·min^(-1),进样量50 μL,经 ODS Hypersil 色谱柱(4.6 mm×250 mm,5 μm)分离,紫外检测,波长为270 nm。结果:在0.25～10 μg·mL^(-1)。浓度范围内,异丙酚线性关系良好(r=0.9992),最低检测浓度为0.02 μg·mL^(-1),定量限为0.1 μg·mL^(-1),方法回收率在95.6%～101.3%之间,日内、日间 RSD 分别为1.15%～3.23%和2.34%～7.96%。结论:本方法检测异丙酚在人肝脏微粒体中的药物浓度较为快速、准确、灵敏、稳定,适用于异丙酚的体外药代动力学研究。

Objective ： To establish an RP - HPLC method for determining the concentrations of propofol in human liver microsomes, and to provide a new way for investigating the metabolism of propofol. Methods：After incubation in human liver microsomes,propofol was extracted by acetonitrile. The HP 1100 HPLC system was adopted,including a quaternary pump, a manual injector, a variable wavelength detector and a fluorescence detector. The mobile phase was a mixture of methanol- acetonitrile- water（ 10： 50： 40） ,and the flow rate was 1.2 mL · min-1.50 μL of sample was injected in and separated by ODS Hypersil column（4. 6 mm × 250 mm,5 μm）. The UV detector was operated at 270 nm. Results： The assay was linear over the range of 0. 25 - 10 μg·mL^-1 for propofol （ r = 0. 9992 ）. The limit of detection was 0. 02 μg·mL^-1 and the limit of quantitation was 0. 1 μg·mL^-1. The method afforded recoveries was 95.6% - 101.3% , and the intra - day and inter - day RSD were 1. 15% - 3.23% and 2. 34% -7.96% ,respectively. Conclusion： The proposed method is rapid, accurate, sensitive, reliable and can be used for pharmacokinetic studies of propofol in vitro.