HPLC-荧光法测定兔血浆中伊曲康唑及其代谢产物羟基伊曲康唑的浓度

Content Determination of Itraconazole and Its Metabolit Hydroxy-itraconazole in Rabbit Plasma by HPLC-Fluorometric Methods

目的：建立可同时测定兔血浆中伊曲康唑（ITZ）及其代谢产物羟基伊曲康唑（HITZ）浓度的高效液相色谱-荧光法。方法：色谱柱为Agilent Eclipse C18，流动相为0．1％三乙胺缓冲液（85％磷酸调pH为2．5）-乙腈（63：37），流速为1．0mL·min^-1，荧光检测激发波长为260nm，发射波长为365nm，柱温为室温。结果：ITZ和HITZ的线性范围分别为7．4～2960（r=0．9991）、5．4～2160ng·mL^-1（r=0．9993），检测限分别为0．518、0．318ng·mL^-1；ITZ、HITz的提取回收率和方法回收率均在80％以上，日内、日间RSD均小于4．47％。结论：所建立的检测方法操作简便、灵敏度高、专一性强、结果可靠，可用于兔血浆中ITZ和HITZ浓度的同时测定和药动学研究。

OBJECTIVE： To establish a high- performance liquid chromatographic （HPLC） - fluorometric method for the concentration determination of itraconazole （ITZ） and its metabolite hydroxy-itraconazole （HITZ） in rabbit plasma simulta- neously. METHODS： Separation was achieved with Agilent Eclipse C18, the mobile phase consisted of acetonitrile （adjusted to pH ： 2.5 with 85% phosphoric acid） -water （63 ： 37）, and the flow rate was 1.0 mL · min^- 1. The excitation and emission wavelengths were 260 nm and 365 nm. The analysis was performed at room temperature..RESULTS： The linear concentration range of ITZ and HITZ were 7.4--296 0 ng · mL^-1（r= 0.999 1）and 5.4--216 0 ng · mL^-1（r = 0.999 3）, respectively. The limit of detection was 0.518 and 0.318 ng · mL^-1, respectively. The extraction and method recovery rate of ITZ and HITZ were more than 80% .The intra- day and inter- day RSD of ITZ and HITZ were less than 4.47%. CONCLUSIONS： The method is simple, sensitive, accurate and reliable for monitoring ITZ and HITZ in rabbit plasma and pharmacokinetic studies.