HPLC荧光法测定高三尖杉酯碱在大鼠体内的分布

Study on Tissue Distribution of Homoharringtonine in Rats by HPLC Using Fluorescence Detection

关键词:  高三尖杉酯碱  高效液相色谱法  荧光检测  组织分布

Key words:  homoharringtonine  HPLC  fluorescence detection  tissue distribution

摘要:

目的 研究高三尖杉酯碱注射液在大鼠体内的组织分布，为该药进行临床研究及合理应用提供依据。方法 生物样品在碱性条件下通过氯仿提取，用Kromasil C₁₈柱(4.6 mm×150 mm，5 μm)，乙腈-10 mmol·L^-1 KH₂PO₄缓冲液(含0.2%三乙胺，用H₃PO₄调pH至2.5)(25∶75)为流动相，荧光检测波长为λex 280 nm，λem 320 nm。结果 绝对回收率为91.27%~102.4%，最低检测限为0.5 ng·mL^-1。SD大鼠单次尾静脉注射高三尖杉酯碱注射液0.5，1，2 h后主要效应器官的浓度分布特点是：C_骨髓>C_心，主要消除器官的浓度分布特点是C_肝>C_肺>C_肾。结论 本法准确，灵敏度高，可用于高三尖杉酯碱的体内过程研究。高三尖杉酯碱在骨髓中有较高浓度分布，应引起重视。

Abstract:

OBJECTIVE To establish an HPLC method for the study of tissue distribution of homoharringtonine in rats. METHODS The biological samples were extracted with chloroform in alkalinity condition. The Kromasil C18 column (4.6 mm×150 mm, 5 mm) was used as analytical column with a mobile phase consisted of acetonitrile-10 mmol·L^-1 phosphate buffer solution (0.2% triethylamine, pH 2.5) (25∶75) at a flow rate of 1.0 mL·min^-1 with fluorescence detector set at λex 280 nm and λem 320 nm. RESULTS The recovery of the method was 91.3%-102.4%, the LOD was 0.5 ng·mL^-1. In main effect tissues, the homoharringtonine concentration was followed as in order C_bm>C_coeur, and that was C_liver>C_lung>C_kidney in eliminated tissues. CONCLUSION The method has a good selectivity and reproducibility and can be used for the investigation of homoharringtonine in biosamples.