高效液相-荧光检测法测定兔血浆中羟基喜树碱的含量

Determination of 10-Hydroxycamptothecin by HPLC with Fluorescence Detection in Rabbit Plasma

目的 建立高效液相-荧光检测法测定免血浆中羟基喜树碱含量的方法学。方法 色谱柱为Discovery C18（15cm×4．6mm，5μm），流动相为柠檬酸缓冲液-乙腈-75nmol·L^-1磷酸二氢钾（70：23：7），75nmol·L^-1磷酸二氢钾中含1％三乙胺。流速为1．oml·min^-1，柱温为40℃，荧光检测波长为λex363nm和λem530nm。结果 该方法的线性范围为13．7656-1957．4468ng·mL^-1（r=0．9993）。提取回收率和方法回收率分别为80．900／00～103．59％．102．72％～108．16％．日内RSD≤7．49％，日间RSD≤9．40％。最低检测限为5．2ng·mL^-1。结论 该方法专属性强，重现性好，操作简便，适用于羟基喜树碱的药代动力学研究和血药浓度监测。

OBJECTIVE A HPLC method with fluorescence detection for determination of 10- Hydroxycamptothecin in rabbit plasma . METHODS The chromatographic condition were as follows a Eclipse Discovery C18 column （4. 6mm × 150mm, 5μm） was eluted with mobile phase citric acid buffer-acetonitrile75nmKH2PO4 （70 ： 23 ： 7, containing 1%triethylamine ）. the column temperature was 40℃, the flow rate was 1.0ml ·min^-1 with the fluorescence detection set at λex 363nm and λem 530nm. RESULTS The method was proved to be linear in the range of 13. 7656- 1957. 4468ng·mL^-1 with correlation coefficients 0. 999343. the average recovery was 102.22%-108.16%, RSD of intra-day and inter-day were no more than 7.49% and 9. 40% . The minimum detection limit was 5.2ng· mL^-1 CONCLUSION The method is simple ,good specificity , reproducibility, can used to investigate the pharma-cokinetics and determinate the concentration of 10- Hydroxycamptothecin.