家兔单硝酸异山梨酯血浆浓度高效液相色谱测定方法

Study on Determination of Isosorbide-5-mononitrate in Animal Plasma by HPLC

建立了动物血浆单硝酸异山梨酯检测的高效液相色谱方法。血浆经高速离心后,以Beckman ODS为色谱柱,流动相为乙腈-水(10∶90,v/v),流速为1.0 mL/min,柱温为35℃,检测波长为220 nm。并用此方法测定4只家兔单剂量口服40 mg单硝酸异山梨酯片后的血药浓度经时过程。结果显示:所建立的测定单硝酸异山梨酯血药浓度HPLC方法在5～500 ng/mL范围内线性关系良好,定量下限为5 ng/mL;高、中、低3个浓度的方法回收率在92%～107%之间,日内精密度和日间精密度约在6%以内。结论本方法简便、快速、准确、可靠,适用于动物血浆中单硝酸异山梨酯浓度的测定。

HPLC method was developed for assay of isosorbide mononitrate（IM） blood level in rabbit.After the centrifugation of blood,the reconstituted samples were analyzed on Beckman ODS column at the wavelength of 220 nm using a mobile phase consisting of acetonitrile-d.d.H2O（10：90,v/v） at a flow rate of 1.0 mL/min and column oven at 35 ℃.IM blood levels in the rabbits exposed to 40-mg IM tablet were assayed.There existed linear relationship between the peak areas in the HPLC system and the concentrations of IM in the spiked samples with the low detection limit of 5 ng/ml.The method recoveries of 92%~107% were found when the samples of high,middle and low levels of interest were loaded.Moreover,the inter-day and intra-day variation coefficients were estimated in 6%.The method was simple,rapid,accurate,and reliable,which was suitable for the determination of IM blood level in rabbit.