固相萃取—高效液相色谱法测定兔血浆中山奈素的含量

兔的血浆样品经过β-葡萄糖醛酸酶和硫酸酯酶酶解、提取、C18固相萃取小柱富集净化后,在Agilent SB-C18柱(250mm×4.6 mm,5μm)上,以甲醇-0.4%磷酸(55∶45)为流动相,流速为1.0 mL.min-1,检测波长为360 nm,对山奈素进行测定的结果表明,血浆样品的最佳酶解条件为:β-葡萄糖醛酸酶和硫酸酯酶的终用量分别为400和20 U.mL-1,酶解时间为2h;血浆中山奈素含量的线性范围为0.019-0.608μg.mL-1,方法回收率为80.23%-84.27%,日内和日间精密度的RSD分别≤7.32%、10.17%.可见,本试验建立的高效液相色谱测定方法可以准确、灵敏地测定兔血浆中山奈素的含量,适用于山奈素血药含量检测和药代动力学研究.

Solid phase extraction-high performance liquid chromatography method for determination of Kaempferol in rabbit plasma

This thesis developed a solid phase extraction-high performance liquid chromatography (SPE-HPLC) method for the determination of Kaempferol in rabbit plasma. In the method, β-Glucuronidase and sulfatase were used to hydrolyze plasma samples before analysis, and Kaempferol in plasma was extracted with a octadecylsilyl C18 SPE column. The mobile phase was methanol. 0.4% phosphoric acid (55∶ 45) with the flow rate of 1 mL·min~(-1), and the detection wavelength was set at 360 nm. The results showed that the optimum enzymatic hydrolysis conditions were β-Glucuronidase concentration 400 U·mL~(-1) and sulfatase concentration 20 U·mL~(-1)for 2 h respectively, the calibration curves for Kaempferol in plasma were liner between 0.019-0.608 g·mL~(-1) the assay recovery was 80.23% to 84. 27%; the intra-day and inter-day precisions (RSD) were all less than 10. 17%. It could be concluded this method had good sensitivity and precision for the determination of Kaempferol in plasma. It was shown to be for pharmacokinetics studies of Kaempferol.