Solid phase extraction-high performance liquid chromatography method for determination of Kaempferol in rabbit plasma | 固相萃取―高效液相色谱法测定兔血浆中山奈素的含量

Chino. 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.[著者文摘]

Inglés. 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.[著者文摘]