RP-HPLC法测定人血浆中黄豆苷元的浓度

目的:建立以反相高效液相色谱法测定人血浆中黄豆苷元浓度的方法。方法:采用甲醇沉淀蛋白法提取血浆中黄豆苷元,色谱柱为Hypersil ODS C18(250mm×4.6mm,5μm),流动相为5%乙酸溶液(用三乙胺调pH值为4)-甲醇(55∶45),检测波长为254nm,流速为1.0mL·min-1,柱温25℃。结果:黄豆苷元血药浓度在1.5～700.0ng·mL-1范围内线性关系良好(r=0.9995),低、中、高浓度(5.0、16.5、231.0ng·mL-1)的平均回收率分别为(96.95±4.36)%、(101.30±3.32)%和(98.73±2.54)%。结论:本法简单、快速、准确,可用于黄豆苷元的血药浓度测定和人体药动学研究。

Determination of the Concentration of Daidzein in Human Plasma by RP-HPLC

OBJECTIVE： To establish a RP-HPLC method for the concentration determination of daidzein in human plasma. METHODS： Daidzein was extracted from plasma by protein precipitation with methanol. The determination was performed on Hypersil ODS C18 （250 mm×4.6mm,5μm） column with 5 % acetic acid （pH adjusted to 4 with triethylamine）-methnol （55： 45） as mobile phase at the flow rate of 1.0 mL·min^-1. The column temperature was set at 25℃ and the detection wavelength was 254 nm. RESULTS： The linear range of daidzein was 1.5-700.0 ng·mL^-1（r= 0.999 5 ）. The recovery rates of low, middle and high concentrations of daidzein （5.0 ng·mL^-1, 16.5 ng·mL^-1, 231.0 ng·mL^-1） were （96.95 ± 4.36）%, （101.30 ± 3.32）% and （98.73 ± 2.54）%, respectively. CONCLUSION： The method is simple, rapid and accurate for content determination and pharmacokinetic research of daidzein.