高效液相色谱法测定新西兰兔血浆中硫辛酸浓度

High-performance liquid chromatography for determining plasma alpha-lipoic acid in New Zealand rabbits

目的建立高效液相色谱法测定新西兰兔血浆中硫辛酸的浓度。方法新西兰兔采血后离心得血浆经酶水解化后,用Baker phenyl固相萃取小柱提取血浆中硫辛酸,采用HPLC法紫外检测器检测。色谱柱为HYPERSIL BDS C18柱(250mm×4.6mm,5μm),流动相为磷酸二氢钾∶乙腈(50∶50,v/v),流速1.0ml/min,检测波长230nm。结果测定方法在硫辛酸浓度为5～100μg/L范围内具有良好线性关系,萃取回收率在77.4%～82.1%之间。日内、日间RSD在1.5%～8.9%之间。新西兰兔单剂量口服硫辛酸片300mg0.5h后测定血浆中硫辛酸浓度为11.08±1.79μg/L。结论本测定方法适用于口服硫辛酸的动物体内硫辛酸含量测定研究。

Objective To establish a method for determining the content of alpha-lipoic acid in New Zealand rabbit plasma. Methods Alpha-lipoic acid in the plasma samples was purified by solid-phase extractor and analyzed on an HYPERSIL C18 column with isocratic mobile phase consisting of potassium dihydrogen phosphate-acetonitrile （50：50, v/v） at a flow rate of 1.0 ml/min and detection wavelength of 230 nm. Results The standard curve was linear in the range of 5-100 μg/L （r=1） and the average recovery was 77.4%-82.1%. The relative standard deviations ofintra-day and inter-day assay were within 1.5%-8.9%. Conclusion The method is sensitive, accurate and simple for determining plasma alpha-lipoic acid levels in New Zealand rabbits.