HPLC法测定人血浆中非诺贝特酸的浓度

目的:建立非诺贝特酸简便快速的血浆样品提取及准确灵敏的高效液相色谱分析方法。方法:血浆样品加地西泮和乙腈,涡旋振荡,高速离心后进样分析。其中,色谱柱为Eclipse XDB C18,流动相为乙腈∶甲醇∶0.04mmol·L-1乙酸钠(冰乙酸调pH值为3.50)=45∶15∶40,检测波长为296nm。结果:乙腈能够去除绝大部分的血浆杂质,不干扰色谱峰;非诺贝特酸、内标地西泮保留时间分别约为4.10、5.40min。非诺贝特酸检测浓度在0.2~20μg·mL-1范围内线性关系良好(r=0.999 7);其低、中、高(0.5、5.0、18.0μg·mL-1)浓度的提取回收率分别为100.53%、102.64%、91.81%,日内RSD均<8%,日间RSD均<7%。非诺贝特酸血浆样品预处理室温放置24h及于—20℃冰箱中贮存15d稳定。结论:本方法简便、快速、准确、重现性好,符合生物样品测定要求,适合非诺贝特临床药动学的研究。

Determination of Fenofibric Acid in Human Plasma by HPLC

OBJECTIVE： To establish a convenient, rapid, accurate and sensitive HPLC method for extracting and analyzing of the blood sample of fenofibric acid. METHODS： Blood sample spiked with diazepam and acetonitrile was subjected to vortex vibration and high speed centrifugation before being analyzed on Eclipse XDB C18. The mobile phase consisted of acetonitrile （45%）, methanol （15%） and sodium acetate （0.04 mmol · L^-1, pH3.50） . The detection wavelength was set at 296 nm. RESULTS： Most of dopants from plasma were removed by acetonitrile and chromatographic peaks were not interfered. The retention time of fenofibric acid and diazepam were about 4.10 min and 5.40 min, respectively. The standard curve was linear in the concentration range 0.2-20 μg · mL^-1（with correlation coecient at 0.999 7）. The average recovery was 100.53% for 0.5μg · mL^-1, 102.64% for 5.0μg · mL^-1, and 91.81% for 18.0μg · mL^-1. The intra - and inter- day assay variability values were less than 8% and 7%, respectively. Pretreated solution of fenofibric acid in human plasma was stable after storage for 24 h under room temperature or after freeze- storing at -20 ℃ for 15 days. CONCLUSION： This method is convenient, rapid, accurate and reproducible, which was consistent with requirement for determining biological specimen and was applicable to the pharmacokinetic study of fenofibric acid.