液相色谱-串联质谱法测定比格犬血浆中淫羊藿苷元

Determination of plasma aglycon of icariin in Beagle dogs by liquid chromatography-tandem mass spectrometry

目的：建立了测定比格犬血浆中淫羊藿苷元浓度的液相色谱-串联质谱法（LC-MS/MS）。方法：血浆样品经酶水解,用液-液萃取法,以乙酸乙酯提取后,以乙腈-5%乙酸（70∶30）为流动相,用Zorbax C8柱分离,流速为0.6 mL.min-1,通过电喷雾离子化四极杆串联质谱,以多反应监测方式（MRM）进行检测。用于定量分析的离子分别为m/z369/313（淫羊藿苷元）和m/z331/315（麦黄酮,内标）。结果：淫羊藿苷元在比格犬血浆浓度测定方法的线性范围为2.5~250 ng.mL-1;日内、日间精密度（RSD）均小于13.3%,准确度（RE）在±5.5%之内。在临床前药动学研究中,应用此法测试了比格犬口服给药后的血药浓度。结论：本方法灵敏度高、专属性强,适合于淫羊藿苷元的临床前药动学研究。

Objective：To establish a specific and sensitive LC-MS/MS method for the determination of plasma aglycon of icariin in Beagle dogs.Methods：Aglycon of icariin and insternal standard tricin were extracted by ethyl acetate from plasma,then separated on a Zorbax C8 column.The mobile phase consisted of acetonitrile and 5% acetic acid（70∶30）.The protonated analytes generated in the positive ion mode were monitored through multiple reaction monitoring（MRM） in an electrospray ionization source.Results：The linear concentration ranges of the calibration curve for aglycon of icariin was 2.5~250ng·mL-1.The lower limit of quantitation was 2.5ng·mL-1.The inter-day and intra-day precision values was below 13.3%,and the accuracy（relative error） was within ±5.5%.Conclusion：The method is suitable for the pharmacokinetic studies of aglycon of icariin with the advantages of high sensitivity and specificity.