【作者】 顾利强； 辛艳飞； 张升； 温磊； 杨士豹； 胡晓玲； 宣尧仙；

【Author】 GU Li-qiang~1,XIN Yan-fei~1,ZHANG Sheng~1,WEN Lei~1,YANG Shi-bao~2,HU Xiao-ling~2,XUAN Yao-xian~1 1 Zhejiang Academy Of Medical Sciences,Department Of New Drug Safety Evaluation Research Center,Hangzhou 310053,Zhejiang,China;2 Jiangsu Xiansheng Drug Research Co.Ltd.,Nanjing 210042,Jiangsu,China

【机构】 浙江省医学科学院新药安全性评价研究中心； 江苏先声药物研究有限公司；

【摘要】 目的:建立Beagle犬血浆中依达拉奉的LC-MS/MS测定方法。方法:血浆样品用两倍乙腈直接沉淀蛋白,13000 rpm离心10 min后取上清液1 mL,加入10%三氯乙酸100μL,混匀后直接进样。以依达拉奉的4-甲基衍生物作为内标,并加入维生素C作为稳定剂。流动相为甲醇:乙酸缓冲液（pH3）=70:30,采用Thermo BDS HYPERSIL C₁₈柱,流速为0.2 mL·min^-1。同时对依达拉奉的m/z=133和内标4-甲基衍生物的m/z=147的碎片离子进行检测。结果:在10-500 ng·mL^-1范围内,依达拉奉在Beagle犬血浆内呈良好的线性关系（r>0.9966）,绝对回收率为32.9%～53.5%,方法回收率为87.4%～110.1%,批内精密度RSD小于14.6%（n=6）,批间精密度RSD小于6.2%（n=3）,定量下限为10 ng·mL^-1（83.2%<方法回收率<117.9%,RSD<8.1%,n=6）。结论:该方法简单、准确,操作迅速方便,适合于Beagle犬血浆中依达拉奉的检测。更多还原

【Abstract】 AIM:To develop an LC-MS/MS method to determine the edaravone in plasma of Beagle dog.METHODS:The plasma sample was precipitated with two partition of acetonitrile.After centrifuged at a speed of 130000 rpm for 10 min,1 mL of supernatant was got and treated with 100μL of trichloracetic acid（TCA）.The mixture was injected into the LC-MS/MS system for analysis after vortexing for several seconds.One of the derivates of the edaravone（4-methyledaravone） was used as internal standard（IS） and Vitamin C was added as stabilizer.The analysis was performed using an Thermo BDS HYPERSIL C₁₈ column（150 mm×4.6 mm,3μm） with a C₁₈ guard column（15 mm×4.6 mm,5μm） at 30℃.The column was isocratically eluted with a mobile phase containing methanol and acetic acid solution of pH3（70:30,v/v） and a flow rate of 0.2 mL·min^-1. The fragment ion m/z=133 of edaravone and m/z=147 of IS was choosed to quantitated to determine the concentration of edaravone. RESULTS:The calibration curve of edaravone in plasma was linear in the range of 10-500 ng·mL^-1.The correlation coefficient（r） for the regression equation of edaravone was in the range of 0.9966- 0.9999.The absolute recovery was in the range of 32.9%-53.5% and the assay recovery was in the range of 87.4%-110.1%with the within-run precision of 14.6%（n=6） and the between-run precision of 6.2%（n=3）.The low limits of quantifications（LLOQ） for edaravone was 10 ng·mL^-1（RSD<8.1%,n=6）.CONCLUSION: The method proved to be simple,accurate and easy to conduct and it is suitable to determine the edaravone in the Beagle’s plasma.更多还原