HPLC法同时测定Beagle犬血浆中茴三硫及其代谢物浓度的方.pdfVIP

HPLC 法同时测定Beagle 犬血浆中茴三硫及其代谢物浓度的方法学研究 1＊ 2# 余佳芮 ，蒋学华 （1.重庆医科大学附属儿童医院，重庆市 400014；2. 四川大学华西药学院，成都市 610041） + 中图分类号 R975.5；R969.1 文献标识码 A 文章编号 1001- 0408（2010）09- 0806- 03 摘 要 目的：考察高效液相色谱法同时测定Beagle 犬血浆中茴三硫（AT ）及其代谢物对羟基苯基三硫酮（ATX ）浓度的方法学。 方法：用乙腈直接沉淀犬血浆样品中的蛋白质，取上清液进样分析。色谱柱为Diamonsil ODS- C18，流动相为甲醇- 水（80∶20），流 速为1.0mL ·min－1，紫外检测波长为346nm 。结果：AT 和ATX 与犬血浆中的内源性物质完全分离且无其它干扰，保留时间分别为 5.5、3.4min 。二者检测浓度线性范围分别为0.254～3.392、0.125～1.664μg ·mL－1（r 分别为0.9999、0.9998）；平均回收率分别为 （93.89±2.39）％、（96.28±2.83）％；日内RSD 分别为0.83％～4.11％、0.55％～1.23％，日间RSD 分别为0.71％～4.51％、1.74％～ 3.92％；最低定量浓度分别为0.254、0.125μg ·mL－1。结论：方法学考察结果表明，所建立方法简便、准确、重复性好，可用于犬血浆 中AT 及ATX 的浓度测定。 关键词 茴三硫；对羟基苯基三硫酮；Beagle 犬；高效液相色谱法；血药浓度 Simultaneous Determination of Anethole Trithione and Its Metabolite in Plasma of Beagle Dogs by HPLC YU Jia-rui （Children ’s Hospital of Chongqing Medical University ，Chongqing 400014，China ） JIANG Xue-hua （West China School of Pharmacy ，Sichuan University ，Chengdu 610041，China ） ABSTRACT OBJECTIVE ：To establish an HPLC method for the determination of anethole trithione （AT ）and its metabolite 4- hydroxy- anethole trithione （ATX ）in plasma of beagle dogs simultaneously. METHODS ：Protein in plasma was deposited direct- ly using acetonitrile and analyzed by supernate. The separation was performed on Diamonsil ODS- C18 column with a mobile phase consisting of methanol and water （80∶20）at a flow rate of 1.0mL ·min－1 with the detection wavelength of 346nm. RESULTS ：AT and ATX were completely separated without interference. The retention time was 5.5min and 3.4 min respectively. The linear range were 0.254~3.392 μg ·mL －1 （r ＝0.999 9）for AT with average recovery of （93.89±2.39）％and 0.125～1.664 μg ·mL －1 for ATX （r ＝0.999 8）with average recovery of （96.28±2.83）％. The intra- day RSD and inter- day RSD of AT were 0.83％～4.11％and 0.71％～4.51％while that of ATX were 0.55％～1.23％and 1.74