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、3.4 min。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0.999 9、0.999 8);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。
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 directly 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 1.0 μg·mL^-1 with the detection wavelength of 346 nm. RESULTS: AT and ATX were completely separated without interference. The retention time was 5.5 rain and 3.4 min respectively. The linear range were 0.25-3.392 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%-44.11% and 0.71% - 4.51% while that of ATX were 0.55 % - 1.23 % and 1.74 % -3.92 %, respectively. The detection limit of AT in human plasma was 0.254μg·mL^-1 and that of ATX was 0.125 μg·mL^-1. CONCLUSION: It is proved that established method is simple, accurate and reproducible for plasma concentration determination of AT and ATX in plasma of beagle dogs.
出处
《中国药房》
CAS
CSCD
北大核心
2010年第9期806-808,共3页
China Pharmacy