摘要
为了研究喹烯酮(Quinocetone,QCT)与草鱼(Ctenopharyngodon idellus)血浆蛋白的结合情况,实验采用超高效液相色谱仪(UPLC)法测定透析袋两侧溶液中喹烯酮的质量浓度,计算QCT的血浆蛋白结合率。血浆样品采用乙腈提取,正己烷脱脂净化。采用ACQUITY UPLC BHE C18分离柱,柱温为30℃,流动相为乙腈-水(30∶70),流速为0.3 mL/min,紫外检测波长为320 nm。本方法的线性范围为0.025~3.0μg/mL,相关指数r2≥0.998。添加水平为0.05、0.1、0.2μg/mL时,平均回收率75%~90%,相对标准偏差均低于15%。结果显示:QCT在质量浓度为0.05、0.2、1μg/mL时与血浆蛋白结合率分别为(81.40±0.80)%、(79.60±0.80)%、(80.54±0.30)%,达到平衡的时间为36 h。喹烯酮的血浆蛋白结合无浓度依赖性,且具有高强度的血浆蛋白结合率。
In order to understand the protein binding of Quinocetone (QCT) with grass carp plasma, a method of ultra- performance liquid chromatography had been developed for determination of the protein binding of Quinocetone with grass carp plasma. After extraction with acetonitrile and defatted with n - hexane, the analytes was separated on ACQUITY UPLC BHE C18 with acetonitrile -water as mobile phase at flow rate of 0. 3 mL/min and column temperature of 30 ℃. The cali- bration curve ranging from 0. 025 - 3.0 μg/mL in fluent phase solvent was used to establish instrument response, with rel- ative standard coefficient beyond 0. 998. The method was validated by analysis of nine replicate plasma samples fortified with three concentration levels of 0. 05 p^g,/L, 0. 1 p^g/L and 0. 2 p,g/L QCT, respectively. Depending on the biological matrix and analyte, relative standard deviations were below 15 % , with average recovery rates around 75 % to 90 % in spiked samples. Finally, the method was successfully used to determine the protein binding of common carp plasma with QCT un- der 4 ~C by equilibrium dialysis method, and the equilibrium time was 36 h and the rate of protein binding were (81.40 ± 0. 8 ) %, ( 79. 60 ± 0. 8 ) %, ( 80. 54 ± 0. 3 ) %, respectively. The protein binding rates of the compounds in the QCT were independent on the area of the concentration which was determinate and the protein binding rates were strength.
出处
《淡水渔业》
CSCD
北大核心
2014年第2期71-76,共6页
Freshwater Fisheries
基金
公益性行业(农业)科研专项资助项目(201203085)
中国水产科学研究院基本科研业务费专项(2012A1005
2013A1004)