铽增敏高效液相色谱柱后衍生荧光检测法同时检测鸡肉中的三种氟喹诺酮类药物残留

Simultaneous Determination of Residues of Three Fluoroquinolones in Chicken Using High Performance Liquid Chromatography with Post-Column Derivatization and Terbium Sensitized Fluorescence

基于氟喹诺酮类药物与铽离子形成配合物后的荧光增强作用,建立了同时检测鸡肉中氟喹诺酮类（FQs）药物环丙沙星、诺氟沙星和恩诺沙星残留的Tb^3＋增敏高效液相色谱（HPLC）柱后衍生荧光检测方法。优化的实验条件如下：流动相为0.05mol/L醋酸/醋酸钠缓冲液（pH6.0）-乙腈（体积比为89∶11）,色谱柱为Hypersil BDS-C18,柱温40℃,流速0.8mL/min;Tb^3＋浓度为8×10^-5mol/L;衍生反应温度40℃,衍生泵流速0.5mL/min;荧光检测激发波长271nm,发射波长545nm。实验结果表明,将上述3种药物以1.0,10.0,50.0,100.0ng/g水平添加到鸡肉后的回收率范围为66.3%-88.0%,相对标准偏差（RSD）均小于15.0%。定量分析的线性范围为0.1-500ng/mL,方法的日内和日间RSD均小于13.0%;最低检出限分别为0.05（环丙沙星）、0.05（诺氟沙星）和0.08（恩诺沙星）ng/g,比前人报道的非衍生高效液相色谱荧光检测法检测FQs药物的灵敏度有极大的提高。该项研究为FQs药物多残留检测提供了灵敏度更高的新方法。

A method of high performance liquid chromatography with post-column derivatization and terbium sensitized fluorescence for simultaneous determination of residues of ciprofloxacin, norfloxacin and enrofloxacin in chicken was established, based on the effect that Tb^3＋ can sensibilize fluorescence of fluoroquinolones. Under the optimal chromatographic condition, three fluoroquinolones were separated on a C18 column with 0. 05 mol/L acetic acid/acetate buffer-acetonitrile （89： 11, v/v） as the mobile phase, and at a flow rate of 0.8 mL/min; then were derivatized with 8 × l0 ^-5mol/L Tb^3＋ at 40℃, and at a flow rate of 0.5 mL/min; finally were detected using a fluorescence detector （λex = 271 nm, λem ： 545 nm）. The recoveries of three fluoroquinolones ranged from 66.3% - 88.0% at the added levels of 1.0, 10.0, 50.0, and 100.0 ng/g, and the relative standard deviations （RSDs） were less than 15.0%. The linear range for quantitative analysis was between 0. 1 and 500 ng/mL. All the RSDs of the inter-day and intra-day analyses were less than 13.0%. The detection limits were 0.05 ng/g for ciprofloxacin and norfloxacin and 0.08 ng/g for enrofloxacin. The method is sensitive for the determination of multi-residues of fluoroquinolones in chicken.