高效液相色谱荧光检测法测定牛奶中4种氟喹诺酮类药物残留

Determination offour Fluoroquinolones Residues in Milk by High Performance Liquid Chromatography with fluorescence detector

为了建立一种同时测定牛奶中4种氟喹诺酮类药物(恩诺沙星、环丙沙星、沙拉沙星、二氟沙星)多残留高效液相色谱检测方法,将牛奶中的残留药物用磷酸-甲醇溶液重复提取2次,正己烷除去脂肪净化后浓缩,流动相溶解后用高效液相色谱-荧光检测器测定。结果表明,4种氟喹诺酮类药物在0.01~1.00μg/mL浓度范围内具有良好的线性关系(R=0.999 9),在0.01,0.05,0.10,0.50μg/mL和1.00μg/mL 5个浓度添加水平上,平均回收率为77%~93%,样品的批内和批间变异系数均值分别为5.29%和7.83%,定量限均为10 ng/mL。环丙沙星、恩诺沙星、沙拉沙星检测限均为5 ng/mL,二氟沙星检测限为8 ng/mL。本研究建立的检测方法简单、快速、灵敏度和回收率高,适用于牛奶中4种氟喹诺酮类药物残留检测。

This study established multi-residue high performance liquid chromatography simultaneous determination enrofloxacin,ciprofloxacin,sarafloxacin and difloxacin in milk. Four kinds of fluoroquinolones in milk were repeatedly extracted twice with phosphoric acid-methanol solution,which was removed the fat from hexane and purified the sample. Then,the liquid phase was dissolved and determined by HPLC-FLD. The four fluoroquinolones have a good linear relationship（ r = 0. 9999） in the range of0. 01-1. 00 μg/mL in the established method. The average recoveries of the sample from spiked milk at the five concentration of0. 01μg/mL,0. 05μg/mL,0. 10μg/mL,0. 50μg/mL,and 1. 00 μg/mL ranged from 77% ~ 93%. The average coefficient of variation between batches and batches of samples was 5. 29% and 7. 83%,respectively and the quantitative limit of 10 ng/mL. The detection limits of ciprofloxacin,enrofloxacin and sarafloxacin were 5 ng/mL and the detection limit of difloxacin was 8 ng/mL. The method is simple,rapid,sensitive and high recovery,which is suitable for the detection of four kinds of fluoroquinolones in milk.