Christodoulou Eleni A, Samanidou Victoria F, Papadoyannis Ioannis N
Laboratory of Analytical Chemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, Greece.
J Sep Sci. 2007 Nov;30(16):2676-86. doi: 10.1002/jssc.200700170.
The aim of this work was to develop an HPLC method for the simultaneous determination of ten quinolones: enoxacin, ofloxacin, norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, oxolinic acid, nalidixic acid, and flumequine, in various tissues of food-producing animals. Separation was achieved on a PerfectSil Target column (250 mm x 4 mm, ODS-3, 5 microm), by MZ-Analysentechnik (Germany), at room temperature. The mobile phase consisted of 0.1% TFA-CH(3)OH-CH(3)CN and was delivered by a gradient program of 35 min. The detection and quantitation was performed on a photodiode array detector at 275 and 255 nm. Caffeine (7.5 ng/microL) was used as the internal standard (IS). Analytes were isolated from tissue samples by 0.1% methanolic TFA solution. SPE, using LiChrolut RP-18 cartridges, was applied for further purification. The extraction protocol was optimized and the final recoveries varied between 92.0 and 107.4%. The method was fully validated according to Commission Decision 2002/657/EC. Limits of quantitation for the examined quinolones extracted from each tissue were much lower than the respective Maximum Residue Levels, ranging between 30 and 50 microg/kg for bovine tissue, between 30 and 55 microg/kg for ovine tissue, and between 40 and 50 microg/kg for porcine tissue.
本研究的目的是开发一种高效液相色谱法，用于同时测定食用动物各种组织中的十种喹诺酮类药物：依诺沙星、氧氟沙星、诺氟沙星、环丙沙星、达氟沙星、恩诺沙星、沙拉沙星、恶喹酸、萘啶酸和氟甲喹。采用德国MZ-Analysentechnik公司的PerfectSil Target柱（250 mm×4 mm，ODS-3，5μm）在室温下进行分离。流动相由0.1%三氟乙酸-甲醇-乙腈组成，通过35分钟的梯度程序输送。在光电二极管阵列检测器上于275和255 nm处进行检测和定量。咖啡因（7.5 ng/μL）用作内标。通过0.1%的甲醇三氟乙酸溶液从组织样品中分离分析物。使用LiChrolut RP-18柱进行固相萃取以进一步纯化。优化了提取方案，最终回收率在92.0%至107.4%之间。该方法根据委员会第2002/657/EC号决定进行了全面验证。从每种组织中提取的所检测喹诺酮类药物的定量限远低于各自的最大残留限量，牛组织中为30至50μg/kg，羊组织中为30至55μg/kg，猪组织中为40至50μg/kg。
