鸡蛋中QNs多残留检测HPLC方法建立.doc

鸡蛋中12种喹诺酮类药物残留检测的HPLC方法建立 廖兰1，2，饶 勇1，3，曾振灵1，曾忠良3 （1华南农业大学兽医学院/广东省兽药研制和安全评价重点实验室，广州510640，2 西南大学荣昌校区，重庆402460；3西南大学药学院，重庆400715） 摘 要：【目的】本文建立并验证了同时检测诺氟沙星、氧氟沙星、培氟沙星、环丙沙星、洛美沙星、达氟沙星、恩诺沙星、沙拉沙星、二氟沙星、噁喹酸、萘啶酸和氟甲喹等12种喹诺酮类药物在鸡蛋中残留的高效液相色谱-荧光检测分析方法。【方法】鸡蛋样品以乙腈-三氯乙酸溶液匀浆和超声提取，经反相聚合物SPE净化和浓缩；样品以乙腈-柠檬酸/乙酸铵缓冲液为流动相、经C18柱HPLC梯度洗脱分离，用荧光检测器程序波长检测。【结果】线性范围除达氟沙星为5.0 μg/kg～160 μg/kg外，其余11种药物为12.5 μg/kg～400 μg/kg。添加6个浓度测得的校正曲线的相关系数除氟甲喹为0.982以外，其它11种药物均大于0.995。添加1/2MRL、MRL、2MRL三个浓度水平三批样品的回收率除萘啶酸为52%外，其它11种药物为68%～85%。批内变异系数＜8%，批间变异系数除氟甲喹1/2MRL为18.0%和恩诺沙星1/2MRL为18.8%外，其它药物均≤13.0%。12种药物的LOD为0.2 μg/kg～4.1 μg/kg，LOQ为0.9 μg/kg～13.5 μg/kg。【结论】建立的鸡蛋中12种喹诺酮类药物残留检测的HPLC方法简便、快速、使用有机溶剂少和适用范围广，特异性、准确度、精密度和灵敏度适合于大量样品的筛选和定量检测。 关键词：喹诺酮类；残留；高效液相色谱；鸡蛋 Development of HPLC Method for Multiresidue Determination of 12 Quinolones in Egg LAO Lan1,2, RAO Yong1,3, ZENG Zhen-ling1, ZENG Zhong-liang3 (1College of Veterinary Medicine, South China Agricultural University/Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation,; Guangzhou 510640, Guangdong, China; 2Rongchang Campus, Southwest University, Rongchang 402460, 3College of Pharmacy, Southwest University, Chongqing 400715, Chongqing China) Abstract: 【Objective】A high performance liquid chromatographic (HPLC) method with fluorescence detection (FLD) was developed and validated for simultaneous determination residues of 12 quinolones (QNs), norfloxacin (NOR), ofloxacin (OFL), ciproflocacin (CIP), peflocacin (PEF), lomefloxacin (LOM), danofloxacin (DAN), enrofloxacin (ENR), sarafloxacin (SAR), difloxacin (DIF), oxolinic acid (OXO), nalidixic acid (NAL) and flumequine (FLU) in egg. 【Method】Egg was extracted by a homogenater and an ultrasonic cleaner in 10% trichloroacetic acid (TCA) in water – acetonitrile (8:2, v/v) and cleared and purified by a solid phase extraction (SPE) cartridges, Strata-X?, based on a modified styrene-divinylbenzene polymer. 12 QNs were separated by C18 column (Hypersil? BDS-C18 ) with acetonitrile-