高效液相色谱荧光检测法同时检测鸡血浆中阿莫西林和氨苄西林残留  

Determination of residues of amoxicillin and ampicillin in chicken plasma by HPLC with fluorescence detection

作　　者：孙礼瑞 赵敏[1,2] 谢恺舟[1,2] 郭辉生[1,2] 李爱华[1,2] 谢星[3] 王剑锋[1,2] 倪海球[1,2] 张跟喜[1,2] 戴国俊[1,2] 王金玉[1,2] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009 [2]江苏省动物遗传繁育与分子设计重点实验室,江苏扬州225009 [3]南京农业大学动物医学院,江苏南京210095

出　　处：《中国兽医学报》2014年第9期1501-1506,共6页Chinese Journal of Veterinary Science

基　　金：国家肉鸡产业技术体系专项资助项目(CARS-42-G23);江苏省科技成果转化专项资助项目(BA2008071);江苏高校优势学科建设工程资助项目;江苏省普通高校研究生科研创新计划资助项目(CXLX12_0935);扬州大学"新世纪人才工程"资助项目

摘　　要：建立高效液相色谱荧光法同时检测鸡血浆中阿莫西林和氨苄西林的残留量。鸡血浆经乙腈提取,饱和二氯甲烷萃取,酸性条件下水杨醛沸水浴衍生化后,以0.01mol/L的磷酸二氢钾溶液（A）和乙腈溶液（B）为流动相,梯度洗脱,流速为1.0mL/min,荧光检测激发波长为354nm,发射波长为445nm。阿莫西林和氨苄西林线性范围均为5.0~5 000.0μg/L,线性关系良好（r=0.999 9,0.999 6）。当阿莫西林和氨苄西林添加水平均为5.0~125.0mg/L时,鸡血浆样品平均回收率均高于74.72%,相对标准偏差（RSD）均低于8.65%,日内相对标准偏差分别低于4.99%和10.60%,日间相对标准偏差分别低于7.98%和11.29%。测定鸡血浆样品中AMO的检测限（LODs）为2.0μg/L（S/N=3）、定量限（LOQs）为5.0μg/L（S/N=10）,AMP的检测限为1.0μg/L（S/N=3）、定量限为2.5μg/L（S/N=10）。该方法操作简便、快速,且结果精确、灵敏、可靠,可适用于鸡血浆中阿莫西林、氨苄西林同时提取和检测。To develop a high performance liquid chromatography with fluorescence detection（HPLC-FLD）method for determination of residues of amoxicillin（AMO）and ampicillin（AMP）in chicken plasma.AMO and AMP were extracted from chicken plasma by acetonitrile and saturated methylene chloride for the reaction with salicylide under acidic and heating conditions.The mobile phase were 0.01mol/L potassium dihydrogen phosphate（A）and acetonitrile（B）,the flow rate was 1.0mL/min.Fluorescence detection was carried out at excitation and emission wavelengths of 354 nm and 445 nm.The calibration curves were linear in the range from 5.0μg/L to5 000μg/L with r=0.999 9and 0.999 6.At spiked level 5.0-125.0mg/L for AMO and AMP,the average recoveries were more than 74.72%,the relative standard deviation（RSD）were less than 8.65%,Inter-day RSD were less than 7.98%and 11.29%,respectively.Inter-day RSD were less than 4.99%and 10.60%,respectively.Limits of detection（LODs）were 2.0μg/L and 1.0μg/L（S/N=3）,respectively.Limits of quantitation（LOQs）were 5.0and 2.5μg/L（S/N=10）,respectively.This HPLC-FLD method is simple,rapid,sensitive and reliable for the determination of residues of amoxicillin and ampicillin in chicken plasma.

关 键 词：高效液相色谱 阿莫西林 氨苄西林 荧光检测法 

分 类 号：S859.79[农业科学—临床兽医学]