【作者】 张晶；

【导师】 王桂琴； 李莉；

【作者基本信息】 宁夏大学 ， 农业硕士（专业学位）， 2018， 硕士

【摘要】 近年来,人类对氟喹诺酮类药物的不合理使用及滥用情况日渐严重,导致其残留量在动物源食品中超标。如果长期食用含有兽药残留超标的畜禽产品,会导致人们的身体出现一系列的问题,如头晕、呕吐和腹泻等症状。因此,对于畜禽产品中该类药物的残留检测,需要制定一种快速、精确的检测方法,以便检测食品中其药物含量,为人类的健康提供保障,也为我国贸易往来提供强大的技术支撑。本研究主要包括3个方面的内容:1.利用酶联免疫吸附法对宁夏地区5个市采集的120份鸡肉和200份鸡蛋样品进行检测,结果表明,所有检测的样品均无氟喹诺酮类药物残留。该方法试验过程简单,可应用于大量样品的快速检测。2.建立固相萃取结合超高效液相色谱技术检测鸡肉中4种氟喹诺酮类药物残留的方法。用磷酸盐溶液对样品进行提取,提取液用C18固相萃取柱净化,流动相洗脱,采用超高效液相色谱仪进行测定。4种药物于0.005 μ g/mL～0.5 μ g/mL线性关系良好,相关系数R均大于0.99994。在浓度为20 μ g/kg、100 μ g/kg、200 μ g/kg添加时,该方法的平均添加回收率在40.0%～94.82%之间,相对标准偏差为1.5%～17.4%,方法检出限为20 μg/kg,定量限为100 μg/kg。按照本研究所建方法,对2017年宁夏地区5个市采集的120份鸡肉样品进行检测,结果均未检测出4种氟喹诺酮类药物残留。3.建立固相萃取结合高效液相色谱-串联质谱技术检测鸡蛋中7种氟唾诺酮类药物残留的方法。用EDTA-Mcllvaine溶液对样品进行提取,正己烷脱脂,提取液经HLB萃取柱净化,利用氮气吹干浓缩,流动相溶解,高效液相色谱-串联质谱定性定量分析。5种药物含量于2.5 μg/kg～100μg/kg内线性关系良好,环丙沙星和噁喹酸含量分别在6μg/kg～100μg/kg和0.25μg/kg～10μg/kg线性关系良好,相对系数均大于0.9959;添加量在100μg/kg、200 μg/kg、500μg/kg(相当于上机浓度 20μg/kg、40μg/kg、100μg/kg)时,此种检测方法平均加标回收率为61.9%～106.2%,相对标准偏差为1.3%～10.1%。该方法中环丙沙星与噁喹酸的检测限是1.2μg/kg、0.05μg/kg,定量限为24μg/kg、1μg/kg,其他5种药物的检测限为0.5 μ g/kg,定量限为10 μ g/kg。按照本研究所建方法,对2017年宁夏地区5个市采集的200份鸡蛋样品进行检测,结果均未检测出7种氟喹诺酮类药物残留。本文建立的两种方法前处理过程简便,方法的灵敏度较高,检测时间较短,且检测出的结果符合国内法规要求,可以应用于动物性食品中氟喹诺酮类药物的残留检测。更多还原

【Abstract】 In recent years,the irrational use and abuse of fluoroquinolones by humans has become increasingly serious,resulting in excessive levels of residues in animal-source foods.If you eat livestock and poultry products that contain excessive drug residues for a long period of time,it can lead to a series of problems such as dizziness,vomiting,and diarrhea.Therefore,for the residue detection of such drugs in livestock and poultry products,it is necessary to develop a rapid and accurate detection technology in order to detect the content of its drugs in foods,provide protection for people’s health,and provide strong technical support for China’s trade.This study mainly includes 3 aspects:1.By using enzyme-linked immunosorbent assay,120 samples of chicken and 200 samples of chicken eggs collected were tested from five cities in Ningxia.The results showed that none of the samples involved in the trial contained quinolones.The method has a simple test procedure and can be applied to the rapid detection of a large number of samples.2.Establish a method for the determination of four fluoroquinolones residue in chicken by solid phase extraction combined with ultra performance liquid chromatography.The sample was extracted with a phosphate solution.The extract was purified with a C18 solid phase extraction column and eluted with a mobile phase.The assay was performed using a high performance liquid chromatograph.The linear relationship between the four drugs at 0.005μg/mL-0.5μg/mL was good,and the correlation coefficient R was greater than 0.99994.At the concentrations of 20μg/kg,100μg/kg and 200μg/kg,the average recovery of the method was between 40.0%and 94.8%with relative standard deviations of 1.5%-17.4%.The method detection limit was 20μg/kg,the quantification limit was 100μg/kg.According to the method established in this study,120 chicken samples collected from 5 cities in Ningxia were tested in 2017.None of the four fluoroquinolones were detected.3.A solid phase extraction combined with high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was developed for the determination of seven fluoroquinolones in eggs.The sample was extracted with EDTA-Mcllvaine solution and degreased with n-hexane.The extract was purified by an HLB extraction column,dried with nitrogen,concentrated by mobile phase,and analyzed by HPLC with tandem mass spectrometry.The linear relationship was good between 5 μg/kg and 100μg/kg for 5 drugs,and the linear relationship between ciprofloxacin and oxolinic acid was 6 μg/kg to 100 μg/kg and 0.25 μg/kg to 10 μg/kg,respectively.The coefficients were all greater than 0.9959.When the amounts were 100μg/kg,200μg/kg,and 500μg/kg(Equivalent to upper machine concentration 20μg/kg,40μg/kg,100μg/kg),the average recoveries were 61.9%～106.2%and the relative standard deviations were 1.3%～10.1%.The detection limits of ciprofloxacin and oxolinic acid in this method were 1.2 pg/kg and 0.05 μg/kg,the quantitation limits were 24 pg/kg and 1 μg/kg.The detection limits of the other five drugs were 0.5 μg/kg and the quantitation limits were 10 μg/kg.According to the method established in this study,200 samples of eggs collected were tested in 5 cities of Ningxia in 2017.None of the 7 fluoroquinolones were detected.The two methods established in this paper have a simple pretreatment process,high sensitivity,and a short detection time.The detection results meet the requirements of domestic regulations and can be applied to the residue detection of fluoroquinolones in animal foods.更多还原