【作者】 夏婉秋；

【导师】 王建平； 王庚南；

【作者基本信息】 河北农业大学 ， 基础兽医学， 2019， 硕士

【摘要】 分子印迹聚合物是一种化学合成性的具有特异性识别能力的高分子材料,它还具有合成成本低、环保、可重复使用等优点。本论文主要内容是合成吩噻嗪类以及苯二氮卓两类药物的分子印迹聚合物,以其为识别元件建立两种化学发光检测法,并结合分子印迹聚合物-固相萃取-超高效液相色法对猪肉中两类药物的残留进行检测。首先,采用计算机模拟技术研究假性模板、4种吩噻嗪类药物以及5种苯二氮卓类药物的三维构象和结构参数,筛选出最佳的模板分子。以2-（三氟甲基）吩噻嗪（TFPZ）和氯硝西泮（CLP）为模板分别合成吩噻嗪类和苯二氮卓类药物的分子印迹聚合物,并对它们进行了结构表征。将它们分别包被在96孔板的不同孔中做为识别元件,然后加入样品液和酶标半抗原进行竞争吸附,最后加入鲁米诺-H₂O₂-IMP诱导发光。结果表明,该竞争化学发光法对4种吩噻嗪类药物和5种苯二氮卓类药物的检测限分别为0.003-0.005 pg/mL和0.001-0.1 pg/mL,两类药物在猪肉中的添加回收率分别为72.3-92.5%和71.6-92%。将它们分别包被在96孔板的不同孔中做为识别元件,然后直接加入样品液进行吸附,最后加入TCPO-H₂O₂-IMZ诱导发光。结果表明,该直接化学发光法对4种吩噻嗪类药物和5种苯二氮卓类药物的检测限分别为0.1-0.6pg/mL和0.1-0.5 pg/mL,两类药物在猪肉中的添加回收率分别为81.2-91.8%和82.4-91.8%。最后合成了一种双模板分子印迹聚合物,制备了一种固相萃取柱对猪肉中的吩噻嗪类和苯二氮卓类药物进行提取净化,用超高效液相色法检测。结果表明,制备的固相萃取柱对4种吩噻嗪药物和5种苯二氮卓类药物的吸附量分别为47.5-48.4μg和37.4-41.5μg,回收率分别为96.5-98%和95.2-99%,制备的固相萃取柱可重复使用50次。该方法对4种吩噻嗪类药物和5种苯二氮卓类药物的检测限分为1.5-7ng/mL和2.5-10 ng/mL,定量限分别为5.0-20 ng/mL和6-32 ng/mL,两类药物的添加回收率分别为85.6-97.9%和85.9-95.7%。本论文以分子印迹聚合物为吸附剂和识别元件建立的样品前处理技术和化学发光检测法,具有操作简单、灵敏度高、选择性强、可循环利用等优点,可用于吩噻嗪类药物和苯二氮卓类药物的快速残留检测。更多还原

【Abstract】 Molecularly imprinted polymers（MIPs）are chemically synthesized polymers with specific recognition.They have the advantages of low cost,environmental-friendly and reusability.The main objectives of this study are to synthesize the molecularly imprinted polymers for phenothiazines and benzodiazepines,establish two chemiluminescence methods,and establish the molecularly imprinted polymers（MIPs）-solid phase extraction（SPE）-ultrahigh performance liquid chromatography method（UPLC）for detection of the two classes of drugs in pork.Firstly,the three-dimensional conformations and structure parameters of pseudotemplate,four phenothiazines and five benzodiazepines were studied by computational simulation technology,and the best template molecule was screened out.Molecular imprinted polymers of phenothiazines and benzodiazepines were synthesized with 2-（trifluoromethyl）phenothiazine（TFPZ）and clonazepam（CLP）as the templates respectively,and their structures were characterized.They were coated in different wells of96-well plate as recognition elements,and then the sample solution and enzyme-labeled hapten were added for competitive adsorption.Finally luminol-H₂O₂-IMP was added to induce luminescence.The results showed that the detection limits of the competitive chemiluminescence method for four phenothiazines and five benzodiazepines were0.003-0.005 pg/mL and 0.001-0.1 pg/mL,respectively.The recoveries of the 9 drugs in pork were in the range of 72.3-92.5%and 71.6-92%,respectively.Secondly They were coated in different wells of 96-well plate as recognition elements,and then the sample solution was directly added for adsorption.Finally TCPO-H₂O₂-IMZ was added to induce luminescence.The results showed that the detection limits of this method for 4phenothiazine drugs and 5 benzotriazine drugs were 0.1-0.6 pg/mL and 0.1-0.5 pg/mL respectively.The recoveries of the 9 drugs in pork were in the range of 81.2-91.8%and82.4-91.8%respectively.Thirdly,a double template molecularly imprinted polymer was synthesized,and a solid phase extraction column was prepared to extract and purify phenothiazines and benzodiazepines in pork,and the extracts were analyzed by UPLC.The results showed that the column showed high adsorptive capacities for four phenothiazines and five benzodiazepines（47.5-48.4μg and 37.4-41.5μg）.The column recoveries were in the range of 96.5-98%and 95.2-99%,respectively.The solid phase extraction column could be reused for 50 times.The detection limits for four phenothiazines and five benzodiazepines were 1.5-7 ng/mL and 2.5-10 ng/mL,and the quantitative limits were5.0-20 ng/mL and 6-32 ng/mL.The recoveries of the 9 drugs were in the range of85.6-97.9%and 85.9-95.7%,respectively.The sample pretreatment technology and chemiluminescence detection method based on molecularly imprinted polymers in this study have the advantages of simple operation,high sensitivity,strong selectivity and reusability.Therefore,these methods can be used for rapid detection of the residenues of phenothiazines and benzodiazepines in meet.更多还原