Study On Residue Detection Of Two Kinds Of Sedative And Hypnotic Drugs

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.