Development Of Analysis Methods For Several Kinds Of Prohibited Drugs In Health-care Products And Animal Products

With theincreasing of chemical substancesin the environment,food safty has become a worldwide challenge and global public health problem.PDE-5inhibitor was widely usedto improvemale erectile dysfunction and treat other diseases.Antiviral drugswere considered as a kind of effective drugsfor human to preventviral infectionsand treat viral diseases.At present,the detection methods of the PDE-5 inhibitorand antiviral drugs mainly include liquid chromatography?LC?and liquid chromatography tandem mass spectrometry?LC-MS/MS?.However,these analysis methods relied on expensive instruments,proficient experts,complicated pre-treatments,etal..Nowadays,more attentions are paid torapiddetection.Comparedwithinstrumentvalidation techniques,immunoassay and molecularly imprinted electrochemical sensor could realize spot quick detection ofPDE-5 inhibitor and antiviral drugs in food samples with the advantages of rapidness and sensitivty.In this study,indirect competitive enzyme-linked immunosorbent assay?ic-ELISA?,colloidal gold immunochromatographic test strip?GICA?and molecularly imprinted electrochemical sensor were estabished for the detectionof PDE-5 inhibitor and antiviral drugs.The main results were as follows:?1?A novel hapten was designed and synthesised for PDE-5 inhibitorbased on molecular simulation,and attached to the proteins through the method of active ester to form the antigen.And then,the broad spectrumantibodies were achieved withgood inhibitory rates under the concentration condition of 1?g/ml PDE-5 inhibitors,such as80%?Sildenafil?,87%?vardenafil?,88%?acedenafil?,70%?mirodenafil?,and so on.After the reaction conditions were optimized by single factor optimization experiment,the ic-ELISA and GICA methodswere established for the detection of a variety of PDE-5 inhibitors in food samples.The ic-ELISA method showed thebroad linear ranges and good sensitivities(IC₅₀)such as0.74-27.68 ng/mL and 4.54 ng/mL for acedenafil,0.97-70.72 ng/mL and 8.32 ng/mL for sildenafil,1.45-22.72 ng/mL and 5.70 ng/mL for vardenafil,0.2-22.8ng/mL and 2.16ng/mL for mirodenafil,and so on.The average recovery ratefor health wine samples was84.9%-116.2%and the coefficient of variations?CV?was 6.37%-14.28%.The detection results showed a good correlation with HPLC-MS/MS?R²=0.9977?.The detection limit of GICA method for acedenafil,sildenafil,vardenafil,mirodenafil was 10,20,5,15ng/mL,respectively.The dectection results of GIC A kept a good consistency with the results of HPLC-MS/MS.?2?Anovel electrochemical sensor for the determination of ribavirin?RIB?was fabricatedby molecularly imprinted technologyby using3-aminobenzeneboronic acid as functional monomer.A molecularly imprinted polymers?MIPs?membrane was synthesized in situon the surface of glassy carbon electrode by the formation of ring ester bondbetween boric acid and vicinal diol.The electrochemical behavior of the membrane was characterized by cyclic voltammetry?CV?and differential pulse voltammetry?DPV?.It was found that the response of peak currents was linear to the concentration of RIB in the range of 5.0×10^-8-1.0×10^-5mol/L with a detection limit of 1.5×10^-8mol/L?S/N=3?under the optimum conditions.The novel electrochemical sensor with high sensitivity,good stability and high reproducibility exihibited the excellent practicalpotential for detectingthe RIB in meat samples.