| Ribavirin,a broad-spectrum antiviral drug,is commonly used in the treatment of viral diseases such as hepatitis C and hemorrhagic fever.It has been widely used in livestock and poultry production for a long time due to lax supervision.These drugs are prone to drug resistance.Long-term use of this product on a large scale can lead to problems such as poisoning,immunosuppression and drug residues,which seriously affect the quality of livestock and poultry products.At present,there are few researches on ribavirin at home and abroad.The existing methods are mainly used analyze instruments for the detection after pretreated by solid phase extraction column,including liquid chromatography,liquid chromatography-mass spectrometry,and so on.Other types of adsorption materials have not been fully developed.Furthermore,innovative detection methods combined with quartz crystal microbalance and electrochemical sensor have not been adequately reported,therefore,it is necessary to explore the new detection technology of ribavirin.For the chromatographic analysis technique,a MSPE-HPLC method for specifically identifying ribavirin was established.First,a zirconium-functionalized Fe3O4 adsorbent material was synthesized by a one-step hydrothermal method,which can bond with the cis-hydroxyl group on ribavirin specifically.Under the optimal experimental conditions,the adsorption properties of the materials were evaluated.It was found that the material has a rapidly adsorb speed to ribavirin and adsorption saturation could reached within 5 min.Next,the adsorption behavior was evaluated by pseudo-second-order kinetic model and the results showed that there was a strong chemical bond between the magnetic material and ribavirin.Under the optimal experimental conditions,the established MSPE-HPLC method has a linear range of 10-200 ?g/L(R2>0.99)and a detection limit of 2.68 ?g/L(S/N=3).Ribavirin was extracted in parallel for 9 times with an RSD of 4.41%.In addition,the reusability of the material was evaluated and it was found that the material could be used 4 times.Finally,the practical applicability of the method was evaluated.After the spike test of three blank samples(chicken liver,egg,shrimp),the recovery ranged from 74.13%to 92.90%.For the sensor detection technology,an inhibitory immunosensor based on quartz crystal microbalance that can specifically recognizes ribavirin was prepared.First,para-aminobenzoic acid was electropolymerized onto the surface of a gold electrode to obtain a carboxyl-modified gold electrode.After activation by EDC/NHS,the carboxyl-modified gold electrode was bound to the ribavirin antigen modified with an amino group,and a sensor that can specifically combined with ribavirin antibody was obtained.Since ribavirin also binds specifically to ribavirin antibodies,there is a competitive relationship between ribavirin and the sensor.When the three coexist,the change in the content of ribavirin causes the change in quantity of ribavirin antibody binding to the sensor.Changes in the amount of ribavirin antibodies,resulting in changes of sensor frequency.Under the optimal conditions of carboxyl modification,the appropriate amount of antigen was selected for combination,and the prepared sensor was characterized by electrochemical means,indicating that the sensor was successfully established.Under the optimal amount of antibody conditions,the linear range of detection of ribavirin was 1-750 ?g/L and the detection limit(IC15)was 2.64 ?g/L.The prepared sensor was used for the detection of ribavirin in the real samples(milk and chicken muscle),and the recovery range was 88.01-94.42%. |
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