Study On Residues And Elimination Of Ribavirin And Its Metabolites In Tissues Of Laying Hens

China has a big production and consumption on poultry products,the egg production ranked first in the world for many years.In poultry breeding process,small scale breeding,large density,the chicken is relatively fragile,easy to infection with avian influenza and other respiratory diseases.Ribavirin has a significant effect in the prevention and treatment of influenza virus.But ribavirin has certain toxic and side effects,long term use may cause the virus to produce different degree of resistance,and induce the emergence of new drug resistant virus.Thereby affecting the prevention and treatment of human viral diseases,causing harm to people's health.Because of the research of ribavirin in the laying hens is still blank,scientific and effective supervision cannot be carried out to face the reality of illicit use.Therefore,it is urgent to carry out the research on the residues of ribavirin in the laying hens,and establish the corresponding detection method.In this study,firstly,the UPLC-MS/MS detection method of ribavirin and its main metabolites,RTCOOH and TCONH2 in the tissues of laying hens was established.Secondly,studies on the residues and elimination of ribavirin in the tissues of laying hens.Lastly,the systematic study of the residues of ribavirin and its main metabolites in eggs.The main contents and results are as follows:1.The establishment of UPLC-MS/MS detection method of ribavirin and its main metabolites in egg layers.In this study,the ultra high performance liquid chromatography-tandem mass spectrometry was used to detectthe residue of ribavirin and its major metabolite in the poultry chicken products,chicken liver and eggs.Aiming at the characteristics of low residueand high conversion rate in tissue,the detection methods of ribavirin and its main metabolite TCONH2 and RTCOOH were established for the first time.Samples were extracted with acetonitrile water(V/V,9:1),N-hexane to remove fat,C18 combined with GCB to remove impurity,Agilent ZORBAX SB-Aq chromatographic column(1.8 mm,3.0*100 mm)separation,determination by ultra high performance liquid chromatography-tandem mass spectrometry.The results showed that:ribavirin,TCONH2 and RTCOOH had a good linear relationship in chicken egg,chicken muscle and chicken liver tissues.In the 0.5?2000 ?g/L range,the correlation coefficient R2>0.99.three drugs recovery respectively were 83.5%?120.6%,75.9%?111.4%and 42.9%?58.9%.Detection limits respectively were 0.54?1.54?g/L,0.04?1.26?g/L and 1.54?4.30?g/L.Through the actual sample test,this method has the advantages of short sample analysis time,simple operation,good separation rate,high recovery rate,and meet the basic requirement of quantitative limit and precision.It can be applied to the routine detection of ribavirin and its main metabolites in the tissues of laying hens.2.Study on the residues and elimination of ribavirin in the tissues of laying hensIn this study,the research object is Beijing Red Layer,with a dose of 30 mg/kg continuous oral feeding,analysis of ribavirin in chicken muscle and liver tissue pharmacokinetics.The results showed that:Ribavirin uptaked quickly,Wide distribution,Tmax are less than 2 h.The drug distribution in different tissues is different,ribavirin has a Large residual in chichen muscle,Cmax was 402.97 ng/g;RTCOOH has a large residual in chichen liver,Cnmax was 9993.10 ng/g.Three kinds of drugs in chicken liver eliminated quickly,the final detection time(Tlast)for 48 h,the chicken muscle Tlast for 120 h.According to the characteristics of Tlast and drug distribution,determine the chicken muscle as target tissue,TCONH2 as the target.3.Study on the residues of ribavirin in eggsIn this study,the research object is Beijing Red Layer,with a dose of 30 mg/kg continuous oral feeding,analysis and study on the residue elimination rule of ribavirin and metabolites TCONH2,RTCOOH in eggs.The results showed that:ribavirin and metabolites entered the egg quickly,TCONH2 has a high residue concentration(1182.50 ng/g),the elimination rate is large.The residual concentrations of ribavirin in eggs were lower than the concentrations of TCONH2 and RTCOOH.Three kinds of drugs have a long residue time,were still detected in eighth days.According to the characteristics of drug residues,the use of ribavirin will bring some security risks in eggs.