Preparation Of Nanofibers Based Kit For Kanamycin Residue Detection In Dairy Products

Kanamycin,an aminoglycoside antibiotic,plays an indispensable role in the prevention and treatment of animal diseases.In recent years,the abuse of kanamycin in animal husbandry has become a global problem,excessive kanamycin will be heavy left in animal-derived foods,which can concentrate in the human body through the food chain,causing great harm to human health.Therefore,it is necessary to strictly control the amount of kanamycin residues in food.This paper focuses on the research of kanamycin residue detection in dairy products.The traditional detection methods have high accuracy and high sensitivity,such as high performance liquid chromatography,microbiological analysis,immunoassay and so on,but detection process complex,time-consuming,costliness,specialized tester requirement,limiting its popularization.With advantages of easy operation,low cost,selectivity,no need for large instruments and the detection results can be distinguished by naked eye,gold nanoparticles based colorimetric method has become one of the most promising detection methods in the rapid detection methods of kanamycin.Until now,many researchers have carried out a lot of research on gold nanoparticles based colorimetric method of kanamycin,but the sensitivity and detection by naked-eye cannot meet the practical application requirements.Relying on the electrospun fiber membrane‘s advantages of large specific surface area,high porosity and controllable bulk density,this paper successfully prepares a nanofiber based on kit which can be used for kanamycin residue detection in commercial dairy products.The main research is as follows:The Polycaprolactone(PCL)fiber membrane was prepared by electrospun,through exploring the effect of concentration of the spinning solution on the morphology of the fiber membrane,the dispersion of fiber diameter and fiber-forming efficiency,12.5% fiber membrane(n-PCL fiber membrane)and 15% fiber membrane(m-PCL fiber membrane)were selected to subsequent modified treatment.The hydrophilic PCL electrospun fiber membrane was obtained by modified treatment with ethanol and Na OH solutions.The effects of ethanol solution pretreatment,concentration of Na OH solution and modified time on the morphology,mechanical properties,fiber morphology,hydrophobicity and wicking properties of fiber membrane were studied systematically.The uniform and dispersed gold nanoparticles were prepared by sodium citrate reduction method,then the gold nanoparticles were coupled with the kanamycin aptamer to form Au NPs@Aptamer.The results of UV-vis spectrophotometry,dynamic particle size,Zeta potential test confirm that gold nanoparticles and aptamer form a stable composites by covalent bond.Different modified PCL fiber membranes used as reaction membrane together with other parts were assembled into strips,and appropriate amount of Au NPs@Aptamer,T-line probes and C-line probes were loaded onto the strips and assemble into kanamycin kits.The results of each PCL fiber based kits showed that n-PCL fiber membrane treated with 4M Na OH solution for 1h had the highest sensitivity,and the detection limit of the naked eye was 0.05 ?g/m L.Thus,n-PCL fiber membrane treated with 4M Na OH solution for 1h was finally optimized as a kit reaction membrane.In addition,the selectivity of the optimal kit and the effectiveness of practical application were investigated.The results show that the kit has good selectivity,and can be used in practical sample detection.