Development Of Upconversion Nanoparticles Based Immunochromatographic Test Strip For The Detection Of Quinolones And Sulfaquinoxaline

In this study, upconversion nanoparticles were chosen as marker. Quinolones and sulfaquinoxaline were chosen as target signal detection. And two fluorescent tags immune chromatography test strip analysis methods were established and applied to the detection of quinolones and sulfaquinoxaline residues in animal-derived foods.Hydrophobic, size-uniformed upconversion nanoparticles (OA-UCNPs) was synthesized by using hydrothermal method. Hydrophilic, size-uniformed upconversion nanoparticles (PAA-UCNPs) were prepared by ligand exchange method at high temperature with using polyacrylic acid (PAA).Norfloxacin complete antigen was realized through the conjugation of norfloxacin and ovalbumin (OVA) under the method of activated ester. Signal probe (PAA-UCNPs-Ab)was prepared through the method of activated ester by the conjugation of norfloxacin monoclonal antibody and PAA-UCNPs. This reaction was conducted under the ratio of PAA-UCNPs: antibody : EDC: NHS=50:3:15:24(mass ratio). The dilution time of the second antibody and the coating antigen, NC membrane variety, handling method of sample pad, pH value and adding amount of working liquid, variety, pH value, concentration of sample buffer,etc. Test strip was assembled under the optimal conditions, experiments were conducted using the method of wet process,the method detection limit is 1.0 ?g/L. Six kinds of actual samples, milk, pork liver, beef, mutton, weever and shrimp were selected to conduct adding-recycling experiment, the sample detection limit is 5 ?g/kg(L).Sulfaquinoxaline complete antigen was realized through the conjugation of sulfaquinoxaline and ovalbumin (OVA) under carbodiimide method. Signal probe (PAA-UCNPs-Ab) was prepared through the method of activated ester by the conjugation of sulfaquinoxaline monoclonal antibody and PAA-UCNPs. This reaction was conducted under the ratio of PAA-UCNPs: antibody: EDC: NHS=50:15:24(mass ratio). Antibody conjugation quantity, the dilution time of the second antibody and the coating antigen, the handling method of sample pad, the varietyof sample buffer, working liquid adding amount, etc. Under the optimal condition to assemble the test strip, using the method of wet process, the method detection limit is 4.0 ?g/L. Milk, pork liver, beef, mutton,weever and shrimp six kinds of actual samples were chosen to conduct adding-recycling experiment, the samples detection limit is 20 ?g/kg(L).The study of immune fluorescence tomography strip analysis method owes advantages of short time-consuming, strong specificity, high sensitivity, weak background interference.Both of them are suitable for detection of quinolones and sulfaquinoxaline residues in animal-derived foods.