Ultraviolet absorber uv-326 detection kit and its preparation method and application
A UV-326, UV absorber technology, applied in measurement devices, color/spectral property measurement, instruments, etc., can solve the problems of expensive laboratory equipment, high detection cost, solvent consumption, etc. Low cost, simple pre-processing effect
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Embodiment 1
[0043] A UV absorber UV-326 detection kit, the main components are as follows:
[0044] 1) Ultraviolet absorber UV-326 specific antigen working solution;
[0045] Prepared by the following method: 0.32g (ie 1.0mmol) of ultraviolet absorber UV-326 with phenolic hydroxyl; 0.075g (ie 1.0mmol) of 3-amino-1-propanol and 0.29g (ie 1.1mmol) of Dissolve triphenylphosphine in 2.0mL of dry tetrahydrofuran (THF), sonicate at room temperature for 5min, add 0.22mL (ie 1.1mmol) diisopropyl azodicarboxylate dropwise during sonication, continue ultrasonic reaction for 20min, after the reaction is completed Use a chromatographic column to separate the hapten of the UV absorber UV-326 that can be coupled to the protein; then mix it with N-hydroxysuccinamide (NHS) and carbodiimide (EDC), and mix it with bovine serum white Proteins were mixed, stirred at room temperature for 1 h, coupled with UV-326 specific antigen prepared by UV absorber, and diluted with concentrated sample diluent (0.01M, pH...
Embodiment 2
[0068] Establishment of an indirect competitive ELISA method.
[0069] Indirect competitive ELISA method was used to detect the competitive inhibition rate of UV-326 monoclonal antibody, the method is as follows:
[0070] 1) Coat a 96-well ELISA plate with 1.5 μg / mL UV-326 specific antigen working solution, 100 μL / well, place in a 4°C refrigerator overnight, and use 250 μL / well blocking solution (5% Skimmed milk powder) was washed with washing liquid after sealing, and patted dry;
[0071] 2) Add UV absorber UV-326 standard solution (concentrations are: 2×10 5 μg / L, 2×10 4 μg / L, 2×10 3 μg / L, 2×10 2 μg / L, 2×10 1 μg / L, 2×10 0 μg / L) or 50 μL of the sample solution to be tested, then add 100 μL of UV-326 specific antibody working solution with a concentration of 3 μg / mL, mix well, and incubate at 37°C for 1 hour;
[0072] 3) After washing and patting dry, add horseradish peroxidase-labeled goat anti-mouse secondary antibody working solution diluted with diluent at a ratio o...
Embodiment 3
[0082] Kit sensitivity, specificity, accuracy.
[0083] 1. Sensitivity determination.
[0084] A standard working curve for the detection of the ultraviolet absorber UV-326 was established using the results of the indirect competitive ELISA method in Example 2. UV absorber UV-326 monoclonal antibody at 2μg / L~2×10 5 Good linearity in the range of μg / L, IC 50 =958.4ng / mL, the lowest detection limit is 5.81ng / mL, and the detection range (between 20% and 80% inhibition) is 49.50ng / mL~1650μg / mL. The detection limit of plastic food packaging products is 2.91ng / cm 2 , the detection limit of food samples is 1.16ng / g, and the detection limit of daily chemical products is 5.81ng / g. 2. Specificity Determination
[0085] Adopt the indirect competition ELISA method of embodiment 2 to measure ultraviolet absorber UV-326 structural analogue (ultraviolet absorber UV-327, ultraviolet absorber UV-234, ultraviolet absorber UV-329, ultraviolet absorber UV-71, ultraviolet absorber agent UV-3...
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