Fluorescent brightening agent CBS-X detection kit and preparation method thereof
A technology of fluorescent whitening agent and detection kit, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to identify the type of fluorescent whitening agent, high detection cost, and solvent consumption, so as to reduce errors and detection costs Low, simple pre-processing effect
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Embodiment 1
[0025] The main components of the fluorescent whitening agent CBS-X detection kit described in the present embodiment are as follows:
[0026] 1) Fluorescent whitening agent CBS-X specific antigen working solution;
[0027] Prepared by the following method: 0.186g, that is, 1.0mmol of 2-sulfobenzaldehyde, 0.20g, that is, 1.0mmol of 2-sulfo-5-hydroxy-benzaldehyde and 0.40g, that is, 1.0mmol of fluorescent The whitening agent CBS-X was dispersed in 30mL of dry DMSO, 3mL of piperidine was added, and the microwave was heated at 500 watts for 5min. After the reaction was completed, it was poured into water, and the solid was filtered with suction to obtain the fluorescent whitening agent CBS-X derivative with phenolic hydroxyl groups; Disperse 0.53g, i.e. 1.0mmol of the fluorescent whitening agent CBS-X derivative with phenolic hydroxyl group and 1.01g, i.e. 10mmol of potassium carbonate in 80mL of dry DMF, stir at room temperature for 30min, add 1.2g, i.e. 5mmol of N -(3-Bromopro...
Embodiment 2
[0050] Establishment of an indirect competitive ELISA method.
[0051] The competitive inhibition rate of fluorescent whitening agent CBS-X monoclonal antibody was detected by indirect competition ELISA method, the method is as follows:
[0052] 1) Coat a 96-well ELISA plate with 2 μg / mL fluorescent whitening agent CBS-X specific antigen, 100 μL / well, place in a 4°C refrigerator overnight, and use 250 μL / well blocking solution (5% skimmed milk powder ) after sealing, wash with washing solution and pat dry;
[0053] 2) Add fluorescent whitening agent CBS-X standard solution (concentration: 1×10 5 μg / L, 1×10 4 μg / L, 1×10 3 μg / L, 1×10 2 μg / L, 1×10 1 μg / L, 1×10 0 μg / L) or 50 μL of sample solution, then add 50 μL of fluorescent whitening agent CBS-X specific antibody working solution with a concentration of 2 μg / mL, mix well, and incubate at 37°C for 1 hour;
[0054] 3) After washing and patting dry, add horseradish peroxidase-labeled goat anti-mouse secondary antibody worki...
Embodiment 3
[0064] Kit sensitivity, specificity, accuracy.
[0065] 1. Sensitivity determination.
[0066] A standard working curve for the detection of fluorescent whitening agent CBS-X was established by using the results of the indirect competitive ELISA method in Example 2. Fluorescent whitening agent CBS-X monoclonal antibody at 1μg / L~1×10 5 Good linearity in the range of μg / L, IC 50 =905.1ng / mL, the minimum detection limit is 0.553ng / mL, and the detection range (between 20% and 80% inhibition) is 4.714ng / mL~1520.113μg / mL. The detection limit of paper and plastic products is 0.277ng / cm 2 , the detection limit of food samples is 0.277ng / g, and the detection limit of daily chemical products is 50ng / g.
[0067] 2. Determination of specificity.
[0068] Adopt the indirect competition ELISA method of embodiment 2 to measure fluorescent whitening agent CBS-X structural analogue (biphenyl, 2-vinylbenzenesulfonic acid sodium salt, 2-styrenebenzenesulfonic acid sodium salt, 2-bibiphenyl ...
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