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Fluorescent brightening agent VBL detection kit and preparation method thereof

A technology of fluorescent whitening agent and detection kit, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to identify the type of fluorescent whitening agent, high detection cost, complicated processing, etc., so as to reduce errors and detection costs. Low, simple preprocessing effect

Active Publication Date: 2015-01-07
GUANGZHOU QUALITY SUPERVISION & TESTING INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the direct ultraviolet light irradiation observation method is only a simple judgment, and cannot identify the type of fluorescent whitening agent; chromatography and chromatography-mass spectrometry have the advantages of high specificity and accurate results, but the disadvantage of these two methods is that the sample The pre-treatment is cumbersome, consumes a lot of solvents, and requires expensive laboratory equipment, and the detection cost is high

Method used

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  • Fluorescent brightening agent VBL detection kit and preparation method thereof
  • Fluorescent brightening agent VBL detection kit and preparation method thereof
  • Fluorescent brightening agent VBL detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The main components of the fluorescent whitening agent VBL detection kit described in the present embodiment are as follows:

[0026] 1) Fluorescent whitening agent VBL specific antigen working solution;

[0027]Prepared by the following method: Disperse fluorescent whitening agent VBL0.87g, namely 1.0mmol, and triethylamine 1.01g, namely 10mmol, in 100mL dry dichloromethane, add 0.5L at zero degrees Celsius, namely 6.46mmol methanesulfonyl chloride as Activate the reagent, stir and react at room temperature for 3.5 hours, the alcoholic hydroxyl group of the fluorescent whitening agent VBL can be converted into a methylsulfonate group; then add the mesylated fluorescent whitening agent VBL to absolute ethanol, and add 20 times the amount of liquid ammonia , reflux and stir for 5 hours to prepare the hapten of the fluorescent whitening agent VBL that can be coupled with the protein; then mix it with N-hydroxysuccinamide (NHS) and carbodiimide (EDC), and then mix it with ...

Embodiment 2

[0050] Establishment of an indirect competition ELISA method.

[0051] The competition inhibition rate of the fluorescent whitening agent 85 monoclonal antibody was detected by the indirect competition ELISA method, and the method was as follows:

[0052] 1) Coat a 96-well ELISA plate with 2 μg / mL fluorescent whitening agent VBL-specific antigen, 100 μL / well, place in a 4°C refrigerator overnight, and block with 300 μL / well blocking solution (5% skimmed milk powder) Then wash with washing liquid and pat dry;

[0053] 2) Add fluorescent whitening agent VBL 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 VBL-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 diluted...

Embodiment 3

[0064] Kit sensitivity, specificity, accuracy.

[0065] 1. Sensitivity determination.

[0066] A standard working curve for the detection of the fluorescent whitening agent VBL was established by using the measurement results of the indirect competition ELISA method in Example 2. Fluorescent whitening agent VBL monoclonal antibody at 1μg / L~1×10 5 Good linearity in the range of μg / L, IC 50 =857.9ng / mL, the lowest detection limit is 0.324ng / mL, and the detection range (between 20% and 80% inhibition) is 2.817ng / mL~1212.182μg / mL. The detection limit of paper and plastic products is 0.162ng / mm 2 , the detection limit of food samples is 0.162ng / g, and the detection limit of daily chemical products is 30ng / g.

[0067] 2. Determination of specificity.

[0068] The indirect competition ELISA method of Example 2 was used to measure the fluorescent whitening agent VBL structural analogues 2-hydroxyethylamino-1,3,5-triazine, 2-anilino-1,3,5-triazine, 2- Amino-4-anilino-6-hydroxyeth...

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Abstract

The invention discloses a fluorescent brightening agent VBL detection kit, belonging to the technical field of additive detection. The kit comprises an enzyme label plate coated by a fluorescent brightening agent VBL specific antigen and a fluorescent brightening agent VBL specific antibody solution, wherein each micropore of the enzyme label plate is coated by the fluorescent brightening agent VBL specific antigen having a concentration of 1-3 mug / mL, the concentration of the fluorescent brightening agent VBL specific antibody is 1-3 mug / mL, and the dosage ratio of the fluorescent brightening agent VBL specific antigen to the fluorescent brightening agent VBL specific antibody solution is 1:1. The invention further discloses a preparation method of the detection kit. The detection kit disclosed herein can continuously detect a plurality of samples at a time when the fluorescent brightening agent VBL is detected, has the characteristics of convenience, rapidness, sensitivity, and low cost, and is suitable for rapidly and accurately detecting fluorescent brightening agent VBL in mass samples.

Description

technical field [0001] The invention relates to an additive detection technology, in particular to an enzyme-linked immunological detection kit for fluorescent whitening agent VBL content and a preparation method thereof. Background technique [0002] Optical Brightener VBL (C.I. Optical Brightener 85), the chemical name is 4,4'-bis[(4-hydroxyethylamino-6-anilino-1,3,5-triazin-2-yl) Amino] stilbene-2,2'-sodium disulfonate, a stilbene triazine-type whitening agent, the fluorescence intensity is 100% ± 5%, the shade is blue light purple, soluble in water, neutral , has excellent level dyeing and bleeding properties, can improve the whiteness and gloss of the substance, and is widely used in paper, plastic products, detergents, textiles, etc. [0003] For a long time, the safety of fluorescent whitening agents has been controversial. Some data show that fluorescent whitening agents are low-toxicity, and some data show that fluorescent whitening agents have a stable molecular s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/545
Inventor 郭新东冼燕萍吴钟玲罗海英吴玉銮蔡玮红王斌陈意光
Owner GUANGZHOU QUALITY SUPERVISION & TESTING INST
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