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A malachite green hapten, artificial antigen and its application in fluorescence quantitative immunochromatography

A technology of malachite green and artificial antigen, which is applied in the field of immunochemistry, can solve the problems of expensive equipment operation, cumbersome, poor sensitivity, etc., and achieve the effect of improving immunogenicity, increasing recognition sites, and increasing potency

Active Publication Date: 2021-04-13
GUANGDONG DAYUAN OASIS FOOD SAFETY TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, gas chromatography and high performance liquid chromatography are mainly used for the detection of malachite green and cryptomalachite green residues, but the equipment is expensive and the operation is cumbersome, so it is not suitable for on-site rapid detection
Although the existing enzyme-linked immunoassay method has high sensitivity and high throughput, it also requires professional operation, and the detection time is long, and it also cannot meet the needs of rapid on-site screening; while the colloidal gold method has high throughput, It has the advantages of simple operation, short detection time and no need for professional and technical personnel to operate, but its poor sensitivity also makes the colloidal gold method unable to meet the requirements of national detection standards

Method used

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  • A malachite green hapten, artificial antigen and its application in fluorescence quantitative immunochromatography
  • A malachite green hapten, artificial antigen and its application in fluorescence quantitative immunochromatography
  • A malachite green hapten, artificial antigen and its application in fluorescence quantitative immunochromatography

Examples

Experimental program
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Effect test

Embodiment 1

[0044]A peacockstone green semi-antigen, the preparation method thereof is as follows:

[0045](1) Take 2.0 g (7.7 mmol) Compound II in a 50 ml round bottom flask, 20 mL of acetone, 2.6 g (19.3 mmol) compound I and 2.1 g (15.4 mmol) potassium carbonate were added, and at 65 ° C for 5 h, reaction After completing, 40 ml of purified water was decompressed, extracted twice with ethyl acetate, and then decompress the solvent, the column chromatography was purified 2.2 g of compound III (ethyl acetate / petroleum ether, 1 / 7, v / v );

[0046](2) 1.2 g (3.3 mmol) compound III is dissolved in 15 ml of anhydrous ethanol, add N, N-dimethylideil 2.0 g (16.5 mmol) and zinc chloride 2.2g (16.5 mmol), 80 ° C back to 26h After the reaction is completed, 40 ml of purified water is reduced, extracted with ethyl acetate, then decompress the solvent, and purify 0.8 g of compound IV (ethyl acetate / petroleum ether, 1 / 4, v / v) );

[0047](3) 0.8 g (1.0 mmol) of compound IV was dissolved in 5 ml of ethanol, an...

Embodiment 2

[0049]A peacockstone green semi-antigen, the preparation method thereof is as follows:

[0050](1) Take 2.0 g (7.7 mmol) of Compound II in 50 ml round bottom flask, 20 mL of acetone, 2.1 g (15.4 mmol) of Compound I and 1.7 g (11.6 mmol) potassium carbonate, reacting at 60 ° C for 2 h, reaction After completing, 40 ml of purified water was added, and 40 ml of purified water was added, ethyl acetate was extracted twice, and then the solvent was removed, and the column chromatography was purified. );

[0051](2) 1.2 g (3.3 mmol) compound III was dissolved in 13 ml of anhydrous ethanol, add N, N-dimethylideil 2.4 g (19.8 mmol) and zinc chloride 2.6 g (19.8 mmol), 85 ° C reflux 15h After the reaction is completed, 40 ml of purified water is reduced, extracted with ethyl acetate, then decompress the solvent, and purify 1.1 g of compound IV (ethyl acetate / petroleum ether, 1 / 4, v / v) );

[0052](3) 1.1 g (1.4 mmol) of compound IV was dissolved in 10 ml of ethanol, and 6 m of lithium hydroxide sol...

Embodiment 3

[0054]A peacockstone green semi-antigen, the preparation method thereof is as follows:

[0055](1) Take 2.0 g (7.7 mmol) Compound II in a 50 ml round bottom flask, 20 ml of acetone, 2.6 g (19.3 mmol) compound I and 2.1 g (15.4 mmol) potassium carbonate were added, and at 65 ° C for 3 h, reaction After completing, 40 ml of purified water was reduced pressure, extracted twice with ethyl acetate, and then decompress the solvent, column chromatography purified 2.1 g of compound III (ethyl acetate / petroleum ether, 1 / 7, v / v );

[0056](2) 1.2 g of Compound III was dissolved in 13 ml of anhydrous ethanol, added to N, N-dimethylideine 2.0 g (16.5 mmol) and zinc chloride 2.2 g (16.5 mmol), 83 ° C for 20 h, and the reaction is completed, Decompression removal, 40 ml of purified water was added, extracted with ethyl acetate, then decompress the solvent, and purified 0.9 g of compound IV (ethyl acetate / petroleum ether, 1 / 4, v / v);

[0057](3) 0.9 g of the compound IV was dissolved in 10 ml of eth...

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Abstract

The invention discloses a malachite green hapten, an artificial antigen and its application in fluorescence quantitative immunochromatography. The structure of the hapten has two malachite green molecules at the same time, so that the space of the hapten in the carrier protein is The position is more prominent, the recognition site is increased, the immunogenicity of the small molecule hapten is effectively improved, and the titer of the artificial antigen is also improved. The malachite green artificial antigen combined with ELISA detection technology has the lowest detection of malachite green The limit is 0.04 μg / L; using the fluorescent quantitative immunochromatography test strip prepared by the artificial antigen of the present invention to cooperate with the detection method of the present invention, the detection of malachite green can be realized quickly and conveniently, and the detection sensitivity can reach 0.2 μg / L.

Description

Technical field[0001]The present invention belongs to the field of immunochemical, and is specifically involved in a peacockstone green semi-antigen, artificial antigen, and its application in fluorescence quantitative immunochromatography.Background technique[0002]Malachite Green, Mg) is also known alkaline green, salt base green, peacock green and China green, is a green crystal with metallic gloss, the chemical name is tetramethyl dialatine, the molecular formula is C23Hide25N2CL, the molecular weight is 364.92, which is soluble in water, methanol and ethanol, and the solution is blue and green. Peacockstone Green is an artificially synthetic triphenylmethane industrial dye, mainly used in industrial colors and cell chemical staining agents such as pottery, textile, leather and food. Peacockstone green has an effect of antibacterial, insecticidal, is a class of antibacterial efficacy in pharmacotic dyes, which is widely used as a insect repellent and insecticide to kill parasite,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07C229/26C07C227/18C07K14/765C07K14/77C07K14/795C07K16/44G01N33/558G01N33/577
CPCC07C227/06C07C227/16C07C227/18C07C229/26C07K14/765C07K14/77C07K14/795C07K16/44C07K19/00G01N33/558G01N33/577
Inventor 李斌石松谢俊平梁巧雯石林花任季玉
Owner GUANGDONG DAYUAN OASIS FOOD SAFETY TECH CO LTD
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