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Malachite green hapten, artificial antigen and application of artificial antigen in fluorescence quantitative immunochromatography

A malachite green and artificial antigen technology, applied in the field of immunochemistry, can solve the problems of long detection time, expensive operation of equipment, cumbersome and other problems, meet the requirements in the process of storage and transportation, improve immunogenicity, and increase recognition sites. Effect

Active Publication Date: 2018-10-12
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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  • Malachite green hapten, artificial antigen and application of artificial antigen in fluorescence quantitative immunochromatography
  • Malachite green hapten, artificial antigen and application of artificial antigen in fluorescence quantitative immunochromatography
  • Malachite green hapten, artificial antigen and application of artificial antigen in fluorescence quantitative immunochromatography

Examples

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Embodiment 1

[0044] A kind of malachite green hapten, its preparation method is as follows:

[0045] (1) Take 2.0g (7.7mmol) of compound II in a 50ml round bottom flask, then add 20ml of acetone, 2.6g (19.3mmol) of compound I and 2.1g (15.4mmol) of potassium carbonate in turn, and react at 65°C for 5h. After completion, the solvent was removed under reduced pressure, 40ml of purified water was added, extracted twice with ethyl acetate, and the solvent was removed under reduced pressure, and purified by column chromatography to obtain 2.2g of compound III (ethyl acetate / petroleum ether, 1 / 7, v / v );

[0046](2) Dissolve 1.2g (3.3mmol) of compound III in 15ml of absolute ethanol, add 2.0g (16.5mmol) of N,N-dimethylaniline and 2.2g (16.5mmol) of zinc chloride, and reflux at 80°C for 26h , after completion of the reaction, remove the solvent under reduced pressure, add 40ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatogr...

Embodiment 2

[0049] A kind of malachite green hapten, its preparation method is as follows:

[0050] (1) Take 2.0g (7.7mmol) of compound II in a 50ml round bottom flask, then add 20ml of acetone, 2.1g (15.4mmol) of compound I and 1.7g (11.6mmol) of potassium carbonate in turn, and react at 60°C for 2h. After completion, the solvent was removed under reduced pressure, 40ml of purified water was added, extracted twice with ethyl acetate, and the solvent was removed under reduced pressure, and purified by column chromatography to obtain 1.9g of compound III (ethyl acetate / petroleum ether, 1 / 7, v / v );

[0051] (2) Dissolve 1.2g (3.3mmol) of compound III in 13ml of absolute ethanol, add 2.4g (19.8mmol) of N,N-dimethylaniline and 2.6g (19.8mmol) of zinc chloride, and reflux at 85°C for 15h , after completion of the reaction, remove the solvent under reduced pressure, add 40ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatog...

Embodiment 3

[0054] A kind of malachite green hapten, its preparation method is as follows:

[0055] (1) Take 2.0g (7.7mmol) of compound II in a 50ml round-bottomed flask, then add 20ml of acetone, 2.6g (19.3mmol) of compound I and 2.1g (15.4mmol) of potassium carbonate in turn, and react at 65°C for 3h. After completion, the solvent was removed under reduced pressure, 40ml of purified water was added, extracted twice with ethyl acetate, and the solvent was removed under reduced pressure, and purified by column chromatography to obtain 2.1g of compound III (ethyl acetate / petroleum ether, 1 / 7, v / v );

[0056] (2) Dissolve 1.2g of compound III in 13ml of absolute ethanol, add 2.0g (16.5mmol) of N,N-dimethylaniline and 2.2g (16.5mmol) of zinc chloride, reflux at 83°C for 20h, and the reaction is complete. Remove the solvent under reduced pressure, add 40 ml of purified water, extract with ethyl acetate, remove the solvent under reduced pressure, and purify by column chromatography to obtain ...

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Abstract

The invention discloses a malachite green hapten, an artificial antigen and an application of the artificial antigen in fluorescence quantitative immunochromatography. The structure of the hapten hastwo malachite green molecule structures at the same time, so a space position of the hapten in a carrier protein is more prominent, recognition sites are increased, the immunogenicity of the small-molecular hapten is effectively improved, and the titer of the artificial antigen is also improved; the malachite green artificial antigen is combined with an ELISA detection technology, and the lowest detection limit of malachite green is 0.04 [mu]g / L; a fluorescence quantitative immunochromatographic test strip prepared by the artificial antigen cooperates with a detection method, malachite green can be detected quickly and conveniently, and the detection sensitivity can reach 0.2 [mu]g / L.

Description

technical field [0001] The invention belongs to the technical field of immunochemistry, and in particular relates to a malachite green hapten, an artificial antigen and an application thereof in fluorescence quantitative immunochromatography. Background technique [0002] Malachite Green (Malachite Green, MG), also known as basic green, base block green, malachite green and Chinese green, is a green crystal with metallic luster, its chemical name is tetramethyldiaminotriphenylmethane, and its molecular formula is C 23 h 25 N 2 Cl, with a molecular weight of 364.92, is easily soluble in water, methanol and ethanol, and its solution is blue-green. Malachite green is a kind of artificially synthesized triphenylmethane industrial dye, which is mainly used as a color agent in pottery, textile, leather and food industries, and as a dye in cell chemistry. Malachite green has antibacterial and insecticidal effects. It is a kind of medicinal dye with strong antibacterial effect. ...

Claims

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Application Information

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Patent Type & Authority Applications(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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