Leuco malachite green hapten, produced antibody and application of the antibody

A cryptomalachite green and hapten technology, applied in the field of immunochemistry, can solve the problems of tedious operation, expensive equipment, and inapplicability for rapid detection, etc., and achieves the effects of reducing pollution, far-reaching application value, and easy post-reaction treatment.

Inactive Publication Date: 2008-08-20
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, gas chromatography and high performance liquid chromatography are mainly used for the detection of malachite green and cryptom

Method used

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  • Leuco malachite green hapten, produced antibody and application of the antibody
  • Leuco malachite green hapten, produced antibody and application of the antibody
  • Leuco malachite green hapten, produced antibody and application of the antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 The synthetic route of the hapten is shown in Figure 5.

[0041] Take 2.50g (20.0mmol) of N,N-dimethylaniline into a 50ml two-necked flask, slowly add 5% Amberlyst 15 Resin, magnetically stir for a few minutes, the magnetic force and stirring time should be selected according to conventional technology, the time is generally 5 ~10min. Under stirring, add 1.50 g of 4-carboxybenzaldehyde or 3-carboxybenzaldehyde, and stir overnight at 80-100°C. After the completion of the reaction, the catalyst was filtered and saturated sodium bicarbonate solution was added to the reaction system to adjust the pH to pH=9.0, and extracted with dichloromethane. The aqueous phase was adjusted to pH=2 with dilute hydrochloric acid, extracted with ethyl acetate, and the organic layer was rotary evaporated under reduced pressure to obtain a crude product. The crude product was purified by reduced pressure silica gel column chromatography to a white solid substance, the structure of which...

Embodiment 2

[0043] Example 2 The synthetic route of the hapten is shown in Figure 6.

[0044] Take 2.50g (20.0mmol) of N,N-dimethylaniline in a 50ml two-necked flask, slowly add 5% Amberlyst 15 Resin, and magnetically stir for several minutes. Under stirring, add 1.53 g of 3-nitrobenzaldehyde or 4-nitrobenzaldehyde. Under nitrogen flow, stir overnight at 80-100°C. After the reaction is completed, the catalyst is filtered, and a yellow solid is obtained by rotary evaporation under reduced pressure.

[0045] Add 3.75g (about 10mmol) of the above crude product in a round bottom flask, add 10ml of 95% ethanol, stir to dissolve it, add 4g (about 70mmol) of iron powder, 20ml of distilled water, and finally add 1ml of concentrated hydrochloric acid, and reflux at 80℃. Stop the reaction at 2h. The water phase was obtained by suction filtration, the organic phase was obtained by extraction with ethyl acetate, and the light yellow crude product was obtained by evaporation under reduced pressure. The ob...

Embodiment 3

[0047] Example 3 The synthetic route of the hapten is shown in Figure 7.

[0048] Take 12.2g of 3-hydroxybenzaldehyde or 4-hydroxybenzaldehyde in a 150ml round bottom flask, dissolve it with 20ml of distilled water, add 4g of NaOH and stir to react. Then, 11.6 g of sodium chloroacetate solution was added dropwise under stirring, and the reaction was stirred at 80-100°C under reflux for 2 to 3 hours. After the reaction is over, add proper amount of concentrated hydrochloric acid to acidify, then add saturated sodium bicarbonate solution to adjust to basicity, extract with ethyl acetate, acidify the aqueous phase with dilute hydrochloric acid until precipitation is precipitated, dry and precipitate under reduced pressure to obtain the intermediate product.

[0049] The intermediate product and N,N-dimethylaniline were added to a 50ml two-necked flask at a molar ratio of 1:2, and 5% Amberlyst 15 Resin was added at the same time. Under nitrogen, the mixture was stirred at 80-100°C ove...

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PUM

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Abstract

The invention discloses an implicit malachite green hapten which retains the characteristic structures of the implicit malachite green, active arms which have different electronic structural properties and different carbon chain length and is suitable for being coupled with macromolecule carriers derive from different chemical sites of the structure of the implicit malachite green, the active arm is R group, aromatic amine, terminal carboxyl group linear chain alkyl ether or amido link with the structure of linear chain terminal carboxyl group; an implicit malachite green artificial antibody is prepared and obtained from implicit malachite green hapten and when being used for detecting the residual quantity of the implicit malachite green and malachite green, the antibody has high specificity and sensitivity and high accuracy, the recovery rate can reach 80 to 110 percent, at the same time, the operation is simple and fast, no complicated pretreatment process is needed, a great amount of samples can be detected at one time, the cost is low, the operation has little requirements to operators so as to be convenient for on-site monitoring and be complementary to conventional methods, therefore, a sensitive, fast, efficient and economical implicit malachite green hapten immunity detecting method is successfully set up.

Description

Technical field [0001] The invention belongs to the technical field of immunochemistry, and specifically relates to a cryptomalachite green hapten and a preparation method thereof, as well as antibodies prepared by using the hapten and applications of the antibodies. Background technique [0002] The residue of illegal drugs in aquatic products is one of the more prominent problems in food safety in recent years, and it is a problem that plagues the world. Since 1997, the Ministry of Agriculture has issued several documents to prohibit the use of nitrofuran, malachite green and other nearly 40 animal (fishing) drugs in the production of animal food. However, due to the lack of legal awareness and related science by some producers and vendors Knowledge, especially driven by economic interests, often illegally use banned drugs in the breeding process. This phenomenon is still common and threatens people’s health seriously. At the same time, it seriously affects the competitiveness ...

Claims

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Application Information

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IPC IPC(8): C07C229/40C07C211/50C07C217/80C07C233/43C07K16/18G01N33/53
Inventor 沈玉栋王宇孙远明雷红涛王弘肖治理徐小艳
Owner SOUTH CHINA AGRI UNIV
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