Preparation method of Alpha-amanita hemolysin molecular imprinting material

A technology of molecular imprinting and amanita toxin, applied in chemical instruments and methods, other chemical processes, etc., can solve the problems of harsh conditions, complex operation, complex structure, etc., and achieve short time, mild reaction conditions and simple experimental operation. Effect

Active Publication Date: 2016-08-24
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most successful application of molecular imprinting technology is the identification of small organic molecules. The successful examples of identifying peptides and proteins are very limited. The reason is the complex structure of biological macromolecules and thermodynamic factors. Another main reason is that peptides and proteins are expensive. It is often difficult to obtain sufficient quantities for the preparation of mol...

Method used

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  • Preparation method of Alpha-amanita hemolysin molecular imprinting material
  • Preparation method of Alpha-amanita hemolysin molecular imprinting material

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1: Preparation of template molecule

[0032] Template molecule of the present invention is N-acetyl tryptophan propionamide, and its preparation method comprises the following steps:

[0033] a. Weigh 4.0 g of N-acetyl tryptophan, measure 100 ml of methanol and add it to a 250 ml three-necked flask to dissolve fully in an ultrasonic environment;

[0034] b. Add 5.0 ml of concentrated sulfuric acid as a catalyst to the mixed solution in the above steps, and reflux for 3 hours in a water bath environment at 60°C to fully esterify N-acetyl tryptophan and methanol;

[0035] c. After the device is completely cooled to room temperature, use 1.0mol / L K 2 CO 3 The solution adjusts the pH of the mixed solution to be 7, and then adds a saturated NaCl solution to allow it to stand for stratification;

[0036] After d, 30min, extract the mixed solution three times with 120ml of dichloromethane;

[0037] e, then use 50ml 1.0mol / L K 2 CO 3 Solution and 60ml of deion...

Embodiment 2

[0043] Example 2: Preparation of surface mercaptolated silica

[0044] Carrier of the present invention is the silicon dioxide of surface mercaptolation, and its preparation method comprises the following steps:

[0045] a. Take 4.0g of silica microspheres, ultrasonically disperse them in 5ml of deionized water, add 120ml of methanol, and ultrasonically disperse for 30min;

[0046] b. Add 120ml of glycerin to the reaction solution obtained in step a of this embodiment, and then sonicate for 10min, transfer the mixed solution into a 500ml three-necked flask, and pass nitrogen gas for 20min to obtain mixed solution 1;

[0047] c. Take 4ml of (3-mercaptopropyl)-triethoxysilane and mix with 40ml of methanol evenly, and pour it into mixed solution 1, start to stir and react, and get mixed solution 2;

[0048] d. Add 10ml of ammonia water to the mixed solution 2, react at 85°C for 1 hour under the protection of nitrogen, and then react for 5 hours after stopping the protection of...

Embodiment 3

[0050] Example 3: Preparation of α-amanitin surface molecularly imprinted material (α-amanitin-MIP)

[0051] The preparation method of the α-amanitin surface molecular imprinting material of the present invention is as follows:

[0052] S1: Add the template molecule prepared in Example 1 and 3-mercaptopropionic acid, ethylene glycol dimethacrylate, and pentaerythritol tetra-3-mercaptopropionate into the solution of dimethyl sulfoxide and stir evenly; The molar ratio of molecule, 3-mercaptopropionic acid and ethylene glycol dimethacrylate is 0.2:1:2; the auxiliary crosslinking agent pentaerythritol tetra-3-mercaptopropionate and crosslinking agent ethylene glycol dimethyl The mol ratio of base acrylate is 9:4;

[0053] S2: Add the mercaptoated silica microspheres obtained in Example 2 into the mixed solution prepared in step S1 of this example, adjust the pH of the reaction solution to about 8 with triethylamine, and polymerize at 40° C. for 6 hours;

[0054] S3: taking out...

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Abstract

The invention provides a preparation method of an Alpha-amanita hemolysin molecular imprinting material, and relates to the field of a separation material. The method comprises the following steps of S1, uniformly mixing a template molecule, a functional monomer, a cross-linking agent and a solvent; S2, performing sulfhydrylation treatment on the carrier surface; S3, adding a carrier obtained through the treatment in the S2 into a mixed solution prepared in the S1, and performing cross-linking polymerization with the functional monomer; S4, eluting the template molecule from the polymerized carrier surface; S5, performing vacuum drying on the molecular imprinting material obtained through treatment in the S4. The preparation method has the advantages that the experiment operation is simple and convenient; nitrogen introduction and oxygen removal are not needed; the reaction can be triggered by regulating the pH value of a reaction system by triethylamine; the reaction condition is mild; the time is short; the method shows great superiority than that of a traditional molecular imprinting preparation method.

Description

technical field [0001] The invention relates to a separation material, in particular to a method for preparing an α-amanitin molecularly imprinted material. Background technique [0002] Amanitin, also known as amanitin, is a polypeptide substance isolated from highly toxic mushrooms, and it is also a class of important biochemical reagents. Amanitin is used in molecular biology, developmental biology, and genetics Science, biochemistry, medicine, biological control and other fields have a wide range of application value. Amanitin is a kind of bicyclic octapeptide. There are 9 kinds of natural amanitin toxins that have been isolated and purified. They are α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amanin, amaninamide, amanullin, amanullinic acid, and proamanullin. The structural formula of α-amanitin is as follows: [0003] [0004] Molecular imprinting is a new type of "artificial receptor" synthesis technology developed from analytical chemistry, polymer materia...

Claims

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Application Information

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IPC IPC(8): C08G75/045C08J9/26B01J20/26B01J20/30
CPCB01J20/268B01J2220/4812C08G75/045C08J9/26C08J2201/0424C08J2381/02
Inventor 滕伟迪梁勇汤又文袁嫣昊谭杰安
Owner SOUTH CHINA NORMAL UNIVERSITY
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