Amanitin Molecularly Imprinted Materials for Solid Phase Extraction of α-amanitin and β-amanitin

A technology of molecular imprinting and amanita toxin, which can be used in analytical materials, material separation, instruments, etc., can solve problems such as limiting the development of SPE, and achieve the effect of improving sensitivity

Active Publication Date: 2019-01-01
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The interaction between the target substance and the adsorbent in traditional SPE is non-specific, and usually requires careful selection of extraction and elution conditions, and the separation of different matrices and analytes require different column packing materials, which limits SPE. further development of

Method used

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  • Amanitin Molecularly Imprinted Materials for Solid Phase Extraction of α-amanitin and β-amanitin
  • Amanitin Molecularly Imprinted Materials for Solid Phase Extraction of α-amanitin and β-amanitin
  • Amanitin Molecularly Imprinted Materials for Solid Phase Extraction of α-amanitin and β-amanitin

Examples

Experimental program
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Effect test

preparation example Construction

[0046] In this embodiment, the preparation method of the amanitin molecularly imprinted material in S1 includes the following steps:

[0047] S11: mixing the template molecule N-acetyl tryptophan propionamide with the functional monomer 3-mercaptopropionic acid, a cross-linking agent, and a solvent;

[0048] S12: subjecting the silica surface to mercapto treatment;

[0049] S13: adding the silicon dioxide obtained in S12 to the mixed solution prepared in S11, and cross-linking and polymerizing the functional monomers and template molecules;

[0050] S14: using an eluent to elute the template molecules from the surface of the polymerized silica, and drying to obtain the imprinted material of amanitin molecules.

[0051]Wherein, the molar ratio of template molecule, functional monomer and crosslinking agent in S11 is preferably 0.2:1:2. The crosslinking agent in S11 is ethylene glycol dimethacrylate, and the solvent is dimethyl sulfoxide. The S11 also includes an auxiliary cr...

Embodiment 1

[0059] Example 1: Preparation of Amanitin Molecularly Imprinted Materials

[0060] This example describes in detail the preparation method of the amanitin molecularly imprinted material for solid phase extraction of α-amanitin and β-amanitin.

[0061] see figure 1 , which is a schematic diagram of the preparation process of the amanitin molecularly imprinted material of the present invention. The preparation method of the amanitin molecularly imprinted material comprises the following steps:

[0062] S11: Combine the template molecule N-acetyl tryptophan propionamide with the functional monomer 3-mercaptopropionic acid, the cross-linking agent ethylene glycol dimethacrylate, the auxiliary cross-linking agent pentaerythritol tetra-3-mercapto propionate and di Methyl sulfoxide solvent mixed. In this embodiment, the molar ratio of the template molecule, the functional monomer, and the crosslinking agent is preferably 0.2:1:2. The molar ratio of the auxiliary crosslinking ag...

Embodiment 2

[0069] Embodiment 2: sample pretreatment

[0070] In the present embodiment, the sample is a mushroom sample, and the specific pretreatment method includes the following steps:

[0071] S21: drying the mushroom sample to constant weight.

[0072] S22: Take 5.0 g of mushroom samples dried to constant weight, add 25 mL of methanol solution with a volume fraction of 50%, place in a shaker and shake at 150 r / min for 24 hours.

[0073] S23: Centrifuge the methanol solution shaken by S22 at 10,000r / min for 10min, and take the supernatant; add 25mL of methanol solution with a volume fraction of 50% to the precipitate obtained by centrifugation, and shake again to take the supernatant; The supernatants were combined, and the methanol was removed by rotary evaporation to obtain the mushroom extract.

[0074] S24: Filter the extract treated in S23 with a 0.45 μm filter membrane to obtain a test solution.

[0075] In this embodiment, the volume of various solvents added in the sampl...

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Abstract

The invention discloses a method for solid-phase extraction of alpha-amanitin and beta-amanitin with an amanitin molecularly imprinted material. The method comprises the following steps that firstly, the amanitin molecularly imprinted material with N-acetyl tryptophan propanamide serving as a template molecule is synthesized; secondly, a sample is pretreated, wherein alpha-amanitin and beta-amanitin in the sample to be detected are extracted, and a solution to be detected is obtained; thirdly, solid-phase extraction is carried out, wherein the amanitin molecularly imprinted material prepared in the first step is added into the solution to be detected, and the amanitin molecularly imprinted material is separated after oscillation; fourthly, alpha-amanitin and beta-amanitin in the amanitin molecularly imprinted material are eluted with a certain volume of an elution solvent, and eluant is collected; fifthly, the concentration of alpha-amanitin and the concentration of beta-amanitin in the eluant are measured with the HPLC. According to the method, the detection method of combining amanitin molecularly imprinted material extraction and the HPLC is established, and alpha-amanitin and beta-amanitin in the complex sample are enriched through the amanitin molecularly imprinted material, so that matrix interference is reduced, the sensitivity, accuracy, precision and selectivity of the method are improved, and the detecting sensitivity on alpha-amanitin and beta-amanitin is greatly improved.

Description

technical field [0001] The invention relates to the field of solid-phase extraction, in particular to a method for solid-phase extraction of amanitin molecularly imprinted materials for α-amanitin and β-amanitin. Background technique [0002] Mass mushroom poisoning incidents occur frequently in our country, and the social concern is extremely high. Mushroom poisoning is mostly caused by eating poisonous Amanita. Amanita toxins are mainly amanita peptides. There are 9 kinds of amanita peptides isolated and identified in my country, among which α-amanitin has the highest content and is the most toxic, followed by β-amanitin. If patients with amanitin poisoning are not diagnosed and rescued in time, they will die of liver failure within 2-8 days. [0003] However, the content of α-amanitin and β-amanitin in mushroom samples is very low, so it is necessary to enrich the trace α-amanitin and β-amanitin components in the sample to reduce matrix interference and improve the sens...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/14G01N30/08C08F292/00
CPCC08F292/00G01N30/02G01N30/06G01N30/08G01N30/14G01N2030/027G01N2030/062C08F222/102
Inventor 陈忻陈晓刚梁勇
Owner FOSHAN UNIVERSITY
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