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Method for preparing aflatoxin B1 aptamer affinity column

An aflatoxin and aptamer technology, which is applied in the fields of affinity chromatography and mycotoxin detection, and can solve problems such as no reports on the application of aptamer affinity column preparation methods.

Active Publication Date: 2015-03-11
INST OF AGRI PROD QUALITY SAFETY & STANDARD JIANGXI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Aptamer affinity chromatography combined with conventional instrumental analysis has become a development direction for mycotoxin analysis, but there is no report on the preparation method and application of aptamer affinity columns

Method used

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  • Method for preparing aflatoxin B1 aptamer affinity column

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Embodiment 1

[0011] Embodiment 1 Preparation of aflatoxin B1 aptamer affinity column:

[0012] 1 Activation: Suspend and swell 60 mg of epoxy-activated agarose microspheres FF in 2 mL of deionized water to form a gel for 1 hour. Wash the gel three times with 60mL deionized water, let it stand for 5min, and remove the washing solution;

[0013] 2 Coupling: Dissolve the amino-modified aflatoxin B1 aptamer in 1 mL of 20.0 mmol / L pH 10.0 phosphate buffer. Suspend the swollen gel in step 1 in the aptamer-containing buffer and shake in a water bath at 37°C for 16 hours. After the reaction, wash the unreacted aptamer with 3mL deionized water, and after standing for 5min, remove the cleaning solution;

[0014] 3 Sealing: Suspend the gel in step 2 into 2mL of 1mol / LpH8.0 ethanolamine solution, shake in a water bath at 50°C for 4 hours, after the reaction, use 3mL of 0.1mol / LpH4.0 acetic acid containing 0.5mol / LNaCl Buffer and 3 mL of pH 8.3 coupling buffer containing 0.5mol / L NaCl are washed alt...

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Abstract

The invention discloses a method for preparing an aflatoxin B1 aptamer affinity column and belongs to the field of affinity chromatography and mycotoxin analysis. The method comprises the following steps: adopting epoxy activated agarose microspheres FF as a solid phase carrier; after the solid phase carrier is coupled with an amino-modified aflatoxin B1 aptamer, closing extra active sites in the carrier, and feeding into micro columns. The prepared aptamer affinity column can be specially combined with aflatoxin B1, the adding standard recovery is 70-110, and the aptamer affinity column can be repeatedly used for at least five times.

Description

technical field [0001] The invention relates to the technical field of affinity chromatography and mycotoxin detection, in particular to a preparation method and application of an aflatoxin B1 aptamer affinity column. Background technique [0002] Aflatoxin B1 is a metabolite produced by Aspergillus flavus and Aspergillus parasiticus. It has carcinogenic, teratogenic and mutagenic effects. It is the most stable mycotoxin found so far. It is not easy to be destroyed by general food processing conditions, so It has laid a huge hidden danger to the food safety of consumers. Countries attach great importance to the status quo of aflatoxin residues in food and have formulated corresponding limit standards. For example, my country stipulates that the allowable amount of aflatoxin in rice and edible oil is (B1 + B2 + G1 + G2) 10 ng / g. [0003] At present, among the analysis methods of aflatoxin B1, high performance liquid chromatography, capillary electrophoresis, liquid chromat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/38B01J20/281G01N1/34
Inventor 廖且根罗林广李伟红胡丽芳
Owner INST OF AGRI PROD QUALITY SAFETY & STANDARD JIANGXI ACAD OF AGRI SCI
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