Visual detection method for aflatoxin B1

A kind of technology of aflatoxin and detection method, which is applied in the field of food safety detection, can solve the problems of being unsuitable for large-scale sample detection, expensive instruments and equipment, and low detection sensitivity, so as to facilitate on-site real-time detection, easy preparation and storage, Simple operation effect

Inactive Publication Date: 2018-05-04
中山市食品药品检验所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Currently, for the detection of aflatoxin B 1 The method mainly contains thin-layer chromatography (TLC), high performance liquid chromatography (HPLC) and enzyme-linked immunoassay (ELISA), and these methods respectively have its advantages and disadvantages, and the equipment that TLC method needs is simple, but detection sensitivity is low; HPLC The method has the advantages of high sensitivity and strong specificity, but it needs complex pretreatment before liquid chromatography separation, which is not suitable for the detection of large quantities of samples and the equipment is expensive; the ELISA method has the advantages of strong specificity and short analysis time. But the stability is poor and the accuracy is not high

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  • Visual detection method for aflatoxin B1
  • Visual detection method for aflatoxin B1
  • Visual detection method for aflatoxin B1

Examples

Experimental program
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Effect test

Embodiment 1

[0049] 1. Synthesis of sulfhydryl-modified aptamer DNA 1 , and the complementary strand DNA paired with the aptamer 2 (purchased from Shanghai Sangon Bioengineering Co., Ltd.).

[0050] (1) Aptamer DNA 1 The sequence is:

[0051] 5'SH-AGCAGCACAGAGGTCAGATGGTGCTATCATGCGCTCAATGGGAGACTTTAGCTGCCCCCACCTATGCGTGCTACCGTGAA-3';

[0052] (2) Complementary strand DNA 2 The sequence is:

[0053] 5'SH-TTCACGGTAGCACGCATAGGTGGGGGCAGCTAAAGTCTCC-3';

[0054] 2. Preparation of gold nanoparticles.

[0055] (1) Soak the used container with aqua regia for 12h, and the aqua regia is prepared by concentrated hydrochloric acid and concentrated nitric acid with a volume ratio of 3:1;

[0056] (2) clean the container soaked in aqua regia with ultrapure water, and dry;

[0057] (3) Add 50mL of ultrapure water with resistivity ≥ 18.2MΩ and 0.5mL, 1% chloroauric acid solution into the container, stir evenly with a rotor, boil for 10min, add 2mL, 1% sodium citrate, which contains 0.05% Citric acid,...

Embodiment 2

[0077] Aflatoxin B in actual samples of peanut butter 1 detection and detection of spiked recovery.

[0078] (1) Test sample pretreatment: Weigh 4.0g of peanut butter and put it in a 50mL centrifuge tube, add 10mL of 70% methanol extraction solution, shake vigorously on the shaker for 10min, let it stand for 30min and filter it with quantitative filter paper, take 0.1mL of the filtrate, Then add 1.9mL ultrapure water for dilution to obtain the solution to be tested for future use.

[0079] (2) Utilize the inventive method to measure aflatoxin B in the sample 1 content, the experimental procedure is the same as in Example 1, and the results are shown in Table 1.

[0080] (3) Aflatoxin B obtained with step (2) 1 The concentration data is the background value, to which different concentrations of AFB are added 1 Standard substance, also utilize the method of the present invention to detect aflatoxin B again 1 content to obtain the detection value. Recovery %=(detection value-...

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Abstract

The invention discloses a visual detection method for aflatoxin B1. The visual detection method comprises the following steps: respectively linking sulfhydrylated aflatoxin B1 aptamer DNA1 and a complementary chain DNA2 of a sulfhydrylated aptamer to 13nm+/-2nm nanogold, so to obtain an aptamer-nanogold probe and a complementary chain-nanogold probe, and carrying out detection by virtue of a competition method, wherein a drone is bonded with the aptamer when a to-be-detected object contains the drone, the aptamer-nanogold probe and the complementary chain-nanogold probe are in a free state, nanogold is easily gathered under a high salt concentration condition, the color of the nanogold is changed from red to blue, when the to-be-detected object does not contain the drone, the nanogold is not gathered under the high salt concentration condition, and the color of the nanogold is still red and is gradually changed from red to blue along the increase of the concentration of the drone; anddetecting an ultraviolet-visible spectrum of the solution at 200nm-800nm, so as to realize the quantitative detection of aflatoxin B1 in the to-be-detected object.

Description

technical field [0001] The invention relates to the fields of nanomaterials and molecular biology, and is used for the detection of food safety. Background technique [0002] Aflatoxins are metabolites produced by Aspergillus flavus and Aspergillus parasiticus, and are mainly divided into aflatoxin B 1 , B 2 , G 1 , G 2 and two other metabolites M 1 , M 2 , where aflatoxin B 1 It is the most toxic and has acute and chronic toxicity, mutagenicity, carcinogenicity and teratogenicity to humans and animals. Its metabolism mainly occurs in the liver, so it has great damage to the liver. Aflatoxins are easily soluble in various organic solvents, such as chloroform, methanol, ethanol, propanol, and dimethylformamide, etc., but insoluble in water, and insoluble in petroleum ether, ether, and hexane. Aflatoxin B 1 The range of contaminated food is very wide, mainly peanuts, corn, rice, wheat, peanut oil and other food crops. The southern part of my country is the most pollute...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N33/53G01N33/543
CPCG01N21/78G01N33/5308G01N33/54346
Inventor 谭贵良刘子雄王乾蕾李向丽郑鸿涛陈坚
Owner 中山市食品药品检验所
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