Method for simultaneously detecting zearalenone and fumonisin B1

A technology of zearalenone and fumonisins, applied in measuring devices, material analysis through electromagnetic means, instruments, etc., can solve problems such as large subjective influence, easy false positives, and easy influence of enzyme activity by operating conditions , to achieve the effect of simple synthesis method, accelerated transfer rate, and reduced risk of poisoning

Active Publication Date: 2020-02-18
SHANGHAI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thin-layer chromatography is low in cost and easy to operate, but semi-quantitative visual inspection is greatly affected by subjectivity; ELISA has strong specificity, simple sample pretreatment, and short analysis time, but the enzyme activity is easily affected by operating conditions and prone to occurrence False positive; high performance liquid chromatography and mass spectrometry have good sensitivity, accuracy and stability, and are suitable for various samples containing complex components. However, this method is only common in scientific research institutes and testing institutions, and is not suitable for grassroots application and promotion, and requires complex pre-processing methods to ensure the accuracy of results, and cannot achieve rapid detection and on-site instant analysis

Method used

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  • Method for simultaneously detecting zearalenone and fumonisin B1
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  • Method for simultaneously detecting zearalenone and fumonisin B1

Examples

Experimental program
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Effect test

Embodiment 1

[0069] Example 1 Preparation of aptasensor

[0070] The first step, preparation of gold nanoparticle dispersion:

[0071] 1mL 10mmol / L HAuCl 4 ·3H 2 O and 37mL of ultrapure water were heated to boiling under stirring, and 1mL of 0.1mol / L sodium citrate solution was quickly added. After the solution turned wine red, the heating was stopped, and it was naturally cooled to room temperature to obtain the desired gold nanoparticle dispersion. liquid.

[0072] The second step, the preparation of CP1-Au-Thi:

[0073] Dilute AP1 to 2 μmol / L with TE buffer (pH 8.0), take 1 mL of AP1 solution and 4 mL of gold nanoparticle dispersion and shake slightly overnight for 8 h, centrifuge at room temperature for 10 min (5000 rpm), remove the supernatant, and remove the precipitate Redisperse in 1mL TE solution. Add 1mL of 10mmol / L Thi solution, shake slightly overnight for 8h, let it stand at room temperature, centrifuge for 10min (5000rpm), remove the supernatant, and redisperse the preci...

Embodiment 2

[0086] Example 2 Using the aptamer sensor of Example 1 to simultaneously detect ZEN and FB in corn 1

[0087] (1) Pretreatment of corn samples: Grind two purchased commercial corn samples (No. 1 and 2), weigh 2 g and add 8 mL of acetonitrile / water / formic acid (79 / 20 / 1, v / v / v) , vortexed for 2min, centrifuged at 4°C for 10min (3000rpm), took the supernatant, added the same volume of acetonitrile / water / formic acid (20 / 79 / 1, v / v / v), passed through a 0.22μm filter membrane, to be tested. Wherein corn samples 1 and 2 were respectively made 3 parallels;

[0088] (2) Detection by CHI660A electrochemical workstation and three-electrode detection device:

[0089] Pipette 100μL sample into 1.9mL 0.01mol / L PBS (pH 6.0) at 4°C to detect ZEN and FB 1

[0090] (3) Use CHI660A electrochemical workstation and three-electrode detection device for detection:

[0091] The saturated calomel electrode is used as the reference electrode, the platinum wire electrode is used as the counter elec...

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Abstract

The invention discloses a method for simultaneously detecting zearalenone (ZEN) and fumonisin B1 (FB1), and also discloses an aptamer sensor for simultaneously detecting the zearalenone and the fumonisin. The aptamer sensor is prepared by the following method: (1) covering a glassy carbon electrode with reducing molybdenum disulfide and a gold nanoparticle composite (rMoS2-Au) nanomaterial, and drying; (2) subsequently dripping a mixed solution of a zearalenone nucleic acid aptamer (AP1) and a fumonisin B1 nucleic acid aptamer (AP2) onto the electrode, incubating for 1-10h at 1-10 mu mol/L, v/v, 1/1, and blocking the same with 1 mg/mL QBSA; and (3) finally, dripping a mixed solution of a self-developed ZEN nucleic acid aptamer partial complementary sequence (CP1-Au-Thi) solution of 5'-endmodified gold nanoparticles and thionine and a FB1 nucleic acid aptamer partial complementary sequence (CP2-Au-FC6S) solution of 5'-end modified gold nanoparticles and ferrocenyl hexanethiol, and incubating for 1-10h at 1-10 mu mol/L, v/v, 1/1 to obtain the required aptamer sensor. The method provided by the invention is suitable for the rapid and simultaneous detection of QEN and FB1 in agricultural products such as corn and feed, thereby effectively preventing the agricultural products with over-standard QEN and FB1 from flowing into the market, and the diet safety of people is guaranteed asa result.

Description

technical field [0001] The invention relates to the field of crop testing and analysis, in particular to a method for simultaneously detecting zearalenone (ZEN) and fumonisin B 1 (FB 1 )Methods. Background technique [0002] Zearalenone (ZEN) is a dihydroxybenzoic acid lactone metabolite with estrogen activity produced by a variety of Fusarium species, which is widely found in corn, wheat, sorghum, rice and other cereals. The structure of ZEN is similar to that of endogenous estrogen, and it can specifically bind to estrogen receptors, thereby inducing a series of reproductive toxicity and teratogenic effects; it can also cause lipid peroxidation in the body, causing damage to the liver and kidneys; In addition, ZEN also has certain toxic effects such as carcinogenicity, immunotoxicity, and teratogenicity. [0003] Fumonisins are mainly a class of secondary metabolites produced by Fusarium moniliforme, Fusarium verticilloides and Fusarium polyphylla under certain temperat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30G01N27/48
CPCG01N27/3278G01N27/3277G01N27/308G01N27/48
Inventor 韩铮聂冬霞赵志辉唐占敏郭文博蒋可秋
Owner SHANGHAI ACAD OF AGRI SCI
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