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Preparation method and application of a fluorescence sensor for detecting ochratoxin A

A fluorescent sensor, ochratoxin technology, applied in chemical instruments and methods, fluorescence/phosphorescence, nano-optics, etc., can solve the problems of limited practical application, technical labor, time-consuming, etc., and achieve fast response, high sensitivity, and stability good sex effect

Active Publication Date: 2019-06-07
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although they have high sensitivity and high precision, these techniques are laborious, expensive, time-consuming and require high-precision instruments or sample preparation, limiting their practical application

Method used

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  • Preparation method and application of a fluorescence sensor for detecting ochratoxin A
  • Preparation method and application of a fluorescence sensor for detecting ochratoxin A
  • Preparation method and application of a fluorescence sensor for detecting ochratoxin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The preparation of a kind of fluorescent sensor that detects ochratoxin A is mainly divided into following four steps (see figure 1 ):

[0045] (1) The first step, the synthesis of ZnCdSe quantum dots

[0046] Dissolve zinc chloride (0.035g, 0.25mmol) and N-acetyl-L-cysteine ​​(0.1253g, 0.76mmol) in 40mL of ultrapure water, and stir for 20 minutes at normal pressure and low temperature (5°C) . After stirring, adjust the pH to 9.7 with sodium hydroxide solution (0.1 mol / L). After the pH was adjusted, cadmium dichloride (0.58mg, 0.0025mmol) was added, then filled with nitrogen and stirred in an ice bath for 15 minutes. Sodium selenium hydride (0.0026 g, 0.025 mmol) was injected into the mixture, and the mixture was filled with nitrogen and stirred in an ice bath for 15 minutes. After the reaction was completed, the solution was transferred to a reaction kettle and reacted in an oven at 200° C. for 65 minutes. The resulting concentration is 4.2×10 -7 mol / L ZnCdSe qua...

Embodiment 2

[0055] Example 2 Determination of OTA in milk by a novel fluorescent sensor

[0056] (1) Preparation of samples to be tested

[0057] Take 1.5ml of milk, add OTA standard, and then add acetonitrile to make the total volume 5mL, and configure samples with concentrations of (2, 6, 30, 60ng / mL).

[0058] (two) the synthesis of ZnCdSe quantum dot fluorescent probe, with embodiment 1

[0059] (3) Synthesis of four-(4-pyridyl) zinc porphyrin nanorod self-assembly solution, same as in Example 1

[0060] (4) Detection of OTA in milk samples

[0061] In a 1.4mL cuvette, add 100uL of milk samples containing different concentrations of OTA, the self-assembly solution of tetrakis-(4-pyridyl) zinc porphyrin synthesized in the 250uL step and 580uL Tris-HCl buffer solution of pH=8, and let it stand for 5 Minutes, then add 70uL of ZnCdSe quantum dots synthesized in step (1), measure the fluorescence emission spectrum at 470-550nm, measure the emission spectrum after 5 minutes, and measure ...

Embodiment 3

[0065] Example 3 Determination of OTA in coffee by a novel fluorescent sensor

[0066] (1) Preparation of samples to be tested

[0067] Weigh 0.5 g of coffee, add mycotoxin standard substance, and then add acetonitrile to make the total volume 5 ml, and prepare samples with concentrations of (2, 6, 30, 60 ng / mL).

[0068] (two) the synthesis of ZnCdSe quantum dot fluorescent probe, with embodiment 1

[0069] (3) Synthesis of four-(4-pyridyl) zinc porphyrin nanorod self-assembly solution, same as in Example 1

[0070] (4) Detection of OTA in coffee samples

[0071] In a 1.4mL cuvette, add 100uL of coffee samples containing different concentrations of OTA, the self-assembly solution of tetrakis-(4-pyridyl) zinc porphyrin synthesized in the 250uL step and 580uL of Tris-HCl buffer solution with pH=8, and let it stand for 5 Minutes, then add 70uL of ZnCdSe quantum dots synthesized in step (1), measure the fluorescence emission spectrum at 470-550nm, measure the emission spectrum...

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Abstract

The invention provides a preparation method and an application of a fluorescence sensor for detecting ochratoxin A. A quantum dot-nano porphyrin fluorescence 'on-off' sensor is formed with a water-soluble ZnCdSe quantum dot as a fluorescent probe and self-assembled nano porphyrin, which is prepared from a tetra-(4-pyridyl)-zincporphyrin-N,N-dimethylformamide solution and dodecyl dimethyl betaine,as a quantum dot fluorescence quencher. The OTA is combined with nano porphyrin to release quantum dots, so that the fluorescence of the quantum dots is recovered, and the quantum dot-nano porphyrin fluorescence "on-off-on" sensor is formed. The addition of other mycotoxins does not cause the change of the fluorescence intensity of the system. The fluorescence sensor realizes high-selectivity detection of OTA, effectively amplifies signals, obtains high-sensitivity and strong-specificity detection results, does not need to depend on expensive large instruments, and is simple in pretreatment and rapid in detection.

Description

technical field [0001] The invention belongs to the technical field of nanomaterial preparation and food safety detection, and in particular relates to a preparation method and application of a fluorescent sensor for detecting ochratoxin A. Background technique [0002] Ochratoxins are mycotoxins that are secondary metabolites of Aspergillus and Penicillium. The types of ochratoxins include ochratoxin A (OTA), ochratoxin B, ochratoxin C and ochratoxin D, among which the most toxic, the most widely distributed, the highest toxin production, the most serious pollution to agricultural products, and human The most closely related to health is ochratoxin A. OTA consists of a dihydroisocoumarin moiety, linked by an amide bond through seven carboxyl groups to an L-β-phenylalanine molecule. OTA has strong nephrotoxicity, hepatotoxicity, immunotoxicity, and potential carcinogenicity, teratogenicity and mutagenicity. OTA-producing molds are widely distributed in nature, resulting i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09K11/88C07D487/22C09K11/06C09K11/07B82Y20/00B82Y40/00B82Y30/00G01N21/64
Inventor 吴永江刘丽蒋可秋栾连军刘雪松
Owner ZHEJIANG UNIV
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