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Method for detecting aflatoxin by naked eye visual detection probe

A technology for detecting probes and aflatoxin, which is applied in the direction of material analysis, measuring devices, color/spectral characteristic measurement, etc. by observing the influence on chemical indicators, and can solve the problems of high detection limit, poor stability, high instrument cost, etc. problem, to achieve the effect of fast detection time, good stability and simple operation

Active Publication Date: 2021-11-26
川北医学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, TLC has low sensitivity, high detection limit, and poor stability, making it difficult to meet increasingly stringent detection requirements; HPLC and HPLC / MS have high instrument costs, complex sample pretreatment and data analysis, and require professional technicians; ELISA is mainly affected by Limited to the limitations of the antibody itself, such as difficulty in storage, easy inactivation, and high cost
Therefore, the existing technology is difficult to be used for point-of-care screening (POCT) of AF, and it is urgent to develop a simple, fast and highly sensitive method for the detection of AF

Method used

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  • Method for detecting aflatoxin by naked eye visual detection probe
  • Method for detecting aflatoxin by naked eye visual detection probe
  • Method for detecting aflatoxin by naked eye visual detection probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Naked-eye visualization probes are used for the detection of AFB1

[0026] (1) Configuration solution 1: containing 10-40mM HEPES buffer solution (pH 7.0), 0.5-1μM probe 1, 5-200mM NaCl, 5-200mM KCl, 0.05%-0.5% Triton-X-100 , 0.5-5 μM Hemin;

[0027] (2) Add 27 μL of the above solution 1 to 13 μL of the AFB1 solution to be tested containing 0 μM and 100 μM to obtain solution A and solution B respectively;

[0028] (3) Configure substrate solution: containing HEPES buffer solution (pH 7.0), H 2 o 2 and ABTS solution;

[0029] (4) Add 10 μL of substrate solution to the above-mentioned solution A and solution B respectively, react, and use a microplate reader to record the absorption value of the solution at 418nm at 1-30min, and take pictures with a mobile phone to record the color change of the solution every minute.

[0030] The result is as figure 2 shown.

Embodiment 2

[0031] Example 2: Establishment of AFB1 concentration-dependent curve

[0032] Using the detection steps in Example 1, the probe 1 or probe 2A+probe 2B combination was used for detection respectively. The AFB1 solutions to be tested are 0, 0.05, 0.5, 1, 5, 10, 15, 20, 50, 80, and 100 μM, respectively. Record the absorption value at 418nm at 15 minutes, and perform fitting, and take pictures of the samples, and finally convert the pictures taken at 15 minutes to G→RGB. The detection result of probe 1 is as follows: image 3 , the combined results of probe 2A+probe 2B are as follows Figure 4 .

Embodiment 3

[0033] Example 3: Detection of AFB1, AFB2, and AFG1 by naked-eye visualization probes

[0034] Using the detection steps in Example 1, wherein the AFB1, AFB2, and AFG1 solutions were 100 μM, the absorbance at 418 nm was recorded for 15 minutes. The result is as Figure 5 shown.

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Abstract

The invention discloses a method for detecting aflatoxin by a naked eye visual detection probe. The method comprises the following steps of preparing a solution 1, wherein the solution 1 contains 10-40 mM of an HEPES buffer solution, 0.5-1 [mu] M of a probe 1, 5-200 mM of NaCl, 5-200 mM of KCl, 0.05%-0.5% of Triton-X-100 and 0.5-5 [mu] M of Hemin; preparing a solution 2, wherein the solution 2 contains AF with different concentrations of 0-100 [mu] M; preparing a solution 3, wherein the solution 3 contains 10 mM to 40 mM of the HEPES buffer solution, 0.5 mM to 5 mM of H2O2 and 0.5 mM to 4 mM of ABTS; and detection: adding 13 [mu] L of the solution 2 into 27 [mu] L of the solution 1, then adding 10 [mu] L of the solution 3, carrying out a reaction, recording the absorption value of the solution at 418 nm within 1-30 minutes by using a microplate reader, and carrying out photographing by using a mobile phone to record the color change condition of the solution every minute. The naked eye visual detection probe is simple to operate, is easy to carry, is low in cost, is free of professional technicians, is short in detection time, good in stability and high in sensitivity, color change can be seen by naked eyes within 5 minutes, and the naked eye visual detection probe can be used as a useful tool for on-site analysis of AF in food and drug samples.

Description

technical field [0001] The invention relates to the technical field of safety detection of food and Chinese medicinal materials, in particular to a method for detecting aflatoxin with a naked-eye visual detection probe. Background technique [0002] Aflatoxins (AF) are a class of highly toxic substances, mainly difuranocoumarin-derived compounds with similar chemical structures produced by the metabolism of Aspergillus flavus and Aspergillus parasiticus. In the process of processing, storage and transportation of food and Chinese medicinal materials, it is easy to mildew and cause aflatoxin pollution, and aflatoxin is resistant to high temperature, difficult to decompose, difficult to remove, and also has strong carcinogenicity, teratogenicity and mutagenicity It is listed as a class I carcinogen by the International Agency for Research on Cancer. In addition, aflatoxins have hepatotoxicity and nephrotoxicity, and can cause acute and chronic poisoning in humans or animals. ...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/31
CPCG01N21/78G01N21/3103Y02A50/30
Inventor 罗荧萍鲁琴芮刘悦
Owner 川北医学院