Colorimetric detection method for aryloxyphenoxypropionate herbicide based on magnetic nanocrystal enzyme preparation

A technology of aryloxyphenoxypropionate and magnetic nano, which is applied in the direction of nanotechnology for sensing, nanotechnology, nanotechnology, etc., and can solve the problems of high cost, complicated preparation process, low sensitivity and stability, etc.

Active Publication Date: 2021-08-03
HUAIBEI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biomacromolecules that have been reported for sensors include DNA, antibody / antigen, and enzyme molecules. Among them, DNA sensors and immunomolecular sensors have low sensitivity and stability, and the preparation process is relatively complicated and costly. Bioenzyme sensors have enzyme The high activity and high specificity of molecules have attracted more and more attention

Method used

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  • Colorimetric detection method for aryloxyphenoxypropionate herbicide based on magnetic nanocrystal enzyme preparation
  • Colorimetric detection method for aryloxyphenoxypropionate herbicide based on magnetic nanocrystal enzyme preparation
  • Colorimetric detection method for aryloxyphenoxypropionate herbicide based on magnetic nanocrystal enzyme preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Preparation of magnetic nanocrystal enzyme preparation QpeH / AOx@mZIF-8

[0040] 1. Magnetic Fe 3 o 4 Synthesis of Nanoparticles (MNPs)

[0041] Add 6.5g FeCl to a 200mL round bottom flask 3 ·6H 2 O and 3.342 g FeSO 4 ·7H 2 O, completely dissolved in 100mL deionized water, heated the system to 50°C and stirred under vacuum for 30min, then quickly added 12.5mL of ammonia water to the system and raised the temperature to 75°C, stirred for 1h with magnetic force, then added 12.5mL of 1.5mol L -1 Trisodium citrate, warm up to 85°C and stir for 1.5h. After the reaction is complete, collect Fe 3 o 4 The nanoparticles were then washed three times with ethanol and deionized water successively, and dried under vacuum at 60 °C for 6 h.

[0042] 2. Extraction and purification of AOPP herbicide hydrolyzing esterase QpeH

[0043] 2a, the extraction of esterase QpeH

[0044] Escherichia coli BL21(DE3) containing the prokaryotic expression plasmid pET-29a-qpeH of t...

Embodiment 2

[0055] Example 2: Characteristic detection of QpeH / AOx@mZIF-8 nanocrystal enzyme preparation

[0056] Such as figure 2 As shown by the scanning electron microscope, QpeH / AOx@mZIF-8 has a regular dodecahedral crystal form, with a rough surface and an average diameter of about 494.5nm ( image 3 ). X-ray diffraction (XRD) ( Figure 4 ) and Fourier transformedinfrared spectroscopy (FT-IR) ( Figure 5 ) detection results showed that MNPs, QpeH and AOx were embedded in ZIF-8 crystals, QpeH / AOx@mZIF-8 had a complete ZIF-8 crystal morphology, and the addition of MNPs, QpeH and AOx did not affect ZIF-8 crystals Formation. Thermal gravimetric analysis (TGA) ( Figure 6 ) results showed that the loading of QpeH and AOx in mZIF-8 was about 8%. N 2 Adsorption test results ( Figure 7 ) shows that the QpeH / AOx@mZIF-8 crystal has typical I-type adsorption-desorption curve characteristics, and its specific surface and pore diameter measured by BET method are 526.2m 2 g -1and 15.2...

Embodiment 3

[0057] Example 3: Performance evaluation of QpeH / AOx@mZIF-8 enzyme sensor for detection of AOPP herbicides

[0058] Disperse 5 mg of QpeH / AOx@mZIF-8 in 1 mL of NaAc solution (0.2 M, pH 4.0), add 1 mL of OPD (1.0 mM) and 3 mL of different concentrations of quizalofop-p-ethyl (QpE), oxaprop-p-ethyl ( FpE) or high-efficiency haloxyfop (HpE) solution, and then incubated at room temperature in the dark for 10 min. After removing QpeH / AOx@mZIF-8 in the reaction solution with a magnet, detect the absorbance value OD at 450nm 450 . Such as Figure 9 As shown, in the wide concentration ranges of 6.7-214.6 μM, 6.7-214.6 μM and 13.4-268.2 μM, the OD of QpeH / AOx@mZIF-8 sensor 450 It has a good linear relationship with the concentration of QpE, FpE or HpE, and obvious color changes can be observed by naked eyes. According to the equation: LOD=3σ / S (where σ represents the standard deviation of the biosensor response value relative to the blank, and S represents the slope value of the ca...

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Abstract

The invention discloses a colorimetric detection method for an aryloxyphenoxypropionate herbicide based on a magnetic nanocrystal enzyme preparation, which comprises the steps of simultaneously embedding magnetic Fe3O4 nanoparticles MNPs, an AOPP herbicide hydrolase QpeH and ethanol oxidase AOx in a ZIF-8 crystal through a one-step synthesis method to prepare the magnetic nanocrystal enzyme preparation QpeH/AOx@mZIF-8, and detecting AOPP herbicide residues in an environmental sample through a colorimetric method by using the magnetic nanocrystal enzyme preparation as a biological enzyme sensor. According to the invention, the characteristics of stability, simplicity and low cost of the nano enzyme MNPs are effectively utilized, and the characteristics of strong specificity and high activity of the biological enzyme (QpeH, AOx) are combined to prepare the QpeH/AOx@mZIF-8 multi-enzyme system sensor, so that the problem that the AOPP herbicide is difficult to accurately, quickly and real-timely detect is effectively solved.

Description

technical field [0001] The invention relates to a colorimetric detection method for aryloxyphenoxypropionate herbicides based on a magnetic nanocrystal enzyme preparation, belonging to the field of biological enzyme sensors. Background technique [0002] Aryloxyphenoxypropionate (AOPP) herbicides are a class of highly active herbicides widely used to control grass weeds. They mainly inhibit the synthesis of fatty acids in cells and destroy the growth of meristems by inhibiting acetyl-CoA carboxylase necrosis of weeds. AOPP herbicide is a new type of dry field stem and leaf treatment agent with high selectivity between grass weeds and dicotyledonous crops. It is widely used in my country and more than 100 countries in the world because of its high efficiency and low toxicity. In 2014, the global sales of AOPP herbicides was about 12.17 billion US dollars, accounting for 4.6% of the total sales of herbicides in the world. At present, the three AOPP herbicides with the larges...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/31G01N1/38B82Y15/00B82Y40/00
CPCG01N21/78G01N21/31G01N1/38B82Y15/00B82Y40/00
Inventor 张辉余婷马恒岩李梦雅
Owner HUAIBEI NORMAL UNIVERSITY
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