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Microorganism-based nitroaromatic pollutant detection method

A technology for the detection of nitroaromatic hydrocarbons, applied in the field of environmental biology, can solve the problems of cumbersome operation, limited use, and bulky volume, and achieve the effects of obvious color development, low cost, and simple operation

Pending Publication Date: 2020-03-13
XIAN INT UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Using these methods for detection requires spectrophotometers, gas chromatography and other instruments for detection and analysis. These instruments have high analysis accuracy, but they are bulky, cumbersome to operate and have a long analysis cycle, which cannot meet the rapid monitoring needs of multiple sampling points on site. Field use is particularly limited

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0049] Embodiment 1 microbial bacterium liquid preparation method

[0050] The preparation of microbial bacterial solution is affected by inoculum size, medium, temperature, pH and incubation time. Generally, the inoculation amount is between 1% and 5%, which can shorten the cultivation time. The medium used for the oligotrophic bacteria XM-1 is a basic medium containing 2,4-dinitrophenol, and the composition of the basic medium is: Na 2 HPO 4 12H 2 O 14.3g, K 2 HPO 4 3g, MnSO 4 ·H 2 O 0.028g, FeSO 4 ·7H 2 O 0.03g, trace elements 1mL, dilute to 1L.

[0051] The composition of LB medium is: tryptone 10g, yeast extract 5g, NaCl 10g, dilute to 1L. The suitable growth conditions for the bacterium are as follows: the pH is 6.5-7.5, the temperature is 25-30° C., and the rotating speed of the shaker is 150-220 r / min. Through the single factor condition experiment, the temperature is 30°C, the medium is the basic medium containing 20% ​​LB medium by volume, and the shaker ...

Embodiment 2

[0052] Embodiment 2 Qualitative detection of 2,4-dinitrophenol

[0053] 2,4-Nitrophenol solid is light yellow granules, and the solution is yellow. Oligotroph XM-1 is a Gram-negative bacterium capable of degrading 2,4-dinitrophenol. Prepared OD 600 Add the bacterial solution of 3.0 to 20mg / L 2,4-dinitrophenol, let it stand at 30°C for 1 hour, take samples regularly, centrifuge to get the supernatant, and compare with the control solution without 2,4-dinitrophenol colors for comparison. It was found that the yellow color of the solution of the experimental group gradually changed to colorless with the prolongation of time. Get the experimental group solution and dilute 10 times, add the chromogenic agent of nitrite ion, find that the solution color of the experimental group becomes purple-red, and the solution color of the matched group is still colorless (the color change is as follows: figure 2 ).

[0054] Qualitative detection experiment:

[0055] Before reaction: 1. ...

Embodiment 3

[0058] Example 3 Semi-quantitative detection of 2,4-dinitrophenol

[0059] 1. Griess reagent is used as the chromogenic agent of nitrite ion. If the system contains nitrite ion, a purple-red substance will be produced after adding Griess reagent for reaction. Preparation of Griess reagent: Liquid A: 0.5 g of p-aminobenzenesulfonic acid, 150 mL of 10% dilute acetic acid, protected from light; liquid B: 0.1 g of α-naphthylamine, 20 mL of distilled water, 150 mL of 10% dilute acetic acid, protected from light and stored at low temperature. When in use, take equal volumes of liquid A and liquid B, put them in a brown bottle and mix them evenly to obtain the nitrite color developer. The specific method is:

[0060] 1), prepare sodium nitrite standard solution

[0061] Accurately weigh 0.10g dry NaNO 2 Dissolve in 100mL water, dilute to 500mL, and make 0.2g / L solution. Accurately pipette 5mL, set the volume to 100mL in a volumetric flask, and make a 10mg / L solution as sodium nit...

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Abstract

A microorganism-based nitroaromatic pollutant detection method includes the following steps: adopting a cell culture plate, adding sample solutions of an experimental group and a control group (the experimental group is a 2, 4-dinitrophenol solution, and the control group is a phosphate buffer solution), respectively adding the sample solutions of the experimental group and the control group intodifferent channels of the cell culture plate; adding a microbial solution into the experimental group, placing the culture plate into an oscillator, oscillating, adding a color developing agent into the experimental group and the control group respectively, standing, and observing the color changes of the experimental group and the control group to judge whether the sample solutions contain nitroaromatic pollutants. Oligotrophic bacterium XM-1 can completely degrade a certain concentration of 2, 4-dinitrophenol in the environment; with this bacterium, the reaction color development of nitriteions generated during the degradation of 2, 4-dinitrophenol is used for qualitative detection of pollutants and semi-quantitative detection of pollutant concentration. The method is simple to operateand is low-cost.

Description

technical field [0001] The invention belongs to the technical field of environmental biology and relates to a method for detecting nitroaromatic pollutants based on microorganisms. Background technique [0002] While the rapid development of the chemical industry has brought convenience to human society, it has also brought pollution to the living environment of human beings that cannot be ignored. Among them, the pollution of organic compounds is particularly serious, especially aromatic compounds, which are stable in nature and have characteristics such as high toxicity, low water solubility and soil adsorption, which enable them to persist in the environment for a long time. Because of the symmetry and stability of the π electron structure in the benzene ring, the aromatic compounds remain in the environment for a long time, causing environmental pollution. As a representative of aromatic compounds, nitrophenol is an important class of chemical raw materials, widely used...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
CPCC12Q1/04C12Q1/045
Inventor 王金佩王琳刘扬杨力侠邢书娟董明清
Owner XIAN INT UNIV
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