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Detection method of toxicity of nanomaterials

A toxic and nano-particle technology, applied in the field of environmental detection, can solve problems such as high price, inaccurate detection, and complicated operation

Inactive Publication Date: 2010-03-17
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above two methods have shortcomings such as inaccurate detection, complicated operation, high price, and poor repeatability.

Method used

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  • Detection method of toxicity of nanomaterials

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Embodiment 1

[0033] This embodiment relates to a method for detecting the toxicity of nanomaterials, including the following steps, wherein the nanomaterials are rutile nano-titanium dioxide.

[0034] 1. Take a tube of 100 microliters of DH5a competent cells, add 1 microliter of green fluorescent protein particles, mix well, and place in an ice bath for 30 minutes;

[0035] 2. Place the tube in a 42°C water bath, heat shock for 90 seconds, and transfer to an ice bath for 2-5 minutes;

[0036] 3. Add 700-800 microliters of non-resistant SOB medium at room temperature to the tube after ice bathing, transfer to a shaker, and shake at 37°C for 1 hour;

[0037] 4. Take 450 microliters of transformed competent cells and add them to the medium resistant to kanamycin. After the medium is completely absorbed, culture overnight at 37°C;

[0038] 5. Select a single colony and identify it by colony PCR;

[0039] 6. Inoculate and incubate at 37°C for 12-16 hours; prepare a black enzyme label plate, a...

Embodiment 2

[0045] This embodiment relates to a method for detecting the toxicity of nanomaterials, including the following steps, wherein the nanomaterials are anatase titanium dioxide. In embodiment 2, the same nanoparticle toxicity detection plate as in embodiment 1 is used to carry out the following steps:

[0046] 1. Take a tube of 100 microliters of DH5a competent cells, add 1 microliter of green fluorescent protein particles, mix well, and place in an ice bath for 30 minutes;

[0047] 2. Place the tube in a 42°C water bath, heat shock for 90 seconds; transfer to an ice bath for 2-5 minutes;

[0048]3. Add 700-800 microliters of non-resistant SOB medium at room temperature to the tube after ice bathing, transfer to a shaker, and shake at 37°C for 1 hour;

[0049] 4. Take 450 microliters of transformed competent cells and add them to the kanamycin-resistant medium. After the medium is completely absorbed, culture at 37°C for 12-16 hours;

[0050] 5. Select a single colony and ident...

Embodiment 3

[0057] This embodiment relates to a method for detecting toxicity of nanomaterials, including the following steps, wherein the nanomaterials are nanometer zinc oxide. In embodiment 3, the same nanoparticle toxicity detection plate as in embodiment 1 is adopted to carry out the following steps:

[0058] 1. Take a tube of 100 microliters of DH5a competent cells, add 1 microliter of green fluorescent protein particles, mix well, and place in an ice bath for 30 minutes;

[0059] 2. Place the tube in a 42°C water bath, heat shock for 90 seconds; transfer to an ice bath for 2-5 minutes;

[0060] 3. Add 700-800 microliters of non-resistant SOB medium at room temperature to the tube after ice bathing, transfer to a shaker, and shake at 37°C for 1 hour;

[0061] 4. Take 450 microliters of transformed competent cells and add them to the kanamycin-resistant medium. After the medium is completely absorbed, culture at 37°C for 12-16 hours;

[0062] 5. Select a single colony and identify ...

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Abstract

The invention relates to a detection method belonging to the technical field of environmental detection, in particular to a method for detecting toxicity of nanomaterials in the environment by adopting Escherichia coli containing green florescent protein plasmids. The method overcomes the interference of a few natural organic matters existing in the environment with the detection of toxicity of the nanomaterials so that the detection of toxicity of the nanomaterials is more convenient and accurate. The specific method selects the Escherichia coli transforming the green florescent protein plasmids as the bacterium for detection and comprises the following steps: 1. using an M9 culture medium to prepare bacterial suspension of the Escherichia coli containing the green florescent protein plasmids; 2. using ultrasonic dispersing nanomaterials to prepare suspension of nano particulate matters; and 3. culturing the nano particulate matters and the bacterial suspension of the Escherichia coliat 37 DEG C, using an enzyme-labeling instrument to detect the change of the florescence intensity of the nano particulate matters in certain period and judging the toxicity of the nano particulate matters to the Escherichia coli according to the difference of the change of the florescence intensity. The invention provides the efficient and stable detection method of toxicity of the nanomaterialsand the method is not likely to be interfered by other factors and is simple and low in cost.

Description

technical field [0001] The invention relates to a method for applying Escherichia coli containing green fluorescent protein particles to detect the toxicity of nanometer materials in the environment, and belongs to the technical field of environmental detection. Background technique [0002] Nanomaterials refer to nanostructured materials with a particle size between 1 and 100 nanometers. Materials at this scale exhibit unique reactivity, optical properties, and electromagnetic properties. There are two sources of nanoparticles, one is natural and the other is man-made. Natural sources have existed for a long time, and the most typical ones are aerosols formed in the atmosphere by dust produced by volcanic eruptions, as well as organic colloids formed by biological effects in water bodies; artificial sources are mainly through some human activities (such as automobile exhaust emissions, the combustion of coal and natural gas and the use of artificially synthesized nanomater...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02G01N21/64C12R1/19
Inventor 沈珍瑶蒋国翔牛军峰王姣杨志峰
Owner BEIJING NORMAL UNIVERSITY
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