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Method for detecting toxicity of 2,4-dinitrotoluene

A technology of dinitrotoluene and a detection method, which is applied in the field of 2,4-dinitrotoluene toxicity detection, can solve the problems of high research cost, low single litter size, difficult to obtain, etc. The effect of low breeding cost and transparent fish body

Active Publication Date: 2018-12-25
WUHAN POLYTECHNIC UNIVERSITY
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  • Abstract
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Benefits of technology

The technology described in this patented method allows researchers to detect harmful chemicals from Zebra flies (Zeus biflagellatus) through their behavioral responses that include behaviors such as eating or drinking them with food containing these substances. This approach makes it easier than other methods like testing blood samples because they don't require expensive equipment nor have long periods of reproductive activity. Additionally, there may be some indicators related to specific organs involved in breathing patterns called olfactory system genes. These systems help scientists better study how pests attack different types of crops - including cereals, vegetables, berries, grains, roots, seeds, nuts, flowers, etc., resulting in improved safety measures against pesticides.

Problems solved by technology

This patents discusses two technical problem: 1） how to accurately determine if certain chemicals or drugs were harmful when exposed to humans' bodies; 2), which requires testing animals with similar levels of these materials over several generations. Current methods require expensive laboratory facilities and take days to produce samples from each generation.

Method used

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  • Method for detecting toxicity of 2,4-dinitrotoluene
  • Method for detecting toxicity of 2,4-dinitrotoluene
  • Method for detecting toxicity of 2,4-dinitrotoluene

Examples

Experimental program
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Embodiment 1

[0069] Embodiment 1 obtains zebrafish embryo

[0070] (1) The wild-type AB zebrafish (purchased from the National Zebrafish Resource Center) and the transgenic zebrafish line Tg(.7apo:GFP) (constructed by the Aquatic Biology Laboratory of Wuhan University of Light Industry) were raised in a recirculating aquaculture system , the breeding water is aerated and heated tap water, the water temperature is controlled at 26-28°C, and the light cycle of 14h light / dark 10h is maintained. During the feeding process, the feeding bait is regularly fed every morning and evening, and the feces and residual bait are removed after feeding. Among them, The feeding bait is small tropical fish feed (purchased from Beijing Crazy Water Plant Company) and red worms. The feeding method of red worms is: mix agar, milk powder and water according to the mass ratio of 0.5:2.5:7, and pour them into the Erlenmeyer flask , sterilized in an autoclave at 121°C for 20 minutes, cooled and poured into a dark ce...

Embodiment 2

[0072] The preparation of the culture fluid of embodiment 2 different 2,4-DNT concentration

[0073] (1) Configuration of acetone stock solution: acetone (analytically pure, Sinopharm Chemical Reagent Co., Ltd.) is mixed with tap water heated by aeration to prepare an acetone solution with a volume concentration of acetone of 10%, as acetone stock solution, placed in Store at 4°C and protected from light for later use;

[0074] (2) Configuration of 2,4-DNT mother liquor: take acetone stock solution, add 2,4-DNT (purity ≥ 98%, Sigma company) to it, and use an ultrasonic crusher to crush it at a frequency of 20-25kHz for 8 minutes , shake well, and prepare 2,4-DNT mother solution with 2,4-DNT concentration of 200mg / mL, and store it at 4°C in the dark for future use;

[0075] (3) According to the preset concentration gradient, mix an appropriate amount of 2,4-DNT mother liquor with aquaculture water to prepare a culture solution containing different 2,4-DNT concentrations, and t...

Embodiment 3

[0076] Example 3 2,4-DNT Exposure Test of Zebrafish Embryos

[0077] The zebrafish embryos obtained in Example 1 were cultured in a recirculating aquaculture system. The recirculating aquaculture system was filled with culture solutions with different 2,4-DNT concentrations, and the water temperature was kept at 26-28°C. Feed the feeding bait, keep the light cycle of 14h light / 10h dark, change the medium every 12h, replace 2 / 3 of the total volume of the culture medium each time, and remove dead individuals in time.

[0078] The test was divided into 8 groups, including a blank group (Control), an acetone cosolvent group (Acetone) and 6 2,4-DNT treatment groups. -DNT), the culture fluid of acetone cosolvent group is the aquaculture water with acetone concentration of 0.05mL / L, and the concentration of 2,4-DNT in the culture fluid of 6 2,4-DNT treatment groups is respectively 2mg / L, 4mg / L, 6mg / L, 8mg / L, 10mg / L and 12mg / L; 4 parallels per group (30 embryos per parallel), includ...

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Abstract

The invention discloses a method for detecting toxicity of 2,4-dinitrotoluene. The method for detecting the toxicity of 2,4-dinitrotoluene includes the following steps that culture solutions containing 2,4-dinitrotolueneare of different concentrations are prepared; zebrafish embryos are correspondingly cultured in the culture solutions containing 2,4-dinitrotoluene of different concentrations, a plurality of groups of five-day toxicity tests are conducted, and in the toxicity test process, the zebrafish embryos hatch into zebrafish larvaes; the death rate of the zebrafish embryos in each groupis observed and recorded in the toxicity test process, the larvae body length, the vitelline sac width, the vitelline sac area and the larvae body surface area of the zebrafish larvaes in each groupas well as the liver and vitelline development status of the zebrafish larvaes in each group are observed and recorded, changes of lipid metabolism related gene expression and oxygen transport relatedgene expression of the zebrafish larvaes in each group are determined, and the toxic effects of 2,4-dinitrotoluene on the zebrafish embryos and larvaes are judged. The method for detecting the toxicity of 2,4-dinitrotoluene improves the visual property and accuracy of toxicity detection of 2,4-dinitrotoluene.

Description

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Claims

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Application Information

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Owner WUHAN POLYTECHNIC UNIVERSITY
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