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Method for detecting toxicity of 2,4-dinitrotoluene

A dinitrotoluene and detection method technology, which is applied in the detection field of 2,4-dinitrotoluene toxicity, can solve the problems of high research cost, low litter size per litter, and difficulty in obtaining it, and achieve low breeding cost, Short breeding cycle and easy to observe effects

Active Publication Date: 2018-09-28
WUHAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, most researchers use rats, hamsters, and mice as test objects for the toxicity research of poisons. The research costs are high, the animal reproduction cycle is long, and the single litter size is low, so it is difficult to obtain homogeneous test individuals. Visually display the pathological changes of living tissues and organs, so it is not conducive to be used as a toxicological model for 2,4-DNT toxicity research

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

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Embodiment 1

[0062] Embodiment 1 Zebrafish feeding and management

[0063] (1) Breed zebrafish (wild type AB adult zebrafish, purchased from the National Zebrafish Resource Center) in an aquarium (20cm×12cm×12cm) with constant temperature circulating water, using filtered and fully aerated tap water 1.5L of culture fluid containing different 2,4-DNT concentrations is contained in the aquarium, and the zebrafish is domesticated in the aquarium for 7 days (during which no fish body dies), and the zebrafish after the domestication is carried out as 2, Zebrafish were used in the test of 4-DNT toxicity detection, and the bait was fed once a day in the morning and evening during the feeding process. The oxygen is greater than 5.0mg / L, the pH is 7.0±0.2, and the light is alternated (light 14h / dark 10h), and the feeding bait is tropical small fish feed (purchased from Beijing Crazy Water Plant Company) and red worms.

[0064] (2) Feeding method of red worms: mix agar, milk powder and water evenly...

Embodiment 2

[0065] The preparation of the culture fluid of embodiment 2 different 2,4-DNT concentration

[0066] (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;

[0067] (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;

[0068] (3) According to the preset concentration gradient, mix an appropriate amount of 2,4-DNT mother liquor with breeding water to prepare a culture solution containing different 2,4-DNT concentrations, and the ...

Embodiment 3

[0069] Example 3 2,4-DNT acute exposure test to zebrafish

[0070] (1) Acetone was selected as the solvent of 2,4-DNT, and the safe concentration of acetone to zebrafish was first determined to be 0.1mL / L through the acetone exposure test, which was used as the solvent control group;

[0071](2) According to the zebrafish feeding and management method provided in Example 1, the zebrafish was placed in an aquarium to raise, and the test was divided into three groups, which were respectively a blank group, an acetone vehicle group and a 2,4-DNT treatment group, The culture solution of the blank group is aquaculture water (does not contain acetone and 2,4-DNT), the culture fluid of the acetone solvent group is the aquaculture water with acetone concentration of 0.1mL / L, and the culture fluid of the 2,4-DNT treatment group For the culture solution containing different 2,4-DNT concentrations, five concentration gradients were set up, namely 2mg / L, 4mg / L, 8mg / L, 12mg / L and 16mg / L. ...

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Abstract

The invention discloses a method for detecting toxicity of 2,4-dinitrotoluene. The method comprises the following steps: preparing culture solutions containing different concentrations of 2,4-dinitrotoluene; respectively feeding the culture solutions containing different concentrations of 2,4-dinitrotoluene to zebrafish adult fish for 96 hours; carrying out multiple groups of acute exposure tests;recording death ratios of various groups of zebrafish and judging the acute toxicity effects of 2,4-dinitrotoluene on the zebrafish; respectively fishing the culture solutions containing the different concentrations of 2,4-dinitrotoluene to the zebrafish adult fish for 5 days; carrying out multiple groups of sublethal exposure tests; taking liver and ovary samples of the various groups of zebrafish and carrying out pathological observation; testing changes of lipid metabolism-related gene expression and oxygen respiratory regulation related gene expression of the various groups of zebrafish;and judging a chronic toxicity effect of 2,4-dinitrotoluene on the zebrafish. According to the method disclosed by the invention, the toxicity detection intuition property and accuracy of 2,4-dinitrotoluene are improved.

Description

technical field [0001] The invention relates to the technical field of toxicology detection, in particular to a detection method for the toxicity of 2,4-dinitrotoluene. Background technique [0002] 2,4-Dinitrotoluene (2,4-DNT) belongs to the family of nitrobenzene compounds and is an intermediate in the production process of polyurethane foam, pesticides, rubber and dyes, and is easily absorbed through the skin. poisoned. In acute poisoning, cyanosis, headache, dizziness, nausea and vomiting, shortness of breath, drowsiness, and even loss of consciousness may occur, which may cause death if not treated in time. Chronic poisoning caused by long-term effects may cause symptoms such as headache, dizziness, fatigue, palpitations, pallor, lip cyanosis, leukocytosis, anemia and jaundice. 2,4-DNT has been included in the list of environmental priority monitoring pollutants in my country and the world because of its acute toxicity and carcinogenic teratogenicity. Such pollutants...

Claims

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Application Information

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IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 熊江林王锐吴灵英刘玉兰
Owner WUHAN POLYTECHNIC UNIVERSITY
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