Method for detecting dioxins in environment

A technology for detecting environment and dioxins, which is applied in biochemical equipment and methods, measuring devices, and microbial measurement/inspection. It can solve the problems of long detection time, high cost, and difficulty in obtaining antibodies. Wide application and low cost effect

Active Publication Date: 2012-06-27
SOUTH CHINA INST OF ENVIRONMENTAL SCI MEP
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  • Abstract
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AI Technical Summary

Problems solved by technology

The toxic effect of dioxin-like chemicals is mainly through the combination with the arylhydrocarbon receptor (AhR) in the body's cells (poland et al., 1982), and the complex formed is transferred into the nucleus and the arylhydrocarbon receptor nucleus. The transposon protein (AhR nuclear translocator, Arnt protein) binds, and the formed DXNs-AhR-Arnt complex can bind to Dioxin Response Elements (DRE), induce specific gene expression (mainly CYP1A1), and cause toxicity effect
Among them, the in vivo detection method uses the living animal to be exposed to the substance to be tested, and detects the index enzyme activity in the body, but the detection time of this method is long and the cost is relatively expensive; the receptor binding experiment is based on the dioxin-Ah receptor Bind and activate its DNA-binding properties, isomorphically detect the amount of dioxin-Ah receptor complexes or complexes bound to DNA to indicate the strength of the dioxin effect of the sample, but antibodies are difficult to obtain; therefore, the above two Neither method is suitable as a rapid screening technique for dioxin-like pollutants
However, the results measured by the current in vitro biological test methods can not truly reflect the biological toxicity of pollutants.

Method used

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  • Method for detecting dioxins in environment
  • Method for detecting dioxins in environment
  • Method for detecting dioxins in environment

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Using the rat hepatoma H4IIE cell EROD enzyme activity induction method to measure the effect of 2,3,7,8-TCDD to draw a standard curve, the method is as follows:

[0036] 1. Cryopreservation: Add 20% serum and 10% DMSO to the DMEM medium to make a cryopreservation solution, add the rat hepatoma H4IIE cell mixture into the cryopreservation tube, suck out the supernatant after centrifugation, and add the cryopreservation solution. Pipette the stored solution evenly, put it in a freezer box and place it in a -80°C refrigerator for gradient cooling for more than 3 hours, and then transfer it to a liquid nitrogen tank for storage.

[0037] 2. Resuscitation: Quickly take out the cryopreservation vial containing the frozen cells from the liquid nitrogen tank, quickly place the vial in a 37°C water bath and shake slowly until it dissolves. Immediately after dissolving, transfer to a 37°C water bath, and shake slowly by hand to keep the temperature constant for 2 minutes. After...

Embodiment 2

[0055] The sample is a soil sample collected in a special zone in the south in September 2010.

[0056] Sample collection and storage methods refer to the requirements of HT / T166-2004 and GB17378.3 respectively. About 1 kg of surface soil samples were collected from a reservoir in the south with a clean stainless steel soil picker, wrapped in tin foil and placed in a sealed bag, sent back to the laboratory in a low-temperature environment, dried and ground with a freeze dryer below -70 °C. Weigh 80 g of the soil sample that has passed through a 200-mesh sieve, and use the traditional Soxhlet extraction method to extract with toluene solvent for 48 hours. The obtained extract is rotary evaporated to 1-2 ml with a rotary evaporator, and then 100 ml of n-hexane is added to dissolve it and continue to spin. Steam to 1-2ml. Dissolve the sample to 20ml with n-hexane, take 5ml of the solution (that is, containing 20g of soil sample), purify the 5ml sample through a multi-stage silic...

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Abstract

The invention relates to a method for detecting dioxins in environment. Rat hepatocyte hepatoma H4IIE cells are used as detection means; the cells are treated with adherent culture for 24h; environment sample organic solvent extract or chemicals dissolved by organic solvent is added into the cells; the cells are further cultured for 72 h; a dosage-effect relation standard curve of EROD enzyme vitality induction of 2,3,7,8-TCDD on H4IIE cells is drafted by determining an EROD enzyme activity and using 2,3,7,8-TCDD as a standard compound, so as to further evaluate toxicity effect of dioxin compound in a sample. The invention has advantages of simple operation, low cost, short detection time and high flux, and can be used as rapid screening means for environment sample and suspected dioxin compound.

Description

technical field [0001] The invention relates to the technical field of environmental monitoring, in particular to a method for detecting dioxins in the environment. Background technique [0002] Dioxins (DXNs) pollutants are a class of chlorinated aromatic compounds with similar chemical structures, physical and chemical properties and biological effects, including polychlorinated dibenzo-p-dioxins (Polychlorinated Dibenzo-p-Dioxins, PCDDs ), Polychlorinated Dibenzofurans (PCDFs) and dioxin like Polychlorinated Biphenyls (dl-PCBs). The toxic effect of dioxin-like chemicals is mainly through the combination with the arylhydrocarbon receptor (AhR) in the body's cells (poland et al., 1982), and the complex formed is transferred into the nucleus and the arylhydrocarbon receptor nucleus. Transposon protein (AhR nuclear translocator, Arnt protein) binds, and the formed DXNs-AhR-Arnt complex can bind to Dioxin Response Elements (DRE), induce specific gene expression (mainly CYP1A1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64G01N21/31C12Q1/02
Inventor 刘芸任明忠张素坤付建平
Owner SOUTH CHINA INST OF ENVIRONMENTAL SCI MEP
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