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Rapid detection method of effect of bromphenol endocrine disrupters

A technology of endocrine disruptors and detection methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, material stimulation analysis, etc. It can solve problems such as bromophenol endocrine disrupting effects, and achieve high sensitivity and sensitive gene expression , The effect of simple sample processing procedures

Inactive Publication Date: 2010-05-05
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

According to the current research analysis, there is no rapid in vitro evaluation method for the endocrine disrupting effect of bromophenol

Method used

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  • Rapid detection method of effect of bromphenol endocrine disrupters
  • Rapid detection method of effect of bromphenol endocrine disrupters
  • Rapid detection method of effect of bromphenol endocrine disrupters

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Embodiment Construction

[0026] A method for evaluating the endocrine disrupting effect of bromophenol, which comprises the following steps:

[0027] 1. H295R cell culture:

[0028] (A), culture conditions: use DMEM / F12 medium, add 1% (v / v) insulin-transferring sodium selenite plus Premix, 2.5% (v / v) Nu-Serum, 100U / ml penicillin, 100μg / ml streptomycin. Culture in a cell incubator at 37°C, 5% (v / v) CO2.

[0029] (B), cell recovery:

[0030] (1) Take out the frozen cells from the liquid nitrogen tank and quickly put them into an enamel tank filled with water at 36°C to 37°C, close the lid and shake from time to time to thaw as soon as possible.

[0031] (2) Take out the cryopreservation tube, wipe and disinfect it with 70% (volume ratio) alcohol, open the cover on the purification table, suck out the cell suspension with a pipette, put it into a centrifuge tube, add 10ml of culture medium, and blow and beat to make the cells Suspended.

[0032] (3) Centrifuge at low speed (500-1000 rpm) for 5 minutes,...

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Abstract

The invention discloses a rapid detection method of the effect of the bromphenol endocrine disrupters, comprising the following steps: A. culturing cells, adopting insulin Fe-Se transferring protein as growth promoter and using carbon filtering serum Nu-Serum which does not contain hormone, and exposing for 46-50 hours; B. selecting the exposure dose to be 0.1, 1 and 10 mu m to ensure 100% of survival rate of cells, and determining the endocrine disrupting effect of bromphenol; C. extracting RNA and carrying out reverse transcription on RNA into cDNA; and D. detecting gene expression variety of enzyme in the synthesizing process of steroid by utilizing fluorescent quantitative PCR technology, therefore, the technology can realize evaluation on the environmental samples. In low-dose exposure, important genes relative to steroid hormone metabolism in H295R cells are influenced; the method has high sensitivity and can detect the dose lower than 0.1 mu m; the sampling processing has simple procedures with no need of exceptionally costly instruments; the invention can be cultured in 96 pore plates with low cost, and a plurality of samples can be detected simultaneously, thus being applicable to the detection of environmental brominated flame retardant endocrine disrupting effect in environment protection departments and scientific research units.

Description

technical field [0001] The invention relates to the technical field of endocrine disrupting detection of environmental compounds, and more particularly to a method for rapidly detecting the endocrine disrupting effect of bromophenol. In order to determine the toxicological effects of the compounds, the method can also be used for environmental protection departments and scientific research units to evaluate the endocrine disrupting effects of persistent organic pollutants and chemicals. Background technique [0002] Bromophenolic compounds are widely used in the production of textiles, polyurethanes, plastics, epoxy resins and paper. Bromophenolic compounds can also be used as additives to produce other brominated flame retardants. At the same time, plastic degradation and the degradation of polybrominated diphenyl ethers (PBDEs) can also generate bromophenolic compounds. For example, tetrabromobisphenol A (TBBPA), the largest amount of brominated flame retardants, can be ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/02G01N21/64
Inventor 周炳升方琪邓军
Owner INST OF AQUATIC LIFE ACAD SINICA
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