Evaluation method for cell metabolism toxicity of organic pollutants

A technology of organic pollutants and cell metabolism, which is applied in the field of evaluation of the toxicity of organic pollutants cell metabolism, can solve the problems of cumbersome test methods, limited information, long analysis cycle, etc., and achieve simple pre-treatment methods, comprehensive information, and analysis short time effect

Inactive Publication Date: 2015-11-04
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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Problems solved by technology

2008, 565-569) However, the traditional cytotoxicity evaluation method has the disadvantages of long analysis period, cumbersome test method, the combination of multiple instruments, and the limited amount of information obtained. Therefore, it is urgent to develop an efficient, fast and informative method. The rapid development of mass spectrometry technology has become the main tool for metabolomics research in recent years. UPLC-QTOF has high sensitivity and can analyze the characteristics of metabolites in a panoramic manner. HPLC-MS/MS exists quantitatively. Precise and wide testing range (Moniqu

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  • Evaluation method for cell metabolism toxicity of organic pollutants
  • Evaluation method for cell metabolism toxicity of organic pollutants
  • Evaluation method for cell metabolism toxicity of organic pollutants

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[0022] In the present invention, HepG2 cells are used as test cells, and a metabolomics technology based on mass spectrometry is established to study the toxic effect of benzopyrene on cells.

[0023] Expose benzopyrene to HepG2 cells, the cell culture conditions are DMEM high-glucose medium, 1% penicillin-streptomycin double antibody, 10% fetal bovine serum, DMSO dissolved benzopyrene after logarithmic phase solution or the same amount of DMSO without benzopyrene, so that the final concentrations of benzopyrene in the solution are 0, 5nM, 500nM respectively, exposed for 24 hours, cultured in a 5% carbon dioxide incubator, and sucked and cultivated for 24 hours Add 10 μL of cell culture solution to 1 ml of methanol and water mixture (volume ratio 80:20), vortex for 2 min, then 15000 r min -1 Centrifuge for 5 minutes, and pass the supernatant through a 0.22 μm syringe filter. In this method, protein removal and sample dilution are completed in one step, and the sampling volume...

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Abstract

The invention provides a method which takes HepG2 cells as testing cells and carries out an exposure treatment experiment of pollutants. The method comprises the following steps: (a) establishing a pre-treatment method for rapidly extracting intracellular and extracellular metabolic products; (b) carrying out panoramic analysis and target analysis on the metabolic products of the cells by using a UPLC-QTOF and HPLC-MS/MS; (c) classifying and screening high-flux metabolites by statistical analysis; and (d) analyzing a high-attention metabolism path of organic pollutant influenced cells by combining a metabolism flow analysis system and a path analysis technology. The pre-treatment method is simple; the UPLC-QTOF is combined with the HPLC-MS/MS so that the intracellular and extracellular metabolic products of the cells can be effectively analyzed; and the high-attention metabolism path of the organic pollutant influenced cells can be directly analyzed by combining statistical treatment, metabolism flow analysis and metabolism path analysis; and the method has the advantages of simplicity in operation, short analysis time, good repeatability, capability of acquiring comprehensive information quantity and the like.

Description

technical field [0001] The invention is a new method for evaluating the toxic effect of environmental pollutants, which is based on mass spectrometry metabolomics technology to study the toxic effect of new organic pollutants on cells of high concern, specifically a method using mass spectrometry analysis technology and metabolomics This method has the advantages of simple pretreatment, short analysis time, good repeatability, and comprehensive information. Background technique [0002] Persistent organic pollutants (POPs) have the characteristics of high toxicity, strong bioaccumulation, and long-distance migration, and have become typical global pollutants. Short chain chlorinated paraffins (SCCPs), hexabromocyclododecanes (HBCDs), Dechlorane Plus (DPs), (tetrabromobisphenol A, TBBPA) and benzo[ a]Pyrene diol epoxides (Benzo[a]pyrene diolepoxides, BPDEs) and other organic pollutants, which have potential carcinogenicity to thyroid, liver, kidney, lung and other organs, ha...

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
Inventor 陈吉平张保琴耿柠波张海军王菲迪
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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