Evaluation method for cell metabolism toxicity of organic pollutants

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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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 (Monique P, Christine V.S, Konstantinos P, et al. Rapid Commun Mass Spectrom, 2003, 17:1297-1311), the metabolic flow analysis system can calculate the corresponding Changes in intracellular metabolites (Jens N, Fozia N, Elmar H. Toxicol Appl Pharm, 2009, 240:327-336), metabolic pathway analysis can use intracellular metabolites to conduct in-depth analysis of metabolic pathways, so organic The combination of mass spectrometry technology, statistical analysis platform and metabolomics software is of great significance for the rapid and efficient study of the toxic effects of environmental pollutants on cells

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