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In-vitro micronucleus detection method for genetic toxicity of water outlet of drinking water disinfection process

A genotoxicity and micronucleus detection technology, applied in the field of water environment research, can solve the problems of quantitative calculation of the contribution rate of non-micronucleus inducing factors, difficulty in distinguishing micronucleus inducing causes, and being easily affected by subjective factors, so as to reduce false positives. Results, improved sensitivity, good reproducibility

Inactive Publication Date: 2014-11-12
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

[0005] Disadvantages of the existing evaluation technology: the traditional micronucleus test microscopic examination method has a long test period and is easily affected by subjective factors; It is difficult to distinguish the cause of micronucleus induction, and it is mostly used in the analysis of standard compounds, without quantitative calculation of the contribution rate of micronucleus induction factors
At present, the pollution index is used to quantitatively evaluate the toxicity of drinking water quality, the toxicity reaction is not intuitive enough, and the classification of toxicity grades is somewhat subjective
In terms of genotoxicity analysis of a large number of environmental samples, the existing micronucleus technology is difficult to meet the demand

Method used

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  • In-vitro micronucleus detection method for genetic toxicity of water outlet of drinking water disinfection process
  • In-vitro micronucleus detection method for genetic toxicity of water outlet of drinking water disinfection process
  • In-vitro micronucleus detection method for genetic toxicity of water outlet of drinking water disinfection process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Evaluation of genotoxicity of effluent from an ozone disinfection process in a drinking water plant in the south

[0058] The in vitro micronucleus test was used to evaluate the genotoxicity of the effluent from the ozone disinfection process of the drinking water plant. For the in vitro micronucleus test procedure, see figure 1 .

[0059] 1. Extraction of effluent extract from drinking water disinfection process of a drinking water plant in the south

[0060] After collecting 10L of water in situ with a brown glass bottle, immediately send it back to the laboratory for filtration through a 0.7μm glass fiber filter membrane to remove suspended solids, and pass through 5mL of n-hexane, 5mL of dichloromethane, 5mL of methanol, 5mL of ultrapure water activated HLB column for solid phase extraction, each HLB column is enriched with 2L of water, the flow rate of the column is controlled at about 10mL / min; after extraction, the adsorption column is dried, and 10mL...

Embodiment 2

[0119] Example 2. Evaluation of genotoxicity of effluent from a drinking water plant chlorination disinfection process in the north

[0120] The evaluation method is the same as in Example 1. At the same exposure concentration, that is, when the exposure concentration is 500mL water sample / mL cell exposure treatment solution, the in vitro micronucleus test results of the chlorination disinfection process of a drinking water plant in the north are shown in Table 3.

[0121] table 3

[0122] sample

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Abstract

An in-vitro micronucleus detection method for genetic toxicity of outlet water of a drinking water disinfection process is provided. The method includes: 1) a step of preparing a solution of an extract of the outlet water of the drinking water disinfection process; 2) a step of subjecting an isolated immortalized human embryonic kidney cell to exposure processing by utilization of the extract solution in the step 1); 3) a step of subjecting the cell processed in the step 2) to EMA staining and SYTOX Green dye staining in sequence; and 4) a step of analyzing with a flow cytometer. The detection method can reduce false positive results. Centrifugation with a pore plate is performed to exclude interference of fragments. The detection method can achieve rapid, micro and high-throughput analysis of the genetic toxicity of water samples of the drinking water disinfection process, and improve the sensitivity of the detection method. Detection results are prone to unification, good in reproducibility, accurate and reliable.

Description

technical field [0001] The invention belongs to the field of water environment research, relates to the detection and analysis of the toxicity of drinking water, in particular to an in vitro micronucleus detection method for the genotoxicity of effluent from a drinking water disinfection process. Background technique [0002] Environmental pollution poses a threat to drinking water quality. Drinking water disinfection technology effectively controls water-borne infectious diseases caused by pathogenic microorganisms in drinking water, and is widely used in the world. However, at the same time of disinfection, the disinfectant inevitably reacts with natural organic matter (NOM, natural organic matters) in water, artificial pollutants, bromine and iodide ions in raw water to form disinfection by-products (disinfection by-products, DBPs). )(Richardson, S.D.; Plewa, M.J.; Wagner, E.D.; Schoeny, R.; DeMarini, D.M., Occurrence, genotoxicity, and carcinogenicity of regulated and em...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCG01N33/5014
Inventor 王子健刘楠楠马梅饶凯锋李娜
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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