Analysis method for lethal toxicity of environmental pollutants to caenorhabditis eleganses

A technology for environmental pollutants and Caenorhabditis elegans, which is applied to the analysis of materials, measuring devices, instruments, etc., and can solve the problems of no reliable scientific mathematical model association between the exposed group and the control group

Inactive Publication Date: 2017-05-31
TONGJI UNIV
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Problems solved by technology

[0014] Although the method of measuring the lethal toxicity of environmental pollutants with C. elegans has been studied, these methods and their results are often simply determined by stereomicroscope. There is no reliable, scientific mathematical model to correlate groups, resulting in more statistically significant data

Method used

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  • Analysis method for lethal toxicity of environmental pollutants to caenorhabditis eleganses
  • Analysis method for lethal toxicity of environmental pollutants to caenorhabditis eleganses

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preparation example Construction

[0049] The preparation method of the synchronous Caenorhabditis elegans culture fluid comprises the steps:

[0050] (a‐1), obtaining the stock solution of Caenorhabditis elegans;

[0051] (a‐2), adding alkaline lysate to the Caenorhabditis elegans stock solution to obtain a liquid containing the eggs of Caenorhabditis elegans; the volume ratio of the Caenorhabd elegans stock solution to the alkaline lysate is 1:7;

[0052] (a‐3), culturing the liquid containing the eggs of C. elegans to obtain a synchronized C. elegans culture solution.

[0053] Specifically, the preparation method of the synchronized Caenorhabditis elegans culture fluid comprises the following steps:

[0054] 1-1. Prepare sterile K-solution, alkaline lysate, nematode growth solid medium (Nematode Growth Medium, NGM), and LB liquid medium (LuriaBertaniMedium) for cultivating Escherichia coli OP50 (E.coli OP50).

[0055] 1-2. Inoculate E.coli OP50 into sterilized LB liquid medium at room temperature, and incu...

Embodiment 1

[0088] This embodiment provides a kind of environmental pollutant (Cu 2+ ) to the analysis method of lethal toxicity of Caenorhabditis elegans, it comprises the steps:

[0089] (1) Inoculate E.coli OP50 into sterilized LB medium at room temperature, and incubate at 37°C for 24 hours;

[0090] (2) Inoculate the cultured E.coli OP50 on the NGM medium, and continue the constant temperature cultivation at 37°C for 24 hours;

[0091] (3) Inoculate Caenorhabditis elegans on the NGM medium on which E.coli OP50 has grown, and culture at a constant temperature of 20° C. for three days;

[0092] (4) Rinse the C. elegans on the surface of the NGM medium with 2 mL of sterile K-solution into a 15 mL graduated centrifuge tube, let it stand for 30 minutes, and discard the supernatant so that the liquid (worm liquid) containing C. elegans remains for about 1.5mL;

[0093] (5), by the volume ratio of worm liquid: alkali lysis solution=1:7, alkali lysis solution is added in the worm liquid, ...

Embodiment 2

[0104] The present embodiment provides a kind of analytical method of environmental pollutant (1-hexyl-3-methylimidazolium bromide) to the lethal toxicity of Caenorhabditis elegans, the difference between the method of the present embodiment and the method of embodiment one is: 1- The stock solution concentration of hexyl‐3‐methylimidazolium bromide (1‐hexyl‐3‐methylimidazolium bromide, [hmim]Br, Damas beta company) is 3.7755×10 ‐2 mol / L, dilution factor For the acute lethal toxicity data of Caenorhabditis elegans, choose the Logit or Weibull function in the APTox software (software registration number 062731, registration number 2006SR15065) to perform nonlinear least square fitting, and obtain the LC50 fitting value and the upper and lower limits of the 95% confidence interval Respectively: 5.962×10 ‐3 mol / L, 5.154×10 ‐3 mol / L, 6.974×10 ‐3 mol / L; All the other parameters are the same as in Example 1.

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Abstract

The invention provides an analysis method for lethal toxicity of environmental pollutants to caenorhabditis eleganses. The analysis method comprises the following steps: diluting synchronized caenorhabditis elegans culture solutions to obtain a caenorhabditis elegans sample solution; diluting an environmental-pollutant stock solution into n environmental-pollutant sample solutions according to a geometric concentration gradient; adding the caenorhabditis elegans sample solution into transparent micropores, correspondingly adding the environmental-pollutant sample solutions with different concentrations, adopting the transparent micropores without adding the environmental-pollutant sample solution as a blank control, and after culture for t0 hours, acquiring the number of normalized survival caenorhabditis eleganses and calculating normalized lethallty rate; and calculating median lethal concentration of the environmental pollutants by curve fitting. According to the analysis method provided by the invention, the lethal toxicity of the environmental pollutants to the caenorhabditis eleganses can be accurately calculated out, more statistic significance is achieved, and more scientific theoretical support can be provided for evaluating the safety problem of the environmental pollutants to the ecological system and the human health.

Description

technical field [0001] The invention belongs to the technical field of environmental pollutant toxicity detection and relates to a method for analyzing the lethal toxicity of environmental pollutants to Caenorhabditis elegans. Background technique [0002] Environmental pollution has become a common problem all over the world, among which environmental pollution caused by environmental pollutants such as organic compounds, heavy metals, and antibiotics is particularly prominent. In recent years, more and more attention has been paid to the damage of environmental pollutants to the environment, the toxic effects on organisms and their mechanisms of action at home and abroad, and a series of methods for detecting the degree of environmental pollution and evaluating the toxicity of poisons have been continuously proposed. [0003] From the perspective of environmental chemical analysis, chemical analysis methods such as high performance liquid chromatography and gas chromatogra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/00
CPCG01N33/00
Inventor 刘树深李恺唐含笑冯利屈锐
Owner TONGJI UNIV
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