Method for evaluating toxic cells jointly polluted by T-2 toxin and metabolites

A metabolite, T-2 technology, applied in biochemical equipment and methods, microbiological determination/inspection, measuring devices, etc., can solve problems such as decreased pregnancy rate in rats, abnormal semen/sperm in mice, etc.

Pending Publication Date: 2021-08-20
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Usually T-2 toxin and HT-2 co-exist in relatively high concentrations, and trace amounts of NEO, T-2triol and T-2tetraol have also been detected in oat, wheat and corn by-products. The combined toxicity of its metabolites has been rarely reported. T-2 toxin can cause abnormal semen / sperm in mice, and decreased pregnancy rate in rats. Toxicity provides another sensitive target

Method used

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  • Method for evaluating toxic cells jointly polluted by T-2 toxin and metabolites
  • Method for evaluating toxic cells jointly polluted by T-2 toxin and metabolites
  • Method for evaluating toxic cells jointly polluted by T-2 toxin and metabolites

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Embodiment Construction

[0022] like Figure 1-2 As shown, this specific embodiment adopts the following technical scheme: a toxic cell evaluation method for joint pollution of T-2 toxin and metabolites, including T-2 toxin, HT-2, NEO, T-2triol, T-2tetraol, fetal Bovine serum (FBS), DMEM medium, HBSS, porcine LCs cells, 0.25% trypsin solution, penicillin-streptomycin (10000Units / mL-10000μg / mL), dimethyl sulfoxide, CCK-8 reagent and Multifunctional fluorescent microplate reader.

[0023] Pig LCs cells were grown in DMEM medium containing 10% FBS and 1% penicillin-streptomycin at a saturated humidity of 5% CO 2 , Routine culture in a 37°C incubator.

[0024]Specifically, pig LCs cells are adherent cells, which can be subcultured every 2 days. When subculturing: suck out the culture medium, add 1-2mL HBSS to wash away the serum, add 2mL 0.25% trypsin solution to digest for 1min, and wait for the cells to become round Immediately add 1mL of complete culture medium to stop digestion when there is a gap,...

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Abstract

The invention discloses a method for evaluating toxic cells jointly polluted by T-2 toxin and metabolites, comprising T-2 toxin, HT-2, NEO, T-2triol, T-2tetraol, fetal bovine serum (FBS), a DMEM culture solution, HBSS, porcine LCs cells, a trypsin solution with the concentration of 0.25%, penicillin-streptomycin (10000Units / mL-10000mu g / mL), dimethyl sulfoxide, a CCK-8 reagent and a multifunctional fluorescence microplate reader. The invention has the following beneficial effects: in order to evaluate the cytotoxic effect of T-2 toxin and four main metabolites thereof on porcine LCs, on the basis of CCK-8 cell viability detection, the combined action type of each toxin is researched by using a combination index method; and a data basis is provided for hazard assessment and risk early warning of combined pollution of the A-type trichothecene mycotoxins and metabolites thereof in food or feed.

Description

Technical field: [0001] The invention belongs to the technical field of toxicity assessment methods, and in particular relates to a toxic cell assessment method for joint pollution of T-2 toxin and metabolites. Background technique: [0002] T-2 toxin is one of the most toxic toxins among type A trichothecenes. People and animals will cause a variety of acute and chronic poisoning, such as vomiting, weight loss, and nausea after eating feed or food contaminated by it. , diarrhea, inflammation and infertility, etc., and T-2 toxin is difficult to volatilize, easily soluble in water, high melting point, strong thermal stability and strong ultraviolet resistance. Therefore, common processing methods in the process of food production, processing, storage, transportation, etc. generally cannot damage it, and it is very easy to pollute various food crops or fruits and vegetables. When humans or animals eat these contaminated food products, they will have different degrees of poiso...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
CPCG01N33/5014
Inventor 杨俊花赵志辉郭文博王建华
Owner SHANGHAI ACAD OF AGRI SCI
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