Biological measuring method for dioxins compound in monitoring environment samples
A dioxin-like, environment-monitoring technology, which is applied in the field of dioxin-like pollutants, can solve problems such as difficult operation of the heart infusion method, contamination of the target tissue, time-consuming and labor-intensive problems, and achieves simple operation, small sample volume, and shortened operation time Effect
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Embodiment 1
[0019] Example 1: Using the primary culture method of grass carp hepatocytes to measure the toxic effect of TCDD to draw a standard curve
[0020] Primary cell culture: Disinfect 1.0Kg of grass carp to be tested, remove the blood from the tail vein, dissect the fish body to take a small piece of liver tissue, and use 0.145mol / L NaCl, 5.4mmol / L KCl, 12mmol / L NaHCO 3 , 5mmol / L Na 2 EDTA, 3mmol / L NaH 2 PO 4 , 1.1mmol / L KH 2 PO 4 , 100mmol / L N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (hereinafter referred to as HEPES) buffer solution (pH7.5) washed to white. Cut the liver tissue into 1mm 3After size, digest with 0.25v% trypsin digestion solution 30 times the weight of the liver for 2 to 3 minutes, and stop the digestion when the cells of the liver tissue are dispersed into small cell clusters; pass through a 100-mesh sieve, transfer the filtrate into a centrifuge tube, Centrifuge at 1000r / min for 2min, pour off the supernatant, and wash the pellet with DMEM / F12 mediu...
Embodiment 2
[0024] Example 2: Using the primary culture method of grass carp liver cells to evaluate the toxicity of dioxin-like pollutants in environmental samples
[0025] River sediment, electrolytic waste residue, floating dust, sediment standard sample, standard soil and chemical products were collected as samples. Using Soxhlet extraction method, adding toluene to extract dioxin-like pollutants in the sample, loading the extracted organic solvent on a silica gel / activated silica gel column to purify the sample, and then eluting the concentrated component with n-hexane , Blow dry in high-purity nitrogen, add DMSO to dissolve and prepare a test sample solution with any concentration within the linear range of the standard curve.
[0026] Utilize the primary culture method of grass carp liver cells described in Example 1 to evaluate the toxicity of dioxin-like pollutants in environmental samples, and the measurement results are compared with those of the H4IIE cell culture method and h...
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