Method for detecting microcystic toxins MC-LR in organism
A technology of microcystin and MC-LR, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of insufficient attention to the severity of shrimp, and achieve reliable detection results, simple operation, and good reproducibility
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Embodiment 1
[0028] Example 1 Detection of Microcystin MC-LR in Macrobrachium rosenbergii
[0029] 1. The specific steps of the detection method
[0030] (1) Extraction of microcystin MC-LR: Weigh 0.1 g of freeze-dried shrimp hepatopancreas sample, grind it in a mortar and put it in a centrifuge tube, add 15 mL of extracting solution (the extracting solution is EDTA -Na 2 Aqueous formic acid, EDTA-Na 2 The concentration of formic acid is 0.01mol / L, the volume percent concentration of formic acid is 5%); after homogenization, it is ultrasonically crushed at 0°C for 5 minutes, then centrifuged at 4000r / min for 10 minutes, and the supernatant is transferred to a clean container and stored in the dark; Add 10mL extract to wash the precipitate, then centrifuge to collect the supernatant, and combine the obtained supernatant; the formic acid and EDTA-Na 2 All are analytically pure;
[0031] (2) Enrichment, separation and purification of microcystin MC-LR: Add the supernatant obtained in st...
Embodiment 2
[0052] Example 2 MC-LR experiment on accumulation of microcystins in Macrobrachium rosenbergii exposed to toxic Microcystis aeruginosa
[0053] A batch of healthy Macrobrachium rosenbergii without microcystin MC-LR was acclimatized in the laboratory for one week, and placed in a concentration of 5×10 7 cells / mL of toxic Microcystis aeruginosa algae liquid for a period of time, and collect shrimp hepatopancreas samples. The collected samples were sealed and stored at -20°C, and then freeze-dried at -46°C for later use. Measure the content of microcystin MC-LR in the shrimp hepatopancreas according to the detection method of embodiment 1, the results are shown in figure 2 . Experimental results show that the method for detecting microcystin MC-LR established according to the present invention can effectively detect microcystin MC-LR accumulated in the hepatopancreas of shrimp.
Embodiment 3
[0054] Example 3 Detection of Microcystin MC-LR in Daphnia magna
[0055] 1. The specific steps of the detection method
[0056] (1) Extraction of microcystin MC-LR: Weigh 0.15 g of freeze-dried Daphnia magna sample, grind it in a mortar, put it in a centrifuge tube, and add 20 mL of extraction solution to the centrifuge tube (the extraction solution is EDTA- Na 2 Aqueous formic acid, EDTA-Na 2 The concentration of formic acid is 0.05mol / L, and the concentration of formic acid is 1% by volume); after homogenization, it is ultrasonically crushed at 4°C for 3 minutes, then centrifuged at 6000r / min for 8 minutes, and the supernatant is transferred to a clean container and stored in the dark; Add 10mL extract to wash the precipitate, then centrifuge to collect the supernatant, and combine the obtained supernatant; the formic acid and EDTA-Na 2 All are analytically pure;
[0057] (2) Enrichment, separation and purification of microcystin MC-LR: Add the supernatant obtained in...
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