Method for detecting poison in leguminosae astragalas herbaceos perennial
A kind of herb, detection method technology, applied in the detection field
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Embodiment 1
[0014] Example 1: Screening of Toxic Extracts of Satawan Virus Strain
[0015] Collect 3 kilograms of the stem part of the diseased plant in Huanxian County, Gansu Province, crush it all, and extract it by cold soaking with 10 liters of 90% ethanol aqueous solution (volume ratio) at room temperature for three times, combine the extracts and distill under reduced pressure Concentrate it and dry it to obtain 200 grams of total extract. After the extract was dissolved in water, it was extracted with an equal volume of petroleum ether, and then the obtained aqueous phase was extracted with an equal volume of ethyl acetate, and then the obtained aqueous phase was extracted with an equal volume of n-butanol to obtain four different poles. Active extract components: petroleum ether component, ethyl acetate component (35g), n-butanol component and aqueous phase component.
[0016] These four components obtained above were subjected to mouse feeding experiments, and it was found that ...
Embodiment 2
[0017] Embodiment 2: Separation and purification of toxic compounds
[0018] The ethyl acetate extract that embodiment 1 obtains is mixed with 35g silica gel (200-300 order), after natural evaporation, carry out column chromatography with 400g silica gel (200-300 order) wet packing column, with sherwood oil / acetic acid Ethyl ester gradient elution, each 200mL as a fraction, to obtain a total of 7 components A ~ G. The elution gradient was as follows: A(30:1), B(15:1), C(8:1), D(4:1), E(2:1), F(1:1), G( Methanol flush). Component D, after TLC detection, combined similar components to obtain nine parts D1-D9, part D9 was recrystallized from petroleum ether-ethyl acetate system to obtain a compound, which was confirmed to be 7,3'-dihydroxy - 2',4'-dimethoxy-isoflavan (DDIF), characterized as follows:
[0019] Physical and spectroscopic data of the compound DDIF: colorless crystals; melting point: 145 °C; [α] D 20 =-18.5° (c=0.5, methanol); ESIMS shows molecular ion peak [M+H...
Embodiment 3
[0020] Embodiment 3: the toxicity experiment of compound DDIF pure product
[0021] The pure DDIF product obtained in Example 2 was injected into mice, and it was found that at a dose of 100 μg, the mice could die.
[0022] DDIF was configured as a 10mg / mL dimethyl sulfoxide standard solution, and stored in a -20°C refrigerator as a test sample. Dilute the test sample into a series of concentrations to administer to Xenopus embryo cells, and observe the effects of different concentrations of compounds on embryo cell division and early embryo development. Embryos were cultured at room temperature for 24 hours, fixed with 3.7% formaldehyde for imaging, and the number of embryonic cells that stopped dividing and died was recorded. Experiments were repeated three times. Through embryonic cell experiments, it is confirmed that the compound DDIF can significantly prevent Xenopus embryonic cell differentiation at 1 μg / mL, see figure 1 .
[0023] Experimental conclusion: DDIF at a...
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