A compound and its application in preventing and controlling Aspergillus flavus and/or aflatoxin contamination
A kind of technology of aflatoxins and compounds, applied in the field of compounds, can solve problems such as morbidity, death, decay, etc., and achieve obvious effects, reduce pollution, and achieve ideal effects
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Embodiment 1
[0051] The extraction and separation of embodiment 1 compound
[0052](1) Take the leaves of the false eagle claw plant, wash them, dry them, and crush them with a high-speed universal grinder.
[0053] (2) Take 500 g of plant leaf powder, add 4 L of absolute ethanol, sonicate for 2 h, filter, evaporate the filtrate to dryness with a rotary evaporator, add a small amount of dichloromethane to dissolve; silica gel (200-300 mesh) column chromatography , with petroleum ether: ethyl acetate (4:1) as the eluent for elution, and the eluent is recycled.
[0054] (3) Take the eluate from silica gel column chromatography and evaporate it to dryness with a rotary evaporator, add a small amount of methanol to dissolve it and separate it with the preparative liquid.
[0055] (4) Methanol: water (50:50) was used as the mobile phase; a Marsh C18 preparative column (50 × 700 mm, 5 μm) was used as the stationary phase.
[0056] (5) Keep the flow rate at 80 mL / min; the detection wavelength i...
Embodiment 2
[0058] Identification of Example 2 Compounds
[0059] (1) Dissolve the compound extracted in Example 1 with methanol, and perform primary mass spectrometry analysis using liquid chromatography-time-of-flight mass spectrometry ( image 3 ), and take the proton adducted ion peak for secondary mass spectrometry analysis ( Figure 4 ).
[0060] (2) According to the experimental results of liquid chromatography-time-of-flight mass spectrometry, it is speculated that the molecular formula of the compound is C 16 h 14 o 6 , the molecular weight is 302.
[0061] (3) Take the compound and use an elemental analyzer for elemental analysis to confirm that there are no N elements and S elements in the molecular structure of the compound, and the ratio of C elements to H elements is consistent with the results of high-resolution mass spectrometry.
[0062] (4) Take the compound and use CD 3 OD as dissolved, proceed 1 H-NMR, 1 H- 1 HCOSY, 13 Nuclear magnetic resonance tests such as...
Embodiment 3
[0065] The minimum inhibitory concentration of embodiment 3 compound
[0066] (1) Add Aspergillus flavus spores to the potato dextrose broth (PDB) solution (2.4 g / 100 mL, pH 5.1) to a final concentration of 10 5 cells / mL, add spore-containing PDB to a 24-well plate, 2 mL per well.
[0067] (2) Add 100 μL, 50 μL, 25 μL, 12 μL, 6 μL, 0, and 100 μL, 50 μL, 25 μL, 12 μL, 6 μL, 0 as the control, two wells for each concentration.
[0068] (3) Incubate at 30°C in the dark for 60 hours, and the antibacterial status is as follows: Figure 12 . Add 25 μL and above volume of compound methanol solution to clarify the microporous culture medium, no growth of Aspergillus flavus was observed, and the minimum inhibitory concentration was 0.02 mg / mL.
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