A kind of steroid compound, its preparation method and its application
A technology of compounds and steroids, applied in the field of steroids, can solve problems such as susceptibility to microorganisms, mildew, decay, and dipping
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Embodiment 1
[0049] Extraction of steroids:
[0050]Take 7 kg of open arrow rhizomes, extract 3 times with 95% ethanol under heat reflux, each time for 3 hours, combine the extracts to recover the solvent under reduced pressure, and obtain the total extract, suspend the total extract with pure water, and use petroleum ether: ethyl acetate (v / v=1:1~3:1) extracted 3 times to remove fat-soluble impurities, the aqueous layer was extracted 5 times with equal volume of ethyl acetate, the upper ethyl acetate layer was combined, concentrated under reduced pressure to extract, 35- Dry to dryness under vacuum at 45°C. Get 70g of ethyl acetate extract, adopt petroleum ether-ethyl acetate system (wherein V (petroleum ether): V (ethyl acetate) = 100:1 / 50:1 / 20:1 / 10: 1 / 8:1 / 4:1 / 2:1 / 1:1 / 1:2 / 1:5 / 1:10 / 0:1) and ethyl acetate-ethanol system (where: V (ethyl acetate) :V (ethanol)=20:1 / 10:1 / 5:1 / 2:1 / 1:1 / 0:1) for gradient elution, collect a bottle every 500mL~600mL, number according to the collection sequence, ...
Embodiment 2
[0096] Penicillium digitatum inhibition test
[0097] In the in vitro inhibitory activity test of the above compounds against Penicillium digitatum, the bacterial strain was Penicillium digitatum (ACCC 30389), and the broad-spectrum fungicide-carbendazim was used as a positive control.
[0098] Antifungal test method: Micro double dilution method:
[0099] The activated Penicillium digitatum was made into a spore suspension, inoculated in a 96-well plate, a positive control, a negative control and a blank control were set, and the absorbance value was measured at 600nm with a microplate reader every 24 hours. After cultivating for 72 hours, calculate Inhibition rate of cell proliferation, and use Graphpad prism software to calculate the half maximal inhibitory concentration (IC50) of the tested samples.
[0100] Cell proliferation inhibition rate=(average OD value of negative control group-average OD value of sample group)÷(average OD value of negative control group-average O...
Embodiment 3
[0105] Inhibition of stoloniferous rhizopus test:
[0106] In vitro inhibitory activity test of compound of the present invention to Rhizopus stolonifera, bacterial strain Rhizopus stolonifera ACCC36973, positive control is broad-spectrum fungicide-carbendazim, common preservatives sodium benzoate and potassium sorbate.
[0107] Antifungal test method: Micro double dilution method:
[0108] Make the spore suspension of the activated Rhizopus stoloniferum, inoculate it in a 96-well plate, set up positive control, negative control and blank control, detect the absorbance value at 600nm with a microplate reader every 24h, after cultivating for 72h, count the cell Proliferation inhibition rate, and use Graphpad prism software to calculate the half maximal inhibitory concentration (IC) of the tested samples 50 ).
[0109] Cell proliferation inhibition rate=(average OD value of negative control group-average OD value of sample group)÷(average OD value of negative control group-ave...
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