Compound Disaspidin BB and application of compound Disaspidin BB in antibiosis
A compound and antibacterial technology, applied in the field of medicine, can solve the problem of very few active ingredients research, and achieve the effect of promoting effective application, clear structure and good application prospect.
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Embodiment 1D
[0049] Example 1 Extraction and identification of Disaspidin BB
[0050] 1. Extraction method
[0051] The method of extracting the compound Disaspidin BB from Dryopteris serrata is as follows figure 1 As shown, it specifically includes the following steps:
[0052] S1. Extract extraction: 10kg dry dry powder of Dryopteris fragrantii was extracted twice with 10 times the volume fraction and 50% concentration of ethanol at room temperature for 2 hours, adjusted to pH 2.4 with hydrochloric acid, and soaked in ethanol The medicinal solution was concentrated and extracted to obtain 1.2 kg of extract;
[0053] S2. Preliminary separation of plant extract: After the sample is dissolved in ethyl acetate and methanol, the sample is mixed with silica gel, and the silica gel column is used for preliminary separation, followed by elution with different ratios of petroleum ether: acetone (100:1, 70: 1,50:1,30:1,10:1,1:1), and the corresponding components were named as components A (96g), B (320g)...
Embodiment 2D
[0068] Example 2 Disaspidin BB's inhibitory effect on fungi
[0069] 1. Experimental method
[0070] Use an inoculating loop to gently scrape the colonies on the surface of the SDA medium, grind the hyphae with a sterile grinder, suspend the dermatophytes in sterile saline, adjust the turbidity to 0.5 McDonnell’s turbidity, and count the spores with a hemocytometer Number and number of short hyphae. Make the concentration of the bacterial solution 1×10 3 CFU / mL to 3×10 3 CFU / mL. That is to use RPMI-1640 liquid medium to dilute the 0.5 McKenzie turbidity bacterial solution 1000 times, and count according to the hemocytometer to obtain the inoculation bacterial solution. The Dryopteris monomer stock solution (20μg / mL) diluted with RPMI-1640 liquid medium was placed in the first column of a 96-well plate, and the culture medium was used for horizontal 2-fold dilution to the 10th column. In the 11th column, 100μL RPMI- 1640 liquid medium, add 200μL RPMI-1640 liquid medium in column ...
Embodiment 3D
[0078] Example 3 Disaspidin BB's inhibitory effect on bacteria
[0079] 1. Experimental method
[0080] Test method: The micro-dilution method is used. This method is often used to test the antibacterial activity of sensitive drugs or new drugs against human pathogens. The operation method is simple and can be used for a large number of antibacterial tests. Generally, a nutrient broth medium is used, and the activated clinically pathogenic strains are inoculated into the medium to prepare a bacterial solution, and the drug to be tested is dissolved in DMSO to make up to 64 mg / mL. Dilute the drug to be tested line by line in a 96-well plate with a two-fold concentration-decreasing dilution method; then add the diluted bacterial solution, leaving two wells as a control, one with only medium and the other with only Add the bacteria solution, shake and mix evenly after the dilution, and then place the 96-well plate in a 35°C constant temperature incubator for 18-24 hours. The lowest c...
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