Broad-spectrum malassezia-resistant natural product composition and application thereof
A Malassezia, natural product technology, applied in the field of broad-spectrum anti-Malassezia natural product composition, can solve the problems of side effects, sensitization, etc., achieve small toxic and side effects, reduce dosage, and broad application prospects Effect
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Embodiment 1
[0023] Example 1: Screening for natural products with anti-Malassezia activity
[0024] 1. Experimental strains (see figure 1 ): (1) Malassezia furfur standard strain (M.furfur) CBS1878, (2) Malassezia globosa standard strain (M.globosa) CBS7966, (3) Malassezia slofi standard strain (M. .slooffiae) CBS7956, (4) Malassezia sympodialis standard strain (M.sympodialis) CBS7222, (5) Malassezia obtusa standard strain (M.obtusa) CBS7876. The five strains were purchased from the Institute of Dermatology, Chinese Academy of Medical Sciences.
[0025] 2. Experimental medium:
[0026] (1) Leeming-Notman medium base (g / L): peptone 10g, glucose 5g, yeast extract powder 1g, ox bile salt 4g, glycerin 1g, glycerol monostearate 0.5g, Tween 60 0.5mL, whole Fat milk powder 10g, chloramphenicol 0.05g, agar 12g, pH 5.6±0.2. Weigh and dissolve in pure water, heat and boil for 1min, add 1.6mL olive oil per 80mL, autoclave at 121°C for 15min, shake well after sterilization, cool to about 50°C, as...
Embodiment 2
[0040] Example 2: Cytotoxicity of Natural Products and Positive Drugs
[0041] Cell line: Hacat (human skin epidermal cells) ATCC-1295, provided by Shanghai Xiangyi Herbal Co., Ltd.
[0042] Experimental materials and reagents: 96-well plate, CO 2 Incubator, alcohol lamp, pipette gun, microplate reader, CCK-8, etc.
[0043] experimental method:
[0044] 1. Collect the logarithmic phase cells, adjust the concentration of the cell suspension, add 90 μL to each well of the 96-well plate, and make the density of the cells to be tested reach (1000-10000) / well.
[0045] Cells were cultured at 37°C for 72 hours in 2.5% CO2.
[0046] 3. Add 10 μL of the substance to be tested at different concentrations, and incubate the culture plate in an incubator for 24 hours.
[0047] 4. Add 10 μL of CCK-8 solution to each well.
[0048] 5. Continue to incubate for 1 hour in the cell culture incubator, and measure the absorbance at 450 nm.
[0049] Interpretation of results:
[0050] Inhib...
Embodiment 3
[0056] Example 3: The effect of the combination of natural products in pairs
[0057] 1. Experimental strains (see figure 1 ): Malassezia furfur standard strain (M.furfur) CBS1878
[0058] Culture medium: with embodiment 1.
[0059] experimental method:
[0060] (1) Liquid sample addition
[0061] ① Dilution of medicinal solution: same as Example 1.
[0062] ②Sampling process: Same as 2.3.1 for single-drug susceptibility plate. For the preparation of the combination drug checkerboard drug sensitivity plate, in the 96-well culture plate, add 96 μL of the culture medium to the 1-10 wells, and add 2 μL of the double concentration of the two drugs in the horizontal and vertical directions, respectively, so that the two drugs are in the 80 Different combinations of drug concentrations are formed in each well, and the drug concentration decreases from the upper left to the lower right. Add 100 μL and 200 μL liquid medium to the 11th and 12th wells respectively, which are growth c...
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