Application of tea saponin and preparation thereof
A technology of tea saponin and inhibitor, which is applied in the field of application of tea saponin and its preparation, can solve the problems that have not been seen in the application of tea saponin, and achieve the effect of convenient material collection and broad application prospects
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Embodiment 1
[0039] Preparation of tea saponin.
[0040] Tea seed cake was used as raw material, 70% ethanol aqueous solution was used as extract, mixed according to the mass ratio of 1:8, heated and refluxed at 85°C for 2 hours; D101 macroporous resin with 4 times the mass of tea seed cake was used After absorbing the extract, it was eluted with 0.2% sodium hydroxide solution until it was colorless; it was washed with distilled water until the pH of the solution was neutral, and then eluted with 90% ethanol water solution until it was colorless, and the eluate of 90% ethanol water solution was collected. Dry to get tea saponin.
Embodiment 2
[0042] Inhibition experiment of tea saponin (Zhejiang Dongfang Tea Technology Co., Ltd.) on Malassezia.
[0043] Experimental strain: Malassezia furfur (Malassezia furfur, ATCC44344);
[0044] Medium: Malassezia solid medium: glucose 40g / L, agar 14g / L, peptone 10g / L, dissolved in distilled water, adjusted to pH 6.8~7.0, autoclaved at 115°C for 20min; Malassezia liquid medium : Glucose 40g / L, dissolved in distilled water, adjusted to pH 6.8~7.0, autoclaved at 115°C for 20min.
[0045] Experimental method: After the test bacteria were subcultured on the Malassezia solid medium, to ensure the purification and viability of the colony, pick a single clone into 50ml Malassezia liquid medium, culture at 30°C for 24h, and use turbidimetric Instrument (BioMérieux) to adjust to the concentration of McFarland No. 4 turbidimetric tube.
[0046] Dilute the bacterial solution with the medium to make the final bacterial concentration 105CFU / mL, then draw 100ul and spread it on the solid me...
Embodiment 3
[0050] Determination of the minimum inhibitory concentration (MIC value) of tea saponin against Malassezia
[0051] Experimental strain: Malassezia furfur (Malassezia furfur, ATCC44344);
[0052] Medium: Malassezia liquid medium: glucose 40g / L, dissolved in distilled water, adjusted to pH 6.8~7.0, autoclaved at 115°C for 20min.
[0053] Experimental method: After the test bacteria were subcultured on the Malassezia solid medium, to ensure the purification and viability of the colony, pick a single clone into 50ml Malassezia liquid medium, culture at 30°C for 24h, and use turbidimetric Instrument (BioMérieux) to adjust to the concentration of McFarland No. 4 turbidimetric tube.
[0054] The tea saponin was directly diluted with Malassezia liquid culture medium to test concentrations, which were 10, 5, 2.5, 1.25, 0.625, 0.3125, 0.156, and 0.078 mg / ml, respectively. Add 100 μL of bacterial solution and 100 μL of medicinal solution to the microwells of a 96-well plate, and set u...
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