Composition with effects of reinforcing barrier, resisting allergy and preserving moisture as well as preparation method and application of composition
A composition and barrier technology, applied in the direction of medical preparations containing active ingredients, drug combinations, cosmetic preparations, etc., can solve complex skin problems, frequent allergic symptoms, etc., to prevent re-aggregation and reduce sensitization , high affinity effect
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[0047] The preparation method is as follows:
[0048] (1) Colostrum preparation: select fresh colostrum, filter to remove impurities, centrifugally degrease, 0.22 μm pore size ceramic membrane cross-flow filtration technology microfiltration to sterilize, collect the filtrate after high-pressure homogenization, add deionized water 20 times, mix well, That is the diluted colostrum. The centrifugation conditions are 3000-5000r / min, 10-30min, below 40°C, and the filtration temperature is below 45°C.
[0049] (2) Preparation of keratin: clean and dry the wool fibers, shred them and add hydrogen peroxide for oxidation pretreatment. The oxidation pretreatment time is 30-40min. Add keratinase to carry out enzymolysis reaction for 100-120min, the amount of keratinase added is 3-5% of the substrate, and the reaction temperature is 60°C. Warm to inactivate the enzyme, the enzymolysis solution is cooled to room temperature and centrifuged, the supernatant is taken, and dried at low tem...
experiment example 1
[0058] Cytotoxicity test:
[0059] MTT assay is an experimental method widely used in the detection of cytotoxicity or cell proliferation by measuring the number of living cells. The detection principle is that succinate dehydrogenase in the mitochondria of living cells can make the water-soluble yellow salt MTT (3- (4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide) is reduced to water-insoluble blue formazan and deposited in cells, while dead cells have no such function. The generated crystalline formazan is generally added to DMSO (dimethyl sulfoxide), and the absorbance is measured after dissolution. The specific experimental method is in a 96-well plate according to 1X10 4 Inoculate 100 μl of DMEM medium containing 10% bovine serum and 100 μl of keratinocytes (HaCaT) at a density of one per well, and replace with serum-free medium after 24 hours of culture. Different concentrations of the composition were added to the serum-free medium for treatment and cultured f...
experiment example 2
[0066] Antisensitivity test:
[0067] Hyaluronidase is a participant in allergic reactions. Studies have shown that hyaluronidase has a strong correlation with inflammation and allergy. Many antiallergic drugs have a strong inhibitory effect on hyaluronidase activity. In vitro hyaluronidase inhibition method uses the hyaluronidase inhibition rate as an index, which can evaluate the anti-sensitivity ability.
[0068] Take 0.1mL of 0.25mmol / L CaCl2 solution and 0.5mL hyaluronidase solution and incubate at 37°C for 20min; add Examples 1-12 and Comparative Examples 1-3, 0.5mL each, and continue to incubate at 37°C for 20min; Add 0.5mL sodium hyaluronate solution at 37°C for 30min, and place it at room temperature for 5min; add 0.1mL of 0.4mol / L NaOH solution and 0.5mL acetylacetone solution, heat in a boiling water bath for 15min, then immediately cool with ice water for 5min; Add 1.0 mL of Ehrlich reagent and dilute with 3.0 mL of absolute ethanol, let stand for 20 min to develo...
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