Application of oligomeric mannuronic acid salts in preparation of drugs and functional foods for delaying skin aging and regulating immunity/resisting inflammation
A technology of polymannuronic acid and functional food, applied in the field of medicine
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Embodiment 1
[0022] Example 1: Extraction, preparation and analysis of oligomannuronate
[0023] Oligomannuronic acid salt among the present invention can adopt conventional technical means in this area to make, and the specific steps in the present embodiment are:
[0024] Prepare alginate (sodium alginate in this example) into a 10wt% aqueous solution, heat it to 80-90°C with 1wt% dilute hydrochloric acid, stir and degrade it for 4-5 hours, neutralize it with 10wt% sodium carbonate aqueous solution after cooling, and then Use 5wt% dilute hydrochloric acid to adjust the pH to about 3.65, centrifuge to collect the precipitate, dissolve it with 2mol / L NaOH, add 3 times the volume of 95wt% ethanol, collect the precipitate, dehydrate with absolute ethanol and dry to obtain polymannuronic acid sodium salt. The sodium salt of polymannuronic acid is prepared into aqueous solutions of different concentrations with pure water, and degraded by the Fenton method to obtain oligomeric mannuronic acids...
Embodiment 2
[0029] Example 2: Effect of oligomannuronate on the survival rate of HaCaT cells induced by UVA
[0030] In this example, the sodium oligomannuronic acid prepared in Example 1 was used. The specific steps are:
[0031] (1) Cell culture: HaCaT cells were inoculated in MEM complete culture medium (containing 100 U / mL penicillin, 100 U / mL streptomycin and 10% FBS), placed at 37°C, 5% CO 2 cultured in an incubator.
[0032] (2) 8,000 cells per well were planted in a 96-well plate, placed in a constant temperature cell incubator and incubated for 12 hours, then added different concentrations of sodium oligomannuronate and placed in a constant temperature cell incubator to continue incubation for 48 hours. After the incubation, replace the medium and give 30J / cm 2 UVA exposure. After placing in a constant temperature cell culture incubator for 12 hours, add 20 μL (5 mg / ml) MTT solution to each well, and incubate for 4 hours in the incubator. After the end, remove the liquid, a...
Embodiment 3
[0038] Example 3: Effect of oligomannuronate on UVA-induced mitochondrial membrane potential of HaCaT cells
[0039]In this example, the sodium oligomannuronic acid prepared in Example 1 was used. The specific experimental steps are:
[0040] (1) Cell culture: HaCaT cells were inoculated in MEM complete culture medium (containing 100 U / mL penicillin, 100 U / mL streptomycin and 10% FBS), placed at 37°C, 5% CO 2 cultured in an incubator.
[0041] (2) 8,000 cells per well were planted in a 96-well plate, placed in a constant temperature cell incubator and incubated for 12 hours, then added different concentrations of sodium oligomannuronate and placed in a constant temperature cell incubator to continue incubation for 48 hours. After the incubation, replace the medium and give 30J / cm 2 UVA exposure. Place in a constant temperature cell incubator and continue to incubate for 12 hours, remove the medium, wash once with PBS solution, add 150 μL of staining working solution, mix ...
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