Radix pseudostellariae active hexacarbaldehyde oligosaccharide as well as preparation method and application thereof
A technology of Taizishen and hexanal aldehyde, which is applied in the directions of organic active ingredients, pharmaceutical formulations, cosmetic preparations, etc., can solve the problem that there is no active hexaldehyde oligosaccharide of Taizi ginseng, and achieve the effect of reducing the level of inflammation-related indicators.
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Embodiment 1
[0068] The extraction of embodiment 1 Radix Pseudostellaria active oligosaccharides
[0069] (1) Take 300 grams of dried Pseudostellariae Radix Pseudostellariae (Guangzhou Zhixin Pharmaceutical Co., Ltd.) and place it in a 4L round-bottomed flask, add 2.4L of 70% ethanol solution by volume and extract three times at 80°C for 1 hour each time. After the extraction, remove the ethanol from the medicinal dregs, add 2.4L deionized water to extract three times at 80°C, each time for 1h, combine the three water extracts, concentrate and spray dry to obtain the water extract of Pseudostellaria Pseudostellaria, with a yield of 31.20±3.89 %.
[0070] (2) Add the above-mentioned Pseudostellariae Pseudostellariae extract into water to make the concentration 1g / 100mL, after ultrasonic dissolution, slowly add absolute ethanol, stir while adding so that the final alcohol concentration is 90% (volume percentage), 4 ° C refrigerator Precipitate in alcohol for 24 hours, centrifuge the solutio...
Embodiment 2
[0073] The structural characterization of the active oligosaccharides of embodiment 2 Radix Pseudostellariae Radix
[0074] (1) Determination of monosaccharide composition and molecular weight: Add 0.5 mL of 4 mol / L trifluoroacetic acid to the active oligosaccharides of Radix Pseudostellariae obtained in Example 1, and make it completely acid-hydrolyzed in an oil bath at 120°C for 2 hours, completely acid-hydrolyzed Add 0.5ml of concentrated ammonia water (mass concentration 25%) to the final sample, shake to make the content dissolve completely, then add 0.5ml of PMP (1-phenyl-3-methyl-5-pyrazole) with a concentration of 0.5mol / L Linone) methanol solution, after carrying out PMP derivatization, through liquid mass analysis; Wherein, liquid mass analysis (UPLC / Q-TOFMS analysis) condition is as follows:
[0075] Mobile phase: aqueous phase (A): 0.05% (v / v) acetic acid, 20 mM ammonium acetate; organic phase (B): acetonitrile.
[0076] Chromatographic column: CORTECS C18 PFP col...
Embodiment 3
[0091] Example 3 Effect of Pseudostellariae Pseudostellariae Active Polysaccharides on LPS-Induced RAW264.7 Cell Inflammation Model
[0092] (1) Cell culture Take out the RAW264.7 cell line (Shanghai Cell Bank, Chinese Academy of Sciences) frozen in liquid nitrogen, quickly place it in a 37°C water bath and shake it until the cells melt, then transfer it to a centrifuge tube that has been added to the culture medium, and set the temperature at 1000rpm Centrifuge for 5min, discard the supernatant, add a small amount of culture medium and gently blow and suck evenly, the culture medium is to contain 10% (v / v) fetal bovine serum, 1% (v / v) double antibody (the final concentration of penicillin is 100μg / mL, streptomycin is 50μg / mL) of 1640 medium, the cell suspension was transferred to the culture dish, at 37 ℃, 5% (v / v) CO 2 , Cultivated under saturated humidity conditions;
[0093] (2) Cultivate RAW264.7 cells into a single cell suspension with 1640 medium containing 10% (v / v) F...
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