Hericium erinaceus large-molecular polysaccharide and preparation method thereof
A high molecular weight, Hericium erinaceus technology, applied in the field of Hericium erinaceus high molecular weight polysaccharide and its preparation, can solve the problems of less research on the large molecular weight part, unfavorable large-scale production and application, complicated operation process, etc., to enhance the immunity of the body. , low cost and simple preparation method
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Embodiment 1
[0030] 1. Water extraction and alcohol precipitation: take the fresh fruiting body of Hericium erinaceus as raw material, cut it into pieces of about 2cm, weigh 2kg of raw material and extract it with boiling water, filter the extract with a 200-mesh filter, and collect it by centrifugation at 4000rpm for 30min Clear, concentrated under reduced pressure until the soluble solid content is 18°Brix (the soluble solid content is measured by a hand-held digital display sugar meter.), after cooling to room temperature, add absolute ethanol until the ethanol mass percentage content is 70%, 4 ℃ Let stand overnight, centrifuge at 10000g for 15min to collect the precipitate.
[0031] 2. Freezing: dissolve the above precipitate with water, centrifuge at 10,000g for 15min to remove impurities, and freeze the supernatant at -20°C. 3. Dissolving: put the above frozen solution in a water bath at 80°C to heat and thaw, centrifuge at 10000g for 20min to collect the precipitate;
[0032] 4. Et...
Embodiment 2
[0038] 1. Water extraction and alcohol precipitation: take the fresh fruiting body of Hericium erinaceus as the raw material, cut it into pieces of about 2cm, weigh 1kg of the raw material and extract it with boiling water, filter the extract with a 200-mesh filter, and collect it by centrifugation at 4000rpm for 30min Clear, concentrated under reduced pressure to a soluble solid content of 18°Brix, cooled to room temperature, added absolute ethanol until the ethanol mass percentage concentration was 60%, stood overnight at 4°C, centrifuged at 10,000g for 15min to collect the precipitate.
[0039] 2. Freezing: dissolve the above precipitate with water, centrifuge at 10,000g for 15min to remove impurities, and freeze the supernatant at -20°C.
[0040] 3. Dissolving: put the above frozen liquid in a 100°C water bath to heat and thaw, centrifuge at 10000g for 20min to collect the precipitate;
[0041] 4. Ethanol precipitation and washing: dissolve and dilute the above precipitate...
Embodiment 3
[0047] Determination of the activity of stimulating macrophages to release NO in vitro
[0048] The RAW264.7 bone marrow macrophage cell strain in this example was purchased from the American National Culture Collection (ATCC number TIB-71 TM );
[0049] In this example, DMEM and RPMI1640 were purchased from GIBCO Company.
[0050] Preparation of test samples
[0051] Accurately weigh the Hericium erinaceus polysaccharide samples prepared in Examples 1 and 2 in a sterilized eppendorf tube, and use PBS buffer solution to prepare a concentration of 5 mg / mL. After fully dissolving, centrifuge at 12,000 rpm / min for 30 min, transfer to a new sterile eppendorf tube under aseptic conditions, and dilute the sample to a final concentration of 50, 200, and 500 μg / mL for use. The negative control was PBS buffer, and the positive control was 10 μg / mL LPS solution.
[0052] Preparation of mouse RAW264.7 macrophages
[0053] with DMEM medium at 37°C, 5% CO 2 After subculture under condi...
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