Ganoderma lucidum polysaccharide and preparation method thereof
A technology of Ganoderma lucidum polysaccharide and Ganoderma lucidum fruiting body is applied in the directions of pharmaceutical formulations, medical preparations containing active ingredients, organic active ingredients, etc. high effect
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Embodiment 1
[0021] 1. Obtaining the residue of Ganoderma lucidum extraction: take red Ganoderma lucidum fruiting bodies as raw materials, crush them into coarse grains, weigh 200g, add 3L of water, soak at room temperature for 30 minutes, heat to boiling, keep for 120 minutes, and filter. Repeat the above steps for the filter residue, and extract once more. After filtering, the obtained residue is dried in a blast dryer at 60°C.
[0022] 2. Ultrafine grinding of Ganoderma lucidum residues: The dried Ganoderma lucidum extraction residues were pulverized for 30 minutes with an ultrafine grinding vibrating mill, and passed through a 200-mesh sieve to collect ultrafine powder.
[0023] 3. Extraction and separation of Ganoderma lucidum polysaccharide: Weigh 150g of the above-mentioned superfine powder of Ganoderma lucidum residue, add 2L of distilled water, heat to boiling at room temperature for 60 minutes, keep boiling slightly for 60 minutes, and centrifuge. Repeat the extraction of the pre...
Embodiment 2
[0029] 1. Obtaining the residue of Ganoderma lucidum extraction: take red Ganoderma lucidum fruiting bodies as raw materials, crush them into coarse grains, weigh 500g, add 6L of water, soak at room temperature for 40 minutes, heat to boiling, keep boiling slightly for 120 minutes, and filter. Repeat the above steps for the filter residue, and extract twice more. After filtering, the obtained residue is dried in a blast dryer at 60°C.
[0030] 2. Ultrafine grinding of Ganoderma lucidum residues: The dried Ganoderma lucidum extraction residues were pulverized for 40 minutes with an ultrafine grinding vibrating mill, and passed through a 200-mesh sieve to collect ultrafine powder.
[0031] 3. Extraction and separation of Ganoderma lucidum polysaccharide: Weigh 400g of the superfine powder of Ganoderma lucidum residue, add 6L of distilled water, heat to boiling at room temperature for 60 minutes, keep boiling slightly for 120 minutes, and centrifuge. Repeat the extraction of the ...
Embodiment 3
[0034] Determination of the activity of Ganoderma lucidum polysaccharide to stimulate macrophages to release NO in vitro:
[0035] 1. Sample preparation: Accurately weigh the Ganoderma lucidum polysaccharide prepared in Example 1 and Example 2 into a sterilized eppendorf tube, and prepare a sample solution with a concentration of 5 mg / mL with sterile PBS. After fully dissolving, centrifuge at 15000r / min for 30min, transfer the supernatant to a new sterile eppendorf tube under aseptic conditions, and dilute the sample to 2mg / mL, 0.5mg / mL, 0.1mg / mL for use.
[0036] 2. Cell culture: Take the RAW264.7 macrophage cell line in the logarithmic growth phase, and use DMEM complete medium at 37 ° C, containing 5% CO 2 The cells were subcultured under conditions, digested with 0.05% trypsin or 5% EDTA solution, and the suspension was centrifuged at 1000rpm / min for 3min to collect the cells and counted for later use.
[0037] 3. Reagent preparation and standard curve drawing
[0038] G...
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