Construction method of ganoderma spore powder polysaccharide fingerprint and standard fingerprint of ganoderma spore powder polysaccharide
A standard fingerprint and Ganoderma lucidum spore powder technology, which is applied in the field of HPLC fingerprint establishment, can solve the problems of not being able to characterize and identify the actual quality of polysaccharides, etc.
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[0046] Example 1. Method for establishing standard fingerprint of Ganoderma lucidum spore powder polysaccharide
[0047] 1. Instruments, reagents and samples
[0048] 1.1 Apparatus
[0049] The Agilent 1100 high performance liquid chromatograph is equipped with a diode array detector (DAD) and an Agilent chromatography workstation.
[0050] 1.2 Reagents and samples
[0051] Monosaccharide standards: glucose (Glc, 99%), mannose (Man, 99%), galactose (Gal, ≥99%) were purchased from Shanghai Chemical Reagent Company; rhamnose (Rha, ≥99%), Fucose (Fuc, ≥99%), glucuronic acid (GlcUA, ≥99%), and galacturonic acid (GalUA, ≥97%) are all products of Sigma-Aldrich, USA; xylose (Xyl, 98%) , Glucosamine (GlcN, 98%), and Galactose (GalN, 99%) are all products of ACROS ORGANICS, USA; Ribose (Rib, >99.0%) and arabinose (Ara, 99%) are all purchased from Xiamen Xinglong Da Biochemical Reagent Co., Ltd. Derivatization reagents: 1-phenyl-3-methyl-5-pyrazolone (PMP), purity 99% (Acros Organics, USA); A...
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[0077] Example 2. Polysaccharide fingerprint analysis of Ganoderma lucidum spore powder sample from a certain place in Shandong
[0078] 1. Extraction of Ganoderma lucidum spore powder polysaccharide
[0079] Accurately weigh 700 mg of the wall-broken Ganoderma lucidum spore powder sample into a 50 mL centrifuge tube, add 30 mL of ultrapure water, sonicate it in a water bath at 60°C for 60 minutes (power 300 watts), and centrifuge after cooling. The residue is washed with a small amount of ultrapure water and centrifuged. The two supernatants were combined, and the ultrapure water was dialyzed for 12 hours, and after concentration under vacuum at 60°C, the ultrapure water was added to make the volume up to 5 mL as the crude polysaccharide aqueous solution for testing.
[0080] 2. Partial acid hydrolysis of Ganoderma lucidum spore powder polysaccharide
[0081] Pipette 100μL of the crude polysaccharide aqueous solution into a 5mL graduated test tube with stopper, add 100μL of 0.5mol / L ...
Example Embodiment
[0088] Example 3. Fingerprint analysis of a sample of unbroken Ganoderma lucidum spore powder from a certain producing area in Jilin
[0089] 1. Extraction of Ganoderma lucidum spore powder polysaccharide
[0090] Accurately weigh 500 mg of the unbroken Ganoderma lucidum spore powder sample into a 50mL centrifuge tube, add 30mL ultrapure water, shake and extract in a 95℃ water bath at a constant temperature for 2 hours, then ultrasonicate it in a water bath at 70℃ for 50min (power 300W), and centrifuge after cooling , The following operations are the same as the extraction steps in Example 1.
[0091] 2. Partial acid hydrolysis of Ganoderma lucidum spore powder polysaccharide
[0092] Pipette 100μL of the crude polysaccharide aqueous solution into a 5mL graduated test tube with stopper, add 100μL of 0.5mol / L TFA, and fill with N 2 Seal the tube and hydrolyze at 100°C for 1h; after cooling, open the lid, vacuum dry at 50°C to remove TFA, and add 200μL of ultrapure water to dissolve the...
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