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.

Active Publication Date: 2012-01-25
焦广俪 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing methods for determining the crude polysaccharide content of Ganoderma lucidum cannot characterize and identify the actual quality of polysaccharides in Ganoderma lucidum spore powder samples

Method used

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  • Construction method of ganoderma spore powder polysaccharide fingerprint and standard fingerprint of ganoderma spore powder polysaccharide
  • Construction method of ganoderma spore powder polysaccharide fingerprint and standard fingerprint of ganoderma spore powder polysaccharide
  • Construction method of ganoderma spore powder polysaccharide fingerprint and standard fingerprint of ganoderma spore powder polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, the establishment method of polysaccharide standard fingerprint of Ganoderma lucidum spore powder

[0047] 1. Instruments, reagents and samples

[0048] 1.1 Instrument

[0049] Agilent 1100 HPLC equipped with diode array detector (DAD) and Agilent chromatography workstation.

[0050] 1.2 Reagents and samples

[0051] Monosaccharide standard products: 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 in the United States; xylose (Xyl, 98%) , glucosamine (GlcN, 98%), and galactosamine (GalN, 99%) are all products of ACROS ORGANICS in the United States; 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...

Embodiment 2

[0077] Example 2, polysaccharide fingerprint analysis of Ganoderma lucidum spore powder sample from a certain origin in Shandong

[0078] 1. Extraction of Ganoderma lucidum spore powder polysaccharide

[0079] Accurately weigh 700 mg of the broken Ganoderma lucidum spore powder sample into a 50 mL centrifuge tube, add 30 mL of ultrapure water, ultrasonicate for 60 minutes in a water bath at 60°C (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, dialyzed with ultrapure water for 12 hours, concentrated in vacuo at 60° C., added ultrapure water to 5 mL as a crude polysaccharide aqueous solution, to be tested.

[0080] 2. Partial acid hydrolysis of Ganoderma lucidum spore powder polysaccharide

[0081] Draw 100 μL of the crude polysaccharide aqueous solution into a 5 mL stoppered graduated test tube, add 100 μL of 0.5 mol / L TFA, fill with N 2 Seal the tube, hydrolyze at...

Embodiment 3

[0088] Example 3, Fingerprint analysis of unbroken Ganoderma lucidum spore powder sample from a place in Jilin

[0089] 1. Extraction of Ganoderma lucidum spore powder polysaccharide

[0090] Accurately weigh 500mg of unbroken Ganoderma lucidum spore powder sample into a 50mL centrifuge tube, add 30mL ultrapure water, oscillate in a water bath at 95°C for 2 hours at a constant temperature, then ultrasonicate for 50 minutes in a water bath at 70°C (power 300 watts), and centrifuge after cooling , the following operations are the same as the extraction steps of Example 1.

[0091] 2. Partial acid hydrolysis of Ganoderma lucidum spore powder polysaccharide

[0092] Draw 100 μL of the crude polysaccharide aqueous solution into a 5 mL stoppered graduated test tube, add 100 μL of 0.5 mol / L TFA, fill with N 2 Seal the tube and hydrolyze at 100°C for 1 hour; after cooling, open the cap, dry in vacuum at 50°C to remove TFA, add 200 μL of ultrapure water to dissolve the residue (hydro...

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Abstract

The invention relates to a construction method of ganoderma spore powder polysaccharide fingerprint and a standard fingerprint of the ganoderma spore powder polysaccharide, belonging to the fingerprint technical field of the traditional Chinese medicine and products thereof and the functional food raw material and products thereof. The construction method in the invention comprises the following steps: extracting ganoderma spore powder polysaccharide; hydrolyzing a part of ganoderma polysaccharide; performing PMP derivatizing reaction on a part of the hydrolyzate of ganoderma polysaccharide; performing reversed phase HPLC fingerprint analysis and determining the standard fingerprint; performing the HPLC analysis on the polysaccharide fractions of ganoderma spore powder samples from fifteen different producing areas and comparing the fingerprints of the ganoderma spore powder samples to determine the common fingerprint characteristics so as to obtain a standard fingerprint. The fingerprint has nineteen common peaks in all, the sum of the peak areas of the common peaks accounts for 98% of the total peak area, and the peak area of each of four fingerprint peaks is more than 5% of thetotal peak area. The method in the invention is stable, has high degree of precision and good repeatability and is easy to grasp; and the quality situation and producing area of the ganoderma spore powder polysaccharide can be grasped from the overall characteristic of the chromatography, and a new scientific method for the quality control and identification of ganoderma spore powder can be provided.

Description

technical field [0001] The invention relates to a method for establishing the HPLC fingerprint of the ganoderma spore powder polysaccharide, and also relates to the standard fingerprint of the ganoderma spore powder polysaccharide obtained by the method. The invention belongs to the technical field of fingerprints of traditional Chinese medicine and its products, functional food raw materials and its products. Background technique [0002] Ganoderma lucidum is a fungus belonging to the Phylum Fungi, Class Basidiomycetes, Polyporaceae, Ganodermaceae, and the genus Ganoderma. Chinese Pharmacopoeia stipulates that Ganoderma lucidum can be used as medicine [ Ganoderma lucidum ( Leyss.ex Fr. ) Karst .] and Zizhi ( Ganoderma sinense Zhao, Xu et Zhang ); Ganoderma lucidum varieties that can be used in health food announced by the State Food and Drug Administration include red lucidum, purple lucidum and Songshan Ganoderma ( Ganoderma tsugae Murr). Ganoderma lucidum spore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02A61K36/074A61P37/02A61P35/00A61P3/06A61P3/10
Inventor 戴军陈尚卫朱松王浩豪
Owner 焦广俪
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