LU503132B1 - Suillus luteus polysaccharide, its preparation method and medical application - Google Patents
Suillus luteus polysaccharide, its preparation method and medical application Download PDFInfo
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- LU503132B1 LU503132B1 LU503132A LU503132A LU503132B1 LU 503132 B1 LU503132 B1 LU 503132B1 LU 503132 A LU503132 A LU 503132A LU 503132 A LU503132 A LU 503132A LU 503132 B1 LU503132 B1 LU 503132B1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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Abstract
The invention discloses a Suillus luteus polysaccharide, a preparation method and medical application thereof, and relates to the field of natural medicine development. The monosaccharide composition of the Suillus luteus polysaccharide comprises fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid. According to the invention, the polysaccharide SIP with novel structure and antioxidant activity is obtained from the natural bacterium Suillus luteus for the first time, and it has strong radical scavenging ability to DPPH free radical, hydroxyl free radical and superoxide anion, and can be used for preparing anti-oxidation related drugs, providing a research foundation for the subsequent development and utilization of Suillus luteus, and having important economic value and market value.
Description
SUILLUS LUTEUS POLYSACCHARIDE, ITS PREPARATION
METHOD AND MEDICAL APPLICATION
The invention relates to the technical field of natural medicine development, and in particular to Swillus luteus polysaccharide, its preparation method and medical application.
Oxidation is an essential process in human life. Usually, some free radicals will be produced in the process of oxidative metabolism, which plays an important role in keeping the environment stable. When the amount of free radicals is too large, lipid and other substances will be oxidized, resulting in dysfunction of cell function and tissue damage, which will lead to heart disease, cancer and other diseases. Therefore, it is a hot topic in various fields to study or search for substances with antioxidant function to reduce or eliminate the existence of free radicals. Therefore, it is of great significance to explore potential drugs with no toxic and side effects, free radical elimination and antioxidant capacity.
Suillus luteus belongs to Basidiomycota, Hymenomycetes, Agaricales, boletaceae and Suillus bovinus. The mushroom is hemispherical, with smooth, glossy and sticky surface, also known as "sticky dough", and is rich in polysaccharides, polyphenols, amino acids and other substances. Swillus luteus is a nutritious and delicious edible mushroom with high nutritional and medicinal value, and is an important natural medicine resource. At present, there are few reports on antioxidant activity of Suillus luteus. Therefore, it is of great significance to develop and utilize its medicinal value.
The purpose of the present invention is to provide a Suillus luteus polysaccharide,
its preparation method and medical application, so as to solve the problems existing in 10909136 the prior art. The Swillus luteus polysaccharide provided by the present invention has an excellent antioxidant capacity and can be used to prepare antioxidant drugs.
To achieve the above object, the present invention provides the following solutions:
The invention provides a Suillus luteus polysaccharide, the monosaccharide composition of the Suillus luteus polysaccharide comprises fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid.
Further, the molar ratio of the fucose, the arabinose, the rhamnose, the galactose, the glucose, the xylose, the mannose, the ribose and the glucuronic acid is 9.68: 0.13: 0.17: 21.51: 42.08: 3.23: 16.63: 3.46: 3.10.
Further, the Suillus luteus polysaccharide has pyranoside bond.
The invention also provides a preparation method of Suillus luteus polysaccharide, which comprises the steps of hot water extraction and ethanol precipitation with Suillus luteus fruiting body as raw material.
Further, the preparation method also comprises the treatment of removing protein by Sevag method.
Further, the preparation method also comprises dialysis impurity removal treatment.
The invention also provides an application of the Suillus luteus polysaccharide in the preparation of antioxidant drugs.
The invention also provides an antioxidant drug, the active ingredient of which comprises the Suillus luteus polysaccharide.
Further, the drug also comprises pharmaceutically acceptable auxiliary materials.
Further, the dosage form of the drug is tablet, capsule, dripping pill, granule, powder, pellet, solution, syrup, emulsion or injection.
The invention discloses the following technical effects:
According to the invention, the polysaccharide SIP with novel structure and antioxidant activity is obtained from the natural bacterium Suillus luteus for the first time, and it has strong radical scavenging ability to DPPH free radical, hydroxyl free radical and superoxide anion, and can be used for preparing anti-oxidation related 10909136 drugs, providing a research foundation for the subsequent development and utilization of Suillus luteus, and having important economic value and market value.
In order to more clearly explain the embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings that need to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained according to these drawings without any creative effort.
FIG. 1 shows the analysis of SIP monosaccharide composition of Suillus luteus; where, A is the ion chromatogram of the standard sample; B is the ion chromatogram of SIP sample;
FIG. 2 shows SIP infrared spectrum analysis of Suillus luteus,
FIG. 3 shows the antioxidant activity of SIP of Suillus luteus in vitro; where, A is the scavenging effect of DPPH free radicals; B is the hydroxyl radical scavenging effect; C is the scavenging effect of superoxide anion.
Now, various exemplary embodiments of the present invention will be described in detail. This detailed description should not be taken as a limitation of the present invention, but should be understood as a more detailed description of some aspects, characteristics and embodiments of the present invention.
It should be understood that the terms mentioned in the present invention are only used to describe specific embodiments, and are not used to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Any stated value or intermediate value within the stated range, and any other stated value or intermediate value within the stated range are also included in the present invention. The upper and lower limits of these 10909136 smaller ranges can be independently included or excluded from the range.
Unless otherwise stated, all technical and scientific terms used herein have the same meanings commonly understood by those of ordinary skill in the field to which this invention relates. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials related to the documents. In case of conflict with any incorporated documents, the contents of this specification shall prevail.
Without departing from the scope or spirit of the present invention, it is obvious to those skilled in the art that many modifications and changes can be made to the specific embodiments of the present specification. Other embodiments obtained from the description of the present invention will be obvious to the skilled person. The description and embodiment of that invention are only exemplary.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open ended terms that mean including, but not limited to.
Embodiment 1
Preparation of Suillus luteus polysaccharide;
Oven-drying the fruiting body of Suillus luteus, pulverizing and sieving with a 100-mesh sieve, extracting Suillus luteus powder with water for 168.83 h at 68.97°C and the ratio of liquid to material of 45.21 mL/g; concentrating the extractive solution to 1/4 of the original concentration, adding absolute ethanol to the final concentration of 75 wt%, standing overnight, centrifuging at 5000 r/min to remove supernatant, precipitating and drying for later use; dissolving the precipitate in deionized water, deproteinizing by Sevag reagent (n-butanol: chloroform =1:4,v/v) twice, and then dialyzing by a 3500 Da cut-off dialysis bag to remove small molecules. After that, concentrating by a rotary evaporator, and then freeze-drying by a freeze dryer to obtain the polysaccharide SIP of Suillus luteus. 10909136
Embodiment 2
Analysis of SIP monosaccharide composition of Suillus luteus polysaccharide 5 Hydrolysis of polysaccharide: accurately weighing 10 mg of Suillus luteus polysaccharide SIP, dissolving in 4 mol/L trichloroacetic acid, shaking thoroughly, and mixing evenly, fully reacting in an electrothermal constant-temperature blast drying oven at 110°C for 6 h. After the reaction, adding absolute ethanol to remove excess trichloroacetic acid, and then drying the sample to obtain the hydrolyzed polysaccharide sample.
Preparation of monosaccharide standard solution and preparation of mixed standard solution: accurately weighing 10 mg of monosaccharide standard solutions of fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid, and dissolving them in purified water to 5 mL to obtain 2 mg/mL monosaccharide standard solution; accurately weighing 10 mg of the monosaccharide standards of fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid again, mixing the six monosaccharides, dissolving them in purified water and fixing the volume to 30 mL, thus obtaining 2 mg/mL of mixed sugar standard solution.
All the prepared sugar solutions are filtered by 0.22 um microporous membrane, and then detected by high performance liquid chromatography. The results are shown in FIG. 1. The results show that the polysaccharide SIP of Suillus luteus is mainly composed of fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid, with the molar percentages of 9.68%, 0.13%, 0.17%, 21.51%, 42.08%, 3.23%, 16.63%, 3.46%, 3.10%.
Embodiment 3
Infrared spectrum analysis of polysaccharide SIP from Suillus luteus;
Accurately weighing 1mg of Suillus luteus polysaccharide SIP, evenly mixing it with 150 mg of potassium bromide, and then performing tabletting treatment, and scanning in the range of 4000-400 cm“! by Fourier transform infrared spectrometer. 7503792
The results are shown in FIG. 2. The results show that the stretching vibration peaks of O-H and C-H are at 3415 cm”! and 2911 cm”! respectively, the absorption peak of
C=0 or C=C in uronic acid is at 1647 cm”!, and the deprotonated COO- absorption peak is at 1410 cm“, the absorption peak of pyranose is at 1000-1200 cm“.
Effect verification
Suillus luteus polysaccharide SIP has the ability to scavenge DPPH free radical, hydroxyl free radical and superoxide anion:
The antioxidant activity of Suillus luteus polysaccharide SIP is demonstrated by measuring its ability to scavenge hydroxyl radical, DPPH radical and superoxide anion, and Vcis used as the positive control.
The method of scavenging DPPH free radical activity: mixing 0.75 mL of 0.1 mM DPPH with 1.5 mL of different concentrations of Suillus luteus polysaccharide
SIP solution (1.0, 2.0, 3.0, 4.0 and 5.0 mg/mL), avoiding light for 30 min, and measuring the absorbance A1 of absolute ethanol at 517 nm; mixing 0.75 mL absolute ethanol with 1.5mL SIP solution of Suillus luteus polysaccharide, avoiding light for 30 min, and then measuring the absorbance A» at 517 nm; mixing 0.75 mL of 0. ImM
DPPH with 1.5 mL of absolute ethanol, avoiding light for 30 min, and then measuring the absorbance Ao at 517nm. According to the following formula, calculating the scavenging capacity of Suillus luteus polysaccharide SIP on DPPH free radical:
Clearance rate (%) = [1-(A1-A2)/Ao] x 100%.
Activity test of scavenging hydroxyl radical: taking ImL of 0.02M phosphate buffer (pH = 7), 0.8mL of 0.15mm methylene blue solution, 0.4mL of 0.01M Fe (ID-EDTA solution, 0.2mL of 7.5mm HO: solution, adding 0.5mL of SIP solutionof
Suillus luteus polysaccharide with different concentrations (1.0, 2.0, 3.0, 4.0 and 5.0mg/mL), diluting to 10 mL with 90% ethanol solution, mixing and reacting for 2 h, and measuring the absorbance at 660 nm. Ao is the absorbance of the initial methylene blue solution, and A1 is the addition of Fenton, Az is the absorbance after adding
Fenton reagent and SIP solution of Suillus luteus polysaccharide. According to the following formula, calculate the scavenging capacity of Suillus luteus polysaccharide
SIP on hydroxyl radical: 7503792
Clearance rate (%) = [1-(Ao-A2)/(Ao-A1)] x 100%.
The method of scavenging superoxide anion activity: taking 4mL of Tris-HCI solution (pH=8.2), adding 0.5mL of SIP solution (1.0, 2.0, 3.0, 4.0 and 5.0mg/mL) of
Suillus luteus polysaccharide, adjusting the volume to 10mL with ethanol, reacting at 48°C for 40 min, and adding 0.3 mL of 3 mM pyrogallol preheated at 48°C, immediately mix well, measuring the absorbance at 319.5 nm, and the absorbance of the blank sample is Ao, and that of the sample after adding the polysaccharide SIP of
Suillus luteus is A1. According to the following formula, calculating the scavenging capacity of Suillus luteus polysaccharide SIP to superoxide anion:
Clearance rate (%)=[1-A1/A0]*100%.
The results of the above three antioxidant experiments are shown in FIG. 3.
Suillus luteus polysaccharide has strong free radical scavenging ability on DPPH free radical, hydroxyl free radical and superoxide anion.
The above-mentioned embodiments only describe the preferred mode of the invention, but do not limit the scope of the invention. On the premise of not departing from the design spirit of the invention, all kinds of modifications and improvements made by ordinary technicians in the field to the technical scheme of the invention shall fall within the scope of protection determined by the claims of the invention.
Claims (10)
1. A Suillus luteus polysaccharide, characterized in that its monosaccharide composition comprises fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, ribose and glucuronic acid.
2. The Suillus luteus polysaccharide according to claim 1, characterized in that the molar ratio of the fucose, the arabinose, the rhamnose, the galactose, the glucose, the xylose, the mannose, the ribose and the glucuronic acid is 9.68: 0.13: 0.17:
21.51: 42.08: 3.23: 16.63: 3.46: 3.10.
3. The Suillus luteus polysaccharide according to claim 1, characterized in that the Suillus luteus polysaccharide has pyranoside bond.
4. A preparation method of Suillus luteus polysaccharide according to any one of claims 1-3, characterized in that the preparation method comprises the steps of hot water extraction and ethanol precipitation with Suillus luteus fruiting body as raw material.
5. The preparation method according to claim 1, characterized in that the preparation method further comprises the treatment of removing protein by Sevag method.
6. The preparation method according to claim 1, characterized in that the preparation method further comprises dialysis impurity removal treatment.
7. An application of Suillus luteus polysaccharide according to any one of claims 1-3 in the preparation of antioxidant drugs.
8. An antioxidant medicine, characterized in that the active ingredient comprises the Suillus luteus polysaccharide according to any one of claims 1 - 3.
9. The medicine according to claim 8, characterized in that the medicine further 10909136 comprises pharmaceutically acceptable auxiliary materials.
10. The medicine according to claim 8, characterized in that the dosage form of the medicine is tablet, capsule, dripping pill, granule, powder, pellet, solution, syrup, emulsion or injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU503132A LU503132B1 (en) | 2022-12-05 | 2022-12-05 | Suillus luteus polysaccharide, its preparation method and medical application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| LU503132A LU503132B1 (en) | 2022-12-05 | 2022-12-05 | Suillus luteus polysaccharide, its preparation method and medical application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| LU503132B1 true LU503132B1 (en) | 2023-06-05 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| LU503132A LU503132B1 (en) | 2022-12-05 | 2022-12-05 | Suillus luteus polysaccharide, its preparation method and medical application |
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| LU (1) | LU503132B1 (en) |
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2022
- 2022-12-05 LU LU503132A patent/LU503132B1/en active IP Right Grant
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Effective date: 20230605 |