CN1398901A - Prepn and use of curcuma oligosaccharide sulfate derivative - Google Patents
Prepn and use of curcuma oligosaccharide sulfate derivative Download PDFInfo
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- CN1398901A CN1398901A CN 01108650 CN01108650A CN1398901A CN 1398901 A CN1398901 A CN 1398901A CN 01108650 CN01108650 CN 01108650 CN 01108650 A CN01108650 A CN 01108650A CN 1398901 A CN1398901 A CN 1398901A
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- curcuma
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- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 54
- -1 curcuma oligosaccharide sulfate derivative Chemical class 0.000 title claims abstract description 41
- 235000014375 Curcuma Nutrition 0.000 title claims description 53
- 244000164480 Curcuma aromatica Species 0.000 title 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 75
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 17
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 16
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 12
- 239000003814 drug Substances 0.000 claims abstract description 10
- 239000000843 powder Substances 0.000 claims abstract description 8
- 239000000706 filtrate Substances 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 5
- UFWWAKOWSBGGCP-UHFFFAOYSA-N pyridine;sulfurochloridic acid Chemical compound OS(Cl)(=O)=O.C1=CC=NC=C1 UFWWAKOWSBGGCP-UHFFFAOYSA-N 0.000 claims abstract description 4
- 238000010992 reflux Methods 0.000 claims abstract description 4
- 241000407170 Curcuma Species 0.000 claims description 52
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 claims description 28
- 229960004756 ethanol Drugs 0.000 claims description 28
- 241000700605 Viruses Species 0.000 claims description 19
- 150000002482 oligosaccharides Chemical class 0.000 claims description 18
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims description 16
- 238000001556 precipitation Methods 0.000 claims description 12
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- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 8
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- 150000003467 sulfuric acid derivatives Chemical class 0.000 claims description 3
- XTHPWXDJESJLNJ-UHFFFAOYSA-N sulfurochloridic acid Chemical compound OS(Cl)(=O)=O XTHPWXDJESJLNJ-UHFFFAOYSA-N 0.000 claims description 3
- 238000011951 anti-virus test Methods 0.000 claims description 2
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- XVMSFILGAMDHEY-UHFFFAOYSA-N 6-(4-aminophenyl)sulfonylpyridin-3-amine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=N1 XVMSFILGAMDHEY-UHFFFAOYSA-N 0.000 claims 1
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- MYKOKMFESWKQRX-UHFFFAOYSA-N 10h-anthracen-9-one;sulfuric acid Chemical compound OS(O)(=O)=O.C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 MYKOKMFESWKQRX-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
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- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 3
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Abstract
The present invention relates to the extraction of a natural matter, and its derivative and its preparation and application. Fragrant solomonseal oligososaccharide is first produced with fragrant solomonseal rhizome powder as raw material and through high-purity ethanol reflux, filtering, leaching the filter residue with ethanol solution, filtering to obtain filtrate, deposition with high-concentration ethanol, dewatering the precipitant with absolute alcohol, acetone and ethyl ether, and stoving to dryness. It is then modified by the chlorosulfonic acid-pyridine process to obtain fragrant solomonseal oligosaccharide sulfate derivative. The derivative may be used in antiviral medicine.
Description
The present invention relates to the extraction of crude substance and the preparation and the application of derivative thereof.
Radix polygonati officinalis Polygonatum odoratum (Mill.) Druce is a kind of traditional Chinese medicine of China, according to " China's book on Chinese herbal medicine " record, its root stock contains radix polygonati officinalis mucopolysaccharide (being made up of fructose, glucose, seminose and galacturonic acid), radix polygonati officinalis Polylevulosan (being made up of fructose and glucose).
Controlling sulfate polyose (sulfated polysaccharides, SPS) or claim Sulfate of polysaccharide (polysaccharides sulfated, PSS) being polyanionic compound, is that certain hydroxyl on the monose molecule is replaced by sulfate radical and a class chemical structure complexity that forms, the polysaccharide derivates that biological activity is various, structure activity relationship is distinct in the polysaccharide macro-molecular chain.Except that the blood coagulation resisting function of having found, find constantly that in recent years controlling sulfate polyose has multiple biological activity, as enhancing body immunologic function, antitumor, antiviral etc., make the research of controlling sulfate polyose become a new focus in the polysaccharide research field.
The harm of virus disease has seriousness and ubiquity, seek and develop new, antiviral makes antiviral activity become the importance that the controlling sulfate polyose biologic activity is studied recently safely and effectively.Find that from 1987 the sulphating dextran has inhibition hiv virus (Human immunodeficiency virus, HIV) since the activity, the research of the antiviral activity aspect of such polysaccharide is very active, it might become the another class hiv inhibitor of continue viral reverse transcriptase activity inhibitor, proteinase inhibitor, therefore, more and more be subject to people's attention through the sulfuric ester relevant research that modify to obtain the antiviral activity newtype drug of deriving about carbohydrate.The xylan sulfuric ester clinic trial of Germany's development has at present obtained good result in the treatment of acquired immune deficiency syndrome (AIDS); And the curdlan sulfuric ester (curdlansulfate) of Japanese scientist development has been carried out I/II phase clinical trial in the U.S., is considered to one of tool potential and actual active polysaccharide formulation of antiretroviral recently.
The mechanism of action of summary controlling sulfate polyose anti-AIDS such as Xiang Daobin mainly contain viral interference to the absorption of host cell, suppress virus antigen expression, suppress plasmodial formation, suppress reverse transcriptase activity, raise immunity etc.And controlling sulfate polyose has been deep into the concrete combining site of HIVgp120 to the interactional interference effect between the CD4 molecule on HIV-1 membrane glycoprotein gp120 and the target cell membrane.According to Suleparoid is the relevant research of the acceptor of mediation virus absorption on the host cell membrane, may combine the absorption of inhibition virus to target cell as the analog of Suleparoid with virus envelope glycoprotein thereby most scholar infers controlling sulfate polyose.Because bibliographical information virus HIV, hsv (Herpessimplex virus, HSV), human cytomegalic inclusion disease virus (Human cytomega lovirus, HCMV) and protease-resistant protein (Proteinase resistant protein, PrP) etc., all will with the Suleparoid effect, this makes controlling sulfate polyose very likely become effect molecular basis New-type wide-spectrum antiviral clearly.
The object of the invention just provides a kind of extraction of curcuma oligosaccharide and the preparation method and the application of curcuma oligosaccharide sulfate derivative in making antiviral of sulfate derivative thereof.
The present invention can be achieved in the following manner: with radix polygonati officinalis root stock dry powder is raw material, adopt high density (〉=70%) alcohol reflux earlier, filter, the ethanol lixiviate of filter residue certain volume (4-20 of radix polygonati officinalis thousand powder doubly) and concentration (30～60%), filter and collect filtrate, use high concentration ethanol (〉=70%) precipitation again, precipitation is dewatered with dehydrated alcohol, acetone, ether, oven dry promptly gets curcuma oligosaccharide, carry out sulfuric ester by chlorsulfonic acid-pyridine method again and derive and modify to obtain the sulfate derivative of curcuma oligosaccharide, this derivative can be applicable to antiviral.
Bulk drug of the present invention select for use " the traditional Chinese medicine radix polygonati officinalis described in the Chinese pharmacopoeia (version in 2000) do not have go mouldy, color and luster is yellowish, do not have assorted assorted piece root, piece undercut sheet, drying is ground into particulate state.Radix polygonati officinalis dry powder carries out reflow treatment with high concentration ethanol (ethanol more than 70%), remove small-molecule substances such as pigment, monose, filter, the retortable recovery ethanol of filtrate, filter residue spends the night with the proper volume (4-20 of radix polygonati officinalis dry powder doubly) and the ethanol lixiviate of concentration (30～60%), nylon cloth filters, filtrate is with 95% ethanol sedimentation of 2-4 times of volume, final concentration is 73～88%, collecting precipitation, with dehydrated alcohol, acetone, ether dehydration, oven dry promptly gets curcuma oligosaccharide respectively for the retortable recovery ethanol of supernatant liquor, precipitation.
The curcuma oligosaccharide of above-mentioned acquisition is that to measure sugared content with anthrone-sulfuric acid process be 80～90% to standard substance with fructose; With the bovine serum albumin is that standard substance Folin-phenol method mensuration protein content is 1～2%; It is 0.5～2% that bariumchloride-gelatin method is measured sulphur content; Thin layer chromatography carries out the monose compositional analysis, and curcuma oligosaccharide is made up of fructose, glucose and seminose; The KBr compressing tablet, Nicolet200 SXV FT-IR Infrared spectroscopy, curcuma oligosaccharide has the characteristic peak of carbohydrate, 3500～3400,2920,2844,1460～1350,1150～1000cm
-1Absorption is arranged, at 937cm
-1And 815cm
-1Near have the absorption peak of the furan nucleus symmetrical stretching vibration of fructose and the vibration of C-H angle, at 810cm
-1And 870cm
-1Near the charateristic avsorption band of seminose is arranged.Entrust country of Dalian Inst of Chemicophysics, Chinese Academy of Sciences chromatogram center to carry out curcuma oligosaccharide matrix-assisted laser desorption-flying time mass spectrum analysis, its molecular weight mainly is distributed in 3000～5000.
Curcuma oligosaccharide with above-mentioned preparation is a raw material, adopts chlorsulfonic acid-pyridine legal system to be equipped with curcuma oligosaccharide sulfate derivative.Under ice bath and agitation condition dropwise with chlorsulfonic acid by a certain percentage (1: 10～1: 2) add in the pyridine, in this sulfur acidizing reagent, add the curcuma oligosaccharide that is dissolved in the methane amide again, the V/W of methane amide and curcuma oligosaccharide (volume/mass) is: 10: 1～50: 1, the V/W of pyridine and curcuma oligosaccharide was: 5: 1.After ice bath and agitation condition reacted certain hour down, with 95% ethanol sedimentation of 2-5 times of volume, the ethanol final concentration was 63～80%.Abandon supernatant liquor, precipitation is used a small amount of dissolved in distilled water, and sodium-acetate is neutralized to neutrality, and with 2-5 times of volume 95% ethanol sedimentation, the ethanol final concentration is 63～80% again.With dehydrated alcohol, acetone, ether dehydration, oven dry promptly gets curcuma oligosaccharide sulfate to precipitation respectively.
The curcuma oligosaccharide sulfate of above-mentioned acquisition is that to measure sugared content with anthrone-sulfuric acid process be 30-70% to standard substance with fructose; With the bovine serum albumin is that standard substance Folin-phenol method mensuration protein content is 0.5-1.5%; Bariumchloride-gelatin method is measured sulphur content and is increased to 3-20%, and its sulfuric ester substitution value is 0.17～2.79; The KBr compressing tablet, Nicolet200 SXV FT-IR Infrared spectroscopy, curcuma oligosaccharide sulfate not only has the absorption peak of curcuma oligosaccharide, has increased 1250cm in addition newly
-1, 810cm
-1Near absorption peak is respectively S=O stretching vibration and C-O-S stretching vibration, 620cm
-1Near spike explanation sulfuric ester exists with sodium-salt form.
The curcuma oligosaccharide sulfate derivative that above method is made is applied to antivirus test, can effectively suppress Vero cytopathy and virus plaque formation that HSV-2 causes, can be developed as broad-spectrum antiviral medicament, be used for the treatment of bleb, influenza, hepatitis and acquired immune deficiency syndrome (AIDS).
The present invention has the following advantages:
1, this invented technology is simple, and required starting material are cheap and easy to get, is used for the curcuma oligosaccharide lixiviate and sedimentary ethanol can distill recovery, can save cost greatly.
2, the derivative of Huo Deing has anti-herpes simplex virus 2 types (Herpes simplex virustype2, HSV-2) activity, can be developed as the herpes medicine, be used for the treatment of other virus disease that simplexvirus property keratitis, genital herpes and simplexvirus cause, improve patient's quality of life.
Thereby 3,, controlling sulfate polyose suppresses the absorption of virus to target cell because may combining with virus envelope glycoprotein as the analog of Suleparoid, and bibliographical information virus HIV, HSV, HCMV and PrP etc., all be to be the acceptor of invasion cell with the Suleparoid, therefore the derivative that obtains very likely is developed as clearly New-type wide-spectrum antiviral of effect molecular basis, is used for the treatment of bleb, influenza, hepatitis and acquired immune deficiency syndrome (AIDS).
The preparation of embodiment 1 curcuma oligosaccharide
200g radix polygonati officinalis dry powder is eluriated 2～3 times with 85% ethanol of certain volume with 4 times of volumes, 85% ethanol boiling reflux 1 hour again, abandons filtrate, and filter residue spends the night with the 50% ethanol lixiviate of 14 times of volumes.Filter residue is abandoned in filtration, filtrate 8500rpm, 4 ℃ centrifugal 30 minutes, get supernatant liquor and spend the night with 95% ethanol sedimentation of 2 times of volumes.Collecting precipitation with dehydrated alcohol, acetone, ether dehydration, gets the 16.77g curcuma oligosaccharide after 50 ℃ of oven for drying.It is 1.17% that bariumchloride-gelatin method is measured sulphur content, and it is 87.88% that anthrone-sulfuric acid process is measured sugared content, and it is 1.77% that Folin-phenol method is measured protein content.The total recovery of this preparation flow is 8.39%.The infrared spectra of curcuma oligosaccharide has the characteristic peak of polysaccharide, and flight time mass spectrum shows that its main molecules amount is distributed in 3000～5000, is made up of 18～31 monosaccharide residues approximately, and thin layer chromatography analysis shows that curcuma oligosaccharide contains fructose, seminose and glucose.
The preparation of embodiment 2 curcuma oligosaccharide sulfates
The 5mL chlorsulfonic acid dropwise adds under ice bath and agitation condition in the 10mL pyridine, and the 2g curcuma oligosaccharide adds in the above-mentioned sulfonated reagent after being dissolved in the 80mL methane amide, and ice bath and agitation condition reacted 6 hours down.In 95% ethanol with 400mL0 ℃ of precooling of above-mentioned reaction solution impouring after reaction finishes, place 4 ℃ of refrigerator precipitations to spend the night.Abandon supernatant, precipitation is used a small amount of dissolved in distilled water, and sodium-acetate is neutralized to neutrality, 95% ethanol sedimentation of 4 times of volumes.Dehydrated alcohol, acetone, ether dehydration get 1.09g curcuma oligosaccharide sulfate product after 50 ℃ of oven for drying.Through the IR spectroscopic analysis, conclusive evidence has sulfate group to replace the hydroxyl of monosaccharide residue.It is 8.47% that bariumchloride-gelatin method is measured sulphur content, calculates and learns that the sulfate group substitution value is 0.588%, and it is 50% that anthrone-sulfuric acid process is measured sugared content, and it is 1.12% that Folin-phenol method is measured protein content.The total recovery of this preparation flow is 39.78%.
Embodiment 3 curcuma oligosaccharide sulfate cytotoxic assay
The Vero cell that is cultured to individual layer in 96 porocyte plates adds the curcuma oligosaccharide sulfate that final concentration is 250～4000 μ g/mL respectively, puts 37 ℃, 5%CO
2Incubator is cultivated, day by day observation of cell growth conditions, after 72 hours, adopt toluylene red viable cell staining to measure the curcuma oligosaccharide sulfate maximum concentration nontoxic to the Vero cell, experimental result shows, below 4000 μ g/mL concentration, curcuma oligosaccharide sulfate does not have tangible toxic action to the Vero cell.
The cytopathy of embodiment 4 curcuma oligosaccharide sulfates suppresses experiment
The Vero cell is cultured to individual layer on 24 porocyte plates, grouping then, 4 every group multiple holes.Cell control group: do not add medicine and virus; Virus control group: infect 100TCID
50HSV-2 virus liquid, do not add medicine; Experimental group: infective virus adds the curcuma oligosaccharide sulfate of different final concentrations simultaneously, investigates the cytopathic restraining effect of Vero that curcuma oligosaccharide sulfate causes HSV-2 in acellular malicious concentration range.Put 37 ℃, 5%CO
2Incubator was cultivated after 72 hours, and the toluylene red staining is measured, and found that concentration can suppress the cytopathy (p＜0.05) that HSV-2 causes well at the curcuma oligosaccharide sulfate of 125～2000 μ g/mL.
Embodiment 5 curcuma oligosaccharide sulfate virus plaques suppress experiment
The Vero cell is cultured to the individual layer grouping on 24 porocyte plates standby.Curcuma oligosaccharide sulfate derivative is diluted to different final concentrations with the bottom solid medium of 90 ℃ of thawings, 43-45 ℃ insulation.Viral dilution to about 30-50PFU/ hole, is added and contains sugared bottom solid medium, add in the corresponding aperture immediately behind the mixing.The cell control group only adds the bottom solid medium; The virus control group only adds and contains viral bottom solid medium; Experimental group adds the bottom solid medium that contains virus and corresponding final concentration curcuma oligosaccharide sulfate.Treat that the bottom solid medium solidifies rearmounted 37 ℃, 5%CO
2Incubator is inverted and was cultivated 2 days, adds the upper strata solid medium that contains 1: 25000 toluylene red, solidifies rearmounted 37 ℃, 5%CO
2Incubator is inverted shading and was cultivated 1 day, observes virus plaque and forms situation.The curcuma oligosaccharide sulfate that found that 200 μ g/mL can suppress the virus plaque formation that HSV-2 causes fully, and the curcuma oligosaccharide sulfate of 100,50,25 μ g/mL has also produced plaque in various degree and formed restraining effect (p＜0.05).
Claims (4)
1, a kind of preparation method of curcuma oligosaccharide sulfate derivative, it is characterized in that: with radix polygonati officinalis root stock dry powder is raw material, adopt high concentration ethanol to reflux earlier, filter, filter residue filters and collects filtrate with the ethanol lixiviate of radix polygonati officinalis dry powder 4-20 30～60% concentration doubly, precipitates with high concentration ethanol again, precipitation is dried and is promptly got curcuma oligosaccharide with dehydrated alcohol, acetone, ether dehydration; Carrying out sulfuric ester by chlorsulfonic acid-pyridine method again derives and modify to obtain the sulfate derivative of curcuma oligosaccharide.
2, the preparation method of curcuma oligosaccharide sulfate derivative according to claim 1 is characterized in that: being used for sedimentary high concentration ethanol concentration is more than 70%; The ethanol lixiviate of 30～60% concentration is spent the night.
3, the preparation method of curcuma oligosaccharide sulfate derivative according to claim 1, it is characterized in that: be raw material with the curcuma oligosaccharide, under ice bath and agitation condition, dropwise chlorsulfonic acid is added in the pyridine in 1: 10～1: 2 ratio, in this sulfur acidizing reagent, add the curcuma oligosaccharide that is dissolved in the methane amide again, the volume/mass of methane amide and curcuma oligosaccharide (V/W) is: 10: 1～50: 1, the V/W of pyridine and curcuma oligosaccharide is: 5: 1, after ice bath and agitation condition react certain hour down, 95% ethanol sedimentation with 2-5 times of volume, the ethanol final concentration is 63～80%, abandon supernatant liquor, the a small amount of dissolved in distilled water of precipitation, sodium-acetate is neutralized to neutrality, and with 2-5 times of volume 95% ethanol sedimentation, the ethanol final concentration is 63～80% again.With dehydrated alcohol, acetone, ether dehydration, oven dry promptly gets curcuma oligosaccharide sulfate to precipitation respectively.
4, make curcuma oligosaccharide sulfate derivative according to claim 1, it is characterized in that: be applied to antivirus test, can effectively suppress Vero cytopathy and virus plaque formation that HSV-2 causes, can be developed as broad-spectrum antiviral medicament, be used for the treatment of bleb, influenza, hepatitis and acquired immune deficiency syndrome (AIDS).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1330670C (en) * | 2004-02-02 | 2007-08-08 | 中南大学湘雅二医院 | Process for extracting polygonatum polysaccharides, preparing process for medical preparation and use thereof |
| CN100363373C (en) * | 2005-01-14 | 2008-01-23 | 庄茅 | Preparation method of isomalto loigosaccharide sulphate (IMOS) |
| CN100404557C (en) * | 2005-12-09 | 2008-07-23 | 武汉理工大学 | Preparation method of umbellate pore fungus polysaccharide sulphate |
| CN103627612A (en) * | 2013-09-30 | 2014-03-12 | 浙江致中和实业有限公司 | White spirit cold-soaking and countercurrent extraction method of polygonatum odoratum polysaccharide in brewing technique and online quality control method thereof |
| CN112125983A (en) * | 2020-09-30 | 2020-12-25 | 华南理工大学 | Water-soluble polygonatum odoratum polysaccharide, sulfated polygonatum odoratum polysaccharide thereof, and preparation method and application of water-soluble polygonatum odoratum polysaccharide |
-
2001
- 2001-07-19 CN CN 01108650 patent/CN1256349C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1330670C (en) * | 2004-02-02 | 2007-08-08 | 中南大学湘雅二医院 | Process for extracting polygonatum polysaccharides, preparing process for medical preparation and use thereof |
| CN100363373C (en) * | 2005-01-14 | 2008-01-23 | 庄茅 | Preparation method of isomalto loigosaccharide sulphate (IMOS) |
| CN100404557C (en) * | 2005-12-09 | 2008-07-23 | 武汉理工大学 | Preparation method of umbellate pore fungus polysaccharide sulphate |
| CN103627612A (en) * | 2013-09-30 | 2014-03-12 | 浙江致中和实业有限公司 | White spirit cold-soaking and countercurrent extraction method of polygonatum odoratum polysaccharide in brewing technique and online quality control method thereof |
| CN103627612B (en) * | 2013-09-30 | 2015-07-29 | 浙江致中和实业有限公司 | The white wine cold soaking of fragrant solomonseal rhizome polyoses, countercurrent extraction method and line Quality Control method thereof in wine-making technology |
| CN112125983A (en) * | 2020-09-30 | 2020-12-25 | 华南理工大学 | Water-soluble polygonatum odoratum polysaccharide, sulfated polygonatum odoratum polysaccharide thereof, and preparation method and application of water-soluble polygonatum odoratum polysaccharide |
| CN112125983B (en) * | 2020-09-30 | 2022-03-18 | 华南理工大学 | Water-soluble polygonatum odoratum polysaccharide, sulfated polygonatum odoratum polysaccharide thereof, and preparation method and application of water-soluble polygonatum odoratum polysaccharide |
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