CN101265302A - Panax japonicus polysaccharides, preparation method and use thereof - Google Patents
Panax japonicus polysaccharides, preparation method and use thereof Download PDFInfo
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- CN101265302A CN101265302A CNA2008100475058A CN200810047505A CN101265302A CN 101265302 A CN101265302 A CN 101265302A CN A2008100475058 A CNA2008100475058 A CN A2008100475058A CN 200810047505 A CN200810047505 A CN 200810047505A CN 101265302 A CN101265302 A CN 101265302A
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- panacis japonici
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- japonici polysaccharides
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Abstract
The invention discloses panax japonicus polysaccharide. The main chain structure thereof is Beta-1 to 4-D glucan and heterosaccharide containing mannan, galactan and xylan branched chain, the weight-average molecular weight of the panax japonicus polysaccharide is 5,000 to 200,000, the intrinsic viscosity at 25 DEG C is 1 to 50ml/g, and the mole ratio of the three elements of C: H: O is 1: 2: 1. The method for preparing the panax japonicus polysaccharide comprises the following steps: after being dried, smashed and skimmed, the panax japonicus is extracted by neutral salt water solution or pure water, and the white panax japonicus polysaccharide which can be easily dissolved in water, be dissolved in DMSO and not be dissolved in methanol, ethanol and acetone is obtained by separation, saponin removal, decolorization, dehydration, protein removal, desalination and drying. The panax japonicus polysaccharide which is prepared by the invention has higher anti-tumor activity and promotion of immune activity, which can be used for preparing drugs or health care products for improving immune function and anti-tumor. The method for preparing the product is simple and easy to operation, and the sources of the needed raw materials are wide.
Description
Technical field
The present invention relates to a kind of Rhizoma Panacis Japonici polysaccharides and its production and use, belong to the Chemistry and Physics of Polymers field, also belong to biology, pharmacy, food and field of health care products.
Background technology
Rhizome of Japanese Ginseng (Rhizoma Panacis Japonici), or claim panax japonicus, Herba Rhodiolae Henryi, Rhizome of Bipinnatifid Ginseng etc., (Panax japonicus C.A.Mey) is the whiplike rhizome of bamboo for Araliaceae platymiscium rhizome of Japanese Ginseng, mainly originates in China middle and south, is China's traditional Chinese medicine.In the domestic and foreign literature the contained chemical constitution study of rhizome of Japanese Ginseng rhizome is only limited to Rhizoma Panacis Japonici saponin (chikusetsu-saponin), ginsenoside (ginsenoside), Panax Notoginseng saponin R at present
2(notoginsenoside R
2) and pseudoginsenoside F
11(pseudo-ginsenoside F
11), Rhizoma Panacis Japonici polysaccharides is also studied.Rhizome of Japanese Ginseng is the local product resource of China, is used as medicine with it, have invigorating blood circulation, stop blooding, analgesia, anti-inflammatory, delay senility and have the help effect of genseng concurrently, and be one of conventional Chinese medicine among the people.But rhizome of Japanese Ginseng directly is used as Chinese medicine, and curative effect is restricted, because contain multiple composition in the rhizome of Japanese Ginseng, the effect of various compositions is also different, wherein polysaccharide just comprises beta-glucan, beta-mannase, α-mannosans and mixed polysaccharide, and change with extraction conditions.
Summary of the invention
The purpose of this invention is to provide a kind of Rhizoma Panacis Japonici polysaccharides and its production and use, not only desired raw material is cheap, and the preparation method is easy, pollution-free, and Rhizoma Panacis Japonici polysaccharides purity height, the biological activity of preparation are good.
A kind of Rhizoma Panacis Japonici polysaccharides, backbone structure is β-1 → 4-D dextran, contains the mixed polysaccharide of mannosans, Polygalactan, xylan side chain, and its weight-average molecular weight is 0.5～200,000,25 ℃ intrinsic viscosity is 1～50mL/g, C: H: O element mol ratio is 1: 2: 1.
Preparing above-mentioned Rhizoma Panacis Japonici polysaccharides is that the rhizome of Japanese Ginseng of earlier drying also being pulverized sloughs fat and small molecular weight impurity with organic solvent, vacuum-drying, get of the aqueous solution or pure water normal temperature down immersion and the stirring extraction of drying back solid with neutral salt, pass through separation, dehydration, decolouring, deproteinated, desalination and take off saponin(e, final drying promptly makes Rhizoma Panacis Japonici polysaccharides.Wherein use supercentrifugal process or/and vacuum filtration method precipitation separation the mixed serum of extraction.Solution concentrates with rotatory evaporator, uses the defoamer froth breaking during with rotatory evaporator vacuum hydro-extraction.With Sevag method deproteinated.Use H
2O
2Decolouring, and be 7～10 with ammoniacal liquor regulator solution pH value.With tap water and distill water dialysis, take off saponin(e with the alcohol reflux method.
Rhizoma Panacis Japonici polysaccharides of the present invention is shown that by Sarcoma 180 tumours and lymphocyte external activity experimental result Rhizoma Panacis Japonici polysaccharides of the present invention has the obvious suppression effect to tumour cell, even surpass the effect (50%) of chemicals 5-fluor-uracil (5-Fu); Lymphocyte there is obvious promotion proliferation function, surpasses lipopolysaccharides (LPS) and canavaline-A (Con-A) effect.This Rhizoma Panacis Japonici polysaccharides has higher anti-tumor activity, and has higher promotion body's immunity, can be used for preparation and improves immunity function and anti-tumor drug or healthcare products.
By gas chromatography-mass spectrum (GC-MS),
1H,
13C and two dimensional NMR spectrum (NMR), infrared spectra (IR), ultimate analysis etc. studies show that this Rhizoma Panacis Japonici polysaccharides structure is β-1 → 4-D dextran main chain, contain the mixed polysaccharide of mannosans, Polygalactan and xylan side chain.Application macromolecular solution theory adopts laser light scattering-size exclusion chromatogram (LLS-SEC), capillary viscosity law technology result of study and transmission electron microscope picture to show and shows that this Rhizoma Panacis Japonici polysaccharides molecular chain is spherical in shape in water.
The present invention utilizes biotechnology, and extraction separation goes out to have the Rhizoma Panacis Japonici polysaccharides of obvious anti-tumor activity and immunity function from rhizome of Japanese Ginseng, simple, the lower cost of method therefor.The present invention utilizes the theoretical and chain conformation of method research Rhizoma Panacis Japonici polysaccharides in solution of Polymer Physics; Prepared Rhizoma Panacis Japonici polysaccharides has higher anti-tumor activity, and has higher promotion body's immunity, can be used for preparation and improves immunity function and anti-tumor drug or healthcare products.
Description of drawings
Fig. 1 is a Rhizoma Panacis Japonici polysaccharides
13The C nmr spectrum.
Fig. 2 is the infrared spectrogram of Rhizoma Panacis Japonici polysaccharides.
Fig. 3 is the laser light scattering-size exclusion chromatography figure of Rhizoma Panacis Japonici polysaccharides.
Fig. 4 is the transmission electron microscope photo of Rhizoma Panacis Japonici polysaccharides.
Fig. 5 is that Rhizoma Panacis Japonici polysaccharides is to Sarcoma180 tumor cell proliferation restraining effect synoptic diagram.
Fig. 6 is the synoptic diagram of different concns Rhizoma Panacis Japonici polysaccharides to the lymphocyte promoter action.
Embodiment
Below in conjunction with concrete example technical scheme of the present invention is described further.
Rhizome of Japanese Ginseng is produced in the enshi of drying and pulverizing refluxed respectively 4～8 hours with ethyl acetate and acetone, slough fat and part small molecular weight impurity, after the vacuum-drying, get dry back solid 100～1000g, soak and stir extraction 5～24 hours, re-extract 2～5 times with 0.1～2M NaCl aqueous solution, 1～10L at 10～30 ℃.Mixed serum supercentrifugal process precipitation separation, upper solution merge the back and concentrate with rotatory evaporator, add micro-n-Octanol when concentrated and remove bubble.Concentrated solution Sevag method deproteinated, adding ammoniacal liquor regulator solution pH value behind the deproteinated is 7～10, adds H again
2O
2Decoloured 1～4 day.Use tap water and distill water dialysis 2～5 days then successively, removed saponin(e in 2～8 hours with alcohol reflux again after the lyophilize, obtain Rhizoma Panacis Japonici polysaccharides sample RPS1.
Gained Rhizoma Panacis Japonici polysaccharides results of elemental analyses sees Table 1.This Rhizoma Panacis Japonici polysaccharides weight-average molecular weight (M
w), molecular weight distribution (M
w/ M
n), all square the rotation radius (＜s of molecule
2 z 1/2) and intrinsic viscosity physico-chemical properties such as ([η]) see Table 2.The gained Rhizoma Panacis Japonici polysaccharides
13The CNMR spectrum is seen Fig. 1, and the IR spectrogram is seen Fig. 2, and LLS-SEC separates spectrogram and sees Fig. 3.The chain form of this Rhizoma Panacis Japonici polysaccharides molecule in the aqueous solution seen Fig. 4, the S-180 tumour cell suppressed external biological activity such as effect and lymphocyte promoter action and sees Fig. 5 and Fig. 6 respectively.
By gas chromatography-mass spectrum (GC-MS),
1H,
13C and two dimensional NMR (NMR), infrared spectra (IR), ultimate analysis show that this Rhizoma Panacis Japonici polysaccharides structure is β-1 → 4-D dextran (glu) main chain, contain the mixed polysaccharide of mannosans, Polygalactan and xylan side chain.Application macromolecular solution theory adopts laser light scattering-size exclusion chromatogram (LLS-SEC), capillary viscosity law technology result of study and transmission electron microscope picture to show and shows that this Rhizoma Panacis Japonici polysaccharides molecular chain is spherical in shape in water.
Show that by Sarcoma 180 tumour cells and lymphocyte external activity experimental result Rhizoma Panacis Japonici polysaccharides of the present invention has the obvious suppression effect to tumour cell, even surpass the effect (50%) of chemicals 5-fluor-uracil (5-Fu); Lymphocyte there is obvious promotion proliferation function, surpasses lipopolysaccharides (LPS) and canavaline-A (Con-A) effect.This Rhizoma Panacis Japonici polysaccharides has higher anti-tumor activity, and has higher promotion body's immunity, can be used for preparation and improves immunity function and anti-tumor drug or healthcare products.
Table 1 Rhizoma Panacis Japonici polysaccharides results of elemental analyses
Table 2 Rhizoma Panacis Japonici polysaccharides molecular dimension and viscosity
| Sample number into spectrum | M w×10 -4 | M w/M n | <s 2> z 1/2(nm) | [η](mL/g) |
| RPS1 | 1.62 | 1.79 | 18.3 | 10.5 |
Claims (6)
1. Rhizoma Panacis Japonici polysaccharides, backbone structure is β-1 → 4-D dextran, contains the mixed polysaccharide of mannosans, Polygalactan and xylan side chain, and its weight-average molecular weight is 0.5～200,000,25 ℃ intrinsic viscosity is 1～50mL/g, C: H: O element mol ratio is 1: 2: 1.
2. the preparation method of the described Rhizoma Panacis Japonici polysaccharides of claim 1, it is characterized in that: after the degreasing of rhizome of Japanese Ginseng drying and crushing, with the aqueous solution or the pure water extraction of neutral salt, to pass through separation, dehydration, decolouring, deproteinated, desalination and take off saponin(e, drying promptly makes Rhizoma Panacis Japonici polysaccharides.
3. according to the preparation method of the described Rhizoma Panacis Japonici polysaccharides of claim 2, it is characterized in that: adopt supercentrifugal process or/and vacuum filtration method precipitation separation is used the rotatory evaporator thickening, use H
2O
2Decolouring with Sevag method deproteinated, is used the dialysis method desalination, takes off saponin(e with the alcohol reflux method.
4. according to the described method for preparing Rhizoma Panacis Japonici polysaccharides of claim 3, it is characterized in that: use the defoamer froth breaking during with rotatory evaporator vacuum hydro-extraction.
5. according to the described method for preparing Rhizoma Panacis Japonici polysaccharides of claim 3, it is characterized in that: use H
2O
2During decolouring, be 7～10 with ammoniacal liquor regulator solution pH value.
6. the described Rhizoma Panacis Japonici polysaccharides of claim 1 is in the medicine of preparation immunotherapy or oncotherapy, the perhaps application in the healthcare products of enhance immunity function.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103059158A (en) * | 2013-01-29 | 2013-04-24 | 江西省蚕桑茶叶研究所 | Method for inhibiting foam generation in tea flower polysaccharide extraction |
| CN103613680A (en) * | 2013-12-05 | 2014-03-05 | 三峡大学 | Alcohol-precipitated panax japonicus polysaccharide, preparation method and application thereof |
-
2008
- 2008-04-29 CN CN2008100475058A patent/CN101265302B/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103059158A (en) * | 2013-01-29 | 2013-04-24 | 江西省蚕桑茶叶研究所 | Method for inhibiting foam generation in tea flower polysaccharide extraction |
| CN103613680A (en) * | 2013-12-05 | 2014-03-05 | 三峡大学 | Alcohol-precipitated panax japonicus polysaccharide, preparation method and application thereof |
| CN103613680B (en) * | 2013-12-05 | 2016-06-01 | 三峡大学 | Alcohol-precipitated panax japonicus polysaccharide, preparation method and application thereof |
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