CN102441012B - Golden mushroom saponifiable extract product with antitumor effect, preparation method and application thereof - Google Patents
Golden mushroom saponifiable extract product with antitumor effect, preparation method and application thereof Download PDFInfo
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Abstract
A golden mushroom saponifiable extract product with an antitumor effect. A preparation method of the saponifiable extract product provided by the invention comprises the following steps of: extracting golden mushroom sporophore by an alcoholic solution, condensing the extract, followed by water dispersion, extracting by using a low-polarity aprotic organic solvent, recovering the low-polarity aprotic solvent, carrying out saponification on an alcoholic solution with the addition of alkali, distilling the saponification liquor and recovering the alcoholic solvent, adding water for dispersion, extracting by using the low-polarity aprotic organic solvent, using hydrochloric acid to acidify the saponification liquor extracted by the low-polarity aprotic organic solvent to pH of 1-2, extracting again by using the low-polarity aprotic organic solvent, and recovering the solvent of the extract to obtain the golden mushroom saponifiable extract product with the antitumor effect. The extract product has an activity for in vitro inhibition of HepG2, SGC, U251 and A549 tumor cell growth.
Description
Technical field:
The present invention relates to JINZHENGU sponifiable extract and preparation method thereof; The invention further relates to the pharmaceutical composition of described sponifiable extract; The invention still further relates to the application of described sponifiable extract in the preparation antitumor drug.
Background technology:
Tumor is one of serious disease of harm humans health in the world today, the experts and scholars of countries in the world seek method and the medicine for the treatment of tumor in active research, especially very active to the research of antitumorigenic substance in edible fungi, about zoopery and the Preliminary Clinical Observation of this respect shows that polysaccharide and some albumen in edible fungi have stronger inhibitory action to the zoografting tumor, immunologic function that can enhancing body alleviates the toxic reaction of radiotherapy, chemotherapy.
JINZHENGU
Flammulina velutipes(Curt.:Fr.) Sing. has another name called money bacterium, Flammulina velutipes (Fr.) Sing, Flammulina velutipes, hair handle money bacterium, the genus Eumycota (
Eumycota), Basidiomycotina (
Basidiomycotina), Hymenomycetes (
Hymenomycetes), Agaricales (
Agaricales), Tricholomataceae (
Tricholomataceae), the flammule Pseudomonas (
Flammulina) or the money Pseudomonas (
Collybia), be one of famous edible fungi in China and foreign countries, be also one of natural health care that extremely sells well on current market.JINZHENGU is distributed widely in the ground such as China, Japan, Russian Siberia and little inferior West Asia and Europe, North America, Australia.In China, North gets Heilungkiang, reach Guangdong in the south, from the east of Fujian, in the vast zone to Sichuan, all have JINZHENGU distribute (
Guo Meiying chief editor, " Chinese JINZHENGU production ", Beijing: Chinese agriculture publishing house, 2000.).
JINZHENGU is not only nutritious as a kind of edible fungi, and is rich in the various active material.From last century the seventies, the state scholars such as Japan, the U.S. have begun JINZHENGU is carried out the basic research of anti-tumor aspect, have caused subsequently international research scholar's broad interest.Studies show that, its active component is mainly protein and polysaccharide composition.Protein-based comprise four kinds of ribosome inactivating proteins such as Flammulin, Velutin, Flammin and Velin, fungal immunomodulatory protein, Flammutoxin, Huogu mushroom essence, fiery mushroom proteinogen, enzyme and agglutinin etc. (
Maruyama H., Ikekawa T., International Journal of Medicinal Mushrooms, 2007,9 (2): 109-122. Kenji, F., Michika, H., Sadaki, A., et al. Biosci. Biotechnol. Biochem., 2008,72 (12): 3107-3113.).Polysaccharide comprises SFA1, extracellular polysaccharide, FVP2, PA
3DE and PA
5DE etc. (
M.Y.K. Leung, K.P. Fung, Y.M. Choy, Immunopharmacology, 1997,35:255-263. K.S. Shin, K.W. Yu, H.K. Lee1, et al, Food Technol. Biotechnol., 2007,45 (3): 319-326.).These compositions have antitumor, and antiviral is pest-resistant, antifungal, and anti-human immunodeficiency virus (HIV) reduces serum cholesterol, and blood pressure lowering improves the extensive pharmacologically actives such as immunity of organisms.Particularly antitumor action is studied at most, and S-180, ehrlich ascites tumor etc. is all had inhibitory action.Its mechanism is considered to immunoregulation effect, namely in the situation that body's immunity is impaired, can strengthen the T cell function, activated lymphocyte and phagocyte, enhancing antibody produces, and can inducing interferon-γ, the generation of tumor necrosis factor-alpha, it is by recovering and improve the immunocompetence of whole body to the inhibition growth of tumor.
In addition, Cao Peirang etc. get 5 kinds and change the platform thing from JINZHENGU submerged fermentation mycelium: soybean isoflavone, and genistein, 6-methoxyl group Dai, adenosine, adenine (
Cao Peirang, Wang Shufang, Xu Guixiang, etc. Chinese herbal medicine, l992,23 (10): 511.).Chen Zhiyi etc. adopt headspace solid-phase microextraction-GC-MSs (HS-SPME-GC-MS) to measure volatile ingredient in JINZHENGU, obtain 5 kinds of esters compositions, 4 kinds of aldehydes compositions, each a kind, alkene, alcohols, furans (
Chen Zhiyi, Liu Xueming, Shi Ying, etc. edible fungi journal, 2009,16 (1), 73-75.).Wang Jinyu etc. adopt steam distillation to extract volatile ingredient in JINZHENGU, use the GC-MS method and isolate 30 components, and identify 6 components such as hyacinthin, phthalic acid isobutyl ester, butyl phthalate, palmitic acid, soft-ethyl ester, linolenic acid (
Wang Jinyu, Chen Kang, woods is encouraged, etc. the time precious traditional Chinese medical science traditional Chinese medicines, 2008,19 (5), 1145-1146.).Lee etc. determine from JINZHENGU C10:0, C14:0, C16:0, C18:0, C18:1, six kinds of fatty acids of C18:2 content (
KJ Lee, IJ Yun, KH Kim, et al, Journal of Food Composition and Analysis, 24 (2011) 175-178.).Yet, have no the pharmacology of above-mentioned substance or the report of bioactivity research.
Kang Jie etc. are studied the chemical composition of golden mushroom mycelium 95% ethanol extraction, obtain altogether more than 20 compounds, comprise 7 steroidals, 3 aminoacid, 3 CYCLIC DIPEPTIDES compounds, 2 flavone, 2 nucleoside compounds, 2 multicomponent alcoholics compounds, 1 sesquiterpene, a ceramide.Wherein 17 compounds are for obtaining from the JINZHENGU fermentation mycelium first.Monomeric compound pharmacological screening result shows: ergosterol and guanosine have the effect of stronger acetylcholine esterase inhibition, and mannitol and ribitol have the stronger effect that improves Memory Problem of Mouse Caused by Scopolamine (
Kang Jie. structure bacterium, sea paint and illiteracy Mulberry chemical composition and bioactivity research [D], Beijing: China Concord Medical Science University, 2006.).
Chinese patent application 03132324.3 " Flammulina velutiper (Fr.) Sing rich in germanium polysaccharide and method for making thereof and purposes ", 200610036622.5 " a kind of high activity Flammukinan-peptide-Fe
2+Sequestration thing and preparation method thereof ", 200810222271.6 " a kind of gold needle mushroom extract and preparation method thereof and application " and 200910010576.5 " preparation method of recombinant gold needle mushroom immunomodulatory protein and application " related to JINZHENGU effect and purposes, but be all about protein in JINZHENGU or polysaccharose substance, do not relate to the sponifiable constituents.
Through consulting domestic and foreign literature, the research of relevant JINZHENGU sponifiable composition is less, particularly aspect antineoplastic agent use, has not yet to see the report of relevant JINZHENGU sponifiable composition antitumor action research and application facet.
Summary of the invention:
One of purpose of the present invention is that a kind of JINZHENGU antitumor sponifiable extract and preparation method thereof is provided.
Two of purpose of the present invention is that a kind of pharmaceutical composition take described sponifiable extract as effective ingredient is provided.
Three of purpose of the present invention is that the application of described sponifiable extract in preparation antitumor and related drugs is provided.
Technical scheme of the present invention is as follows:
the sponifiable extract that has antitumor action in a kind of JINZHENGU, it is that the JINZHENGU sporophore extracts through alcoholic solution, concentrated extracting solution, with extracting with low polar non-proton organic solvent after aqueous dispersion, after reclaiming low polar non-solute, add alkali alcosol and carry out saponification, saponification liquor adds aqueous dispersion after reclaiming alcoholic solvent, through low polar non-proton organic solvent extraction, saponification liquor after low polar non-proton organic solvent extraction with hcl acidifying to pH=1-2, again with low polar non-proton organic solvent extraction, extract reclaims the sponifiable extract that has antitumor action in the JINZHENGU that obtains after solvent.
A kind ofly prepare the method that above-mentioned JINZHENGU has the sponifiable extract of antitumor action, it comprises the steps:
With the alcohol reflux of 70%-95%, cooling after step 1. is pulverized the JINZHENGU sporophore, filter, concentrated extracting solution gets extractum;
Step 2. adds aqueous dispersion with the extractum of step 1, with low polar non-proton organic solvent extraction, after the extract distillating recovering solvent, adds the alcoholic solution reflux saponification of alkali, and is cooling;
Step 3. is with the saponification liquor Distillation recovery ethanol of step 2, extract with low polar non-proton organic solvent after adding water, saponification liquor after extraction adds hydrochloric acid and is acidified to pH=1-2, with low polar non-proton organic solvent extraction, extract namely gets the sponifiable extract that has antitumor action in JINZHENGU after reclaiming solvent again.
Concrete operation can be:
A kind of method for preparing the sponifiable extract that has antitumor action in above-mentioned JINZHENGU, it comprises the steps:
Step 1. is pulverized the sporophore of JINZHENGU, extracts three times with the alcoholic solution of the 70%-95% of 1～10 times of crude drug amount, and merge extractive liquid, reclaims solvent and gets extractum;
Step 2. extractum adds aqueous dispersion, and with the low polar non-proton organic solvent extraction of 1～10 times of volume, after the extract distillating recovering solvent, the alcoholic solution reflux that adds concentration and be the alkali of 1mol/L is carried out saponification;
Step 3. saponification liquor adds aqueous dispersion after reclaiming ethanol, low polar non-proton organic solvent extraction through 1～10 times of volume, saponification liquor after extraction with hcl acidifying to pH=1～2, with low polar non-proton organic solvent extraction, extract namely gets the sponifiable extract that has antitumor action in JINZHENGU after reclaiming solvent again.；
The method that has the sponifiable extract of antitumor action in above-mentioned JINZHENGU, described low polar non-proton organic solvent is ether or ethyl acetate.
Saponification of the present invention refers to the ester hydrolysis reaction under base catalysis, espespecially the hydrolysis of oils and fats.
The present invention further provides the application of sponifiable extract in preparation treatment antitumor drug in the JINZHENGU.Test as activity index carries out anti-tumor activity to suppress human liver cancer cell (HepG2), people's gastric adenocarcinoma cells (SGC), human glioma cells (U251), human lung adenocarcinoma cell (A549), human cervical carcinoma cell (HeLa) and human colon cancer cell (Lovo) multiplication capacity.Experimental result shows, when drug level is 100 μ gmL
-1Perhaps 125 μ gmL
-1Time is when extending to 72 h from 24 h, the sponifiable extract is more and more higher to the suppression ratio of tumor cell, reach maximum during 72 h, suppression ratio is respectively 68.49%(HepG2), 71.40%(SGC), 61.57%(U251), 76.87%(A549), 0.31%(HeLa) and 44.20%(Lovo).Medicine is better than HeLa and Lovo to the proliferation inhibition rate of HepG2, SGC, U251 and A549.
Beneficial effect:
1, advantage of the present invention and effect are to adopt the conventional methods such as solvent extraction, extraction, extract from JINZHENGU and obtained the new compositions useful with antitumor action-sponifiable extract, this compositions useful the experiment proved that have vitro inhibition HepG2, SGC, the activity of U251 and A549 growth of tumour cell.
2, the present invention has expanded the raw material channel of antitumor drug, has enlarged the purposes of JINZHENGU, makes JINZHENGU develop into the new raw material of antitumor drug from bread and cheese, can significantly improve the added value of JINZHENGU; Simultaneously, find that for screening from edible fungi new antitumor drug provides new approaches.
The specific embodiment:
Below listed embodiment help those skilled in the art to understand better the present invention, but do not limit the present invention in any way.
The preparation of embodiment 1 JINZHENGU sponifiable extract
Bright JINZHENGU sporophore 5000 g are pulverized, and with 95% alcohol reflux 2 hours, amount of alcohol was 3 times of JINZHENGU weight, residue after filtration repeats to extract 1 time, filter, merge secondary ethanol extract 34 L, reclaim ethanol extremely near without the alcohol flavor, continue to be concentrated into 600 ml, add isopyknic extracted with diethyl ether 4 times, merge ether extraction liquid 2400 ml, anhydrous sodium sulfate dehydration is dry, reclaim ether, approximately get ether extract 10 g.Ether extract adds 95% alcoholic solution of 1.0 mol/L potassium hydroxide, and agitating heating refluxes, and saponification liquor adds appropriate distilled water diluting after removing ethanol, uses extracted with diethyl ether non-saponifiable matter 4～5 times; The water saponification liquor that will extract after non-saponifiable matter with concentrated hydrochloric acid transfers to pH=1～2, with extracted with diethyl ether saponifiable matter 3～4 times, merges ether extracted liquid, is washed till neutrality through distilled water, after anhydrous sodium sulfate dehydration, filters, and reclaims ether and namely gets sponifiable extract 20 mg.
The preparation of embodiment 2 JINZHENGU sponifiable extracts
With the lower air blast oven dry of 50 ℃ of bright JINZHENGU sporophore, pulverize, with 70% alcohol reflux 2 hours, amount of alcohol was 10 times of JINZHENGU weight, the residue after filtration repeats to extract 1 time, the merging ethanol extract; Reclaim ethanol, the extraction 4 times that adds diethyl ether merges ether extraction liquid, and anhydrous sodium sulfate dehydration is dry, reclaims ether, namely gets ether extract.Ether extract adds 95% alcoholic solution of 1.0 mol/L potassium hydroxide, and agitating heating refluxes, and saponification liquor adds appropriate distilled water diluting after removing ethanol, uses extracted with diethyl ether non-saponifiable matter 4～5 times; The water saponification liquor that will extract after non-saponifiable matter with concentrated hydrochloric acid transfers to pH=1～2, with extracted with diethyl ether saponifiable matter 3～4 times, merges ether extracted liquid, is washed till neutrality through distilled water, after anhydrous sodium sulfate dehydration, filters, and reclaims ether and namely gets the sponifiable extract.
Embodiment 3 anti-tumor activity experiments
The JINZHENGU that obtains take embodiment 1 or embodiment 2 not the saponification thing as tested medicine, Human Hepatocarcinoma Cell line in vitro HepG2, fetching is counted the cell of trophophase, after trypsinization, 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 1.
Table 1 JINZHENGU sponifiable extract anti-tumor activity experiment (HepG2)
Embodiment 4 anti-tumor activity experiments
The JINZHENGU that obtains take embodiment 1 or embodiment 2 not the saponification thing as tested medicine, vitro culture of human gastric adenocarcinoma cells SGC, fetching is counted the cell of trophophase, after trypsinization, 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 2.
Table 2 JINZHENGU sponifiable extract anti-tumor activity experiment (SGC)
Embodiment 5 anti-tumor activity experiments
The JINZHENGU that obtains take embodiment 1 or embodiment 2 not the saponification thing as tested medicine, the vitro culture of human glioma cell of U251, fetching is counted the cell of trophophase, after trypsinization, 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 3.
Table 3 JINZHENGU sponifiable extract anti-tumor activity experiment (U251)
Embodiment 6 anti-tumor activity experiments
The JINZHENGU that obtains take embodiment 1 or embodiment 2 not the saponification thing as tested medicine, the In vitro culture human A549 cell lines, fetching is counted the cell of trophophase, after trypsinization, 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 4.
Table 4 JINZHENGU sponifiable extract anti-tumor activity experiment (A549)
Embodiment 7 anti-tumor activity experiments
The saponification thing is not as tested medicine take JINZHENGU that embodiment 1 or embodiment 2 obtain, and vitro culture of human cervical cancer cell HeLa, fetching are counted the cell of trophophase, after trypsinization, and 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 5.
Table 5 JINZHENGU sponifiable extract anti-tumor activity experiment (HeLa)
Embodiment 8 anti-tumor activity experiments
The JINZHENGU that obtains take embodiment 1 or embodiment 2 not the saponification thing as tested medicine, In vitro culture human colon cancer cell Lovo, fetching is counted the cell of trophophase, after trypsinization, 1500 rmin
-1Centrifugal 5 min are resuspended in cell in culture medium, and it is 2 * 10 that cell concentration transfers to
4Individual/mL is inoculated in 96 porocyte culture plates, every hole 100 μ L, the placement cell culture incubator (37 ℃, 5%CO2) middle 24 h that cultivate.Add afterwards tested medicine 20 μ L, it is 0.5%DMSO that negative control group adds final concentration, and each group is all established 3 multiple holes.After dosing 24 h, 48h, 72 h, every hole adds 5 mgmL respectively
-1MTT reagent 20 μ L continue to be placed in 37 ℃ and cultivate 4 h.Every hole adds DMSO 100 μ L, and the vibration mixing is measured each hole at the absorbance at wavelength 570 nm places.Calculate cell proliferation inhibition rate: suppression ratio (%)=(1-experimental group absorbance/matched group absorbance) * 100%.It the results are shown in Table 6.
Table 6 JINZHENGU sponifiable extract anti-tumor activity experiment (Lovo)
Claims (5)
1. has the sponifiable extract of antitumor action in a JINZHENGU, it is characterized in that: it is that the JINZHENGU sporophore extracts through alcoholic solution, concentrated extracting solution, with extracting with low polar non-proton organic solvent after aqueous dispersion, after reclaiming low polar non-solute, add alkali alcosol and carry out saponification, saponification liquor adds aqueous dispersion after reclaiming alcoholic solvent, through low polar non-proton organic solvent extraction, saponification liquor after low polar non-proton organic solvent extraction with hcl acidifying to pH=1-2, again with low polar non-proton organic solvent extraction, extract reclaims the sponifiable extract that has antitumor action in the JINZHENGU that obtains after solvent.
2. one kind prepares the method that JINZHENGU claimed in claim 1 has the sponifiable extract of antitumor action, it is characterized in that it comprises the steps:
With the alcohol reflux of 70%-95%, cooling after step 1. is pulverized the JINZHENGU sporophore, filter, concentrated extracting solution gets extractum;
Step 2. adds aqueous dispersion with the extractum of step 1, with low polar non-proton organic solvent extraction, after the extract distillating recovering solvent, adds the alcoholic solution reflux saponification of alkali, and is cooling;
Step 3. is with the saponification liquor Distillation recovery ethanol of step 2, extract with low polar non-proton organic solvent after adding water, saponification liquor after extraction adds hydrochloric acid and is acidified to pH=1-2, with low polar non-proton organic solvent extraction, extract namely gets the sponifiable extract that has antitumor action in JINZHENGU after reclaiming solvent again.
3. preparation JINZHENGU according to claim 2 has the method for the sponifiable extract of antitumor action, it is characterized in that: described low polar non-proton organic solvent is ether or ethyl acetate.
4. the application of sponifiable extract in the preparation antitumor drug that has antitumor action in JINZHENGU claimed in claim 1.
5. have the application of sponifiable extract in the preparation antitumor drug of antitumor action in JINZHENGU according to claim 4, it is characterized in that: described antitumor drug is: suppress human liver cancer cell HepG2 medicine, suppress people's gastric adenocarcinoma cells SGC medicine, suppress human glioma cells U251 medicine, suppress the human A549 cell lines medicine, suppress human cervical carcinoma cell HeLa medicine or suppress human colon cancer cell Lovo medicine.
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| CN1887912A (en) * | 2006-07-21 | 2007-01-03 | 华南理工大学 | High activity golden mushroom polysaccharide-peptide-Fe chelate and its prepn |
| CN101143002A (en) * | 2007-09-03 | 2008-03-19 | 江苏省江大绿康生物工程技术研究有限公司 | Deep liquid fermentation method for preparing selenium-rich gold needle mushroom crude polysaccharide powder |
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| CN1887912A (en) * | 2006-07-21 | 2007-01-03 | 华南理工大学 | High activity golden mushroom polysaccharide-peptide-Fe chelate and its prepn |
| CN101143002A (en) * | 2007-09-03 | 2008-03-19 | 江苏省江大绿康生物工程技术研究有限公司 | Deep liquid fermentation method for preparing selenium-rich gold needle mushroom crude polysaccharide powder |
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