CN107602721A - The method that acidic buffer solution extracts tremella polysaccharides - Google Patents
The method that acidic buffer solution extracts tremella polysaccharides Download PDFInfo
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- CN107602721A CN107602721A CN201711021626.0A CN201711021626A CN107602721A CN 107602721 A CN107602721 A CN 107602721A CN 201711021626 A CN201711021626 A CN 201711021626A CN 107602721 A CN107602721 A CN 107602721A
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- tremella polysaccharides
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- 150000004676 glycans Chemical class 0.000 title claims abstract description 138
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 138
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 138
- 241001506047 Tremella Species 0.000 title claims abstract description 96
- 238000000034 method Methods 0.000 title claims abstract description 57
- 239000007853 buffer solution Substances 0.000 title claims abstract description 46
- 230000002378 acidificating effect Effects 0.000 title claims abstract description 29
- 239000000284 extract Substances 0.000 title description 33
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 71
- 239000007788 liquid Substances 0.000 claims abstract description 70
- 238000001556 precipitation Methods 0.000 claims abstract description 64
- 238000000605 extraction Methods 0.000 claims abstract description 63
- 241000233866 Fungi Species 0.000 claims abstract description 45
- 239000000843 powder Substances 0.000 claims abstract description 26
- 239000007787 solid Substances 0.000 claims abstract description 13
- 238000000926 separation method Methods 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 66
- 239000000243 solution Substances 0.000 claims description 61
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical group [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 25
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 24
- 238000005119 centrifugation Methods 0.000 claims description 14
- 239000000872 buffer Substances 0.000 claims description 7
- -1 disodium hydrogen Chemical class 0.000 claims description 6
- 229910052739 hydrogen Inorganic materials 0.000 claims description 6
- 239000001257 hydrogen Substances 0.000 claims description 6
- 230000003139 buffering effect Effects 0.000 claims description 4
- 239000008236 heating water Substances 0.000 claims description 4
- 238000002386 leaching Methods 0.000 claims description 4
- 239000001509 sodium citrate Substances 0.000 claims description 4
- 235000005979 Citrus limon Nutrition 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 3
- 235000019800 disodium phosphate Nutrition 0.000 claims description 3
- 239000001488 sodium phosphate Substances 0.000 claims description 3
- 244000248349 Citrus limon Species 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000011084 recovery Methods 0.000 abstract description 22
- 238000003809 water extraction Methods 0.000 abstract description 5
- 239000006228 supernatant Substances 0.000 description 28
- 239000002904 solvent Substances 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 14
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 14
- 239000002244 precipitate Substances 0.000 description 10
- 230000003111 delayed effect Effects 0.000 description 9
- 238000000638 solvent extraction Methods 0.000 description 8
- 239000003795 chemical substances by application Substances 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- CBMPTFJVXNIWHP-UHFFFAOYSA-L disodium;hydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].OP([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O CBMPTFJVXNIWHP-UHFFFAOYSA-L 0.000 description 6
- 208000035199 Tetraploidy Diseases 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 description 4
- 239000008280 blood Substances 0.000 description 3
- 238000005265 energy consumption Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000007981 phosphate-citrate buffer Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002893 slag Substances 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000007974 sodium acetate buffer Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- TWNIBLMWSKIRAT-RWOPYEJCSA-N (1r,2s,3s,4s,5r)-6,8-dioxabicyclo[3.2.1]octane-2,3,4-triol Chemical compound O1[C@@]2([H])OC[C@]1([H])[C@@H](O)[C@H](O)[C@@H]2O TWNIBLMWSKIRAT-RWOPYEJCSA-N 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 206010053567 Coagulopathies Diseases 0.000 description 1
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
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- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
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- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
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- Polysaccharides And Polysaccharide Derivatives (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a kind of method of acidic buffer solution extraction tremella polysaccharides, comprise the following steps:In a heated condition, white fungus powder is extracted using acidic buffer solution, obtains extracting feed liquid;The extraction feed liquid is subjected to separation of solid and liquid, liquid component is obtained, contains tremella polysaccharides in the liquid component;The liquid component is subjected to alcohol precipitation, what is obtained is precipitated as tremella polysaccharides.The recovery rate of the method for the invention tremella polysaccharides can reach 31.57%, be 3 ~ 4 times of traditional water extraction, greatly improve the recovery rate of tremella polysaccharides.
Description
Technical field
The invention belongs to plant polyose extractive technique field, and in particular to a kind of acidic buffer solution extraction tremella polysaccharides
Method.
Background technology
White fungus, also known as tremella, it is a kind of higher fungus, the function with nourishing Yin and moistening lung, QI invigorating and blood.White fungus is more
Sugar is the principle active component of white fungus, is a kind of acid heteroglycan, from white fungus (Tremella fuciformisBerk) son is real
Obtained in body, its backbone structure is that side chain is by glucuronic acid and xylose by the mannosan of α-(1 → 3) glucosides key connection
Composition.Tremella polysaccharides has regulation immunologic function, and anti-blood is fastened, and extends the clotting time, and hypoglycemic, reducing blood lipid, auxiliary suppress swollen
Knurl, anti-aging and radioresistance etc. act on.Japanese scholars etc. extracted with Hot water extraction first from Tremella fructification obtain it is more
Sugar substance, hereafter, the problem of how domestic and foreign scholars are with regard to improve polysaccharide material yield, is constantly studied and is inquired into.
How further to improve the recovery rate of tremella polysaccharides is the key for realizing tremella polysaccharides industrialized production.At present, may be used
Using hot water extraction method, ultrasound assisted extraction method, microwave―assisted extraction, to extrude assisted extraction method, enzyme process extraction, alkali leaching
The methods of formulation, extracts tremella polysaccharides.
The cell membrane of Tremella fructification cell has the compact texture that double-deck protein and polysaccharide are combined so that makes merely
The method extracted with hot water is difficult to destroy cell wall structure, fully discharges fruitbody polysaccharide composition.If add ultrasonic wave hair
The supplementary instrument extraction tremella polysaccharides such as raw instrument, microwave reaction system or pressure cooker, the expense of additional instruments input is added, and
And high energy consumption.Although enzyme extraction method more gently but can face the increase of extract solution viscosity, filtration difficulty, protein in extraction process
Increase Deng impurity, enzymatic reagent cost the problems such as.
The content of the invention
In view of this, high it is an object of the invention to provide a kind of recovery rate, energy consumption is low, and the low tremella polysaccharides of cost carries
Take method.
In order to realize foregoing invention purpose, the present invention provides following technical scheme:
The present invention provides a kind of method of acidic buffer solution extraction tremella polysaccharides, comprises the following steps:
1)In a heated condition, white fungus powder is extracted using acidic buffer solution, obtains extracting feed liquid;
2)Extraction feed liquid is subjected to separation of solid and liquid, liquid component is obtained, contains tremella polysaccharides in liquid component;
3)Liquid component is subjected to alcohol precipitation, centrifuged, what is obtained is precipitated as tremella polysaccharides.
Step 1)Described in acidic buffer solution be citric acid-sodium citrate buffer, NaAc_HAc buffer solution
Or disodium hydrogen phosphate-citric acid solution.The concentration of the citric acid-sodium citrate buffer is 0.1 mol/L, pH models
Enclose for pH 3.0 ~ 6.6, be made up of citric acid and sodium citrate;The concentration of the NaAc_HAc buffer solution is 0.2mol/
L, pH scope are pH 3.6 ~ 5.8, are made up of acetic acid and sodium acetate;Disodium hydrogen phosphate-the citric acid solution be then by
0.1mol/L citric acid solution and 0.2mol/L disodium phosphate soln are formulated, and pH scopes are pH 2.2 ~ 6.0.
Step 1)The material-water ratio of middle extraction is 1:30 ~ 1:70 g/mL.
Step 1)In mode of heating be heating water bath, bath temperature is 80 ~ 100 DEG C, and water bath time is 2 ~ 6 h.
Step 2)The mode of middle separation of solid and liquid is centrifugation, and the rotating speed of centrifugation be 7000 ~ 9000 rpm, centrifugation time for 5 ~
10min。
Step 3)Middle alcohol precipitation uses ethanol solution, and the volumetric concentration of ethanol solution is 85 ~ 99%;Liquid group during alcohol precipitation
It is 1 to divide with the volume ratio of ethanol solution:3~5.
Step 3)The time of middle alcohol precipitation is 8 ~ 14h.
Step 3)Rotating speed during middle centrifugation is 8000 ~ 11000 rpm, and centrifugation time is 5 ~ 10 min.
Beneficial effects of the present invention:Tremella polysaccharides extracting method of the present invention, it is molten to extract using acidic buffer solution
Agent, traditional water extraction is replaced, alcohol precipitation is carried out after extraction and obtains tremella polysaccharides.It is molten in acidic buffer solution using tremella polysaccharides
Xie Du is more than the principle of solubility in the hot water.Method provided by the invention can reach to the recovery rate of tremella polysaccharides
31.57%, the recovery rate of the method for the invention is 3 ~ 4 times of traditional water extraction, greatly increases the extraction of tremella polysaccharides
Rate.In addition, the present invention does not need other extra supplementary instruments or reagent, such as ultrasonic wave or subcritical state, energy consumption is low, into
This is relatively low.
Embodiment
The present invention provides a kind of method of acidic buffer solution extraction tremella polysaccharides, comprises the following steps:1)In water-bath bar
Under part, white fungus powder is extracted using acidic buffer solution, obtains extracting feed liquid;The acidic buffer solution includes lemon
Acid-sodium citrate buffer, NaAc_HAc buffer solution and disodium hydrogen phosphate-citric acid solution;The water-bath
Temperature is 80 ~ 100 DEG C, and time of water-bath is 2 ~ 6h, material-water ratio 1:30 ~ 1:70 g/mL;2)Extraction feed liquid is subjected to solid-liquid
Separation, obtains liquid component, the liquid component includes tremella polysaccharides;3)Liquid component is subjected to alcohol precipitation, centrifuges, obtains
It is precipitated as tremella polysaccharides.
In the present invention, the source of the white fungus powder is not limited, is crushed after being dried using commercially available white fungus, sieves and obtain
.Described dries using the conventional solarization drying method in this area, can specifically be dried naturally using daylight.The crushing is excellent
Choosing is carried out using crusher for Chinese herbal medicine, and the mesh number of the sieving is preferably 60 ~ 100 mesh, more preferably 80 mesh;The silver
The granularity of ear powder is preferably 150 ~ 250 μm, more preferably 200 ~ 220 μm.
The present invention mixes white fungus powder with acidic buffer solution acquisition extraction feed liquid after white fungus powder is obtained.In the present invention
The acidic buffer solution is citric acid-sodium citrate buffer, NaAc_HAc buffer solution or disodium hydrogen phosphate-lemon
Lemon acid buffering solution;The concentration of the citric acid-sodium citrate buffer is 0.1 mol/L, and pH scopes are pH 3.0 ~ 6.6,
It is made up of citric acid and sodium citrate;The concentration of the NaAc_HAc buffer solution is 0.2mol/L, and pH scopes are pH 3.6
~ 5.8, it is made up of acetic acid and sodium acetate;Disodium hydrogen phosphate-the citric acid solution is then molten by 0.1mol/L citric acid
Liquid and 0.2mol/L disodium phosphate soln are formulated, and pH scopes are pH 2.2 ~ 6.0.
The quality of heretofore described white fungus powder and the volume ratio of acidic buffer solution are preferably 1:30~70 g/mL;It is more excellent
Elect 1 as:35~45 g/mL;Most preferably 1:40 g/mL.
The present invention carries out heating water bath to extraction feed liquid.The temperature of the water-bath be 80 ~ 100 DEG C, preferably 90 ~ 100
DEG C, more preferably 100 DEG C;The water bath time is 2 ~ 6 h, preferably 4 ~ 5 h;Most preferably 5 h.The water-bath
Using the conventional water bath equipment in this area, without other particular determinations.Specifically water-bath can be used in embodiments of the present invention
Pot.During the heating water bath, acidic buffer solution extracts to the tremella polysaccharides in white fungus powder, obtains extraction material
Liquid, the tremella polysaccharides that the extraction feed liquid includes white fungus powder solid residue and is dissolved in system.
The extraction feed liquid is carried out separation of solid and liquid by the present invention, collects liquid component.In the present invention, the separation of solid and liquid
Mode preferably centrifuges;The rotating speed of the centrifugation is preferably 7000 ~ 9000 rpm, more preferably 8000 rpm;It is described
Centrifugation time is preferably 5 ~ 10 min, more preferably 8 min.The effect of separation of solid and liquid of the present invention is by white fungus powder
Solid residue insoluble in acidic buffer solution removes, in favor of the alcohol precipitation of follow-up tremella polysaccharides.
The present invention carries out alcohol precipitation after liquid component is obtained, by the liquid component, obtains alcohol precipitation feed liquid, the alcohol jetsam
Liquid includes tremella polysaccharides and the alcohol precipitation liquid phase separated out, and what is obtained is precipitated as tremella polysaccharides.In the present invention, the alcohol precipitation examination
Agent is preferably ethanol solution;The volumetric concentration of the ethanol solution is preferably 85 ~ 99%, more preferably 95%;The liquid
Component and the volume ratio of ethanol solution are preferably 1:3 ~ 5, more preferably 1:4.
In an embodiment of the present invention, the alcohol precipitation be specially the liquid component is mixed with ethanol solution, stand into
Row precipitation.The present invention is not particularly limited to the mixed method, as long as mixing can be realized.Heretofore described alcohol precipitation
Time be preferably 8 ~ 14 h, more preferably 11 ~ 13 h, most preferably 12 h;Heretofore described precipitation is to pass through
The hydrogen bond that ethanol is destroyed in liquid component, solubility of the tremella polysaccharides in liquid component is reduced, so that polysaccharide precipitation comes out.
The present invention carries out the second separation of solid and liquid after alcohol precipitation feed liquid is obtained, to the alcohol precipitation feed liquid, obtains tremella polysaccharides.
In the present invention, second solid-liquid separation method preferably centrifuges;The rotating speed of the centrifugation is preferably 8000 ~ 11000
Rpm, more preferably 9500 ~ 10500 rpm, most preferably 10000rpm;The centrifugation time is preferably 5 ~ 10 min, more
Preferably 8min.The present invention collects solid precipitation, the tremella polysaccharides as extracted after the second separation of solid and liquid.
The calculation formula of heretofore described tremella polysaccharides recovery rate is:
。
In the present invention, the tremella polysaccharides aqueous solution is obtained after the tremella polysaccharides that said extracted obtains being dissolved in into water.Use phenol
Polyoses content in the sulfuric acid process measure tremella polysaccharides aqueous solution, calculate the recovery rate of tremella polysaccharides.
The extracting method of tremella polysaccharides provided by the invention is described in detail with reference to embodiment, but can not
They are interpreted as limiting the scope of the present invention.
Embodiment 1
Commercially available white fungus is dried, crushed 90 mesh sieves.The citric acid-sodium citrate that 40ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 100 DEG C of hot water and extracts, after 5h, feed liquid will be extracted 8min is centrifuged with 8000rpm rotating speed, gone
Except residue, the volumetric concentration that 4 times of volumes are added in supernatant is 90% ethanol solution, 12h is precipitated, with 10000rpm rotating speed
8min is centrifuged, removes supernatant, obtains tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
15.79mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 315.8mg, polysaccharide extract rate 31.58%, are compareed using water as leaching
The recovery rate of extraction solvent extraction tremella polysaccharides is 7.55%, improves 318.3%.
Embodiment 2
Commercially available white fungus is dried, crushed 80 mesh sieves.The citric acid-sodium citrate that 40ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 100 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 5min is centrifuged with 7500rpm rotating speed, gone
Except residue, the volumetric concentration that 4.5 times of volumes are added in supernatant is 95% ethanol solution, precipitates 11h, with turning for 11000rpm
Speed centrifugation 6min, removes supernatant, obtains tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
14.35mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 287.0mg, polysaccharide extract rate 28.70%, are compareed using water as leaching
The recovery rate of extraction solvent extraction tremella polysaccharides is 7.35%, improves 290.5%.
Embodiment 3
Commercially available white fungus is dried, crushed 80 mesh sieves.The citric acid-sodium citrate that 60ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 80 DEG C of hot water and extracts, after 5h, feed liquid will be extracted 7min is centrifuged with 9000rpm rotating speed, removed
Residue, the volumetric concentration that tetraploid product is added in supernatant is 90% ethanol solution, precipitates 10h, with 10000rpm rotating speed from
Heart 10min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
5.10mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 102.0mg, polysaccharide extract rate 10.20%, are compareed using water as extraction
The recovery rate of solvent extraction tremella polysaccharides is 7.25%, improves 40.69%.
Embodiment 4
Commercially available white fungus is dried, crushed 70 mesh sieves.The citric acid-sodium citrate that 30ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 7min is centrifuged with 7000rpm rotating speed, removed
Residue, the volumetric concentration that tetraploid product is added in supernatant is 90% ethanol solution, precipitates 9h, with 11000rpm rotating speed from
Heart 5min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
8.16mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 162.2mg, and polysaccharide extract rate 16.32%, control water is that extraction is molten
The recovery rate of agent extraction tremella polysaccharides is 6.22%, improves 162.4%.
Embodiment 5
Commercially available white fungus is dried, crushed 60 mesh sieves.The citric acid-sodium citrate that 60ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 100 DEG C of hot water and extracts, after 3h, feed liquid will be extracted 6min is centrifuged with 8000rpm rotating speed, gone
Except residue, the volumetric concentration that three times volume is added in supernatant is 99% ethanol solution, 10h is precipitated, with 9000rpm rotating speed
9min is centrifuged, removes supernatant, obtains tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
13.34mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 266.8mg, and polysaccharide extract rate 26.68%, control water is extraction
The recovery rate of solvent extraction tremella polysaccharides is 5.62%, improves 374.7%.
Embodiment 6
Commercially available white fungus is dried, crushed 70 mesh sieves.The citric acid-sodium citrate that 50ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 90 DEG C of hot water and extracts, after 6h, feed liquid will be extracted 8min is centrifuged with 9000rpm rotating speed, removed
Residue, the volumetric concentration that tetraploid product is added in supernatant is 95% ethanol solution, precipitates 8h, with 10000rpm rotating speed from
Heart 10min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
12.88mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 257.6mg, and polysaccharide extract rate 25.76%, control water is extraction
The recovery rate of solvent extraction tremella polysaccharides is 7.95%, improves 224.0%.
Embodiment 7
Commercially available white fungus is dried, crushed 80 mesh sieves.The citric acid-sodium citrate that 50ml pH 3.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 90 DEG C of hot water and extracts, after 2h, feed liquid will be extracted 5min is centrifuged with 7000rpm rotating speed, removed
Residue, the volumetric concentration that 3 times of volumes are added in supernatant is 99% ethanol solution, precipitates 12h, with 11000rpm rotating speed from
Heart 7min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
4.72mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation is 94.4mg, and polysaccharide extract rate 9.44%, control water is extraction solvent
The recovery rate for extracting tremella polysaccharides is 3.67%, improves 157.2%.
Embodiment 8
Commercially available white fungus is dried, crushed 100 mesh sieves.70ml pH 3.0 citric acid-sodium citrate is added to 1g white fungus powder
After cushioning liquid, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 7min is centrifuged with 8000rpm rotating speed, gone
Except residue, the volumetric concentration that 4 times of volumes are added in supernatant is 99% ethanol solution, precipitates 8h, with 11000rpm rotating speed from
Heart 7min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
12.19mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 243.8mg, and polysaccharide extract rate 24.38%, control water is extraction
The recovery rate of solvent extraction tremella polysaccharides is 7.32%, improves 233.1%.
Embodiment 9
Commercially available white fungus is dried, crushed 80 mesh sieves.50ml pH 2.2 disodium hydrogen phosphate-citric acid is added to 1g white fungus powder
After cushioning liquid, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 8min is centrifuged with 9000rpm rotating speed, gone
Except residue, the volumetric concentration that 5 times of volumes are added in supernatant is 90% ethanol solution, 14h is precipitated, with 11000rpm rotating speed
5min is centrifuged, removes supernatant, obtains tremella polysaccharides precipitation.
Check experiment be with water replace disodium hydrogen phosphate-citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
12.45mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 249.0mg, and polysaccharide extract rate 24.90%, control water is extraction
The recovery rate of solvent extraction tremella polysaccharides is 7.31%, improves 240.6%.
Embodiment 10
Commercially available white fungus is dried, crushed 90 mesh sieves.The Acetic acid-sodium acetate buffering that 50ml pH 3.6 is added to 1g white fungus powder is molten
After liquid, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 10min is centrifuged with 7000rpm rotating speed, removed residual
Slag, the volumetric concentration that 4 times of volumes are added in supernatant is 95% ethanol solution, precipitates 10h, is centrifuged with 10000rpm rotating speed
10min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace Acetic acid-sodium acetate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
6.13mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 122.6mg, and polysaccharide extract rate 12.26%, control water is that extraction is molten
The recovery rate of agent extraction tremella polysaccharides is 7.31%, improves 67.72%.
Embodiment 11
Commercially available white fungus is dried, crushed 80 mesh sieves.The Acetic acid-sodium acetate buffering that 50ml pH 5.8 is added to 1g white fungus powder is molten
After liquid, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 8min is centrifuged with 8000rpm rotating speed, removed residual
Slag, the volumetric concentration that 3 times of volumes are added in supernatant is 95% ethanol solution, precipitates 12h, is centrifuged with 10000rpm rotating speed
8min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace Acetic acid-sodium acetate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
9.20mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 184.0mg, and polysaccharide extract rate 18.40%, control water is that extraction is molten
The recovery rate of agent extraction tremella polysaccharides is 7.31%, improves 151.7%.
Embodiment 12
Commercially available white fungus is dried, crushed 70 mesh sieves.The citric acid-sodium citrate that 50ml pH 5.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 5min is centrifuged with 8000rpm rotating speed, removed
Residue, the volumetric concentration that tetraploid product is added in supernatant is 95% ethanol solution, precipitates 12h, with 10000rpm rotating speed from
Heart 5min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
6.98mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 139.6mg, and polysaccharide extract rate 13.96%, control water is that extraction is molten
The recovery rate of agent extraction tremella polysaccharides is 7.31%, improves 90.97%.
Embodiment 13
Commercially available white fungus is dried, crushed 90 mesh sieves.The citric acid-sodium citrate that 50ml pH 6.0 is added to 1g white fungus powder is delayed
After rushing solution, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 6min is centrifuged with 8500rpm rotating speed, removed
Residue, the volumetric concentration that three times volume is added in supernatant is 99% ethanol solution, precipitates 10h, with 11000rpm rotating speed from
Heart 8min, supernatant is removed, obtain tremella polysaccharides precipitation.
Check experiment be with water replace citric acid-sodium citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
8.50mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 170.0mg, and polysaccharide extract rate 17.0%, control water is that extraction is molten
The recovery rate of agent extraction tremella polysaccharides is 7.31%, improves 132.6%.
Embodiment 14
Commercially available white fungus is dried, crushed 90 mesh sieves.50ml pH 3.0 disodium hydrogen phosphate-citric acid is added to 1g white fungus powder
After cushioning liquid, it is placed in water-bath in 90 DEG C of hot water and extracts, after 4h, feed liquid will be extracted 6min is centrifuged with 8500rpm rotating speed, gone
Except residue, the volumetric concentration that three times volume is added in supernatant is 99% ethanol solution, 10h is precipitated, with 11000rpm rotating speed
8min is centrifuged, removes supernatant, obtains tremella polysaccharides precipitation.
Check experiment be with water replace disodium hydrogen phosphate-citrate buffer solution be extraction solvent, method is same as above.
The tremella polysaccharides precipitation of acquisition is all dissolved in 20ml water, determining polyoses content in solution with Phenol sulfuric acid procedure is
12.27mg/ml, the quality for calculating polysaccharide in polysaccharide precipitation are 245.3 mg, and polysaccharide extract rate 24.53%, control water is extraction
The recovery rate of solvent extraction tremella polysaccharides is 7.31%, improves 235.6%.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (9)
- A kind of 1. method of acidic buffer solution extraction tremella polysaccharides, it is characterised in that comprise the following steps:1)In a heated condition, white fungus powder is extracted using acidic buffer solution, obtains extracting feed liquid;2)Extraction feed liquid is subjected to separation of solid and liquid, liquid component is obtained, contains tremella polysaccharides in liquid component;3)Liquid component is subjected to alcohol precipitation, centrifuged, what is obtained is precipitated as tremella polysaccharides.
- 2. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 1)Middle institute It is citric acid-sodium citrate buffer, NaAc_HAc buffer solution or disodium hydrogen phosphate-lemon to state acidic buffer solution Acid buffering solution.
- 3. the method for acidic buffer solution extraction tremella polysaccharides according to claim 2, it is characterised in that the lemon The concentration of acid-sodium citrate buffer is 0.1 mol/L, pH 3.0 ~ 6.6;The concentration of the NaAc_HAc buffer solution It is 0.2mol/L, pH 3.6 ~ 5.8;Disodium hydrogen phosphate-the citric acid solution is then the citric acid solution by 0.1mol/L It is formulated with 0.2mol/L disodium phosphate soln, pH 2.2 ~ 6.0.
- 4. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 1)Middle leaching The material-water ratio carried is 1:30 ~ 1:70 g/mL.
- 5. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 1)In Mode of heating is heating water bath, and bath temperature is 80 ~ 100 DEG C, and water bath time is 2 ~ 6 h.
- 6. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 2)In it is solid The mode of liquid separation is centrifugation, and the rotating speed of centrifugation is 7000 ~ 9000 rpm, and centrifugation time is 5 ~ 10min.
- 7. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 3)Middle alcohol Heavy to use ethanol solution, the volumetric concentration of ethanol solution is 85 ~ 99%;The volume of liquid component and ethanol solution during alcohol precipitation Than for 1:3~1:5.
- 8. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 3)Middle alcohol The heavy time is 8 ~ 14 h.
- 9. the method for acidic buffer solution extraction tremella polysaccharides according to claim 1, it is characterised in that step 3)In from The rotating speed during heart is 8000 ~ 11000 rpm, and centrifugation time is 5 ~ 10 min.
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