CN106397625B - A method of extracting Radix Panacis Quinquefolii polysaccharide from American Ginseng fibrous root - Google Patents
A method of extracting Radix Panacis Quinquefolii polysaccharide from American Ginseng fibrous root Download PDFInfo
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Abstract
The method that the present invention relates to a kind of to extract Radix Panacis Quinquefolii polysaccharide from American Ginseng fibrous root, extracts using American Ginseng fibrous root as raw material, and step is to be followed successively by crushing；Alkali leaching；Complex enzyme zymohydrolysis；Ultrasonic extraction method；Concentration；Alcohol precipitation；Vacuum drying.The present invention breaches traditional water and mentions or the method for alcohol extracting Radix Panacis Quinquefolii polysaccharide, using a certain amount of concentration aqueous slkali promote Radix Panacis Quinquefolii polysaccharide particle inside and between physical-chemical reaction carry out alkali extraction take, and cellulose-binding enzyme acts on the broken wall of plant cell, and protease specificity degradation, using complex enzyme hydrolysis and ultrasonic technique assisted extraction, simple process, it is easy to operate, the recovery rate and yield of Radix Panacis Quinquefolii polysaccharide are significantly improved, the industrialized production extracted for Radix Panacis Quinquefolii polysaccharide provides some theoretical reference foundations.
Description
Technical field
The present invention relates to a kind of extracting method more particularly to a kind of sides that Radix Panacis Quinquefolii polysaccharide is extracted from American Ginseng fibrous root
Method.
Background technique
American Ginseng (Panax quinquefolius L.) is the dry root of araliaceae ginseng plant, and perennial herb is planted
Object, also known as Western ginseng, American ginseng, American ginseng, panacis quinquefolii radix etc., originate in the ground such as Canada, the U.S., France.80 years 20th century
Dai Qi, China's large area are introduced a fine variety, and plantation is concentrated mainly on the ground such as three provinces in the northeast of China, Shandong, Beijing at present, and China has become generation at present
The third big country of American Ginseng, and maximum American Ginseng country of consumption in the world are produced in boundary.American Ginseng has similar with ginseng
Function, but it is different from ginseng because cool in nature, there is the effect of boosting qi and nourishing yin, clearing heat and promoting fluid, cure mainly fever due to yin deficiency, dry tongue
The dry, diseases such as quench one's thirst.Radix Panacis Quinquefolii polysaccharide (PPQ) is that one kind that content is most in American Ginseng has the substance of special bioactivity, mesh
Before the ingredient separated have sucrose, panose, maltose, glucose, fructose, sorbose, galacturonic acid, galactolipin,
Arabinose, xylose, rhamnose etc..It is related studies have shown that Radix Panacis Quinquefolii polysaccharide can not only enhance gulping down for body reticuloendothelial system
Bite function, moreover it is possible to increase the nospecific immunity and cellular immune function of body.With immunological regulation, antitumor, hypoglycemic, anti-
The effects of radiation, prevention influenza and infection of the upper respiratory tract.
It is mentioned currently, being mainly that Hot water extraction, water extraction and alcohol precipitation method etc. are traditional for Radix Panacis Quinquefolii polysaccharide Study on extraction
Method is taken, through looking into, " Radix Panacis Quinquefolii polysaccharide peptide prepares producer for the Chinese patent of existing Patent No. CN201110157090.1
Method ", comprising the following steps: with American Ginseng stem, leaf, fibrous root it is raw material, is extracted with water, solid-liquid ratio 1:10, at 60 DEG C, extraction time
2h filters merging filtrate, vacuum concentration filter is dense at 10:1, and 75% alcohol precipitation, centrifuging and taking supernatant recovered alcohol simultaneously mentions twice
Saponin(e is taken, then elutes sediment, recovered alcohol with 95% edible alcohol；It is dissolved in water to sediment by 1:10,1% flavor is added
Protease hydrolyzes 4h, enzyme deactivation at 55 DEG C, PH 5.6, and filtering vacuum concentration is spray-dried, linen Radix Panacis Quinquefolii polysaccharide is made
Peptide, the method use traditional water extract-alcohol precipitation extracting method and single enzymatic hydrolysis auxiliary process, that there are effective component extraction rates is low,
The disadvantages of yield is low, cumbersome, time-consuming.Therefore, the preparation process for optimizing Radix Panacis Quinquefolii polysaccharide has important research and opens
Send out application value.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of simple process, easy to operate and recovery rate are high from the West
Join the method that Radix Panacis Quinquefolii polysaccharide is extracted in fibrous root.
The technical scheme of the invention to solve the technical problem is: a kind of extract American Ginseng from American Ginseng fibrous root
The method of polysaccharide, it is characterised in that the following steps are included:
1) it crushes: after dry American Ginseng fibrous root is crushed with pulverizer, it is spare to cross 35~45 meshes；
2) alkali soaks: 85~95 DEG C of 2~4% sodium hydroxides of mass concentration are added in the American Ginseng fibrous root powder after above-mentioned sieving
It is impregnated in solution, filters to obtain filter residue, filter residue volatilizes；
3) complex enzyme zymohydrolysis: it is 0.9~1.1% complex enzyme that enzyme bottom mass ratio, which is added, in the resulting filter residue of step 2), in perseverance
It is digested 50~70 minutes on warm oscillator；
4) ultrasonic extraction method: distilled water is added in the filter residue after step 3) enzymatic hydrolysis, ultrasonic extraction 2~4 times, is collected by centrifugation
Filtrate；
5) it is concentrated: above-mentioned resulting filtrate decompression is concentrated；
6) alcohol precipitation: dehydrated alcohol will be added after the filtrate cooling after concentration, stands overnight, precipitating is collected after centrifugation；
7) it is dried in vacuo: resulting precipitating being dried in vacuo, target product Radix Panacis Quinquefolii polysaccharide is obtained.
As an improvement, the solid-liquid ratio of American Ginseng fibrous root powder and sodium hydroxide solution in the step 2 be 1g:20ml~
1g:25ml, alkali leaching time are 2~3 hours.
Preferably, the temperature of the sodium hydroxide solution be 90 DEG C, mass percent 3%, American Ginseng fibrous root powder with
The solid-liquid ratio of sodium hydroxide solution is 1g:25ml, and alkali soaks the time 3 hours.
Preferably, the complex enzyme of the step 3) is cellulase, pectase, dextranase, fruit juice enzyme, Papain
Two kinds in enzyme, bromelain or subtilopeptidase A, the mass ratio of two kinds of enzymes is 1:1, complex enzyme zymohydrolysis material in complex enzyme
Liquor ratio is 1g:8ml~1g:12ml, and the temperature of constant temperature oscillator is 45~50 DEG C.
Further preferably, the complex enzyme zymohydrolysis solid-liquid ratio is 1g:10ml, enzymolysis time 1 hour, constant temperature oscillator temperature be
50℃。
Preferably, the filter residue of the step 4) and the solid-liquid ratio of distilled water are 1g:25ml~1g:30ml, ultrasonic extraction
Power is 45~60W, and ultrasonic extraction 3 times, each time is 15~30min.
Further preferably, the solid-liquid ratio of the filter residue and distilled water is 1g:25ml, and ultrasonic extraction power is 60W, when extracting every time
Between be 20min.
It being further improved, the temperature of the reduced pressure of the step 5) is 50~60 DEG C, and pressure is 0.07~0.09MPa,
The relative density of concentration is 1.25~1.30.
It is further improved, the final volume percentage of the ethyl alcohol of the step 6) reaches 70~80%, and it is small to stand 11~13
When.
Finally, the vacuum drying temperature of the step 7) is 70~80 DEG C, pressure is 0.08~0.09MPa.
Compared with the prior art, the advantages of the present invention are as follows:
(1) aqueous slkali of a certain amount of concentration promote inside Radix Panacis Quinquefolii polysaccharide particle and between physical-chemical reaction, and make
The dissolution of polysaccharide faster and more inside particle is obtained, to improve the recovery rate of polysaccharide；
(2) the main component cellulose of degrading plant cell wall that can be specific using cellulase, makes the cell of plant
Wall rupture, polysaccharide easily increase juice from intracellular release, improve polysaccharide yield；
(3) there is decomposition to the protein to dissociate in plant cell using protease, keeps its loosely organized；
(4) it can also make the protein to dissociate in glycoprotein and proteoglycans hydrolysis using protease, reduce the knot to raw material
With joint efforts, be conducive to the leaching of polysaccharide；
(5) using papain be it is a kind of it is acid, neutral, under alkaline environment can decomposing protein protease,
With wider substrate specificity；
The present invention breaches traditional water and mentions or the method for alcohol extracting Radix Panacis Quinquefolii polysaccharide, using alkaline leaching combination double enzymolysis and super
Sound auxiliary process extracts, and triple technique superpositions greatly improve the recovery rate of Radix Panacis Quinquefolii polysaccharide, present invention process letter
It is single, it is easy to operate, the recovery rate and yield of Radix Panacis Quinquefolii polysaccharide are significantly improved, the industrialized production extracted for Radix Panacis Quinquefolii polysaccharide provides
Some theoretical reference foundations.
Specific embodiment
Present invention is further described in detail with reference to embodiments.
Embodiment 1
After the American Ginseng fibrous root that 100g has been dried is crushed with pulverizer, it is spare to cross 40 meshes.By solid-liquid ratio 1:25 (g:
Ml it) is added 3% soaking with sodium hydroxide 3 hours of 90 DEG C, filters to obtain filter residue, filter residue volatilizes；Resulting filter residue is 1 by solid-liquid ratio:
It is 1% complex enzyme (cellulase: papain=1:1) that enzyme bottom mass fraction, which is added, in 10 (g:ml), is shaken in 50 DEG C of constant temperature
It swings and is digested 1 hour on device；The filter residue of enzymatic hydrolysis by solid-liquid ratio 1:28 (g:ml) be added distilled water, 60W ultrasonic extraction three times, every time
20min is centrifuged back collection filtrate；Resulting filtrate material is concentrated under reduced pressure, and the temperature of reduced pressure is 60 DEG C, and pressure is 0.07~
0.09MPa, the relative density of concentration are 1.25；Dehydrated alcohol is added after concentrate is cooling, the final volume score of ethyl alcohol reaches
80%, 12 hours are stood, precipitating is collected after centrifugation；After resulting precipitating vacuum drying, target product Radix Panacis Quinquefolii polysaccharide is obtained, is obtained
Rate 13.55%；Vacuum drying temperature is 70~80 DEG C, and pressure is 0.08~0.09MPa.
Embodiment 2
After the American Ginseng fibrous root that 100g has been dried is crushed with pulverizer, it is spare to cross 40 meshes.By solid-liquid ratio 1:20 (g:
Ml it) is added 2% soaking with sodium hydroxide 2 hours of 90 DEG C, filters to obtain filter residue, filter residue volatilizes；Resulting filter residue is 1:8 by solid-liquid ratio
(g:ml) enzyme bottom mass fraction is added is 1% complex enzyme (pectase: papain=1:1), in 45 DEG C of constant temperature oscillator
Upper enzymatic hydrolysis 1 hour；The filter residue of enzymatic hydrolysis by solid-liquid ratio 1:25 (g:ml) be added distilled water, 45W ultrasonic extraction three times, each 15min,
It is centrifuged back collection filtrate.Resulting filtrate decompression concentration, the temperature of reduced pressure are 50 DEG C, and pressure is 0.07~0.09MPa, concentration
Relative density be 1.30；Dehydrated alcohol is added after concentrate is cooling, the final volume score of ethyl alcohol reaches 70%, and it is small to stand 12
When, precipitating is collected after centrifugation；After resulting precipitating vacuum drying, target product Radix Panacis Quinquefolii polysaccharide, yield 11.17% are obtained；Very
The dry temperature of sky is 70~80 DEG C, and pressure is 0.08~0.09MPa.
Embodiment 3
After the American Ginseng fibrous root that 100g has been dried is crushed with pulverizer, it is spare to cross 40 meshes；By solid-liquid ratio 1:23 (g:
Ml it) is added 4% soaking with sodium hydroxide 3 hours of 90 DEG C, filters to obtain filter residue, filter residue volatilizes；Resulting filter residue is 1 by solid-liquid ratio:
It is 1% complex enzyme (cellulase: bromelain=1:1) that enzyme bottom mass fraction, which is added, in 12 (g:ml), is shaken in 50 DEG C of constant temperature
It swings and is digested 1 hour on device.The filter residue of enzymatic hydrolysis by solid-liquid ratio 1:30 (g:ml) be added distilled water, 50W ultrasonic extraction three times, every time
25min is centrifuged back collection filtrate；The concentration of resulting filtrate decompression, the temperature of reduced pressure are 55 DEG C, pressure is 0.07~
0.09MPa, the relative density of concentration are 1.25；Dehydrated alcohol is added after concentrate is cooling, the final volume score of ethyl alcohol reaches
75%, 12 hours are stood, precipitating is collected after centrifugation；After resulting precipitating vacuum drying, target product Radix Panacis Quinquefolii polysaccharide is obtained, is obtained
Rate 12.32%；Vacuum drying temperature is 70~80 DEG C, and pressure is 0.08~0.09MPa.
Embodiment 4 (comparative example)
After the American Ginseng fibrous root that 100g has been dried is crushed with pulverizer, it is spare to cross 40 meshes；By solid-liquid ratio 1:25 (g:
Ml it) is added 90 DEG C 3% soaking with sodium hydroxide 3 hours, filters to obtain filter residue, filter residue volatilizes；Filter residue is added by solid-liquid ratio 1:25 (g:ml)
Enter water, three times, each 20min is centrifuged back collection filtrate to 60W ultrasonic extraction；Resulting filtrate material is concentrated under reduced pressure, the temperature of reduced pressure
Degree is 60 DEG C, and pressure is 0.07~0.09MPa, and the relative density of concentration is 1.25；Dehydrated alcohol, second is added after concentrate is cooling
The final volume score of alcohol reaches 80%, stands 12 hours, precipitating is collected after centrifugation；After resulting precipitating vacuum drying, obtain
Target product Radix Panacis Quinquefolii polysaccharide, yield 7.15%；Vacuum drying temperature is 70~80 DEG C, and pressure is 0.08~0.09MPa.
It can be seen that the present embodiment does not have enzymolysis step, ultrasonic wave extraction is directly used after alkali leaching, final Radix Panacis Quinquefolii polysaccharide
Yield is lower.
Embodiment 5 (comparative example)
After the American Ginseng fibrous root that 100g has been dried is crushed with pulverizer, it is spare to cross 40 meshes；By solid-liquid ratio 1:25 (g:
Ml it) is added 3% soaking with sodium hydroxide 3 hours of 90 DEG C, filters to obtain filter residue, filter residue volatilizes；Resulting filter residue is 1 by solid-liquid ratio:
It is 1% complex enzyme (cellulase: papain=1:1) that enzyme bottom mass fraction, which is added, in 10 (g:ml), is shaken in 50 DEG C of constant temperature
It swings and is digested 1 hour on device；Distilled water is added by solid-liquid ratio 1:28 (g:ml) in the filter residue of enzymatic hydrolysis, and 100 DEG C of water-baths flow back 2 times, every time
2.5 hours are centrifuged back collection filtrate；Resulting filtrate material is concentrated under reduced pressure, and the temperature of reduced pressure is 60 DEG C, and pressure is 0.07~
0.09MPa, the relative density of concentration are 1.25；Dehydrated alcohol is added after concentrate is cooling, the final volume score of ethyl alcohol reaches
80%, 12 hours are stood, precipitating is collected after centrifugation；After resulting precipitating vacuum drying, target product Radix Panacis Quinquefolii polysaccharide is obtained, is obtained
Rate 10.77%；Vacuum drying temperature is 70~80 DEG C, and pressure is 0.08~0.09MPa.
The method that the present embodiment uses water-bath refluxing extraction after enzymatic hydrolysis, it can be seen that compared with using ultrasonic wave extraction
Compared with yield is declined.
Claims (9)
1. a kind of method for extracting Radix Panacis Quinquefolii polysaccharide from American Ginseng fibrous root, it is characterised in that the following steps are included:
1) it crushes: after dry American Ginseng fibrous root is crushed with pulverizer, it is spare to cross 35~45 meshes；
2) alkali soaks: 85~95 DEG C of 2~4% sodium hydroxide solutions of mass concentration are added in the American Ginseng fibrous root powder after above-mentioned sieving
Middle immersion, filters to obtain filter residue, and filter residue volatilizes；
3) complex enzyme zymohydrolysis: it is 0.9~1.1% complex enzyme that enzyme bottom mass ratio, which is added, in the resulting filter residue of step 2), is shaken in constant temperature
It swings and is digested 50~70 minutes on device；
4) ultrasonic extraction method: distilled water is added in the filter residue after step 3) enzymatic hydrolysis, ultrasonic extraction power is 45~60W, ultrasound
It extracts 2~4 times, each time is 15~30min, and filtrate is collected by centrifugation；
5) it is concentrated: above-mentioned resulting filtrate decompression is concentrated；
6) alcohol precipitation: dehydrated alcohol will be added after the filtrate cooling after concentration, stands overnight, precipitating is collected after centrifugation；
7) it is dried in vacuo: resulting precipitating being dried in vacuo, target product Radix Panacis Quinquefolii polysaccharide is obtained；
The complex enzyme of the step 3) is cellulase, pectase, dextranase, papain, bromelain or withered grass bar
Two kinds in mycoproteinase, the mass ratio of two kinds of enzymes is 1:1 in complex enzyme, and complex enzyme zymohydrolysis solid-liquid ratio is 1g:8ml~1g:
12ml, the temperature of constant temperature oscillator are 45~50 DEG C.
2. according to the method described in claim 1, it is characterized by: the American Ginseng fibrous root powder and sodium hydroxide of the step 2)
The solid-liquid ratio of solution is 1g:20ml~1g:25ml, and the alkali leaching time is 2~3 hours.
3. according to the method described in claim 2, it is characterized by: the temperature of the sodium hydroxide solution be 90 DEG C, quality hundred
Divide than being 3%, the solid-liquid ratio of American Ginseng fibrous root powder and sodium hydroxide solution is 1g:25ml, and alkali soaks the time 3 hours.
4. according to the method described in claim 1, it is characterized by: the complex enzyme zymohydrolysis solid-liquid ratio is 1g:10ml, when enzymatic hydrolysis
Between 1 hour, the temperature of constant temperature oscillator be 50 DEG C.
5. according to the method described in claim 1, it is characterized by: the filter residue of the step 4) and the solid-liquid ratio of distilled water are
1g:25ml~1g:30ml, ultrasonic extraction 3 times.
6. according to the method described in claim 5, it is characterized by: the solid-liquid ratio of the filter residue and distilled water be 1g:25ml, surpass
It is 60W that sound, which extracts power, and each extraction time is 20min.
7. according to the method described in claim 1, it is characterized by: the temperature of the reduced pressure of the step 5) is 50~60
DEG C, pressure is 0.07~0.09MPa, and the relative density of concentration is 1.25~1.30.
8. according to the method described in claim 1, it is characterized by: the final volume percentage of the ethyl alcohol of the step 6) reaches
70~80%, stand 11~13 hours.
9. according to the method described in claim 1, it is characterized by: the vacuum drying temperature of the step 7) is 70~80
DEG C, pressure is 0.08~0.09MPa.
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| CN106749747A (en) * | 2017-03-13 | 2017-05-31 | 哈尔滨商业大学 | The method that Radix Panacis Quinquefolii polysaccharide is extracted using zymolysis technique |
| CN113577239B (en) * | 2021-07-30 | 2022-11-08 | 山东第一医科大学(山东省医学科学院) | American ginseng glycopeptide and preparation method and application thereof |
| CN115260334B (en) * | 2022-09-13 | 2023-04-07 | 河北象大合众生物科技有限公司 | Compound extraction process of mulberry leaf polysaccharide |
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