CN108250320A - A kind of low ash content ganoderan extract and preparation method thereof - Google Patents
A kind of low ash content ganoderan extract and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of low ash content ganoderan extracts and preparation method thereof, and the preparation method includes the following steps：(1) acid carries：Ganoderma lucidum medicinal material after water intaking rinsing, is dried, crushed into coarse powder, adds in aqueous hydrochloric acid at reflux extraction twice, and filtering, merging filtrate is concentrated under reduced pressure, obtains concentrate；(2) alcohol precipitation：The ethyl alcohol of concentration 95% is slowly added into concentrate, it is stirring while adding, it stands at room temperature, filters, obtain filter residue 1；(3) it washs：With ethanol solution washing step (2) filter residue 1, it is repeated twice, obtains filter residue 2；(4) it purifies：It with water dissolution step (3) filter residue 2, stands, centrifuges at room temperature, centrifuged supernatant crosses gel-network precipitation method column chromatography, collects lower column liquid, is dried in vacuo after reduced pressure, crushes, obtains ganoderan extract.Ganoderma polyoses content is high in ganoderan extract of the present invention, and content of ashes is low；Present invention process is easy, and production process is environmentally protective, is suitble to industrialized production.
Description
Technical field
The invention belongs to field of plant extraction, and in particular to a kind of low ash content ganoderan extract and preparation method thereof.
Background technology
Ganoderma lucidum is the rare Chinese medicine of China, have nourishing and fit keeping function, strengthen the body resistance to consolidate the constitution, macrobiosis of making light of one's life by commiting suicide the effect of, genus polyporus bacterium
Class fungi, the main active component of water extract is polysaccharide.In recent years, it is more and more research shows that, ganoderma lucidum polysaccharide has
Strongly antitumor, antiviral, anti-aging, the effect of anti-oxidant, hypoglycemic and immunocompetence.
One of the principle active component of ganoderma lucidum polysaccharide as ganoderma lucidum, is gradually paid attention in recent years by researcher.At present,
There are many relevant reports about ganoderma lucidum polysaccharide extraction process, such as：
Ganoderma lucidum fruitbody is ground into glossy ganoderma powder by prior art CN101805414A, then uses microwave radiation technology hot water successively
Extraction method, ultrasonic wave auxiliary hot water extraction method extract, the extracting solution of gained is concentrated, impurity elimination, concentration, alcohol precipitation and it is dry after
Obtain water-soluble ganoderma lucidum polysaccharide, polyoses content 5.46%；Ganoderma lucidum residue is extracted using alkali extraction method again, gained carries
Alkali solubility ganoderma lucidum polysaccharide is obtained after taking neutralized liquid, dialysis, concentration, impurity elimination, alcohol precipitation and drying process, polyoses content is
10.74%；
After prior art CN103073651A uses ultrasonic wave combination Enzymatic Extraction ganoderma lucidum polysaccharide, ganoderma lucidum to crush, by carrying
It takes, ultrafiltration, concentration and ethanol precipitation separation acquisition ganoderma lucidum polysaccharide, ganoderma lucidum polysaccharide recovery rate is 3.14~4.21%；
Prior art CN101434660 uses microwave and ultrasonic wave synergic extraction machine, is extracted by solvent of water；It has extracted
It is filtered after, concentrates alcohol precipitation, ganoderma lucidum polysaccharide, recovery rate 2.93% are obtained after vacuum freeze drying；
Prior art CN101367881 use ultrasonic equipment assisted extraction, by alcohol precipitation, organic solvent (absolute ethyl alcohol,
Acetone, ether) washing be dried to obtain ganoderma lucidum polysaccharide sample afterwards three times, purity of polysaccharide has reached 47.88%.
Ash content be substance in high temperature sintering, occur a series of physical and chemical change, last organic principle volatilizees loss,
And inorganic constituents (mainly inorganic salts and oxide) left behind, these residues are known as ash content.It is in product it is inorganic into
Divide an index of total amount, be control food or the important evidence of health products finished product or semi-manufactured goods quality.Ash content includes water solubility
Ingredient, water insoluble active ingredient and sour insoluble composition.When extracting ganoderma lucidum polysaccharide, grey branch is extracted together, and is formed
Enrichment, causes content of ashes in ganoderan extract excessively high, influences the quality of final products.The prior art does not have also for spirit
Sesame polyoses extract carries out the report of de-ash.
Invention content
The technical problems to be solved by the invention are to overcome the technological deficiency of background technology, and it is more to provide a kind of low ash content ganoderma lucidum
Sugar extract and preparation method thereof.Ganoderma polyoses content is high in ganoderan extract produced by the present invention, and content of ashes is low,
Middle ganoderma polyoses content >=60.8%, ash content≤1.2%；Production technology of the present invention is easy, easily operated, while production process is green
Colour circle is protected, and gel rubber material repeated multiple times can utilize, and is suitble to industrialized production.
Technological means is used by the present invention solves above-mentioned technical problem：
A kind of preparation method of low ash content ganoderan extract, includes the following steps：
(1) acid carries：Ganoderma lucidum medicinal material after water intaking rinsing, is dried, crushed into coarse powder, adds in aqueous hydrochloric acid at reflux extraction two
Secondary, filtering, merging filtrate is concentrated under reduced pressure, obtains concentrate；
(2) alcohol precipitation：A concentration of 95% ethanol solution is slowly added into concentrate, it is stirring while adding, it stands at room temperature,
It filters, obtains filter residue 1；
(3) it washs：With ethanol solution washing step (2) filter residue 1, it is repeated twice, obtains filter residue 2；
(4) it purifies：It with water dissolution step (3) filter residue 2, stands, centrifuges at room temperature, centrifuged supernatant crosses gel-type sun
Ion exchange resin column chromatographs, and collects lower column liquid, is dried in vacuo after reduced pressure, crushes, and obtains low ash content ganoderma lucidum polysaccharide extraction
Object.
Preferably, in the step (1), the drying temperature is 38~42 DEG C, and drying time is 10~12h.
Preferably, in the step (1), the mesh number of the coarse powder is 20~30 mesh.
Preferably, in the step (1), the pH of the aqueous hydrochloric acid solution is 5~6.
Preferably, in the step (1), the addition aqueous hydrochloric acid at reflux extracts twice, wherein first time hydrochloric acid water
The addition of solution is 10 times (v/m) of ganoderma lucidum medicinal material, and the addition of second of aqueous hydrochloric acid solution is 8 times of (v/ of ganoderma lucidum medicinal material
m)。
Preferably, in the step (1), the addition aqueous hydrochloric acid at reflux extraction twice, carries wherein flowing back for the first time
The time taken is 2h, and the time of second of refluxing extraction is 1.5h.
Preferably, in the step (1), the filtering is carried out using 300 mesh filter clothes.
Preferably, in the step (1), the vacuum degree during reduced pressure is -0.07~-0.09MPa, temperature 60
~80 DEG C.
Preferably, in the step (1), the volume of the concentrated liquid is 1/5~1/10 (v/m) of ganoderma lucidum medicinal material.
Preferably, in the step (2), the addition of a concentration of 95% ethanol solution is the volume of the concentrated liquid
4~6 times amount (v/v).
Preferably, in the step (2), the time of repose is 2~5h.
Preferably, in the step (3), the ethanol solution concentration is 70~80%.
Preferably, in the step (3), the ethanol solution addition is 1~2 times (v/m) of ganoderma lucidum medicinal material amount.
Preferably, in the step (4), the addition of the water is 2~3 times (v/m) of ganoderma lucidum medicinal material.
Preferably, in the step (4), the time of repose is 2~5h.
Preferably, in the step (4), the gel-network precipitation method is 732 types or Dowex50 types.
Preferably, in the step (4), the gel-network precipitation method dosage exchanges tree for gel type cation
Fat: ganoderma lucidum medicinal material=4~5: 1 (v/m).
Preferably, in the step (4), the flow velocity during column chromatography is 0.5~1BV/h.
Preferably, in the step (4), the vacuum degree during reduced pressure is -0.07~-0.09MPa.
A kind of low ash content ganoderan extract, is prepared using the above method.
In above-mentioned technical proposal, the ethanol solution concentration is ethyl alcohol mass fraction.
The basic principle of the present invention：
(1) by adding in hydrochloric acid when step (1) acid of the present invention carries, target product ganoderma lucidum polysaccharide can not only be made preferably molten
Go out, moreover it is possible to part of ash such as metal oxide-type ash content be made to be transformed into ionic condition, convenient for the processing of subsequent technique；
(2) the concentrate main component after step (1) acid of the present invention carries is ganoderma lucidum polysaccharide, triterpene compound, lipid, egg
Bletilla grades in the ash of ionic state, in step (2) alcohol precipitation by add in a concentration of 95% ethanol solution so that filter residue ingredient
Refined that (filter residue ingredient further includes a small amount of triterpenes chemical combination similar to ganoderma lucidum polysaccharide dissolubility in addition to ganoderma lucidum polysaccharide
Object, albumen and ash grade impurity)；
(3) during step (3) washing of the present invention filter residue is washed by using a concentration of 70~80% ethanol solution twice, it can
Larger impurity is differed with ganoderma lucidum polysaccharide dissolubility so that filter residue ingredient is further refined, while can also be incited somebody to action to wash away
The ash content that a part dissolves in ethanol solution elutes away；
(4) ganoderma lucidum polysaccharide similar for dissolubility of centrifuged supernatant main component obtained by step (4) of the present invention, triterpenes
Object, albumen and water-soluble ass etc. are closed, due to the difference of physicochemical property between these substances, such as ganoderma lucidum polysaccharide and triterpene compound
And molecular weight has differences between albumen, can be detached by molecular sieve principle, meanwhile, in sample aqueous solution is in cation
The ash content of state can by cationic exchange resin adsorption, therefore, the present invention using 732 types or Dowex50 type gel-type sun from
Sub-exchange resin column layer purifies ganoderma lucidum polysaccharide, comprehensively utilizes its molecular sieve principle and cation exchange principle, is prepared into
To low ash content, the ganoderan extract of high ganoderma polyoses content.
By analyze above it is found that acid of the present invention carry, alcohol precipitation, washing, purifying four steps it is all linked with one another, it is progressive, collaboration make
With, the ganoderan extract of low ash content, high ganoderma polyoses content is finally prepared, any increase and decrease test procedure of the present invention
Technical solution can not obtain the ganoderan extract of technique effect of the present invention.
Compared with prior art, technical scheme of the present invention has the following advantages that：
(1) ganoderma polyoses content is high in ganoderan extract produced by the present invention, and content of ashes is low, wherein ganoderma lucidum polysaccharide
Content >=60.8%, ash content≤1.2%；
(2) present invention carries out refluxing extraction, extraction using aqueous hydrochloric acid solution as Extraction solvent using ganoderma lucidum fruitbody as raw material
Liquid filtering and concentrating adds in a concentration of 95% ethanol solution of 4 times of amounts of concentrate, is taken out after standing 2h at room temperature to certain volume
Filter, filter residue are washed twice using suitable a concentration of 80% ethanol solution, and the filter residue after washing adds in suitable water dissolution, mistake
Gel-network precipitation method collects lower column liquid, is concentrated and dried, you can obtain low ash content ganoderan extract；The present invention
Production technology is easy, easily operated, while production process is environmentally protective, and gel rubber material repeated multiple times can utilize, and is suitble to industrialization
Production.
Description of the drawings
Fig. 1 is that ganoderan extract made from the embodiment of the present invention 3 and comparative example 1 makees the removing of DPPH free radicals
With；
Fig. 2 is removing of the ganoderan extract to hydroxy radical (OH) made from the embodiment of the present invention 3 and comparative example 1
Effect；
Fig. 3 is the reducing power of ganoderan extract made from the embodiment of the present invention 3 and comparative example 1；
Fig. 4 is the reducing power of positive control Vc.
Specific embodiment
In order to better understand the content of the present invention, it is described further with reference to specific embodiments and the drawings.Ying Li
Solution, these embodiments are only used for that the present invention is further described rather than limits the scope of the invention.In addition, it should also be understood that,
After having read present disclosure, person skilled in art makes the present invention some nonessential changes or adjustment, still belongs to
In protection scope of the present invention.
Ganoderma lucidum fruitbody medicinal material described in Examples 1 to 3 and comparative example 1 originates in Longquan, Zhejiang Province.
Polyoses content, product yield, yield advantage calculation formula are as follows described in Examples 1 to 3 and comparative example 1：
Ash content calculation formula described in Examples 1 to 3 and comparative example 1 is as follows：
In formula：The content of ash content, % in X- samples；m1The quality of crucible and ash content, g；m2The quality of crucible, g；m3Earthenware
The quality of crucible and sample, g.
Embodiment 1
Ganoderma lucidum fruitbody medicinal material after appropriate potable water rinse under the conditions of 38~42 DEG C is dried into 10~12h, is ground into
The coarse powder of 20 mesh, weighs 500g, and the aqueous hydrochloric acid at reflux for adding in pH=5 extracts twice, and the quantity of solvent of addition is respectively medicinal material
Amount 10 times, 8 times (v/m), extraction time is respectively 2h and 1.5h, 300 mesh filter-cloth filtering after extraction, merging filtrate ,-
The volume of the concentrated liquid is concentrated under reduced pressure under 0.07Mpa as 50ml.The ethyl alcohol that 300ml a concentration of 95% is slowly added into concentrate is molten
Liquid, it is stirring while adding, it is filtered after standing 2h at room temperature, retains filter residue.The ethyl alcohol that filter residue is added to 500ml a concentration of 80% is molten
In liquid, 15min is stood after stirring 30min, is filtered, retains filter residue, repeated washing is twice.It is precipitated with the water dissolution of 1L, after standing
Centrifugation retains supernatant, and supernatant crosses 732 gel-network precipitation method column chromatographies, resin demand: medicinal material amount=5: 1, stream
Fast 1BV/h, collects lower column liquid, and lower column liquid -0.09Mpa is crushed after being dried in vacuo after being concentrated under reduced pressure.Use Phenol sulfuric acid procedure
It is 61.4% to detect sample ganoderma polyoses content, and ash content is detected as 0.8%.
Embodiment 2
Ganoderma lucidum fruitbody medicinal material after appropriate potable water rinse under the conditions of 38~42 DEG C is dried into 10~12h, is ground into
The coarse powder of 30 mesh, weighs 500g, and the aqueous hydrochloric acid at reflux for adding in pH=6 extracts twice, and the quantity of solvent of addition is respectively medicinal material
Amount 10 times, 8 times (v/m), extraction time is respectively 2h and 1.5h, 300 mesh filter-cloth filtering after extraction, merging filtrate ,-
0.08Mpa is concentrated under reduced pressure into the volume of the concentrated liquid as 100ml.The ethyl alcohol that 400ml a concentration of 95% is slowly added into concentrate is molten
Liquid, it is stirring while adding, it is filtered after standing 4h at room temperature, retains filter residue.Filter residue is added to the ethyl alcohol of 1000ml a concentration of 70%
In solution, 30min is stood after stirring 30min, is filtered, retains filter residue, repeated washing is twice.It is precipitated, stood with the water dissolution of 1L
After centrifuge, retain supernatant, supernatant crosses 732 type gel-network precipitation method column chromatographies, resin demand: medicinal material amount=4:
1, flow velocity 0.5BV/h, collect lower column liquid, and lower column liquid -0.08Mpa is crushed after being dried in vacuo after being concentrated under reduced pressure.Use phenol
Sulfuric acid process detection sample ganoderma polyoses content is 63.2%, and ash content is detected as 1.2%.
Embodiment 3
Ganoderma lucidum fruitbody medicinal material after appropriate potable water rinse under the conditions of 38~42 DEG C is dried into 10~12h, is ground into
The coarse powder of 20 mesh, weighs 500g, and the aqueous hydrochloric acid at reflux for adding in pH=5 extracts twice, and the quantity of solvent of addition is respectively medicinal material
Amount 10 times, 8 times (v/m), extraction time is respectively 2h and 1.5h, 300 mesh filter-cloth filtering after extraction, merging filtrate ,-
0.08Mpa is concentrated under reduced pressure into the volume of the concentrated liquid as 50ml.The ethyl alcohol that 300ml a concentration of 95% is slowly added into concentrate is molten
Liquid, it is stirring while adding, it is filtered after standing 2h at room temperature, retains filter residue.The ethyl alcohol that filter residue is added to 500ml a concentration of 80% is molten
In liquid, 15min is stood after stirring 30min, is filtered, retains filter residue, repeated washing is twice.It is precipitated, stood with the water dissolution of 1.5L
After centrifuge, retain supernatant, supernatant crosses Dowex50 type gel-network precipitation method column chromatographies, resin demand: medicinal material amount
=5: 1, flow velocity 1BV/h, collect lower column liquid, and lower column liquid -0.08Mpa is crushed after being dried in vacuo after being concentrated under reduced pressure.Use benzene
Phenol sulfuric acid method detection sample ganoderma polyoses content is 60.8%, and ash content is detected as 1.1%.
Comparative example 1
Ganoderma lucidum fruitbody medicinal material after potable water rinse dry 10~12h under the conditions of 38~42 DEG C in right amount, is ground into 20
Purpose coarse powder weighs 500g, adds in drinking water refluxing extraction twice, the amount for adding in water is respectively 10 times of medicinal material amount, 8 times of (v/
M), extraction time is respectively 2h and 1.5h, and 300 mesh filter-cloth filtering, merging filtrate, -0.09Mpa are concentrated under reduced pressure into after extraction
The volume of the concentrated liquid is 50ml.The ethanol solution of 300ml a concentration of 95%, stirring while adding, room temperature are slowly added into concentrate
It is filtered after lower standing 1.5h, retains filter residue.Filter residue is added in the ethanol solution of 500ml a concentration of 80%, after stirring 30min
30min is stood, is filtered, retains filter residue, repeated washing is twice.It is precipitated with the water dissolution of 1.5L, is centrifuged after standing, retain supernatant
Liquid, supernatant cross Dowex50 type gel-network precipitation method column chromatographies, resin demand: medicinal material amount=5: 1, flow velocity 1BV/
H, collects lower column liquid, and lower column liquid -0.09Mpa is crushed after being dried in vacuo after being concentrated under reduced pressure.Sample is detected using Phenol sulfuric acid procedure
Product ganoderma polyoses content is 54.2%, and ash content is detected as 4.4%.
Polyoses content and content of ashes are shown in Table 1 in ganoderan extract made from Examples 1 to 3 and comparative example 1.
Polyoses content and content of ashes in ganoderan extract made from 1 Examples 1 to 3 of table and comparative example 1
| Product | Polyoses content % | Product yield % | Yield advantage % | Ash content % |
| Embodiment 1 | 61.4 | 1.13 | 69.4 | 0.8 |
| Embodiment 2 | 63.2 | 1.20 | 75.8 | 1.2 |
| Embodiment 3 | 60.8 | 1.17 | 71.1 | 1.1 |
| Comparative example 1 | 54.2 | 1.01 | 67.7 | 4.4 |
First, ganoderma polyoses content detection method (phend-sulphuric acid) of the present invention：
1. the preparation of reference substance solution：
The glucose control product that precision weighs 105 DEG C of dryings to constant weights are appropriate, add water that the solution that every 1ml contains 100 μ g is made,
To obtain the final product.
2. the preparation of standard curve：
It is accurate respectively to draw reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, it puts in 25ml tool plug test tubes,
Benefit adds water to 2.0ml, is separately added into 5% phenol solution 1mL, mixing, and rapid vertical addition sulfuric acid 7.0mL shakes up, puts boiling water bath
Middle colour developing 2min takes out.10min in ice-water bath is put, is taken out, using corresponding reagent as blank, according to UV-VIS spectrophotometry
(one annex VA of Chinese Pharmacopoeia version in 2010) measures absorbance at 490nm wavelength, a concentration of using absorbance as ordinate
Abscissa draws standard curve.
3. the preparation of test solution：
This product about 1g is taken, it is accurately weighed, it puts in 100mL measuring bottles, adds water 20mL ultrasonic dissolutions and shake frequently, treat sample point
Dissipate uniformly without caking after, then add 40mL water, ultrasonic 30min is diluted with water to scale, shakes up, be placed at room temperature for 1h after cooling.Essence
Close measurement supernatant 2mL vertically adds in ethyl alcohol (a concentration of 95% ethanol solution) 10ml, shakes up, and 4 DEG C of placement 12h take out,
4000r/min centrifuges 20min, and carefully incline supernatant, and centrifuge tube is gently tipped upside down on filter paper, after no longer liquid is flowed down, then
Appropriate amount of water ultrasonic dissolution precipitation is added in, is fully transferred in 100mL volumetric flasks, is diluted with water to scale, shake up.On precision measures
State solution 2mL, the method under the preparation of sighting target directrix curve from " being separately added into 5% phenol solution 1mL ", is measured and inhaled in accordance with the law
Luminosity, from standard curve read test solution in DEXTROSE ANHYDROUS weight (mg), calculate to get.
2nd, ganoderma lucidum polysaccharide total ash
Reference《Chinese Pharmacopoeia》Four 2302 Ash determination method of the general rule detections of version in 2015.
3rd, ganoderma lucidum polysaccharide antioxidation activity in vitro
3.1 experiment ganoderma lucidum polysaccharide raw materials
Ganoderma lucidum polysaccharide 1：Ganoderma lucidum polysaccharide sample (pH5~6HCl aqueous solutions refluxing extraction) prepared by case study on implementation 3；Ganoderma lucidum is more
Sugar 2：Ganoderma lucidum polysaccharide sample (drinking water refluxing extraction) prepared by comparative example 1.
3.2 DPPH radicals scavengings effect measures
Difference accurately draw the mass concentration that has been configured for 0,0.2,0.4,0.6,0.8 and 1.0mg/mL ganoderma lucidum polysaccharide 1,
Ganoderma lucidum polysaccharide 2, positive control Vc sample solution 2mL be added in the tool plug teat glass of each dried and clean, be protected from light condition
Lower that the DPPH ethanol solutions that 2mL mass fractions are 0.004% are separately added into each test tube, mixing is anti-in dark surrounds
30min is answered, repeats experiment 3 times, results are averaged to measure absorbance value at 517nm in wavelength after reaction.Clearance rate
Calculation formula is as follows：
In formula：A0Add 2mL DPPH ethanol solution absorbance values for 2mL pure water；A1Add 2mL DPPH second for 2mL solution to be measured
Alcoholic solution absorbance value；A2Add 2mL ethanol solution absorbance values for 2mL solution to be measured.
3.3 hydroxy radicals (OH) scavenging effect measures
Respectively by the FeSO of 1mL 9mmol/L4Salicylic acid-ethanol solution of solution and 2mL 9mmol/L is gradually added to
In the tool plug teat glass of each dried and clean, mixing, then be separately added into the mass concentration that 2mL has been configured be 0,1.0,2.0,
3.0th, the ganoderma lucidum polysaccharide 1 of 4.0 and 5.0mg/mL, ganoderma lucidum polysaccharide 2, positive control Vc solution, are finally separately added into 2mL
The H of 8.8mmol/L2O2Solution starts reaction, reacts 1h at room temperature, goes out to measure its absorbance value at wavelength 510nm.Experiment weight
3 times multiple, results are averaged.Clearance rate calculation formula is as follows：
In formula：A1To add in the polysaccharide solution and H of different quality concentration2O2The absorbance of solution；A2To add in equal volume
H2O substitutes H2O2The absorbance of solution；A0To add in isometric H2O substitutes polysaccharide solution and H2O2The absorbance of solution.
The measure of 3.4 reducing powers
The ganoderma lucidum polysaccharide 1 and spirit that the mass concentration being configured is 0,0.2,0.4,0.6,0.8 and 1.0mg/mL are drawn respectively
2 sample solution of sesame polysaccharide is added to respectively with positive control Vc solution (concentration is respectively 0,20,40,60,80 and 100 μ g/mL) 1mL
In the tool plug teat glass of a dried and clean, respectively into each test tube add in 2.5mL phosphate buffer (pH 6.6,
0.2mol/L) with 1% potassium ferricyanide solutions of 2.5mL, it is mixed evenly, water-bath 20min, is then separately added into 2.5mL again at 50 DEG C
Mass fraction is 10% solution of trichloroacetic acid, and centrifuge speed is to centrifuge 10min, then use liquid-transfering gun respectively under 3000r/min
Precision draws the supernatant of 2.5mL, is separately added into the 0.1%FeCl that 2.5mL pure water and 2.5mL now match3Solution is uniformly mixed,
After being placed at room temperature for 30min, the absorbance value of solution is measured at wavelength 700nm.Experiment is repeated 3 times, and results are averaged.
4th, anti-oxidant experimental result
4.1 DPPH radicals scavengings act on
The ability that the clearance rate of DPPH free radicals provides hydrogen with antioxidant is related.
Ganoderma lucidum polysaccharide 1 and ganoderma lucidum polysaccharide 2 have the scavenging effect of certain DPPH free radicals as seen from Figure 1, and clear
Removing solid capacity is relatively close together, and when polysaccharide concentration is 1.0mg/ml, ganoderma lucidum polysaccharide 1 is with ganoderma lucidum polysaccharide 2 to DPPH free radicals
Clearance rate is respectively 87.3%, 85.5%.
4.2 hydroxy radicals (OH) scavenging effect
Ganoderma lucidum polysaccharide 1 has scavenging effect with ganoderma lucidum polysaccharide 2 to hydroxy radical (OH) as seen from Figure 2, removes energy
Power is relatively close together, when polysaccharide concentration is 5.0mg/ml, ganoderma lucidum polysaccharide 1 and removing of the ganoderma lucidum polysaccharide 2 to DPPH free radicals
Rate is respectively 91.2%, 93.4%.
4.3 reducing power
Ganoderma lucidum polysaccharide 1 and ganoderma lucidum polysaccharide 2 have reducing power to a certain extent, and reducing power phase as seen from Figure 3
To close, when polysaccharide concentration is 1.0mg/ml, the absorbance value of ganoderma lucidum polysaccharide 1 and ganoderma lucidum polysaccharide 2 is respectively 0.343,
0.342, but reducing power is less than positive control Vc.
4.4 it summarizes
From above ganoderma lucidum polysaccharide obtain antioxidation in vitro experimental result can be seen that embodiment 3 and comparative example 1 ganoderma lucidum it is more
Sugar-like product all have antioxidant activity to a certain extent, and the antioxidant activity of the two does not have apparent difference, this shows
Extraction process of the present invention will not generate ganoderma lucidum polysaccharide bioactivity big influence, therefore, using preparation method of the present invention
Compared with the prior art the ganoderan extract added value being prepared greatly improves.
Above description is not the limitation to invention, and the present invention is also not limited to the example above.The common skill of the art
Art personnel are in the essential scope of invention, and the variations, modifications, additions or substitutions made should also belong to the scope of protection of the present invention.
Claims (10)
1. a kind of preparation method of low ash content ganoderan extract, which is characterized in that include the following steps：
(1) acid carries：Ganoderma lucidum medicinal material after water intaking rinsing, is dried, crushed into coarse powder, adds in aqueous hydrochloric acid at reflux extraction twice,
Filtering, merging filtrate are concentrated under reduced pressure, obtain concentrate；
(2) alcohol precipitation：A concentration of 95% ethanol solution is slowly added into concentrate, it is stirring while adding, it stands, takes out at room temperature
Filter, obtains filter residue 1；
(3) it washs：With ethanol solution washing step (2) filter residue 1, it is repeated twice, obtains filter residue 2；
(4) it purifies：It with water dissolution step (3) filter residue 2, stands, centrifuges at room temperature, centrifuged supernatant crosses gel type cation
Exchanger resin column chromatography is collected lower column liquid, is dried in vacuo after reduced pressure, crushes, obtains low ash content ganoderan extract.
A kind of 2. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(1) in, the mesh number of the coarse powder is 20~30 mesh.
A kind of 3. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(1) in, the pH of the aqueous hydrochloric acid solution is 5~6.
A kind of 4. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(1) in, twice, wherein the addition of first time aqueous hydrochloric acid solution is ganoderma lucidum medicinal material for the addition aqueous hydrochloric acid at reflux extraction
10 times (v/m), the addition of second of aqueous hydrochloric acid solution are 8 times (v/m) of ganoderma lucidum medicinal material；The time of first time refluxing extraction is
2h, the time of second of refluxing extraction is 1.5h.
A kind of 5. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(1) in, the filtering is carried out using 300 mesh filter clothes；The vacuum degree during reduced pressure is -0.07~-0.09MPa, temperature
It is 60~80 DEG C.
A kind of 6. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(2) in, the addition of a concentration of 95% ethanol solution is 4~6 times of amounts (v/v) of the volume of the concentrated liquid.
A kind of 7. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(3) in, the ethanol solution concentration is 70~80%；The ethanol solution addition is 1~2 times (v/m) of ganoderma lucidum medicinal material amount.
A kind of 8. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the step
(3) in, gel-network precipitation method is 732 types or Dowex50 types.
A kind of 9. preparation method of low ash content ganoderan extract as described in claim 1, which is characterized in that the gel
Type cation exchange resin dosage is gel-network precipitation method: ganoderma lucidum medicinal material=4~5: 1 (v/m).
10. a kind of low ash content ganoderan extract, is prepared using such as claim 1~9 any one the method.
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