CN109180825A - Pilose antler mushroom polyoses extract and the preparation method and application thereof - Google Patents
Pilose antler mushroom polyoses extract and the preparation method and application thereof Download PDFInfo
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- CN109180825A CN109180825A CN201810729885.7A CN201810729885A CN109180825A CN 109180825 A CN109180825 A CN 109180825A CN 201810729885 A CN201810729885 A CN 201810729885A CN 109180825 A CN109180825 A CN 109180825A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A61P39/06—Free radical scavengers or antioxidants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
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Abstract
A kind of pilose antler mushroom polyoses extract, it is prepared as follows to obtain: after pilose antler massee fruiting bodies dry product is crushed, it is impregnated through 95% ethyl alcohol, using distilled water as solvent after degreaser drying, high speed shear is extracted at solid-liquid ratio 1:15~25,9500~14500rpm of revolving speed, 1~4min of extraction time, extracting solution centrifugation discards precipitating, supernatant is collected, after the concentration of gained supernatant, through being classified alcohol precipitation, centrifugation, freeze-drying obtain pilose antler mushroom polyoses extract；Gained pilose antler mushroom polyoses extract of the invention has apparent antioxidant activity, ABTS free radical, DPPH free radical can be effectively removed, and there is certain reducing power, can be applied to prepare novel antioxidant, and pilose antler mushroom safety, it is natural, have no toxic side effect, there is applications well prospect.
Description
(1) technical field
The present invention relates to a kind of pilose antler mushroom polyoses extracts and the preparation method and application thereof.
(2) background technique
Pilose antler mushroom is also known as Lyophyllum decastes (Lyophyllum decastes.), is under the jurisdiction of Basidiomycetes, Agaricales, dried mushroom
Section is the wild excellent fungi of large size for eating medicine dual-purpose, nutriment rich in, based on fresh food, bacterial context is fertile from pleat umbrella category
Thick exquisiteness, A sweety scent assails the nostrils, the crisp cunning of mouthfeel, delicious flavour, and drying rear nutrition and mouthfeel are constant, are referred to as in Europe
"Friedchi cken mushroom".Research shows that crude protein, amino acid content are higher in pilose antler massee fruiting bodies, fat content
It is low, but also contain trace element zinc, selenium and the copper and a large amount of vitamin B beneficial to human body1, vitamin B2, vitamin
B6, vitamin B12And niacin, it has very high nutritive value.Meanwhile its fructification can be used as a kind of traditional drug,
Main component is pilose antler mushroom polysaccharide, and existing research proves that polysaccharide isolated from decastes destroying angel has anti-swell
Tumor, anti-oxidant, reducing blood sugar and blood lipid effect, health-care effect are obvious.
Currently, extracting the common method of polysaccharide has hot water extraction, ultrasonic wave assisted extraction, multiplex-enzyme extraction etc..Tradition side
Method Hot water extraction is although easy to operate but overlong time；Ultrasonic wave assisted extraction shortens extraction time, however also needs several small
When；Multiplex-enzyme extraction mild condition, impurity easily remove, but enzyme itself is easy inactivation and higher cost.High speed shear technology is as close
One kind for growing up homogenizes technology over year, is widely used in natural product extraction, separation and modified aspect, main by negative
The various external forces such as pressure, shearing, high velocity impact rapidly diffuse into effective component in raw material in solvent, reach in a short time
Dissolution equilibrium.(Li Li, Liu Yewei, Zhao Jianxi wait high speed shear technology to be crushed bee pollen form cole cell wall technique [J] to Li Li
Food Science, 2012,33 (12): 97-101) etc. using high speed shear technology be crushed bee pollen form cole cell wall, make pollen broken wall
Rate reaches 99% or more.
The biological function of polysaccharide depends on its complicated and multiplicity structural property, such as monosaccharide composition, molecular weight, branch degree
Deng can influence the bioactive functions of polysaccharide, make it have a variety of different important physiology such as antitumor and immunological regulation
Function；With the potentiality being applied in health food or drug as important active constituent, thus also attract more and more
Food and medicine field experts and scholars concern.A large amount of research has shown that the lipid-loweringing of the fructification of fungus, mycelium polysaccharides
Hypoglycemic, anti-oxidant, anticancer bioactivity clearly, and have had a large amount of fungal resources to be used as drug and serve to face
Bed, the development and utilization of fungal resources are increasingly becoming a new hot spot of drug research.The bacterium is by feat of safety, natural, nontoxic pair
The advantages of effect, increasingly causes to pay close attention to, popular among consumers at home and abroad, there is wide development prospect.
Pilose antler mushroom polysaccharide is one of very important active constituent in pilose antler mushroom, the immunological regulation shown with it, protect liver,
The important physiological function such as antitumor, anti-oxidant is closely related.The existing multiple biological activities about pilose antler mushroom be reported with
Its polysaccharide component is related, and existing research proves that polysaccharide isolated from pilose antler massee fruiting bodies has antitumor, hypoglycemic, drop
A series of effects such as blood lipid, anti-oxidant.(Jia Ning Lyophyllum decastes Polysaccharides on Mice Immune Function and antioxygen are turned into Jia Ning
With the whole nation research [C] Gansu Agriculture University animal institute .Vol15 veterinary pathology, animal Pathological Physiology academic meeting paper
Collection, Gansu, 2009,367.) the pilose antler plant mushroom fruit body polysaccharide that is obtained with traditional water extraction and alcohol precipitation method can remove it is negative in Mice Body
Ion radical can enhance the immunocompetence of mouse, to improve anti-oxidant, anti-aging product.
(3) summary of the invention
The present invention provides a kind of pilose antler mushroom polyoses extract and preparation method thereof, which, which has, is removed freely
The activity of base can be used for preparing novel antioxidant.
Technical scheme is as follows:
A kind of pilose antler mushroom polyoses extract, is prepared as follows to obtain:
(1) after crushing pilose antler massee fruiting bodies dry product, volume fraction is added to by solid-liquid ratio 1:3~6 (preferably 1:4, g:mL)
It in the aqueous solution of 95% ethyl alcohol, impregnates 8~16h (preferably 12h), filter residue is collected in filtering, and naturally dry obtains pilose antler mushroom skimmed milk
End；
The pilose antler mushroom is from pleat umbrella category, Lyophyllum decastes (Lyophyllum decastes.)；
(2) take pilose antler mushroom defatted seed flour obtained by step (1) that distilled water is added to carry out high speed shear extraction, extracting solution centrifugation
(10000rpm, 10min) discards precipitating, collects supernatant；
The conditional parameter that the high speed shear is extracted are as follows: solid-liquid ratio 1:15~25 (g:mL), 9500~14500rpm of revolving speed,
1~4min of extraction time；It is preferred that solid-liquid ratio 1:25 (g:mL), revolving speed 11500rpm, extraction time 1min；
(3) supernatant obtained by step (2) is concentrated into 1/7~1/12 (preferably 1/10) of original volume, it is thick to obtain pilose antler mushroom
Polysaccharide concentrate (is denoted as LDS)；
(4) aqueous solution of 95% ethyl alcohol of volume fraction is added into pilose antler mushroom Thick many candies concentrate obtained by step (3) and makes
It obtains ethyl alcohol final concentration and reaches 30%~80%, stand 8~16h (preferably 12h), be centrifuged (10000rpm, 10min), gained precipitating
It is freeze-dried after removal residual ethanol, obtains the pilose antler mushroom polyoses extract；
Wherein, the method for gained precipitating removal residual ethanol are as follows: redissolve precipitating plus water, evaporating solvent under reduced pressure is residual to remove
Ethyl alcohol is stayed, repetitive operation is to without obvious ethyl alcohol smell.
Further, the operating method of the step (4) are as follows:
The aqueous solution of 95% ethyl alcohol of volume fraction is added into pilose antler mushroom Thick many candies concentrate obtained by step (3) and makes second
Alcohol final concentration reaches 30%, stands 12h, is centrifuged (10000rpm, 10min), isolated primary sedimentation and a supernatant；To
The aqueous solution of 95% ethyl alcohol of volume fraction is added in supernatant and ethyl alcohol final concentration is made to reach 60%, stands 12h, centrifugation
(10000rpm, 10min), isolated secondary precipitation and secondary supernatant；Volume fraction 95% is added into secondary supernatant
The aqueous solution of ethyl alcohol simultaneously makes ethyl alcohol final concentration reach 80%, stands 12h, is centrifuged (10000rpm, 10min), separates and collects three
Secondary precipitating；
By gained primary sedimentation, secondary precipitation, three times precipitate respectively plus water redissolve, evaporating solvent under reduced pressure is to remove residual second
Alcohol, to without obvious ethyl alcohol smell, freeze-drying respectively obtains three kinds of pilose antler mushroom polyoses extracts, is successively denoted as repetitive operation
LDS30、LDS60、LDS80。
Antioxidation in vitro test is carried out to pilose antler mushroom polyoses extract produced by the present invention, the results showed that, it mentions in this way
It takes gained pilose antler mushroom polyoses extract that there is apparent antioxidant activity, can effectively remove ABTS free radical, DPPH free radical,
And there is certain reducing power, can be applied to prepare novel antioxidant.
Compared with existing technology (hot water extraction, ultrasonic wave assisted extraction, multiplex-enzyme extraction etc.), beneficial effect of the present invention
It is mainly reflected in:
Conventional hot water's extraction is easy to operate but overlong time；Though ultrasonic wave assisted extraction is unlike conventional hot water's extraction
Between it is too long, but also need several a few houres；Multiplex-enzyme extraction method extraction conditions are mild, impurity easily removes, but enzyme itself be easy inactivation and at
This is higher；
The method of the present invention high speed shear extractive technique, which mainly passes through the various external forces such as shearing, high velocity impact, to be made in raw material
Effective component rapidly diffuses into solvent, can reach dissolution equilibrium within a short period of time, and with separative efficiency height, rate is fast, sets
The advantages that standby simple, pollution-free, water extract-alcohol precipitation is assisted, the yield of pilose antler mushroom polysaccharide can be significantly improved；
The pilose antler mushroom polyoses extract antioxidant activity of the method for the present invention preparation is high, has applications well prospect, and pilose antler
Mushroom by feat of safety, it is natural, have no toxic side effect the advantages of cause more and more concerns, it is popular among consumers at home and abroad.
(4) Detailed description of the invention
Fig. 1: pilose antler mushroom polyoses extract preparation technology flow chart；
Fig. 2: the reducing power curve graph of LDS30, LDS60, LDS80 component in embodiment 2；
Fig. 3: the ABTS free radical scavenging activity curve graph of LDS30, LDS60, LDS80 component in embodiment 3；
Fig. 4: the DPPH free radical scavenging activity curve graph of LDS30, LDS60, LDS80 component in embodiment 4.
(5) specific embodiment
Preferably to explain the purpose of the present invention, preparation method, come below with specific embodiment to technical side of the invention
Case is described further, and but the scope of the present invention is not limited thereto.
1 high speed shear of embodiment extracts pilose antler mushroom polysaccharide
Dry pilose antler massee fruiting bodies dry product is weighed, through 4 times of 95% ethyl alcohol soaked overnight after crushing, next day is collected by filtration
Filter residue volatilizes reagent and obtains pilose antler mushroom defatted seed flour, for use；
Using pilose antler mushroom polyoses content as index, different feed liquid ratio, revolving speed, extraction time three in high speed shear extraction have been probed into
Influence of a factor to pilose antler mushroom polyoses content, as a result such as table 2 obtains shown；
Polyoses content (%)=total sugar content (%)-content of reducing sugar (%)
| Tested number | Solid-liquid ratio mL/g | Revolving speed r/min | Time min | Polysaccharide extract rate (%) |
| 1 | 15 | 9500 | 1 | 22.53 |
| 2 | 15 | 11500 | 2 | 22.34 |
| 3 | 15 | 14500 | 4 | 22.23 |
| 4 | 20 | 9500 | 2 | 23.15 |
| 5 | 20 | 11500 | 4 | 22.85 |
| 6 | 20 | 14500 | 1 | 22.97 |
| 7 | 25 | 9500 | 4 | 23.16 |
| 8 | 25 | 11500 | 1 | 23.71 |
| 9 | 25 | 14500 | 2 | 23.42 |
It show that optimal high speed shear extracts the condition of pilose antler mushroom polysaccharide using pilose antler mushroom polyoses content as index by table 1, that is, expects
Liquor ratio is 1:25mL/g, revolving speed 11500r/min, time 1min, with this condition by repeating verification experimental verification recovery rate point three times
Not Wei 23.84,23.81,23.80, show that this prioritization scheme is feasible；
Pilose antler mushroom defatted seed flour 100g is taken, high speed shear is extracted in optimal conditions, and extracting solution is with 10000r/min rate
It is centrifuged 10min, precipitating is discarded and collects supernatant；Supernatant rotary evaporation is concentrated into 1/10 that volume is original volume, obtains pilose antler
Mushroom Thick many candies concentrate LDS；
Being slowly added to 95% ethyl alcohol into pilose antler mushroom Thick many candies concentrate LDS makes concentration of alcohol reach 30%, stands overnight,
10000r/min is centrifuged 10min, precipitation and separation I and supernatant I；Continuously adding 95% ethyl alcohol in upward clear liquid I keeps ethyl alcohol in I dense
Degree reaches 60%, stands overnight, is ibid centrifuged 10min, precipitation and separation II and supernatant II；It is continuously added in upward clear liquid I I again
95% ethyl alcohol makes concentration of alcohol in II reach 80%, stands overnight, and is ibid centrifuged 10min, precipitation and separation III；It will centrifugation gained
Precipitating I, II, III add water to redissolve respectively, and revolving removal residual ethanol, to without obvious ethyl alcohol smell, freeze-drying can for repetitive operation
The polysaccharide crude extract for obtaining three components, is respectively designated as LDS30, LDS60, LDS80；
Embodiment 2 pilose antler mushroom Thick many candies LDS30, LDS60, LDS80 --- reducing power measurement
Example 1 gained pilose antler mushroom Thick many candies LDS30, LDS60, LDS80, by Thick many candies LDS30, LDS60, LDS80
It is made into the aqueous solution of 0,0.2,0.4,0.6,0.8,1.0,1.2mg/mL respectively, for use；
Take 0,0.2,0.4,0.6,0.8,1.0, each 1.0mL of LDS30 of 1.2mg/mL in test tube, be added
The phosphate buffer solution of the pH=6.6 of 1.0mL0.2mol/L and the potassium ferricyanide solution of 1.0mL1%, vortex oscillation mix,
The TCA solution of 1.0mL10% is added later, 1.0mL0.1%FeCl is added after cooling by 50 DEG C of insulation reaction 20min3Solution, instead
10min is answered, light absorption value A is measured at 700nmx.It replaces LDS30 to make blank control with isometric distilled water, surveys light absorption value A0,
VCAs positive control；LDS60, LDS80 are ibid operated.Reducing power formula is as follows:
Reducing power=Ax- Ao
Ax: the light absorption value of sample；Ao: blank control group light absorption value
The result of measurement as shown in Fig. 2, the reducing power of component LDS30, LDS60, LDS80 all have concentration dependent, and
It is increased as sample concentration rises, reducing power is bigger, shows that its oxidation resistance is stronger.
Embodiment 3 pilose antler mushroom Thick many candies LDS30, LDS60, LDS80 --- ABTS radicals scavenging Scavenging activity measurement
Measure 8.0mL7mmol/L ABTS solution mixed with the potassium persulfate solution of 141 μ L 140mmol/L after in 4 DEG C
Under be protected from light 12-16h.It is standby to OD=0.70 ± 0.02 with the phosphate buffer dilution stock solution of 10mmol/LpH=7.4
With；
Take 0,0.2,0.4,0.6,0.8,1.0, each 1.0mL of LDS30 of 1.2mg/mL in test tube, 3.0mLABTS is added
Free-atom aqueous solution is protected from light 5min at 30 DEG C after mixing, and light absorption value A is measured at 734nmi；Made with 1.0mL distilled water empty
White control measures light absorption value A under the conditions of0；It uses 3.0mL distilled water to replace ABTS free-atom aqueous solution as sample controls group, surveys
Light absorption value Aj, VCAs positive control；LDS60, LDS80 are ibid operated.ABTS radicals scavenging Scavenging activity formula is as follows:
Ai: sample light absorption value；Aj: 3.0mL distilled water replaces ABTS free-atom aqueous solution to measure light absorption value；A0Blank control is inhaled
Light value.
For the result of measurement as shown in figure 3, within the scope of a certain concentration, the ABTS free radical of LDS30, LDS60, LDS80 are clear
Removing solid capacity is positively correlated with its concentration, shows that pilose antler massee fruiting bodies Thick many candies have preferable ABTS free radical scavenging ability.
Embodiment 4 pilose antler mushroom Thick many candies LDS30, LDS60, LDS80 --- DPPH radicals scavenging Scavenging activity measurement
Take 0,0.2,0.4,0.6,0.8,1.0, each 2.0mL of LDS30 of 1.2mg/mL in test tube, be separately added into
The DPPH- ethanol solution of 1.0mL0.1mmol/L, mixes well, and constant temperature is protected from light 1h under the conditions of being sealed in 25 DEG C,
Light absorption value A is measured at 517nmi；It replaces LDS30 to make blank control with 2.0mL distilled water, surveys light absorption value A under equal conditions0；Sample
Control replaces DPPH- ethanol solution with 1.0mL dehydrated alcohol, surveys light absorption value A under equal conditionsj, VCAs positive control；
LDS60, LDS80 are ibid operated.DPPH radicals scavenging Scavenging activity formula is as follows:
Ai: sample light absorption value；Aj: 1.0mL dehydrated alcohol replaces DPPH- ethanol solution to measure light absorption value；A0: blank
Compare light absorption value
The result of measurement as shown in figure 4, show that component LDS30, LDS60, LDS80 all have antioxidant activity by Fig. 4, and
As the increase antioxidant activity of concentration enhances.
Claims (9)
1. a kind of pilose antler mushroom polyoses extract, which is characterized in that the pilose antler mushroom polyoses extract is prepared as follows to obtain:
(1) it after crushing pilose antler massee fruiting bodies dry product, is added to by solid-liquid ratio 1:3~6 in the aqueous solution of 95% ethyl alcohol of volume fraction,
8~16h is impregnated, filter residue is collected in filtering, and naturally dry obtains pilose antler mushroom defatted seed flour；
(2) take pilose antler mushroom defatted seed flour obtained by step (1) that distilled water is added to carry out high speed shear extraction, it is heavy that extracting solution centrifugation discards
It forms sediment, collects supernatant；
The conditional parameter that the high speed shear is extracted are as follows: solid-liquid ratio 1:15~25,9500~14500rpm of revolving speed, extraction time 1
~4min；
(3) supernatant obtained by step (2) is concentrated into the 1/7~1/12 of original volume, obtains pilose antler mushroom Thick many candies concentrate；
(4) aqueous solution of 95% ethyl alcohol of volume fraction is added into pilose antler mushroom Thick many candies concentrate obtained by step (3) and makes second
Alcohol final concentration reaches 30%~80%, stands 8~16h, centrifugation, and gained precipitating is freeze-dried after removing residual ethanol, obtains institute
State pilose antler mushroom polyoses extract.
2. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (1), the solid-liquid ratio is 1:4.
3. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (1), the time of the immersion is
12h。
4. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (2), what the high speed shear was extracted
Conditional parameter are as follows: solid-liquid ratio 1:25, revolving speed 11500rpm, extraction time 1min.
5. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (3), the supernatant is concentrated into original
The 1/10 of volume.
6. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (4), the time of the standing is
12h。
7. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that in step (4), gained precipitating removal residual second
The method of alcohol are as follows: redissolve precipitating plus water, evaporating solvent under reduced pressure is to remove residual ethanol, and repetitive operation is to without obvious ethyl alcohol smell
?.
8. pilose antler mushroom polyoses extract as described in claim 1, which is characterized in that the operating method of the step (4) are as follows:
The aqueous solution of 95% ethyl alcohol of volume fraction is added into pilose antler mushroom Thick many candies concentrate obtained by step (3) and makes ethyl alcohol whole
Concentration reaches 30%, stands 12h, centrifugation, isolated primary sedimentation and a supernatant；Volume is added into a supernatant
The aqueous solution of 95% ethyl alcohol of score simultaneously makes ethyl alcohol final concentration reach 60%, stands 12h, centrifugation, isolated secondary precipitation and
Secondary supernatant；The aqueous solution of 95% ethyl alcohol of volume fraction is added into secondary supernatant and ethyl alcohol final concentration is reached
80%, 12h is stood, centrifugation is separated and collected and precipitated three times；
By gained primary sedimentation, secondary precipitation, three times precipitate respectively plus water redissolve, evaporating solvent under reduced pressure to remove residual ethanol,
To without obvious ethyl alcohol smell, freeze-drying respectively obtains three kinds of pilose antler mushroom polyoses extracts for repetitive operation.
9. pilose antler mushroom polyoses extract as described in claim 1 is preparing the application in antioxidant.
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| CN110218264A (en) * | 2019-06-26 | 2019-09-10 | 西华师范大学 | A kind of Lyophyllum decastes granulose and its preparation method and application |
| CN110218264B (en) * | 2019-06-26 | 2021-04-23 | 西华师范大学 | Lyophyllum nuciferum polysaccharide and preparation method and application thereof |
| CN111825778A (en) * | 2020-07-22 | 2020-10-27 | 贵州省贵福菌业发展有限公司 | Method for extracting polysaccharide and preparing velvet antler mushroom cultivation base material by taking morchella esculenta dregs as raw material |
| CN113061194A (en) * | 2021-03-09 | 2021-07-02 | 中国科学院微生物研究所 | Preparation method and application of Lyophyllum decastes fruiting body polysaccharide |
| CN113061194B (en) * | 2021-03-09 | 2022-03-18 | 中国科学院微生物研究所 | Preparation method and application of Lyophyllum decastes fruiting body polysaccharide |
| CN113100368A (en) * | 2021-04-29 | 2021-07-13 | 安徽大学 | Antioxidant lotus leaf beverage and preparation method thereof |
| CN115975068A (en) * | 2022-12-26 | 2023-04-18 | 上海市农业科学院 | Velvet antler mushroom polysaccharide rich in beta-glucan and preparation method thereof |
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Application publication date: 20190111 |