CN107337741A - A kind of preparation method and application of beautiful flower bud Shorthairy Antenoron polyoses extract - Google Patents
A kind of preparation method and application of beautiful flower bud Shorthairy Antenoron polyoses extract Download PDFInfo
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- CN107337741A CN107337741A CN201710460111.4A CN201710460111A CN107337741A CN 107337741 A CN107337741 A CN 107337741A CN 201710460111 A CN201710460111 A CN 201710460111A CN 107337741 A CN107337741 A CN 107337741A
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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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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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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- 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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
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Abstract
The present invention relates to a kind of preparation method of beautiful flower bud Shorthairy Antenoron polyoses extract, after preparation method includes raw material degreasing, precipitate to obtain Thick many candies by water extraction, alcohol again, Thick many candies slough albumen, pigment by dialysis, ion-exchange chromatography, small-molecule chemical composition is removed, the beautiful flower bud Shorthairy Antenoron polyoses extract of higher degree is made.The content of beautiful flower bud Shorthairy Antenoron polysaccharide accounts for beautiful flower bud Shorthairy Antenoron extract weight percentage more than 74% with glucose meter, the content of the polysaccharide in extract of the present invention.
Description
Technical field
The present invention relates to pharmaceutical technology field, and in particular to a kind of beautiful flower bud Shorthairy Antenoron Polyose extraction with antifatigue effect
The preparation method and its usage of thing.
Background technology
Fatigue is a kind of common physiological phenomenon, and its process causes organism physiology biochemical function to change for excessively work or motion
Become and cause body work or locomitivity temporary transient reduction, actually fatigue be prevent body occur failure function and it is caused
A kind of aversion response.Fatigue can cause a series of consequences such as lassitude, absent minded, resistance decline, seriously
Operating efficiency or sports achievement are influenceed, therefore research delays or the measure of dispelling fatigue tool is of great significance.Existing market
It is upper that there is certain effect in terms of delay fatigue and dispelling fatigue for antifatigue chemicals, but long-term use of, tool
There is certain additive and dependence, therefore limit its clinical practice.Therefore, extracted from natural plant resource safely, have
Effect, the antifatigue composition without harmful composition have turned into the study hotspot of anti-fatigue medicament in recent years.
Beautiful flower bud Shorthairy Antenoron is the beautiful flower bud Shorthairy Antenoron of orchid family Anoectochilus Blume plantAnoectochilus lylei Rolfe ex
Downie fresh or dry herb.The existing research on beautiful flower bud Shorthairy Antenoron is less, has no that it has the phase of antifatigue effect
Close report.
The content of the invention
One of the object of the invention is to provide a kind of beautiful flower bud Shorthairy Antenoron for having definite anti-fatigue active, having no toxic side effect
The preparation method of polyoses extract.
The second object of the present invention be by above-mentioned beautiful flower bud Shorthairy Antenoron polyoses extract be made various anti-fatigue medicament preparations or
Prepare the bulk drug of other anti-fatigue medicaments.
The above-mentioned purpose of the present invention is achieved through the following technical solutions：
A kind of preparation method of beautiful flower bud Shorthairy Antenoron polyoses extract, after beautiful flower bud Shorthairy Antenoron raw medicinal material degreasing, then extract by water,
Alcohol precipitates to obtain Thick many candies, and Thick many candies slough albumen, pigment by dialysis, ion-exchange chromatography, remove small-molecule chemical composition,
The beautiful flower bud Shorthairy Antenoron polyoses extract of higher degree is made.
It includes step in detail below：
（1）Beautiful flower bud Shorthairy Antenoron medicinal material is flowed back with high concentration ethanol and removes oil-soluble impurities；
（2）Residue adds water heating extracting, filtrate concentration, adds ethanol alcohol precipitation, centrifuging to precipitate；
（3）Precipitation is washed respectively with absolute ethyl alcohol, acetone successively；
（4）Precipitation after washing is dialysed after being dissolved with distilled water；
（5）Dialyzate is concentrated into certain volume, and pigment and protein in polysaccharide are sloughed using cation exchange resin processes；
（6）By the eluent concentration except depigmentaton and protein, ethanol alcohol precipitation is added, centrifuges to obtain polysaccharide precipitation；
（7）Polysaccharide precipitation is washed respectively with absolute ethyl alcohol, acetone successively, is dried, is produced beautiful flower bud Shorthairy Antenoron polyoses extract.
In order to reach more preferable extraction effect, it is preferred that
Step（1）Middle 80~95% alcohol reflux degreasing 2~3 times by beautiful flower bud Shorthairy Antenoron with 10~14 times of weight, every time backflow
1.5~3h；
Step（2）The middle distillation water heating extracting 2~3 times that residue is added to 10~16 times of weight, extracts 2~3h every time, and merging carries
Liquid is taken, extract solution is concentrated, the volume mass ratio of concentrate and crude drug is 1~3：1, add 95% ethanol alcohol and be sink to alcohol content
For 80%, 12h is stood, precipitation must be collected by centrifuging；
Precipitation is washed respectively with the absolute ethyl alcohol of 2~4 times of weight, acetone successively in step (3), (7)；
Precipitation in step (4) is redissolved with the distilled water of 10~50 times of weight, with the dialysis of retention relative molecular weight more than 3500
24~72h of bag dialysis.
Polysaccharide dialyzate was collected into post liquid, then by JK008 types, 732 type cationic ion-exchange resins in step (5)
With the distillation water elution of 3 times of column volumes, flow velocity is 2 BV/h.
Step（6）Eluent is concentrated, the volume mass ratio of concentrate and crude drug is 1~3：1, add 95% ethanol alcohol
It is 80% to be sink to alcohol content, stands 12h, centrifuging to precipitate.
Precipitation uses absolute ethyl alcohol successively, and acetone washs respectively, dries, and the beautiful flower bud Shorthairy Antenoron polysaccharide for obtaining de- albumen and pigment carries
Take thing.
Monose constitutive molar ratio is mannose in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention：Galacturonic acid：Glucose：
Galactolipin：Arabinose=0.15:0.06:0.78:0.38:0.10, the content of polysaccharide is with glucose meter, the beautiful flower bud Shorthairy Antenoron
The content of total starches accounts for more than the 74% of extract weight percentage in polyoses extract.
The advantage of the invention is that：
The beautiful flower bud Shorthairy Antenoron polyoses extract of this purifying shows significantly extend the mice burden swimming time through pharmacodynamic experiment, dropped
Serum urea nitrogen, serum lactic content after low mouse swimming, improve oxidation resistance of the mouse under stress situation, improve glycogen
Former and muscle glycogen content, has obvious antifatigue effect.
Brief description of the drawings
Fig. 1 is according to the efficient of the monosaccharide derivatives of the beautiful flower bud Shorthairy Antenoron polyoses extract of an embodiment of the invention
Liquid chromatogram（HPLC）Figure.
Embodiment
The invention will now be further described with reference to specific embodiments, advantages of the present invention and feature will be with description and
It is apparent.But these embodiments are only exemplary, do not form any restrictions to the scope of the present invention.Art technology
Personnel should be understood that without departing from the spirit and scope of the invention can be to the details and form of technical solution of the present invention
Modify or replace, but these modifications and replacement are each fallen within protection scope of the present invention.
Embodiment 1
Beautiful flower bud Shorthairy Antenoron medicinal material 2kg is taken, with the ethanol heating and refluxing extraction 3 times, each 2h of 12 times of weight 80%, filtering, by filter residue
Drying, residue adds the distillation water heating extracting 3 times of 12 times of weight, every time 2 h, filtering, merges extract solution, extract solution is concentrated,
The volume mass of concentrate and crude drug ratio is 2：1, ethanol to the alcohol content for adding 95% is 80%, stands 12 h, centrifuges, receives
Collection precipitation, precipitation is measured into absolute ethyl alcohol, acetone with 4 times and washed respectively, then by drying precipitate, produce beautiful flower bud Shorthairy Antenoron Thick many candies
Extract.Beautiful flower bud Shorthairy Antenoron Thick many candies are dissolved in water, the bag filter that supernatant retention relative molecular weight is more than 3500 is saturating
48h is analysed, then by dialyzate by JK008 type cationic ion-exchange resins, with the distillation water elution of 3 times of column volumes, flow velocity 2
BV/h.Water elution is collected, eluent is concentrated, the volume mass ratio of concentrate and crude drug is 2：1, add 95% ethanol
It is 80% to alcohol content, stands 12 h, precipitation is collected in centrifugation, and precipitation is washed, done respectively with 4 times of amount absolute ethyl alcohols, acetone successively
It is dry, obtain the beautiful flower bud Shorthairy Antenoron polyoses extract of de- albumen and pigment.Polysaccharide contains in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention
Amount accounts for beautiful flower bud Shorthairy Antenoron polyoses extract percentage by weight as 78.38% with glucose meter, the content of the polysaccharide.
Embodiment 2
Beautiful flower bud Shorthairy Antenoron medicinal material 2kg is taken, with the ethanol heating and refluxing extraction 3 times, each 2h of 10 times of amounts 95%, filtering, filter residue is dried in the air
Dry, residue adds the distillation water heating extracting 3 times of 10 times of amounts, and 2 h, filtering, merges extract solution, extract solution is concentrated every time, concentrates
The volume mass of liquid and crude drug ratio is 1：1, ethanol to the alcohol content for adding 95% is 80%, stands 12 h, centrifugation, it is heavy to collect
Form sediment, precipitation is measured into absolute ethyl alcohol, acetone with 4 times and washed, then by drying precipitate, produce beautiful flower bud Shorthairy Antenoron crude polysaccharide extract.
Beautiful flower bud Shorthairy Antenoron Thick many candies are dissolved in water, the bag filter dialysis 48h that supernatant retention relative molecular weight is more than 3500, so
Afterwards by dialyzate by 732 type cationic ion-exchange resins, with the distillation water elution of 3 times of column volumes, flow velocity is 2 BV/h.Collect water
Eluent, eluent is concentrated, the volume mass ratio of concentrate and crude drug is 1：1, ethanol to the alcohol content for adding 95% is
80%, 12 h are stood, precipitation is collected in centrifugation, and precipitation is washed with 4 times of amount absolute ethyl alcohols, acetone, dries, obtain de- albumen and color successively
The beautiful flower bud Shorthairy Antenoron polyoses extract of element.The content of polysaccharide is with glucose meter, institute in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention
The content for stating polysaccharide accounts for beautiful flower bud Shorthairy Antenoron polyoses extract weight percent as 74.42%.
The beautiful flower bud Shorthairy Antenoron polyoses extract of embodiment 3（Extract in embodiment 1）Antifatigue experiment
Male ICR mouse is taken, body weight 18-22g, is randomly divided into blank control group, gadol extract group（Positive drug group）, group
Beautiful flower bud Shorthairy Antenoron polyoses extract low dose group, beautiful flower bud Shorthairy Antenoron polyoses extract middle dose group, beautiful flower bud Shorthairy Antenoron Polyose extraction
Thing group, every group of 8 mouse.After successive administration 14 d, last dose 1h, each group mouse carries out swimming with a load attached to the body power and exhausts experiment respectively.
Each load in mouse tail root its body weight 10% galvanized wire as weight, be put into glass jar, depth of water 35cm, water temperature (25 ±
0.5) DEG C, ensure that mouse can not use tail contact bottom in water.When can not be emerged in mouse head submerged, 10s
Time, this time is mouse swimming time.Taken a blood sample after 20 min of swimming rest are stopped, according to kit specification
Determine mice serum urea nitrogen（BUN）, lactic acid（BLA）Content；Mouse liver and gastrocnemius are taken, liver is determined by kit specification
Dirty superoxide dismutase（SOD）, MDA（MDA）, hepatic glycogen and gastrocnemius muscle glycogen content.Experimental result is shown in Table 1,
Table 2.
Influence of the beautiful flower bud Shorthairy Antenoron polyoses extract of table 1 to mouse swimming time, serum BUN and LDH（N=8,）
Note：*p<0.05, * *p<0.01, compared with blank control group
Influence of the beautiful flower bud Shorthairy Antenoron polyoses extract of table 2 to mouse liver SOD, MDA, hepatic glycogen and muscle glycogen（N=8,）
Note：*p<0.05, * *p<0.01, compared with blank control group
Experimental result：From table 1, table 2, beautiful flower bud Shorthairy Antenoron polyoses extract low dosage and high dose can significantly extend mouse and bear
Weight swimming time, reduces BUN the and LDH contents in mice serum, improves SOD contents in hepatic tissue, reduces MDA water in hepatic tissue
It is flat, increase hepatic glycogen and muscle glycogen content, it is indicated above that beautiful flower bud Shorthairy Antenoron polyoses extract is with significantly anti-tired in the present invention
Work is used.
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, it should all belong to the covering scope of the present invention.
Claims (5)
- A kind of 1. beautiful flower bud Shorthairy Antenoron polyoses extract, it is characterised in that：The monose constitutive molar ratio of the polyoses extract is sweet dew Sugar：Galacturonic acid：Glucose：Galactolipin：Arabinose=0.15:0.06:0.78:0.38:0.10, the beautiful flower bud Shorthairy Antenoron The content of total starches accounts for more than the 74% of extract weight percentage in polyoses extract.
- A kind of 2. preparation method of beautiful flower bud Shorthairy Antenoron polyoses extract as claimed in claim 1, it is characterised in that：Beautiful flower bud gold thread After blue raw medicinal material degreasing, then extract by water, alcohol precipitates to obtain Thick many candies, Thick many candies take off by dialysis, ion-exchange chromatography Deproteinized, pigment, small-molecule chemical composition is removed, beautiful flower bud Shorthairy Antenoron polyoses extract is made.
- 3. the preparation method of beautiful flower bud Shorthairy Antenoron polyoses extract according to claim 2, it is characterised in that：Including following step Suddenly：（1）Beautiful flower bud Shorthairy Antenoron medicinal material is flowed back with high concentration ethanol and removes oil-soluble impurities；（2）Residue adds water heating extracting, filtrate concentration, adds ethanol alcohol precipitation, centrifuging to precipitate；（3）Precipitation is washed respectively with absolute ethyl alcohol, acetone successively；（4）Precipitation after washing is dialysed after being dissolved with distilled water；（5）Dialyzate is concentrated into certain volume, is sloughed using weak acid and weak base type anion and cation exchange resin series process in polysaccharide Pigment and protein；（6）By the eluent concentration except depigmentaton and protein, ethanol alcohol precipitation is added, centrifuges to obtain polysaccharide precipitation；（7）Polysaccharide precipitation uses absolute ethyl alcohol successively, and acetone washs respectively, produces beautiful flower bud Shorthairy Antenoron polyoses extract.
- 4. according to the method for claim 3, it is characterised in that：Step（1）It is middle by beautiful flower bud Shorthairy Antenoron with 10~14 times of weight 80~95% alcohol reflux degreasing 2~3 times, flow back 1.5~3h every time；Step（2）It is middle that residue is added into 10~16 times of weight Water heating extracting 2~3 times is distilled, extracts 2~3h every time, merges extract solution, extract solution is concentrated, the body of concentrate and crude drug Product mass ratio is 1~3：1, it is 80% to add 95% ethanol alcohol and be sink to alcohol content, and after standing 12h, centrifuging to precipitate；Step (3), （7）It is middle to wash precipitation respectively with the absolute ethyl alcohol of 2~4 times of weight, acetone successively；Step（4）In precipitation with 10~50 times The distilled water of weight redissolves, with 24~72h of bag filter dialysis of retention relative molecular weight more than 3500；By polysaccharide in step (5) Dialyzate collected post liquid, and was then 2 BV/h with the distillation water elution of 3 times of column volumes, flow velocity by cationic ion-exchange resin； Step（6）Eluent is concentrated, the volume mass ratio of concentrate and crude drug is 1~3：1, add 95% ethanol alcohol and be sink to containing alcohol Measure as 80%, after standing 12h, centrifuging to precipitate.
- 5. application of the beautiful flower bud Shorthairy Antenoron polyoses extract in anti-fatigue medicament is prepared as claimed in claim 1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111603538A (en) * | 2020-06-30 | 2020-09-01 | 华侨大学 | Application and preparation method of anoectochilus formosanus extract |
| CN112843177A (en) * | 2021-02-24 | 2021-05-28 | 中国人民解放军海军军医大学 | Application of anoectochilus formosanus extract in preparation of medicine or health-care product for treating or relieving pain |
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Cited By (3)
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| CN112843177A (en) * | 2021-02-24 | 2021-05-28 | 中国人民解放军海军军医大学 | Application of anoectochilus formosanus extract in preparation of medicine or health-care product for treating or relieving pain |
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