CN107337741B - 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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- CN107337741B CN107337741B CN201710460111.4A CN201710460111A CN107337741B CN 107337741 B CN107337741 B CN 107337741B CN 201710460111 A CN201710460111 A CN 201710460111A CN 107337741 B CN107337741 B CN 107337741B
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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 methods of beautiful flower bud Shorthairy Antenoron polyoses extract, after preparation method includes raw material degreasing, it is extracted using water, alcohol precipitates to obtain Thick many candies, Thick many candies slough albumen, pigment by dialysis, ion-exchange chromatography, small-molecule chemical ingredient is removed, the beautiful flower bud Shorthairy Antenoron polyoses extract of higher degree is made.The content of beautiful flower bud Shorthairy Antenoron polysaccharide is in extract of the present invention with glucose meter, 74% or more the content Zhan Lilei Shorthairy Antenoron extract weight percentage of the polysaccharide.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to a kind of beautiful flower bud Shorthairy Antenoron Polyose extraction with antifatigue effect
The preparation method and its usage of object.
Background technique
Fatigue is a kind of common physiological phenomenon, and process is that excessively labour or movement causes organism physiology biochemical function to change
Become and lead to body labour or the temporary reduction of locomitivity, actually fatigue prevents body from failure function occurs and generates
A kind of aversion response.Fatigue can lead to a series of consequences such as lassitude, absent minded, resistance decline, seriously
Working efficiency or sports achievement are influenced, therefore the measure that research delays or eliminate fatigue has a very important significance.Existing market
It is upper to have the effect of certain, but long-term use for antifatigue chemicals in delay fatigue and in terms of eliminating fatigue, have
There is certain additive and dependence, therefore limits its clinical application.Therefore, extracted from natural plant resource safely, have
Effect, the research hotspot that anti-fatigue medicament in recent years is had become without the antifatigue ingredient for the ingredient that is harmful to the human body.
Beautiful flower bud Shorthairy Antenoron is the beautiful flower bud Shorthairy Antenoron of orchid family Anoectochilus Blume plantAnoectochilus lylei Rolfe ex
The fresh or dry herb of Downie.The existing research about beautiful flower bud Shorthairy Antenoron is less, has no its phase with antifatigue effect
Close report.
Summary of the invention
One of the object of the invention is to provide a kind of with exact anti-fatigue active, the beautiful flower bud Shorthairy Antenoron having no toxic side effect
The preparation method of polyoses extract.
The second object of the present invention is to by above-mentioned beautiful flower bud Shorthairy Antenoron polyoses extract be made various anti-fatigue medicament preparations or
Prepare the bulk pharmaceutical chemicals of other anti-fatigue medicaments.
Above-mentioned purpose of the 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, using water
Extraction, alcohol precipitate to obtain Thick many candies, and Thick many candies slough albumen, pigment by dialysis, ion-exchange chromatography, remove small molecule
It studies point, the beautiful flower bud Shorthairy Antenoron polyoses extract of higher degree is made.
Itself comprising the following specific steps
(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, and ethyl alcohol alcohol precipitation is added in filtrate concentration, is centrifuged to obtain precipitating；
(3) precipitating is successively washed with dehydrated alcohol, acetone respectively；
(4) precipitating 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) it will be concentrated except the eluent of depigmentaton and protein, ethyl alcohol alcohol precipitation be added, is centrifuged to obtain polysaccharide precipitation；
(7) polysaccharide precipitation is successively washed with dehydrated alcohol, acetone respectively, dry to get beautiful flower bud Shorthairy Antenoron polyoses extract.
In order to reach better extraction effect, it is preferred that
By 80~95% alcohol reflux degreasing 2~3 times of 10~14 times of weight of beautiful flower bud Shorthairy Antenoron in step (1), every time
Flow back 1.5~3h；
Residue is added to distillation water heating extracting 2~3 times of 10~16 times of weight in step (2), extracts 2~3h every time, is closed
And extracting solution, extracting solution is concentrated, the volume mass ratio of concentrate and crude drug is 1~3:1, and 95% ethyl alcohol alcohol of addition, which is sink to, to be contained
Alcohol amount is 80%, stands 12h, is centrifuged to collect precipitating；
Precipitating is successively washed with the dehydrated alcohol of 2~4 times of weight, acetone respectively in step (3), (7)；
Precipitating in step (4) is redissolved with the distilled water of 10~50 times of weight, with 3500 or more relative molecular weight of retention
24~72h of bag filter dialysis.
Polysaccharide dialyzate was collected into column liquid, then by JK008 type, 732 type cation exchange resins in step (5)
With the distillation water elution of 3 times of column volumes, flow velocity is 2 BV/h.
Eluent is concentrated step (6), and the volume mass ratio of concentrate and crude drug is 1~3:1, and 95% ethyl alcohol alcohol is added
Being sink to alcohol content is 80%, stands 12h, is centrifuged to obtain precipitating.
Precipitating successively uses dehydrated alcohol, and acetone washs respectively, dry, and the beautiful flower bud Shorthairy Antenoron polysaccharide for obtaining de- albumen and pigment mentions
Take object.
Monosaccharide constitutive molar ratio is mannose: galacturonic acid: glucose in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention:
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 74% or more of extract weight percentage in polyoses extract.
The present invention has the advantages that
When the beautiful flower bud Shorthairy Antenoron polyoses extract of this purifying shows significantly extend mice burden swimming through pharmacodynamic experiment
Between, serum urea nitrogen, serum lactic content after mouse swimming are reduced, oxidation resistance of the mouse under stress situation is improved, mentions
High hepatic glycogen and muscle glycogen content have apparent antifatigue effect.
Detailed description of the invention
Fig. 1 is the efficient of the monosaccharide derivatives of beautiful flower bud Shorthairy Antenoron polyoses extract according to embodiment of the present invention
Liquid chromatogram (HPLC) figure.
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent.But examples are merely exemplary for these, and it is not intended to limit the scope of the present invention in any way.Art technology
Personnel should be understood that without departing from the spirit and scope of the invention can details and form to technical solution of the present invention
It modifies or replaces, but these modifications and replacement are fallen within the protection scope of the present invention.
Embodiment 1
Beautiful 2 kg of flower bud Shorthairy Antenoron medicinal material is taken, with ethyl alcohol heating and refluxing extraction 3 times of 12 times of weight 80%, 2 h, is filtered every time,
Filter residue is dried, residue adds distillation water heating extracting 3 times of 12 times of weight, every time 2 h, filtering, combined extract, by extracting solution
The volume mass ratio of concentration, concentrate and crude drug is 2:1, and ethyl alcohol to the alcohol content for being added 95% is 80%, stands 12 h, from
The heart collects precipitating, precipitating is washed respectively with 4 times of amount dehydrated alcohols, acetone, then by drying precipitate to get beautiful flower bud Shorthairy Antenoron
Crude polysaccharide extract.Beautiful flower bud Shorthairy Antenoron Thick many candies are dissolved in water, supernatant retain relative molecular weight be 3500 or more it is saturating
Bag dialysis 48h is analysed, then by dialyzate by JK008 type cation exchange resin, with the distillation water elution of 3 times of column volumes, stream
Speed is 2 BV/h.Water elution is collected, eluent is concentrated, the volume mass ratio of concentrate and crude drug is 2:1, is added 95%
Ethyl alcohol to alcohol content be 80%, stand 12 h, centrifugation, collect precipitating, precipitating successively washed respectively with 4 times of amount dehydrated alcohols, acetone
It washs, it is dry, obtain the beautiful flower bud Shorthairy Antenoron polyoses extract of de- albumen and pigment.Polysaccharide in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention
Content with glucose meter, the content Zhan Lilei Shorthairy Antenoron polyoses extract weight percent of the polysaccharide is 78.38%.
Embodiment 2
Beautiful 2 kg of flower bud Shorthairy Antenoron medicinal material is taken, with ethyl alcohol heating and refluxing extraction 3 times of 10 times of amounts 95%, 2 h, is filtered every time, will
Filter residue dries, and residue adds distillation water heating extracting 3 times of 10 times of amounts, every time 2 h, filtering, and combined extract is dense by extracting solution
The volume mass ratio of contracting, concentrate and crude drug is 1:1, and ethyl alcohol to the alcohol content for being added 95% is 80%, stands 12 h, is centrifuged,
Precipitating is collected, precipitating is mentioned with 4 times of amounts dehydrated alcohols, acetone washing, then by drying precipitate to get beautiful flower bud Shorthairy Antenoron Thick many candies
Take object.Beautiful flower bud Shorthairy Antenoron Thick many candies are dissolved in water, supernatant retains the bag filter that relative molecular weight is 3500 or more and dialyses
48h, then by dialyzate by 732 type cation exchange resins, with the distillation water elution of 3 times of column volumes, flow velocity is 2 BV/h.
Water elution is collected, eluent is concentrated, the volume mass ratio of concentrate and crude drug is 1:1, and 95% ethyl alcohol is added to alcohol-containing
Amount is 80%, stands 12 h, and precipitating is collected in centrifugation, and precipitating is dry successively with 4 times of amounts dehydrated alcohols, acetone washing, obtains de- albumen
With the beautiful flower bud Shorthairy Antenoron polyoses extract of pigment.The content of polysaccharide is in the beautiful flower bud Shorthairy Antenoron polyoses extract of the present invention with glucose
Meter, the content Zhan Lilei Shorthairy Antenoron polyoses extract weight percent of the polysaccharide are 74.42%.
The antifatigue experiment of 3 beautiful flower bud Shorthairy Antenoron polyoses extract (extract in embodiment 1) of embodiment
Male ICR mouse is taken, weight 18-22g is randomly divided into blank control group, gadol extract group (positive drug
Group), the beautiful flower bud Shorthairy Antenoron polyoses extract low dose group of group, beautiful flower bud Shorthairy Antenoron polyoses extract middle dose group, beautiful flower bud Shorthairy Antenoron it is more
Sugar extract 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 respectively and exhausts
Experiment.Each load in mouse tail root its weight 10% galvanized wire as weight, be put into glass jar, depth of water 35cm, water temperature (25
± 0.5) DEG C, guarantee that mouse cannot use tail to contact bottom in water.When being unable to emersion water in mouse head submerged, 10s
The time in face, this time are mouse swimming time.It takes a blood sample after stopping 20 min of swimming rest, illustrates according to kit
Book measures mice serum urea nitrogen (BUN), lactic acid (BLA) content；Mouse liver and gastrocnemius are taken, is measured by kit specification
Liver Superoxide Dismutase Activity (SOD), malonaldehyde (MDA), the content of hepatic glycogen and gastrocnemius muscle glycogen.Experimental result is shown in Table
1, table 2.
The beautiful flower bud Shorthairy Antenoron polyoses extract of table 1 to mouse swimming time, serum BUN and LDH influence (n=8,)
Note: *p< 0.05, * *p< 0.01, compared with blank control group
The beautiful flower bud Shorthairy Antenoron polyoses extract of table 2 to mouse liver SOD, MDA, hepatic glycogen and muscle glycogen influence (n=8,)
Note: *p< 0.05, * *p< 0.01, compared with blank control group
Experimental result: by table 1, table 2 it is found that beautiful flower bud Shorthairy Antenoron polyoses extract low dosage and high dose can significantly extend it is small
Mouse walking weight load reduces BUN the and LDH content in mice serum, improves SOD content in hepatic tissue, reduces in hepatic tissue
MDA is horizontal, increases hepatic glycogen and muscle glycogen content, it is indicated above that beautiful flower bud Shorthairy Antenoron polyoses extract has obviously in the present invention
Antifatigue effect.
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
Claims (5)
1. a kind of beautiful flower bud Shorthairy Antenoron polyoses extract, it is characterised in that: the monosaccharide 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 74% or more of extract weight percentage in polyoses extract.
2. a kind of preparation method of beautiful flower bud Shorthairy Antenoron polyoses extract as described in claim 1, it is characterised in that: beautiful flower bud gold thread
After blue raw medicinal material degreasing, precipitate to obtain Thick many candies using water extraction, alcohol, Thick many candies are de- by dialysis, ion-exchange chromatography
Deproteinized, pigment remove small-molecule chemical ingredient, beautiful flower bud Shorthairy Antenoron polyoses extract are 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
It is rapid:
(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, and ethyl alcohol alcohol precipitation is added in filtrate concentration, is centrifuged to obtain precipitating；
(3) precipitating is successively washed with dehydrated alcohol, acetone respectively；
(4) precipitating 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；
(6) it will be concentrated except the eluent of depigmentaton and protein, ethyl alcohol alcohol precipitation be added, is centrifuged to obtain polysaccharide precipitation；
(7) polysaccharide precipitation successively uses dehydrated alcohol, and acetone is washed respectively to get beautiful flower bud Shorthairy Antenoron polyoses extract.
4. according to the method described in claim 3, it is characterized by: by 10~14 times of weight of beautiful flower bud Shorthairy Antenoron in step (1)
80~95% alcohol reflux degreasing 2~3 times, every time flow back 1.5~3h；Residue is added into 10~16 times of weight in step (2)
2~3h is extracted in distillation water heating extracting 2~3 times every time, and extracting solution is concentrated combined extract, the body of concentrate and crude drug
Product mass ratio is 1~3:1, and it is 80% that 95% ethyl alcohol alcohol, which is added, and is sink to alcohol content, after standing 12h, is centrifuged to obtain precipitating；Step (3),
(7) precipitating is successively washed with the dehydrated alcohol of 2~4 times of weight, acetone respectively in；Precipitating in step (4) is with 10~50 times
The distilled water of weight redissolves, with 24~72h of bag filter dialysis of 3500 or more relative molecular weight of retention；By polysaccharide in step (5)
Dialyzate collected column liquid by cation exchange resin, and then with the distillation water elution of 3 times of column volumes, flow velocity is 2 BV/h,
Collect water elution；Eluent is concentrated step (6), and the volume mass ratio of concentrate and crude drug is 1~3:1, is added 95%
It is 80% that ethyl alcohol alcohol, which is sink to alcohol content, after standing 12h, is centrifuged to obtain precipitating.
5. beautiful flower bud Shorthairy Antenoron polyoses extract as described in claim 1 is preparing the application in anti-fatigue medicament.
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| CN111603538B (en) * | 2020-06-30 | 2021-04-27 | 华侨大学 | Application 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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| CN101580554B (en) * | 2009-07-01 | 2011-08-03 | 韩金光 | Anoectochilus roxburghii polyose, applications thereof in pharmacy and preparation method of anoectochilus roxburghii polyose |
| CN105315381A (en) * | 2014-07-22 | 2016-02-10 | 漳州市亚热带园艺技术有限公司 | Anoectochilus roxburghii polysaccharide extraction and preliminary purifying method and preparation method for oral liquid of anoectochilus roxburghii polysaccharide |
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