CN110483656A - A kind of preparation of radix tetrastigme polysaccharide antibacterial bacteriostatic mouthwash - Google Patents
A kind of preparation of radix tetrastigme polysaccharide antibacterial bacteriostatic mouthwash Download PDFInfo
- Publication number
- CN110483656A CN110483656A CN201910850486.0A CN201910850486A CN110483656A CN 110483656 A CN110483656 A CN 110483656A CN 201910850486 A CN201910850486 A CN 201910850486A CN 110483656 A CN110483656 A CN 110483656A
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- CN
- China
- Prior art keywords
- radix tetrastigme
- polysaccharide
- ultrapure water
- chloroform
- antibacterial
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q11/00—Preparations for care of the teeth, of the oral cavity or of dentures; Dentifrices, e.g. toothpastes; Mouth rinses
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
Abstract
The invention belongs to develop the application of the antibacterial mouthwash of natural products, and in particular to and it has arrived Chinese herbal medicine radix tetrastigme polysaccharide and has prepared antibacterial medicines, the application in mouthrinse product.It using radix tetrastigme aerial part as raw material, extracts in accordance with the following steps: (1) using 50 DEG C of extraction 5h of ethyl alcohol, discarding supernatant liquid, using 50 DEG C of extraction 5h of ultrapure water, separate to obtain supernatant, 65 DEG C of concentrations are added 95% ethyl alcohol and stay overnight, precipitated.(2) it is dissolved and is precipitated with ultrapure water, with water: chloroform: the mixing of n-butanol=8:4:1 ratio is stirred the 20 minutes protein and lipid removed in mixture, repeated to no precipitating；(3) ultrapure water is dialysed two days, and radix tetrastigme polysaccharide is lyophilized to obtain.Radix tetrastigme polysaccharide of the present invention, can significantly kill bacterium on of short duration drug treating time, and maintain bacterium that can not be proliferated for a long time, have good antibacterial and bacteriostasis, be a to be suitably applied in quick sterilization, drug antibacterial for a long time.
Description
Technical field
The invention belongs to herbal plant resource development and utilization, and in particular to arrive a kind of radix tetrastigme polysaccharide antibacterial and bacteriostasis
Purposes.
Background technique
Radix tetrastigme (Tetrastigma hemsleyanum DielsEt Gilg) also known as tetratigma hemsleyanum also known as gold thread
Hoist, hemsley rockvine root are grown on the areas such as dark and damp hillside, mountain valley, are mainly distributed on China Zhejiang, Fujian, Guangxi, Yunnan etc.
Ground.Radix tetrastigme herb can be used as medicine, and be famous one of the Chinese herbal medicine in China, have significant medical value, such as treat viral
Meningitis, bronchitis, children with high fever induced convulsions, prevention and treating cancer etc..
Oral hygiene focuses on controlling bacterial plaque, elimination dirt and swill, enhances physiological stimulation, makes oral cavity and tooth
Jaw system has the good environment of a cleaning health, to play its physiological function, safeguards oral health.Due to current diet
Diversified and some undesirable growths for being accustomed to easily leading to oral bacteria, will lead to dental plaque, saprodontia, the diseases such as canker sore
Disease, especially saprodontia are listed in the main disease of third place in the world kind, are only second to coronary heart disease and cancer, and many sufferers are to tooth
The ignorance of disease leads to have to when treatment of late stage cutting off dental nerve or whole tooth is pulled out.The people's badly damaged in this way
Dental health.
The present invention compared radix tetrastigme polysaccharide to Gram-negative bacteria, Escherichia coli (E.coli) and gram-positive bacteria,
The inhibitory effect of staphylococcus aureus (S.aureus), it is desirable to provide one kind is preparing anti-inflammatory, antibacterial medicine based on radix tetrastigme polysaccharide
Application in object.
Summary of the invention
Due to the diversified and some undesirable growth for being accustomed to easily leading to oral bacteria of current diet, tooth will lead to
Plaque, saprodontia, the diseases such as canker sore, especially saprodontia are listed in the main disease of third place in the world kind, be only second to coronary heart disease and
Cancer, and many sufferers lead to have to when treatment of late stage cutting off dental nerve or whole tooth to the ignorance of odontopathy
It pulls out.The dental health of the people badly damaged in this way.And the mouthwash being prepared at present using natural plant extracts as raw material
Bacteria breed can not often be inhibited in a short time.
In order to solve the above technical problems, the present invention can be realized by following steps:
A kind of preparation method of radix tetrastigme polysaccharide, specifically includes the following steps:
(1) the radix tetrastigme powder 250g for weighing 60 dry mesh is extracted 5 hours with 50 DEG C of 1L dehydrated alcohol, discards supernatant liquid；
It is extracted 5 hours with 50 DEG C of 1L ultrapure water, suction filtration obtains filtrate, and the 1/4 of original volume is concentrated under 65 DEG C of bath temperatures；It is added 4
95% ethyl alcohol of times volume precipitates left undisturbed overnight；5000 revs/min of centrifugation collection in 20 minutes precipitatings, obtain slightly more after freeze-dried
Sugar；
(2) radix tetrastigme Thick many candies 20g is weighed, is dissolved in 1000mL ultrapure water, after ultrasonic dissolution is complete, is proportionally added into chloroform
And n-butanol, 20min is sufficiently stirred, 5000r/min is centrifuged 20 min, discards organic layer and albumin layer, in triplicate, until nothing
Until protein white precipitate generates；
(3) using 8000-14000Da bag filter ultrapure water dialysis 48h, freeze-drying obtains radix tetrastigme essence polysaccharide.
The adding proportion of step (2) chloroform and n-butanol is to count by volume: polysaccharide: chloroform: n-butanol=8:4:
1。
Further, the above-mentioned radix tetrastigme polysaccharide being prepared is applied to preparation to inhibit in Dental Plaque mouthwash, system
Preparation Method are as follows: the mouthwash of final concentration of 0.05-2.0wt% is configured to distilled water.
The present invention has the advantages that radix tetrastigme polysaccharide of the present invention, can show on of short duration drug treating time
The kill bacterium of work, and maintain bacterium that can not be proliferated for a long time, there is good antibacterial and bacteriostasis, be a suitable application
In quick sterilization, drug antibacterial for a long time, applied to can effectively inhibit bacterial growth in mouthwash, in shorter administration
Between (5 min) each concentration can effectively inhibit bacterial growth in 2 hours.When meeting administration required for oral bacteriostasis model
Between it is short, scavenging is good, duration long feature.
Detailed description of the invention
Fig. 1 is radix tetrastigme polysaccharide to Escherichia coli function and effect figure.
Fig. 2 is that radix tetrastigme polysaccharide acts on scanning electron microscope diagram to bacterium.
Fig. 3 is based on oral antimicrobial model radix tetrastigme polysaccharide to Escherichia coli effect；Wherein a-e is respectively radix tetrastigme polysaccharide
Aqueous solution culture bacterium solution 5 min, 10 min, 15 min, 20 min, the fungistatic effect of 25 min.
Specific embodiment
Embodiment 1 prepares radix tetrastigme polysaccharide
(1) the radix tetrastigme powder 250g for weighing 60 dry mesh is extracted 5 hours with 50 DEG C of 1L dehydrated alcohol, discards supernatant liquid；
It is extracted 5 hours with 50 DEG C of 1L ultrapure water, suction filtration obtains filtrate, and the 1/4 of original volume is concentrated under 65 DEG C of bath temperatures；It is added 4
95% ethyl alcohol of times volume precipitates left undisturbed overnight；5000 revs/min of centrifugation collection in 20 minutes precipitatings, obtain Thick many candies after freeze-dried
26.33 grams；
(2) radix tetrastigme Thick many candies 20g is weighed, is dissolved in 1000mL ultrapure water, after ultrasonic dissolution is complete, is proportionally added into chloroform
And n-butanol, 20min is sufficiently stirred, 5000r/min is centrifuged 20 min, discards organic layer and albumin layer, in triplicate, until nothing
Until protein white precipitate generates；The adding proportion of chloroform and n-butanol is to count by volume: polysaccharide: chloroform: n-butanol=
8:4:1；
(3) using 8000-14000Da bag filter ultrapure water dialysis 48h, freeze-drying；Obtain radix tetrastigme Thick many candies 0.984
g。
2 radix tetrastigme polysaccharide bacteriostatic experiment of embodiment
Escherichia coli and staphylococcus aureus are cultivated to logarithmic growth phase (1*106A/mL), according to bacterium solution: culture medium (volume
Than)=1:1000 using sterile LB medium dilutes bacterium, prepare radix tetrastigme polysaccharide solution be (2.0wt%, 1.0 wt %, 0.5
Wt %, 0.1 wt %, 0.05 wt %) 200 μ L, 200 μ L are added and dilute bacterium solution, test 600 nm of [email protected] result is shown in figures
1.Radix tetrastigme polysaccharide can effectively inhibit bacterial number on shorter action time as seen from Figure 1, in two hours
Effectively.
Bacterium pattern after observing administration 1h using scanning electron microscope.Experimental result is shown in Fig. 2, can be seen that by Fig. 2 a
Cylindrical bar pattern is presented in normal E. coli, and scale meets described in document at 3 μm or so, and Fig. 2 b can be observed obviously
The phenomenon that bacterium pattern is changed, and has purulence swollen and fracture.Radix tetrastigme polysaccharide using dilution bacterium solution and above-mentioned concentration is water-soluble
Liquid cultivates 5 min, 10 min, 15 min, 20 min, 25 min respectively.Centrifugal separating cell and culture solution, use are sterile later
1 × PBS of grade washing is added sterile LB medium afterwards three times and cultivates OD600 nm light absorption value on test different time.Experimental result is shown in
Fig. 3.By Fig. 3 it can be seen that each concentration radix tetrastigme polysaccharide can effectively inhibit bacterial growth, in shorter administration time (5
Min) each concentration can effectively inhibit bacterial growth in 2 hours.It is short to meet administration time required for oral bacteriostasis model,
Scavenging is good, the feature of duration length.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.All within the spirits and principles of the present invention, made any to repair
Change, equivalent replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of preparation method of radix tetrastigme polysaccharide, which is characterized in that specifically includes the following steps:
(1) the radix tetrastigme powder 250g for weighing 60 dry mesh is extracted 5 hours with 50 DEG C of 1L dehydrated alcohol, discards supernatant liquid；
It is extracted 5 hours with 50 DEG C of 1L ultrapure water, suction filtration obtains filtrate, and the 1/4 of original volume is concentrated under 65 DEG C of bath temperatures；It is added 4
95% ethyl alcohol of times volume precipitates left undisturbed overnight；5000 revs/min of centrifugation collection in 20 minutes precipitatings, obtain slightly more after freeze-dried
Sugar；
(2) radix tetrastigme Thick many candies 20g is weighed, is dissolved in 1000mL ultrapure water, after ultrasonic dissolution is complete, is proportionally added into chloroform
And n-butanol, 20min is sufficiently stirred, 5000r/min is centrifuged 20 min, discards organic layer and albumin layer, in triplicate, until nothing
Until protein white precipitate generates；
(3) using 8000-14000Da bag filter ultrapure water dialysis 48h, freeze-drying obtains radix tetrastigme essence polysaccharide.
2. preparation method according to claim 1, which is characterized in that the addition ratio of step (2) chloroform and n-butanol
Example is to count by volume: polysaccharide: chloroform: n-butanol=8:4:1.
3. a kind of radix tetrastigme polysaccharide that the method as described in claim 1 is prepared is inhibiting the application in Dental Plaque.
4. application according to claim 3, which is characterized in that using radix tetrastigme polysaccharide as solute, be configured to end with distilled water
Concentration is the mouthwash of 0.05-2.0wt%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910850486.0A CN110483656A (en) | 2019-09-10 | 2019-09-10 | A kind of preparation of radix tetrastigme polysaccharide antibacterial bacteriostatic mouthwash |
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| CN201910850486.0A CN110483656A (en) | 2019-09-10 | 2019-09-10 | A kind of preparation of radix tetrastigme polysaccharide antibacterial bacteriostatic mouthwash |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111514295A (en) * | 2020-05-29 | 2020-08-11 | 中玺(泉州)科技有限公司 | Preparation method of radix tetrastigme polysaccharide modified sulfur-doped carbon nitride nanosheet material |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107778376A (en) * | 2017-11-27 | 2018-03-09 | 福州大学 | A kind of preparation method and applications of radix tetrastigme polysaccharide |
| CN108686055A (en) * | 2018-07-26 | 2018-10-23 | 杭州金域医学检验所有限公司 | Application of the Lemna paucicostata in anti-Staphylococcus aureus drug |
-
2019
- 2019-09-10 CN CN201910850486.0A patent/CN110483656A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107778376A (en) * | 2017-11-27 | 2018-03-09 | 福州大学 | A kind of preparation method and applications of radix tetrastigme polysaccharide |
| CN108686055A (en) * | 2018-07-26 | 2018-10-23 | 杭州金域医学检验所有限公司 | Application of the Lemna paucicostata in anti-Staphylococcus aureus drug |
Non-Patent Citations (1)
| Title |
|---|
| XIAO CHEN等: ""Antibacterial mechanism of Tetrastigma hemsleyanum Diels et Gilg"s polysaccharides by metabolomics based on HPLC/MS"", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111514295A (en) * | 2020-05-29 | 2020-08-11 | 中玺(泉州)科技有限公司 | Preparation method of radix tetrastigme polysaccharide modified sulfur-doped carbon nitride nanosheet material |
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Application publication date: 20191122 |
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