CN110317279A - A kind of Asiatic polysaccharide, the purposes as plant source antiseptic agent and the application in cosmetics - Google Patents
A kind of Asiatic polysaccharide, the purposes as plant source antiseptic agent and the application in cosmetics Download PDFInfo
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- CN110317279A CN110317279A CN201910762107.2A CN201910762107A CN110317279A CN 110317279 A CN110317279 A CN 110317279A CN 201910762107 A CN201910762107 A CN 201910762107A CN 110317279 A CN110317279 A CN 110317279A
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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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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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
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/524—Preservatives
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Abstract
The invention discloses a kind of Asiatic polysaccharide, the purposes as plant source antiseptic agent and the applications in cosmetics.Asiatic polysaccharide is prepared via a method which: being weighed centella herb, is successively used petroleum ether, 95% alcohol reflux degreasing, filtering, after filter residue volatilizes solvent, distillation water heating extracting, filtering are used, filtrate is concentrated into 1/5 volume, filters again, and the dehydrated alcohol of 4 times of volumes is added, it stirs evenly, stands alcohol precipitation, filter, precipitating successively is washed with dehydrated alcohol, acetone, ether, redissolved, repeated alcohol precipitation 1 time, Sevag method removing protein with distilled water, dialysis, freeze-drying.The experiment results show that Asiatic polysaccharide provided by the invention all has excellent inhibitory activity to bacterium and fungi, and to cutaneous safety height, it will not cause skin allergy, it may be used as plant source antiseptic agent, and then can be used for cosmetics anti-corrosion, have both natural, effective, mild non-stimulated advantage.
Description
Technical field
The invention belongs to cosmetic fields, are related to application of the Asiatic polysaccharide in cosmetics, and in particular to a kind of accumulated snow
Grass polysaccharide, the purposes as plant source antiseptic agent and the application in cosmetics.
Background technique
The study found that nature is permitted extract natural antirot substance in various plants, have to bacterium and/or fungi
There is significant inhibiting effect.These natural antirot substances include alkaloid, flavones, cumarin, saponin(e etc..
Because the side effect of natural antirot substance is usually smaller, safer to skin, so being developed from plant natural
Anti- humus is used as preservative and is added to the hot spot for having become cosmetics preservative exploitation in cosmetics.
Centella is the drying herb of umbelliferae centella, containing there are many triterpenes effective components, including centella
Glycosides, madecassoside, asiatic acid and brahmic acid etc., Clinical and experimental study show that centella can effectively promote
Into union of wounded skin, and the pathologic scar formed to excessive healing is inhibited.And modern pharmacological studies have shown that
Centella can not only promote cell growth, accelerating wound healing and repair skin injury, also have anti-oxidant, antiulcer, disappear
The effects of scorching and protection nerve cell.So asiaticosid, madecassoside, asiatic acid and hydroxyl product in centella
The small molecule compounds such as snow oxalic acid are widely used in makeup kind.
Asiatic polysaccharide is the macromolecular compound in centella, there is not yet its report with antibacterium and fungi activity
Road.
Summary of the invention
An object of the present disclosure is to provide a kind of Asiatic polysaccharide, and second, which is designed to provide the Asiatic polysaccharide, is used as
The purposes of plant source antiseptic agent, third are designed to provide application of the Asiatic polysaccharide in cosmetics.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A kind of Asiatic polysaccharide, is prepared via a method which: weighing centella herb, successively uses 60~90 DEG C of boiling ranges
Petroleum ether, 95% alcohol reflux degreasing, filtering after filter residue volatilizes solvent, use distillation water heating extracting, filtering, filtrate concentration
It to 1/5 volume, filters again, the dehydrated alcohol of 4 times of volumes is added, stirs evenly, stand alcohol precipitation, filter, successively with anhydrous second
Alcohol, acetone, ether washing precipitating, are redissolved with distilled water, are repeated alcohol precipitation 1 time, filtered, successively washed with dehydrated alcohol, acetone, ether
Precipitating is washed, Sevag method removing protein is dialysed, freeze-drying.
Preferably, the solid-liquid ratio of defatting step is 1；5.
Preferably, two kinds of solvent difference degreasings reflux 3h.
Preferably, the solid-liquid ratio of aqueous extraction step is 1；10.
Preferably, water mentions 2 times, each 2h.
Preferably, it is dialysed with the bag filter of molecular cut off 5000u.
Preferably, it dialyses for 24 hours.
Above-mentioned Asiatic polysaccharide is used as the purposes of plant source antiseptic agent.
Above-mentioned Asiatic polysaccharide is used as the purposes of cosmetics preservative.
Technical effect:
The present invention provides a kind of Asiatic polysaccharides, the experiment results show that Asiatic polysaccharide provided by the invention is to bacterium
Excellent inhibitory activity is all had with fungi, and to cutaneous safety height, skin allergy will not be caused, it is anti-to may be used as plant source
Rotten agent, and then can be used for cosmetics anti-corrosion, have both natural, effective, mild non-stimulated advantage.
Detailed description of the invention
Fig. 1 is the SEM figure that embodiment 1 prepares polysaccharide, intermolecular to deposit from SEM figure it can be seen that the polysaccharide regularity is not strong
In a large amount of random intersection constructions, branch is more.
Specific embodiment
Essentiality content of the present invention is introduced below with reference to embodiment, but the scope of the present invention is not limited with this.
Embodiment 1: the extraction of Asiatic polysaccharide and antiseptic property test
One, experimental material
Centella herb is purchased from Bozhou Chinese Medicinal Materials Markets, cleans, air-dries, to crushed 60 meshes spare.
For examination bacterium bag include bacterium and fungi, in which: bacterium include Escherichia coli, proteus, Pseudomonas aeruginosa, streptococcus,
Staphylococcus aureus, bacillus subtilis, fungi include black-koji mould and saccharomycete.Bacterium uses beef extract-peptone culture
Base, black-koji mould use potato dextrose agar, and saccharomycete uses yeast extract peptone glucose agar medium.
Two, experimental method
1, the preparation and qualitative detection of Asiatic polysaccharide
Centella herb is weighed, successively uses 60~90 DEG C of boiling ranges according to the solid-liquid ratio of 1:5 (1kg herb uses 5L solvent)
Petroleum ether, 95% ethyl alcohol (volumn concentration) respectively reflux degreasing 3h, filtering, after filter residue volatilizes solvent, according to the material of 1:10
Liquor ratio uses 85 DEG C of distilled water heating extraction 2 times, each 2h, and combined extract filters, and filtrate is concentrated into 1/5 volume, again mistake
Filter, is added the dehydrated alcohol of 4 times of volumes, stirs evenly, and stands alcohol precipitation 12h, filters, is successively washed with dehydrated alcohol, acetone, ether
It washs precipitating each 2 times, is redissolved, repeated alcohol precipitation 1 time with distilled water, filtered, successively wash precipitating each 2 with dehydrated alcohol, acetone, ether
Secondary, Sevag method removing protein is dialysed for 24 hours with the bag filter that molecular cut off is 5000u, freeze-drying.
Molish reaction: the polysaccharide 1g for taking the above method to be prepared is dissolved in 20mL distilled water and polysaccharide solution is made, and takes 1mL
Polysaccharide solution adds 1mL alpha-Naphthol in test tube, then plus the 1mL concentrated sulfuric acid, there is purplish red colour circle to occur, Molish reacting positive.
Phenolsulfuric acid reaction: taking the above-mentioned polysaccharide solution of 1mL in test tube, add 1mL phenol, then plus the 5mL concentrated sulfuric acid, occur
Yellowish-brown compound, phenolsulfuric acid reacting positive.
Fig. 1 is that the SEM of above-mentioned polysaccharide schemes, and regularity is not strong, and intermolecular have a large amount of random intersection constructions, branch compared with
It is more.
2, inhibiting effect of the Asiatic polysaccharide to bacterium and fungi
The preparation of 2.1 polysaccharide solutions and bacteria suspension
The Asiatic polysaccharide for weighing the preparation of embodiment 1 is configured to the solution of various concentration with distilled water dissolution, then by than dilute
It releases to other concentration.It is placed in slant medium for examination bacterium, for 24 hours in 37 DEG C of activation, der Pilz is in 28 DEG C of activation 48h, ferment for bacterium class
Female bacterium activates for 24 hours in 30 DEG C.Essence chooses lawn in sterile water, is prepared into 1 × 106~1 × 108The bacteria suspension of CFU/mL.
2.2 bacteriostasis qualitative detections
Sterilising medium is slowly poured into cooled and solidified in sterile petri dish under aseptic condition, takes 0.2mL for examination respectively
Bacteria suspension is spread evenly across on solid plate.The circular filter paper piece of the diameter 12mm made of sterilizing filter paper is put into 10mg/mL
Polysaccharide solution in impregnate 1h, media surface is gently placed in after dry, the culture medium of inoculated bacteria is placed in 37 DEG C of inversion and trains
It supports for 24 hours, the culture medium of inoculated aspergillus niger bacterium is placed in 28 DEG C of inversion culture 48h, and the culture medium of inoculation yeast bacterium is placed in 30 DEG C of inversions
Culture is for 24 hours.It is feminine gender without inhibition zone, having inhibition zone is the positive, and measures antibacterial circle diameter with crossing method.Parallel behaviour
Make 3 parts.It is equipped with 1 part of blank control for not containing polysaccharide simultaneously.
Testing result is as shown in table 1.
The antibacterial circle diameter of 1 10mg/mL Asiatic polysaccharide of table
| Strain name | Antibacterial circle diameter (mm) |
| Escherichia coli | 18.5±1.4 |
| Pseudomonas aeruginosa | 19.2±1.6 |
| Proteus | 20.8±1.4 |
| Streptococcus | 19.6±1.2 |
| Staphylococcus aureus | 18.1±1.3 |
| Bacillus subtilis | 18.8±1.5 |
| Black-koji mould | 25.5±1.4 |
| Saccharomycete | 24.8±1.6 |
| Blank control | 12 (remarks: filter paper diameter, no inhibition zone) |
The testing result shows that Asiatic polysaccharide all significantly inhibits bacterium and fungi.
The measurement of 2.3 minimal inhibitory concentrations (MIC)
Asiatic polysaccharide is configured to the solution of 0.5,0.25,0.125,0.0625,0.0313mg/mL with distilled water, point
1mL is not pipetted into different culture mediums, mixes well polysaccharide solution with culture medium, it is spare after solidification is cooling.It takes respectively
0.2mL is spread evenly across on solid plate for examination bacteria suspension, and the culture medium of inoculated bacteria is placed in 37 DEG C of inversion cultures for 24 hours, inoculation
The culture medium of black-koji mould is placed in 28 DEG C of inversion culture 48h, and the culture medium of inoculation yeast bacterium is placed in 30 DEG C of inversion cultures for 24 hours.
"-" represents sterile length of being born, and "+" representative has bacterium colony growth, and the minimum concentration of no bacterium colony is minimal inhibitory concentration (MIC).
Testing result is as shown in table 2.
The testing result of 2 minimum inhibitory concentration of table (MIC)
The testing result shows that Asiatic polysaccharide is 0.25mg/mL to the minimum inhibitory concentration (MIC) of bacterium, to fungi
Minimum inhibitory concentration (MIC) be 0.125mg/mL, bacterium and fungi are all significantly inhibited.
Embodiment 2: scratch test (patch test)
Tested material is Asiatic polysaccharide prepared by embodiment 1.
Referring to " cosmetics safety technical specification " (2015 editions) 30 healthy volunteers of selection as subject, wherein male 15
Name, female 15.After taking a shower, tested material 0.025g is taken to be placed in spot tester cell, it will be added with the low sensitization of the spot tester of tested material
Adhesive tape is pasted on the back or forearm song side of subject, is gently pressed with palm and is allowed to equably be pasted on skin, continued for 24 hours.
After removing tested material spot tester 30min (after impression disappearance after), for 24 hours with 48h press 3 standard sight skin of table
Reaction, and record observation result and reference table 3 evaluated.
3 skin closure type patch test dermoreaction grade scale of table
As a result 30 healthy volunteers are feminine gender, negative rate 100%.
The experiment results show that Asiatic polysaccharide provided by the invention all has excellent inhibitory activity to bacterium and fungi,
And to cutaneous safety height, skin allergy will not be caused, may be used as plant source antiseptic agent, and then it is anti-to can be used for cosmetics
Corruption has both natural, effective, mild non-stimulated advantage.
Claims (9)
1. a kind of Asiatic polysaccharide, which is characterized in that be prepared via a method which: weigh centella herb, successively using 60~
The petroleum ether of 90 DEG C of boiling ranges, 95% alcohol reflux degreasing, filtering after filter residue volatilizes solvent, use distillation water heating extracting, mistake
Filter, filtrate are concentrated into 1/5 volume, filter again, the dehydrated alcohol of 4 times of volumes is added, stirs evenly, and stand alcohol precipitation, filter, according to
It is secondary to wash precipitating with dehydrated alcohol, acetone, ether, with distilled water redissolve, repeat alcohol precipitation 1 time, filter, successively with dehydrated alcohol,
Acetone, ether washing precipitating, Sevag method removing protein are dialysed, freeze-drying.
2. Asiatic polysaccharide according to claim 1, it is characterised in that: the solid-liquid ratio of defatting step is 1；5.
3. Asiatic polysaccharide according to claim 2, it is characterised in that: two kinds of solvent difference degreasings reflux 3h.
4. Asiatic polysaccharide according to claim 1, it is characterised in that: the solid-liquid ratio of aqueous extraction step is 1；10.
5. Asiatic polysaccharide according to claim 4, it is characterised in that: water mentions 2 times, each 2h.
6. Asiatic polysaccharide according to claim 1, it is characterised in that: dialysed with the bag filter of molecular cut off 5000u.
7. Asiatic polysaccharide according to claim 6, it is characterised in that: dialysis is for 24 hours.
8. the purposes that any Asiatic polysaccharide of claim 1~7 is used as plant source antiseptic agent.
9. the purposes that any Asiatic polysaccharide of claim 1~7 is used as cosmetics preservative.
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| CN201910762107.2A CN110317279B (en) | 2019-08-19 | 2019-08-19 | Herba Centellae polysaccharide, application of herba Centellae polysaccharide as plant source antiseptic, and application of herba Centellae polysaccharide in cosmetics |
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| CN201910762107.2A CN110317279B (en) | 2019-08-19 | 2019-08-19 | Herba Centellae polysaccharide, application of herba Centellae polysaccharide as plant source antiseptic, and application of herba Centellae polysaccharide in cosmetics |
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| CN113456796A (en) * | 2020-03-31 | 2021-10-01 | 无限极（中国）有限公司 | Anti-aging composition and preparation method and application thereof |
| CN115558036A (en) * | 2022-10-23 | 2023-01-03 | 爱生泽(上海)生物科技有限公司 | Preparation method of centella asiatica polysaccharide and application of centella asiatica polysaccharide in cosmetics |
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| CN113456796A (en) * | 2020-03-31 | 2021-10-01 | 无限极（中国）有限公司 | Anti-aging composition and preparation method and application thereof |
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| CN112656707A (en) * | 2020-12-29 | 2021-04-16 | 中华全国供销合作总社南京野生植物综合利用研究所 | Extraction method of centella asiatica polysaccharide extract, centella asiatica polysaccharide extract and application thereof |
| CN115558036A (en) * | 2022-10-23 | 2023-01-03 | 爱生泽(上海)生物科技有限公司 | Preparation method of centella asiatica polysaccharide and application of centella asiatica polysaccharide in cosmetics |
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