CN109806205A - A kind of FRUCTUS TERMINALIAE IMMATURUS extract and its antibacterial application - Google Patents
A kind of FRUCTUS TERMINALIAE IMMATURUS extract and its antibacterial application Download PDFInfo
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- CN109806205A CN109806205A CN201910282146.2A CN201910282146A CN109806205A CN 109806205 A CN109806205 A CN 109806205A CN 201910282146 A CN201910282146 A CN 201910282146A CN 109806205 A CN109806205 A CN 109806205A
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- fructus terminaliae
- terminaliae immaturus
- extract
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- bactericidal composition
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Abstract
The present invention relates to a kind of FRUCTUS TERMINALIAE IMMATURUS extracts and application thereof.Specifically, the purposes of the antibacterial agent of the purposes the present invention relates to FRUCTUS TERMINALIAE IMMATURUS extract as antibacterial agent, especially beauty or bactericidal composition.Moreover, it relates to a kind of bactericidal composition, it includes FRUCTUS TERMINALIAE IMMATURUS extract of the invention and optional benzoic acid or its cosmetically acceptable soluble-salt and/or optional ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salts.The invention further relates to the preparation methods of the FRUCTUS TERMINALIAE IMMATURUS extract.
Description
Invention field
In general, the present invention relates to a kind of FRUCTUS TERMINALIAE IMMATURUS extract and its antibacterial applications.FRUCTUS TERMINALIAE IMMATURUS extract of the invention
It is particularly suitable for being used as antibacterial agent in beauty or cosmetic product.The invention further relates to a kind of bactericidal compositions, and it includes the present invention
FRUCTUS TERMINALIAE IMMATURUS extract and optional benzoic acid or its cosmetically acceptable soluble-salt and/or optional ethylenediamine tetrem
Acid or its cosmetically acceptable soluble-salt.The bactericidal composition is especially suitable for beauty or cosmetic product.In addition, this hair
The bright method for further relating to prepare the FRUCTUS TERMINALIAE IMMATURUS extract.
Background technique
The dosage form of presently commercially available most of cosmetics or cosmetics belongs to water-soluble, and moisture content is high, and containing a large amount of
The nutriments such as carbohydrate, protein or lipid, thus the good nutrient of microorganism and growing environment are provided.These cosmetics or change
Cosmetic use process or preserving type is improper is easy for cooperating microorganisms and causes to pollute.Product is once by the dirt of pathogen
Dye, it is rotten in addition to product may be caused because of active ingredient degradation, it is also possible to which that direct infection user seriously endangers consumer
Health.
Chemical preservative can efficiently control the growth of microorganism, prevent microorganism from polluting to cosmetic product, commonly use
In the chemical preservative of cosmetics and food include: imidazolidinyl urea, imidazolidine -2,4- diketone, isothiazolinone, hydroxy benzenes first
The chemical preservatives such as acetoacetic ester, benzoic acid and its derivative.However, these chemical preservatives, which can have, for example leads to chafing
Equal side effects have some with more neurotoxicity and carcinogenesis.Therefore, these chemical preservatives are not joyous by consumer institute
It meets, while being also the emphasis that supervision department examines, this usually influences the marketing of product.Since preservative or antibacterial agent are being changed
Occupy critically important role in cosmetic or cosmetics formula, therefore develops new natural antibacterial agent and bactericidal composition to cosmetics
Industry has urgent need and actual meaning.
FRUCTUS TERMINALIAE IMMATURUS is Chinese medicine name, also known as myrobalan, is the drying young fruit of combretaceae plant myrobalan.Pharmacopeia is recorded, and west is green
Fruit has clearing heat and detoxicating and other effects, has been applied to the symptoms such as sphagitis, tonsillitis.Chinese patent application publication No.
CN108467417A describes a kind of FRUCTUS TERMINALIAE IMMATURUS extract, passes through the FRUCTUS TERMINALIAE IMMATURUS that will be crushed successively through petroleum ether, ethyl acetate
Extraction, gained ethyl acetate portion are separated through normal phase column dichloromethane-acetone-formic acid system gradient elution again, the extraction of acquisition
Object, which has, inhibits monoamine oxidase B activity, can be used for treating the nervous system disease.Chinese patent application publication No.
CN102266360A describes a kind of FRUCTUS TERMINALIAE IMMATURUS extract, by extracting the dry FRUCTUS TERMINALIAE IMMATURUS crushed with ethyl alcohol, then with just
Hexane extraction, then water-methanol gradient elution is used, the extract of acquisition can be used for treating with alpha-glucosaccharase enzyme inhibition activity
Diabetes.Chinese patent application publication No. CN104623245A is described in a kind of clearing heat and promoting fluid comprising FRUCTUS TERMINALIAE IMMATURUS extract
Medicine chewable tablets.Chinese patent application publication No. CN105029702A describes a kind of three Extraction of acetic acid object of FRUCTUS TERMINALIAE IMMATURUS and its in cigarette
In application.However, being used as antibacterial agent about FRUCTUS TERMINALIAE IMMATURUS extract currently without any document or report, especially it is used as makeup
The antibacterial agent of product or cosmetics.
Summary of the invention
The purpose of the present invention may include providing a kind of natural antibacterial agent, and especially from fruit, (such as crushing is dry
Dry fruit) in extract natural antibacterial agent.Such antibacterial agent is without the scorching even Nervous toxicity of the sensitization of chemical antibacterial agent and cause
Property and carcinogenic equal side effects, be therefore particularly suitable for local application or dermal administration, such as beauty or cosmetic use.It is mentioned when naturally
When object being taken to be used for beauty or cosmetic product, the extract is also required not significantly affect physical property (such as the color, gas of product
Taste and stability), it otherwise will affect consumer and the purchase of product be intended to, product market value is caused to decline.Mesh of the invention
May also include a kind of method for preparing such natural extract is provided.In addition, the purpose of the present invention may also include offer one
Bactericidal composition of the kind comprising the natural antibacterial agent and beauty or make-up composition.
Technical problem of the invention is solved by providing a kind of FRUCTUS TERMINALIAE IMMATURUS extract, which passes through herein
It is prepared by the ad hoc approach (such as specific extract and elution process) of description.Since the method for preparing the extract is different from now
There is the preparation method in technology, therefore ingredient in FRUCTUS TERMINALIAE IMMATURUS extract of the present invention and its content are different from west in the prior art
Fructus Canarii extract.
In general, it there is provided herein a kind of FRUCTUS TERMINALIAE IMMATURUS extract, is extracted by water and ethanol water elutes to make
It is standby.The extract is to various bacteriums (including gram-positive bacterium and gram are with negative bacteria) and fungi (including yeast etc.)
All have and show inhibitory effect, therefore there is the bacteriostatic activity of more wide spectrum.In addition, FRUCTUS TERMINALIAE IMMATURUS of the invention is extracted for beauty
Appearance/cosmetics physical property does not constitute negative effect, and has safety to human skin.In addition, offer of the invention
Bactericidal composition there is broader spectrum of bacteriostatic activity, and the content of chemical antibacterial agent contained therein can further subtract
It is few.
In one aspect, the present invention relates to a kind of FRUCTUS TERMINALIAE IMMATURUS extract, the extract passes through the side included the following steps
Method preparation: (1) FRUCTUS TERMINALIAE IMMATURUS through fragmentation is extracted with water to obtain water extract, and (2) make the water extract pass through macroporous absorption
Resin, and then eluted with ethanol water.In one aspect, FRUCTUS TERMINALIAE IMMATURUS extract of the invention can in solution form or
Lyophilized form (such as freeze-dried powder).
On the other hand, the present invention relates to a kind of method for preparing FRUCTUS TERMINALIAE IMMATURUS extract, the method includes following steps
It is rapid: (1) FRUCTUS TERMINALIAE IMMATURUS through fragmentation is extracted with water to obtain water extract, and (2) make the water extract pass through macroporous absorbent resin,
And it is then eluted with ethanol water, to obtain the extract.
In above-mentioned steps (1), it can be used equipment or method (such as pulverizer) well known in the art by FRUCTUS TERMINALIAE IMMATURUS fragment
Change (such as crushing), wherein degree of grinding may be, for example, at least 4 mesh, preferably at least 10 mesh, preferably at least 20 mesh, such as 30-200
Mesh, such as 40 mesh.In step (1), after water is added to the FRUCTUS TERMINALIAE IMMATURUS of fragmentation, heating and refluxing extraction is carried out, wherein heating
Time is at least 30 minutes, preferably at least 1 hour, 1.5-10 hours more preferable, such as 2 hours.
In one aspect, the above method can comprise the further steps of: (1 ') by the water extract obtained from step (1) carry out from
The heart, wherein the supernatant to acquisition carries out subsequent step.In the step (1 '), can by centrifugation obtain sediment again into
The extraction process of row step (1), and the water extract of acquisition is centrifuged, obtain supernatant again and is closed with supernatant before
And it is used for subsequent step.In this aspect, at least 1 time repetition can be carried out to the sediment to extract, such as 2-3 repetition mentions
It takes.
In above-mentioned steps (2), the water extract is made by macroporous absorbent resin to be to make active ingredient adsorption to the tree
The process of rouge.Chromatography or chromatography technology, preferred liquid phase chromatography or chromatography are used in this process.The macroporous absorption tree used
Rouge can be low pole macroporous absorbent resin, more preferable low pole styrene tyle macroporous adsorption resin, most preferably AB-8 macroporous absorption
Resin.The macroporous absorbent resin is the form of large pore resin absorption column.In the step (2), eluted with ethanol water
It is to elute active constituent from the resin to obtain the process of eluent.In the step, 5%- is used
50% ethanol water, the ethanol water of more preferable 8%-40%, the ethanol water of more preferable 10%-30%, such as
5%, 10%, 15%, 20%, 25%, 30%, 35%, 40% or 45% ethanol water.In the step, stream is collected
The eluent of the ethanol water through macroporous absorbent resin.
In one aspect, the above method can comprise the further steps of: in the eluent that (3) will be obtained from step (2)
Ethyl alcohol removal.In the step (3), ethyl alcohol can be removed with any method known in the art, such as use rotary evaporation
Instrument.
In one aspect, the above method can comprise the further steps of: (4) will obtain from the step (3) eluent carry out it is dense
Contracting and optional constant volume, to obtain the extract in solution form.
In one aspect, the above method can comprise the further steps of: (5) will obtain FRUCTUS TERMINALIAE IMMATURUS extract from step (4)
Freeze-drying process, to obtain the extract in freeze-dried powder form.
In the above-mentioned methods, the FRUCTUS TERMINALIAE IMMATURUS extract is with 30g/L-300g/L, preferably 50g/L-200g/L, more preferably
80g/L-150g/L, more preferable 100g/L FRUCTUS TERMINALIAE IMMATURUS/final FRUCTUS TERMINALIAE IMMATURUS extract amount obtained from FRUCTUS TERMINALIAE IMMATURUS.For example,
After the FRUCTUS TERMINALIAE IMMATURUS of 100g is by fragmentation and comprising the processing of above-mentioned steps (1)-(4) method, the FRUCTUS TERMINALIAE IMMATURUS of 1L is finally obtained
Extract, this corresponds to FRUCTUS TERMINALIAE IMMATURUS/final FRUCTUS TERMINALIAE IMMATURUS extract amount of 100g/L.
In one aspect, the present invention relates to a kind of bactericidal compositions, and it includes FRUCTUS TERMINALIAE IMMATURUS extracts of the present invention.?
In this aspect, the bactericidal composition can further include benzoic acid or its cosmetically acceptable soluble-salt.The benzene
Formic acid can be existed in the bactericidal composition by soluble form or the form of ion.For example, the bactericidal composition can
Further include sodium benzoate or Potassium Benzoate, preferred sodium benzoate.On the other hand, the bactericidal composition can be further
Include ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt.The ethylenediamine tetra-acetic acid can in the bactericidal composition
Exist by by the form of soluble or in the form of ion.For example, the bactericidal composition can further include ethylenediamine tetra-acetic acid
Sodium salt or ethylene diamine tetraacetic acid sylvite, preferably disodium ethylene diamine tetraacetate.In one aspect, the composition may include the present invention
FRUCTUS TERMINALIAE IMMATURUS extract, benzoic acid or its cosmetically acceptable soluble-salt and ethylenediamine tetra-acetic acid or its cosmetically may be used
The soluble-salt of receiving.In one aspect, the composition also may include solvent, preferably water.In one aspect, the composition
It can be made of above-mentioned antibacterial components and solvent (such as water).
In above-mentioned bactericidal composition, calculated based on the bactericidal composition, the weight percent of the FRUCTUS TERMINALIAE IMMATURUS extract
Than that can be 0.1%-20%, preferably 0.5%-10%, more preferable 1%-5%, more preferable 1.25%-2%, such as 1.25%,
1.5%, 1.75% or 2%.It in the bactericidal composition, is calculated based on the bactericidal composition, the benzoic acid or its beauty
The weight percent of acceptable soluble-salt can be 0.01%-2%, preferably 0.05%-1%, more preferable 0.075%- in appearance
0.75%, more preferable 0.1%-0.5%, such as 0.1%, 0.2%, 0.3%, 0.4% or 0.5%.In the bactericidal composition
In, it is calculated based on the bactericidal composition, the weight hundred of the ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt
Point than can be 0.025%-1%, more preferable 0.05%-0.5%, more preferable 0.075%-0.35%, preferably 0.1%-0.3%,
More preferable 0.125%-0.2%, such as 0.1%, 0.2%, 0.25% or 0.35%.
In one aspect, the present invention relates to above-mentioned FRUCTUS TERMINALIAE IMMATURUS extract or bactericidal compositions to be used as part or dermal administration
The purposes of the antibacterial agent of product (such as beauty product or cosmetic product).In one aspect, the present invention relates to a kind of beauty or changes
Composition for cosmetics, it includes above-mentioned FRUCTUS TERMINALIAE IMMATURUS extract or bactericidal compositions.Preferably, the beauty or make-up composition are bases
Beauty or make-up composition in water, such as washing lotion (shin moisturizer), lotion (such as moisturizing emulsion), Essence etc..It is described to be based on
The beauty of water or make-up composition are easier to the pollution by microorganism, therefore antibacterial agent of the invention is especially suitable for such
Composition.
It in the composition of above-mentioned local application or dermal administration, is calculated based on the composition, the FRUCTUS TERMINALIAE IMMATURUS extracts
The weight percent of object can be 0.1%-20%, preferably 0.5%-10%, more preferable 1%-5%, more preferable 1.25%-2%, example
Such as 1.25%, 1.5%, 1.75% or 2%.In the composition of the local application or dermal administration, it is based on the composition
Calculate, the benzoic acid or its cosmetically acceptable soluble-salt weight percent can be 0.01%-2%, preferably
0.05%-1%, more preferable 0.075%-0.75%, more preferable 0.1%-0.5%, such as 0.1%, 0.2%, 0.3%, 0.4%
Or 0.5%.In the composition of the local application or dermal administration, calculated based on the composition, the ethylenediamine tetrem
Acid or its cosmetically acceptable soluble-salt weight percent can be 0.025%-1%, more preferable 0.05%-0.5%, more
It is preferred that 0.075%-0.35%, preferably 0.1%-0.3%, more preferable 0.125%-0.2%, such as 0.1%, 0.2%, 0.25%
Or 0.35%.The composition of the above-mentioned local application or dermal administration can be beauty or make-up composition.
According to some embodiments, beauty or make-up composition are any form for Traditional skin external preparation, packet
It includes but is not limited to beautifying face such as washing lotion (shin moisturizer), lotion, creme and cosmetic pack；Beauty of refining the make-up such as refines the make-up in advance,
Foundation cream, cheek coloring, lipstick, lip frost, eye shadow, eyeliner, mascara and opacifier；Body beauty；Fragrance beauty；Skin is clear
Clean dose of remover of such as refining the make-up, facial cleansing agent and body shampoo；Aerosol, muffin, creme, ointment, emulsion, essence, bubble
Foam, gel, washing lotion, mousse, paste, ointment (sticking patch), pen, slurries, solution, towelette, facial mask, body film or spray.Example
Such as, the beauty or make-up composition can be solution or suspension, such as solution or suspension based on water.
In one aspect, FRUCTUS TERMINALIAE IMMATURUS extract of the invention or bactericidal composition are suitable for acid or weakly acidic product,
Preferably pH4.0-6.0.
It is also contemplated that other than beauty or make-up composition, FRUCTUS TERMINALIAE IMMATURUS extract of the invention or bactericidal composition
It can be used for the composition of other local applications, the composition of dermal administration or oral cavity composition, such as local application or skin
The drug of application, including such as externally used solution agent, lotion, liniment, ointment, emplastrum, paste or patch；Such as oral cavity is clear
Clean product or teeth cleaning product, including toothpaste, tooth powder, mouthwash, oral spray or dental floss etc..
Term " antibacterial agent " as described herein or " antimicrobial " may be used interchangeably, and refer to bacterium, yeast and mould
The growth and breeding of the microorganisms such as bacterium are inhibited or even kill the substance of the microorganism.In the present invention, antibacterial agent
It may include FRUCTUS TERMINALIAE IMMATURUS extract or bactericidal composition.It include: gram by the bacterium that antibacterial agent of the present invention inhibits or kills
Positive bacteria such as staphylococcus aureus and bacillus subtilis and gramnegative bacterium such as Friedlander's bacillus,
Escherichia coli and pseudomonas aeruginosa.It include Candida albicans by the yeast that antibacterial agent of the present invention inhibits or kills.It is of the invention
Antibacterial agent inhibits or the mould of killing includes aspergillus niger.
FRUCTUS TERMINALIAE IMMATURUS extract or bactericidal composition as described herein may be used as preservative." preservative " of the present invention
Refer to the substance for food, drug, particularly makeup/cosmetics etc. to be added, to postpone the microorganism in these industrial products
Corruption caused by growth.When being used as preservative, it may be desirable to which this preservative is for various microorganisms (including bacterium, yeast
And mould) inhibitory effect with wide spectrum.
Term described in text " cosmetically acceptable soluble-salt " refer to provided by concentration when being applied to skin pair
Human body be it is safe, do not interfere the activity of other compositions in composition usually and be soluble salt in the composition.The salt
It can be sodium salt, sylvite etc..
For beauty or make-up composition, there is antibacterial agent the bacteriostatic activity of wide spectrum to be particularly advantageous, and thus may be used
To reduce the addition of other types antibacterial agent.At the same time, it must also be considered that broad spectrum antimicrobial agent is from for beauty or makeup combination
Side effect will not be generated when object to skin or human body, therefore natural antibacterial agent is obviously more advantageous relative to chemical antibacterial agent.It is existing
The problem of technology is to find a kind of not only wide spectrum and will not generate side effect, (such as protects with beauty/makeup group condition appearance simultaneously
Hold the stability, homogeneity and good appearance of product) antibacterial agent.Such technical problem is by providing FRUCTUS TERMINALIAE IMMATURUS of the invention
It extracts and bactericidal composition is addressed.
The invention further relates to following embodiments:
1. a kind of FRUCTUS TERMINALIAE IMMATURUS extract, the extract is prepared by method comprising the following steps:
(1) FRUCTUS TERMINALIAE IMMATURUS through fragmentation is extracted with water to obtain water extract, and
(2) make the water extract by macroporous absorbent resin, and then eluted with ethanol water.
2. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 1, wherein the macroporous absorbent resin is low pole macroporous absorbent resin.
3. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 2, wherein the macroporous absorbent resin is the styrene tyle macroporous suction of low pole
Attached resin.
4. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 2, wherein the macroporous absorbent resin is AB-8 macroporous absorbent resin.
5. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 1 or 2, wherein the ethanol water includes the ethyl alcohol of 5%-50%.
6. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 1 or 2, wherein the ethanol water includes the ethyl alcohol of 8%-40%.
7. the FRUCTUS TERMINALIAE IMMATURUS extract of embodiment 1 or 2, wherein the ethanol water includes the ethyl alcohol of 10-30%.
8. a kind of bactericidal composition, it includes the FRUCTUS TERMINALIAE IMMATURUS extracts of any one of embodiment 1-7.
9. the bactericidal composition of embodiment 8, also includes:
Benzoic acid or its cosmetically acceptable soluble-salt, and/or
Ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt.
10. the bactericidal composition of embodiment 9, wherein the cosmetically acceptable soluble-salt of benzoic acid is benzoic acid
Sodium.
11. the bactericidal composition of embodiment 9, wherein the cosmetically acceptable soluble-salt of ethylenediamine tetra-acetic acid is second
Edetate disodium.
12. the bactericidal composition of any one of embodiment 8-11, wherein being calculated based on the bactericidal composition, the west
The weight percent of Fructus Canarii extract is 0.5%-10%.
13. the bactericidal composition of any one of embodiment 8-11, wherein being calculated based on the bactericidal composition, the west
The weight percent of Fructus Canarii extract is 1%-5%.
14. the bactericidal composition of any one of embodiment 8-11, wherein being calculated based on the bactericidal composition, the west
The weight percent of Fructus Canarii extract is 1.25%-2%.
15. the bactericidal composition of any one of embodiment 8-11, wherein the FRUCTUS TERMINALIAE IMMATURUS extract is with 50-300g/L
FRUCTUS TERMINALIAE IMMATURUS/final FRUCTUS TERMINALIAE IMMATURUS extract amount obtained from FRUCTUS TERMINALIAE IMMATURUS.
16. the bactericidal composition of any one of embodiment 8-11, wherein the FRUCTUS TERMINALIAE IMMATURUS extract is with 80-200g/L
FRUCTUS TERMINALIAE IMMATURUS/final FRUCTUS TERMINALIAE IMMATURUS extract amount obtained from FRUCTUS TERMINALIAE IMMATURUS.
17. the bactericidal composition of any one of embodiment 8-11, wherein the FRUCTUS TERMINALIAE IMMATURUS extract is with 100g/L
FRUCTUS TERMINALIAE IMMATURUS/final FRUCTUS TERMINALIAE IMMATURUS extract amount is obtained from FRUCTUS TERMINALIAE IMMATURUS.
18. the bactericidal composition of embodiment 12, wherein being calculated based on the bactericidal composition, the benzoic acid or its beauty
The weight percent of acceptable soluble-salt is 0.05%-1% in appearance；And/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt weight percent be 0.075%-
0.5%.
19. the bactericidal composition of embodiment 12, wherein being calculated based on the bactericidal composition, the benzoic acid or its beauty
The weight percent of acceptable soluble-salt is 0.075%-0.75% in appearance；And/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt weight percent be 0.1%-0.3%.
20. the bactericidal composition of embodiment 12, wherein being calculated based on the bactericidal composition, the benzoic acid or its beauty
The weight percent of acceptable soluble-salt is 0.1%-0.5% in appearance；And/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt weight percent be 0.125%-
0.2%.
21. the FRUCTUS TERMINALIAE IMMATURUS extract of any one of embodiment 1-7 or the antibacterial group of any one of embodiment 8-20
Close the purposes that object is used as the antibacterial agent of beauty or cosmetic product.
22. the composition of a kind of part or dermal administration, it includes the FRUCTUS TERMINALIAE IMMATURUS extracts of any one of embodiment 1-7
Or the bactericidal composition of any one of embodiment 8-20.
23. the composition of embodiment 22 is beauty or make-up composition.
24. the composition of embodiment 22 or 23, wherein calculated based on the composition of the part or dermal administration, it is described
The weight percent of FRUCTUS TERMINALIAE IMMATURUS extract is 1%-5%.
25. the composition of embodiment 22 or 23, wherein calculated based on the composition of the part or dermal administration, it is described
The weight percent of FRUCTUS TERMINALIAE IMMATURUS extract is 1.25%-2%.
26. the composition of embodiment 22 or 23, it includes
The benzoic acid or its cosmetically acceptable soluble-salt that weight percent is 0.075%-0.75%, and/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt that weight percent is 0.1%-0.3%,
Wherein the weight percent is calculated based on the composition of the part or dermal administration.
27. the composition of embodiment 22 or 23, it includes
The benzoic acid or its cosmetically acceptable soluble-salt that weight percent is 0.1%-0.5%, and/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt that weight percent is 0.125%-0.2%,
Wherein the weight percent is calculated based on the composition of the part or dermal administration.
Brief description
30% ethanol eluate of Fig. 1 FRUCTUS TERMINALIAE IMMATURUS and various concentration EDTA-2Na combination are to pseudomonas aeruginosa and aspergillus niger
Antibacterial reaction.
After the processing of various concentration sodium benzoate, growth curve moves back Fig. 2 staphylococcus aureus, grows the effect that slows down.
Fig. 3 staphylococcus aureus is after anti-corrosion/bactericidal composition A, B, C processing of embodiment 7, growth retardation phenomenon.
After the processing of various concentration sodium benzoate, growth curve moves back Fig. 4 Escherichia coli, grows the effect that slows down.
Fig. 5 Escherichia coli are after anti-corrosion/bactericidal composition A, B, C processing of embodiment 7, growth retardation phenomenon.
The absorption spectrum of the various antibacterial agents of Fig. 6.Absorbance: absorbance, Wavelength: wavelength.
Specific embodiment
Unless otherwise stated, all scientific terms used herein are all generally understood with persons skilled in the art
Meaning it is consistent.Exemplary method and material is described below, its equivalent can be used.All publications being mentioned above and its
Its bibliography all passes through reference and is incorporated integrally into herein.
There is provided embodiment below is in order to which the present invention is further explained.Following embodiment is not intended to limit this with any reason
The range of invention.
Embodiment
The preparation of 1-FRUCTUS TERMINALIAE IMMATURUS of embodiment extract and its freeze-dried powder
A. FRUCTUS TERMINALIAE IMMATURUS extract-ethanol eluate preparation
1. taking 100g FRUCTUS TERMINALIAE IMMATURUS, 40 mesh are crushed to pulverizer, distilled water is added according to mass body buildup liquor ratio 1:10, adds
Circumfluence distillation 2h.
2.3000r/min is centrifuged 10min, collects supernatant.Sediment repeats to extract twice, merges supernatant, obtains FRUCTUS TERMINALIAE IMMATURUS
Water extract.
3. AB-8 macroreticular resin column purification on is eluted with the ethyl alcohol that mass fraction is 10%, 20% and 30% respectively,
The elution fraction obtained respectively under each concentration ethanol is collected, eluent Rotary Evaporators are removed into ethyl alcohol.
4. the concentration of FRUCTUS TERMINALIAE IMMATURUS eluent is settled to 1L, it is placed on -80 DEG C of refrigerators and saves.
B. the preparation of FRUCTUS TERMINALIAE IMMATURUS water extract
After above-mentioned steps 1 and 2, FRUCTUS TERMINALIAE IMMATURUS water extract is settled to 1L.
C. FRUCTUS TERMINALIAE IMMATURUS extract-freeze-dried powder preparation
The FRUCTUS TERMINALIAE IMMATURUS extract obtained by above step 1-4 is further lyophilized, the freeze-drying of FRUCTUS TERMINALIAE IMMATURUS extract is obtained
Powder.
The detection of 2-antibacterial agent of embodiment SGT
According to microbial growth curve, we can determine whether enter the time of exponential phase from the lag period, that is, when starting growth
Between (Start Growing Time, SGT).Microorganism amount of survival after antibacterial agent is added can be reduced, and cell is also by one
The destruction for determining degree, causes the lag period to extend, and SGT increases (i.e. growth curve moves to right).The microorganism that joined trial drug
The SGT of growth curve can know composition for the inhibitory effect of microorganism compared with the SGT of blank control group.
The FRUCTUS TERMINALIAE IMMATURUS water extract prepared respectively using embodiment 1 is detected for staphylococcus aureus and Escherichia coli
SGT, penicillin/streptomycin are as a result as follows as positive control.
Rejection ability of the FRUCTUS TERMINALIAE IMMATURUS water extract of 1. various concentration of table for staphylococcus aureus
Rejection ability of the FRUCTUS TERMINALIAE IMMATURUS water extract of 2. various concentration of table for Escherichia coli
Embodiment 3-uses the antibacterial activity of survival Counting alive microbial FRUCTUS TERMINALIAE IMMATURUS extract
Using test strain shown in table 1 in the 4th general rule 1121 " inhibitory effect detection method " of Chinese Pharmacopoeia (2015 editions)
It is cultivated with according to the condition of culture in table.Each strain is divided into 4 parts and is cultivated under sterile working, until strain connects bacterium
It measures up to 105-106cfu/mL.The test strain for examination is added for cultivating 2 days, 7 days in sample sheet according to the requirement of general rule table 2-2
And 14 days.Using Plating (including tilt-pour process and rubbing method) --- the 4th general rule 1105 of Chinese Pharmacopoeia (2015 editions) calculates
The average colony number of each test group.
Aqueous extracts (without chromatographic isolation), 10% ethyl alcohol of the FRUCTUS TERMINALIAE IMMATURUS of the method preparation described respectively using embodiment 1
Eluent, 20% ethanol eluate, 30% ethanol eluate and cosmetics often use chemical preservative GLP, the 0.1%-0.5% to be
Average additional amount, detects their inhibitory activity to representative bacterium (including Gram-positive and negative bacteria) and yeast.Knot
Fruit is as shown in table 3.
Table 3. compares FRUCTUS TERMINALIAE IMMATURUS extract and bacteriostasis of the traditional chemical preservative on different strain using Plating
In addition, the Aqueous extracts of FRUCTUS TERMINALIAE IMMATURUS, 10% ethanol eluate and 30% ethanol eluate are for the antibacterial of Escherichia coli
Ability be also it is similar, reach ++ rank, and positive control GLP is +++.
Inventors have found that with ethyl alcohol, to be eluted the color that will effectively remove FRUCTUS TERMINALIAE IMMATURUS water extract, (Examples below 8 is tested
Card), while remaining the bacteriostasis to different strain.The experiment shows that FRUCTUS TERMINALIAE IMMATURUS water extract and ethanol eluate are not only right
Bacterium (including G+And G-) there is inhibitory activity, and also there is inhibitory activity to fungi (such as Candida albicans).This shows west
Fructus Canarii extract can be used as the antibacterial agent of wide spectrum for beauty/cosmetic product.
The fungistatic effect that 4-FRUCTUS TERMINALIAE IMMATURUS of embodiment extract is combined with disodium ethylene diamine tetraacetate
Disodium ethylene diamine tetraacetate (EDTA-2Na) is usually used as stabilizer in cosmetics.Inventors have found that using west
The freeze-dried powder of Fructus Canarii extract (30% ethanol eluate) combines the broad spectrum activity that can enhance antibacterial with EDTA-2Na.Using following
The combination of ingredient detects their inhibitory effects to pseudomonas aeruginosa and aspergillus niger.
Table 4. includes the composition of FRUCTUS TERMINALIAE IMMATURUS extract and EDTA-2Na
In upper table 4, the freeze-dried powder of 1.75g is equivalent to 30% ethanol eluate of 20g (or mL), and the freeze-dried powder of 1.05g
It is equivalent to the eluent of 12.5g (or mL).
As a result as shown in Figure 1, the composition comprising FRUCTUS TERMINALIAE IMMATURUS extract and EDTA-2Na can effectively inhibit verdigris false single
The growth of born of the same parents bacterium and aspergillus niger.
The fungistatic effect that embodiment 5- FRUCTUS TERMINALIAE IMMATURUS extract is combined with sodium benzoate
Benzoic acid and sodium benzoate is common chemical preservative, it is expected that its dosage in cosmetics can be reduced while can
To retain the fungistatic effect of preservative.
The aqueous sodium benzoate solution for preparing 0.1%, 0.2%, 0.3%, 0.4% and 0.5% respectively uses 2 institute of embodiment
The method stated detects them respectively to the SGT of staphylococcus aureus and Escherichia coli.In addition, to these aqueous sodium benzoate solutions
30% ethanol eluate of FRUCTUS TERMINALIAE IMMATURUS of 12.5mg/ml is added, equally detects their SGT.As a result it see the table below.
The SGT of the individual aqueous sodium benzoate solution of table 5.
The SGT of individual 30% ethanol eluate of aqueous sodium benzoate solution+FRUCTUS TERMINALIAE IMMATURUS of table 6.
The usage amount of chemical preservative sodium benzoate can be largely reduced using the FRUCTUS TERMINALIAE IMMATURUS extract of 12.5mg/ml.It is another
Aspect, the usage amount of FRUCTUS TERMINALIAE IMMATURUS extract itself can also be reduced using the sodium benzoate of low concentration, to reach satisfied
Fungistatic effect.The effect of table 5 and 6 in conjunction with the embodiments 2 and 3 experimental result be able to demonstrate that sodium benzoate and FRUCTUS TERMINALIAE IMMATURUS ethanol elution
Liquid has synergistic effect at antibacterial aspect.
The fungistatic effect of 6-bactericidal composition of embodiment
Antibacterial/anticorrosive composite of the invention is prepared according to following component and proportion.
Antibacterial/anticorrosive composite of 7. different components content of table
Small beaker is added in the water of 30% ethanol eluate of FRUCTUS TERMINALIAE IMMATURUS, sodium benzoate, disodium ethylene diamine tetraacetate and surplus
In, be warming up to 40-60 DEG C, stirring to it is complete it is molten after be down to room temperature to obtain the final product；Or continue to filter, it dispenses.
1. experimental strain
Experimental strain includes: staphylococcus aureus, Escherichia coli, bacillus subtilis, pseudomonas aeruginosa, white thought
Pearl bacterium and aspergillus niger.
2. Odontothrips loti measures the step of inhibition zone:
A. bacterium is activated, and secondary culture is to OD595For 0.3-0.5, it is diluted to 106cfu/ml.It is uniformly coated on respectively
On LB agar plate；By fungi activation culture, and it is diluted to 105Cfu/ml is uniformly coated on rose-bengal agar plate respectively
On.
B. composition A, B, C are dissolved separately in distilled water, are settled to 100mL.
C. Oxford cup is placed on planar surface, 100 μ L bacteria inhibiting composition solution is separately added into each Oxford cup, put down
Row three.
D. smoothly move into constant incubator in, 37 DEG C stationary culture 16 hours.With the transparent inhibition zone of vernier caliper measurement
Diameter.
3. measurement result
Experimental result such as following table, passes through the fungistatic effect of Odontothrips loti detection combination object, the results showed that composition A, B, C points
It is other that all there is preferable inhibitory effect to bacterium, yeast and mould.When sodium benzoate independent role, golden yellow grape cannot be inhibited
Coccus and Escherichia coli grow on plate, fail to form transparent inhibition zone；When FRUCTUS TERMINALIAE IMMATURUS, EDTA-2Na and sodium benzoate compound
In use, playing inhibiting effect to the growth of the microorganisms such as staphylococcus aureus and Escherichia coli, formed around Oxford cup
Transparent antibacterial circle diameter is greater than 8mm.
Inhibitory effect of the anti-corrosion/bactericidal composition of the invention of table 8. for various bacteriums, yeast and mould
The above result shows that bactericidal composition of the invention has the bacteriostatic activity of a wider spectrum.
Embodiment 7-determines the fungistatic effect of bactericidal composition according to growth curve
0.1%, 0.3% and 0.5% aqueous sodium benzoate solution is determined and by embodiment 6 according to the raw curve of microorganism
The growth curve of staphylococcus aureus and Escherichia coli in the presence of composition A, B and C of preparation.50 μ g/ml kanamycins are made
For positive control.
1. the drafting of microbial growth curve includes the following steps:
A. 10 μ L microbial bacteria suspensions are taken, until 37 DEG C are cultivated 16 hours in 5mL LB culture medium.
B. it takes within second day the bacteria suspension of 10uL to be transferred to 5mL respectively and contains trial drug, blank control and positive control (50
μ g/ml kanamycins) in LB culture medium, 37 DEG C are cultivated 16 hours.
C. third day is centrifuged all culture solutions for including trial drug, blank control and positive control by 5,000 RPM
Processing 5 minutes, outwells supernatant.It is configured to bacteria suspension again with 5mL fresh LB respectively again.It repeats by centrifugal treating extremely
Again the step of being configured to bacteria suspension is twice to ensure that drug ingedient is removed in culture medium.
D. take 200 μ L bacteria suspensions into 96 orifice plates, light absorption value under every 15 minutes reading 595nm.By light absorption value and time point
Draw growth curve of the microorganism in different tests drug, blank control and positive control.
2. measurement result
Growth curve lag it is more, indicate drug or composition it is stronger to the damage capability of microorganism, illustrate the drug or
The antibacterial anti-corrosion effect of composition is better.Fig. 2 is after sodium benzoate acts solely on staphylococcus aureus 16 hours, and growth is bent
The hysteresis of line.Sodium benzoate dosage includes 0.1%, 0.3% and 0.5%.Fig. 3 is FRUCTUS TERMINALIAE IMMATURUS, EDTA-2Na and benzoic acid
Sodium collective effect is after staphylococcus aureus 16 hours, growth curve stagnation behavior.Fig. 4 is that sodium benzoate acts solely on greatly
After enterobacteria 16 hours, the hysteresis of growth curve.To Fig. 5 be FRUCTUS TERMINALIAE IMMATURUS, EDTA-2Na and sodium benzoate collective effect in
After Escherichia coli 16 hours, growth curve stagnation behavior.To sum up, FRUCTUS TERMINALIAE IMMATURUS and sodium benzoate, which are applied in combination, can enhance to micro- life
The inhibiting effect of object.
Embodiment 8-antibacterial agent of the present invention quantitative measurement
In the case where antibacterial agent is applied to makeup/beauty product, need to antibacterial agent application in the product into
Row assessment.Other than considering the Costco Wholesale of antibacterial formula material, if the ingredient in antibacterial agent will lead to product and physics occur
The variation (such as color and smell variation) of property, it will the purchase for influencing consumer is intended to, and product market value is caused to decline.
1. color measurement
The absorption spectrum of slave 200nm to the 700nm of the various antibacterial agents of the present invention is detected using spectrophotometer.Detection resists
Microbial inoculum is as follows:
1) FRUCTUS TERMINALIAE IMMATURUS water extract (20mg/ml)
2) 30% ethanol eluate of FRUCTUS TERMINALIAE IMMATURUS water extract (20mg/ml)
3) 30% ethanol eluate of FRUCTUS TERMINALIAE IMMATURUS water extract (20mg/ml)+0.125%EDTA-2Na+0.1% sodium benzoate
As a result as shown in Figure 6.From fig. 6, it can be seen that FRUCTUS TERMINALIAE IMMATURUS water extract after 30% ethanol elution (antibacterial agent 2) and
3)) obvious than (antibacterial agent 1) before elution) absorptance reduction.It is wherein most obvious with wavelength 420nm to 700nm (visible light), show
Antibacterial agent 2) and perceived color 3) obviously turn light.Since color will affect beauty/cosmetics perception, antibacterial agent 2) and
3) it is suitable for the product.
2. smell is tested
Smell test assessment uses forced-choice method (forced according to international standard BS EN13725:2003's
Choice) Lai Jinhang odorousness is analyzed.
All formula odorousnesses are 25 to 47OUE/M3, average out to 34OUE/M3(LOD=6OUE/M3)
Smell release rate calculating=(34X0.01M on an hourly basis3/s/0.7101g)X1000/3600
=0.133OU/mg/M3/h
All formula (antibacterial agents 1), 2) and 3)) odorousness be far below the grass view industrial odor of European Union 2010 and discharge
Rate is lower than 10OU/mg/M3The standard of/h.
Embodiment 9-includes the cosmetic formulations and its estimation of stability of bactericidal composition of the present invention
1. moisturizing emulsion formula
9. moisturizing emulsion formula of table
2. estimation of stability
For the stability applied in cosmetics of antiseptic composition prepared by research embodiment 6, matching with table 9 respectively
Side prepares moisturizing emulsion, and number is A, B, C respectively.It stores one month under normal temperature conditions, observes its stability.As a result it sees below
Table.
Table 10. includes estimation of stability of the bactericidal composition of the present invention in beauty/make-up composition
10 stability test of table include as the result is shown the cosmetics of antibacterial agent of the present invention under various conditions, be able to maintain
One, stablize.
To the antibacterial agent 2 in embodiment 8) Stability Determination similar with 3) progress, as a result same display is comprising described anti-
The cosmetics of microbial inoculum are stable and uniform in one month.
The inhibitor effectiveness of 10-cosmetics of embodiment is tested
In order to test practical anti-corrosion ability of the bactericidal composition of the present invention in cosmetics, to the guarantor comprising the composition
Wet emulsion formulations have carried out inhibitor effectiveness test.Recommend with reference to U.S.'s cosmetics, toilet articles and essence association (CTFA) anti-
Rotten System Effectiveness evaluation method carries out 28 days singles and connects the experiment of bacterium challenge.
1. experimental strain: bacterium (Escherichia coli, staphylococcus aureus, pseudomonas aeruginosa)；Mould (aspergillus niger) and
Saccharomycete (Candida albicans).It is inoculated with using Mixed Microbes.
2. test specimen: being formulated preparing cosmetics by embodiment 9, every 100g cosmetics contain antibacterial/anticorrosive composite A respectively
Or B or C, it is A, B, C that prepared cosmetics are numbered respectively.
3. sample inoculation: weigh each 2 parts of the cosmetics (A containing composition, B, C respectively) of embodiment 9 respectively, 100 grams every part,
It is separately added into 0.2mL bacterial suspension and 0.2mL fungi suspension, is uniformly mixed, makes bacterial concentration 106CFU/g, fungi are dense
Degree is 104CFU/g.Sample is saved in (22.5 ± 2.5 DEG C) of room temperature, and respectively in 0,7,14,21,28 day detection cosmetics
Clump count, to judge the inhibitor effectiveness of cosmetics.
Comparative example is without FRUCTUS TERMINALIAE IMMATURUS, and other components are identical as A.
4. experimental result
Table 11
As seen from the above table, this law invention bactericidal composition A, B, C is used in moisturizing emulsion, can add bacterium-by single
Preservation challenge test in 28 days.
The assessment of 11-consumer safety of embodiment
Cutaneous sensibility test: 18-35 years old voluntary participation researcher is divided into two groups no less than 10 ages, respectively
For experimental group and placebo.Experiment is with the progress of double blind mode, the beauty/change for the antibacterial agent of the present invention that joined various concentration
Cosmetic and blank control and positive control carry out skin sensitivity test with volunteer, and one month by a definite date.Dduring test, certainly
Hope person must not coat other skin-protection products on the skin of test.It is tested on the inside of the left and right of volunteer.Test article first by
Aseptic filter paper absorbs, then with the gauze of subsensitivety and immobilization with adhesive tape position.The skin surface of volunteer's hand medial surface changes will
It can be with cameras record.Reaction of the volunteer to product is collected with way of questionnaires after being completed.
After all volunteers's (including experimental group and placebo) coat the cosmetics containing antibacterial agent of the present invention, do not have
The red and swollen trace of appearance, which is itched, waits allergic reactions.
Claims (10)
1. a kind of FRUCTUS TERMINALIAE IMMATURUS extract, the extract is prepared by method comprising the following steps:
(1) FRUCTUS TERMINALIAE IMMATURUS through fragmentation is extracted with water to obtain water extract, and
(2) make the water extract by macroporous absorbent resin, and then eluted with ethanol water.
2. the FRUCTUS TERMINALIAE IMMATURUS extract of claim 1, wherein the macroporous absorbent resin is low pole macroporous absorbent resin, more preferably
Low pole styrene tyle macroporous adsorption resin, most preferably AB-8 macroporous absorbent resin.
3. the FRUCTUS TERMINALIAE IMMATURUS extract of claims 1 or 2, wherein the ethanol water includes the ethyl alcohol of 5%-50%, preferably
The ethyl alcohol of 8%-40%, the ethyl alcohol of more preferable 10-30%.
4. a kind of bactericidal composition, it includes the FRUCTUS TERMINALIAE IMMATURUS extracts of any one of claim 1-3.
5. the bactericidal composition of claim 4, also includes:
Benzoic acid or its cosmetically acceptable soluble-salt, preferred sodium benzoate and/or ethylenediamine tetra-acetic acid or its cosmetically
Acceptable soluble-salt, preferably disodium ethylene diamine tetraacetate.
6. the bactericidal composition of claim 4 or 5, wherein calculated based on the bactericidal composition, the FRUCTUS TERMINALIAE IMMATURUS extract
Weight percent is 0.5%-10%, preferably 1%-5%, more preferable 1.25%-2%.
7. the bactericidal composition of claim 4 or 5, wherein the FRUCTUS TERMINALIAE IMMATURUS extract is with 50-300g/L, preferably 80-200g/
L, the FRUCTUS TERMINALIAE IMMATURUS of more preferable 100g/L/final FRUCTUS TERMINALIAE IMMATURUS extract amount is obtained from FRUCTUS TERMINALIAE IMMATURUS.
8. the bactericidal composition of claim 6, wherein calculated based on the bactericidal composition, the benzoic acid or its cosmetically may be used
The weight percent of the soluble-salt of receiving is 0.05%-1%, preferably 0.075%-0.75%, more preferable 0.1%-0.5%；
And/or
The ethylenediamine tetra-acetic acid or its cosmetically acceptable soluble-salt weight percent be 0.075%-0.5%, it is excellent
Select 0.1%-0.3%, more preferable 0.125%-0.2%.
9. the FRUCTUS TERMINALIAE IMMATURUS extract of any one of claim 1-3 or the bactericidal composition of any one of claim 4-8 are used
Help the purposes of the perhaps antibacterial agent of cosmetic product.
10. the composition of a kind of local application or dermal administration, it includes the FRUCTUS TERMINALIAE IMMATURUS extracts of any one of claim 1-3
Or the bactericidal composition of any one of claim 4-8, the composition of the local application or dermal administration is preferably beauty
Or make-up composition.
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