CN108703931A - A kind of composition and preparation method thereof with moisture-keeping function - Google Patents

Abstract

The invention belongs to daily chemical product field, a kind of composition with moisture-keeping function and preparation method thereof and skin care item are disclosed.Composition of the present invention with moisture-keeping function is made of dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis, and the dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are (1~4):(1~4):(1~4):(1~4).The dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis proportion it is shared, moistening effect be better than each one-component.Composition of the present invention can be applied to the external preparation for skin skin care item of the various dosage forms such as gel, mildy wash, face cream, lotion.

Description

A kind of composition and preparation method thereof with moisture-keeping function

Technical field

The invention belongs to daily chemical product fields, and in particular to a kind of composition with moisture-keeping function and preparation method thereof with Skin care item.

Background technology

Skin divides epidermis, corium and subcutaneous fat.The outermost layer of epidermis is cuticula, due to the suction of cuticula itself The effect of natural moisturizing factor, that is, amino acids, lactate and carbohydrate contained in water, barrier function and cuticula etc., So that cuticula is kept certain water content, the moistening of skin, normal keratoderma is maintained to usually contain the water of 10%-30% Point, to maintain the softness and elasticity of skin.But with advancing age, keratoderma moisture can gradually decrease, and work as When the moisture of keratoderma is less than 10%, skin just will appear dry, tight, coarse and furfur etc..In general, people Moisture in body skin can change with factors such as age, season, environment, health status, when water content in skin When relatively low, not only the dark and gloomy influence of pachylosis is beautiful, can also accelerate the aging of skin, or even there is also trace and itch, is rubescent, piercing Bitterly, the symptoms such as sensitivity.So keeping the moisture in skin not only for the beautiful health for more importantly maintaining skin.And The skin moment is all in direct contact with external environment, if not being protected, more or less has water shortage, directly affects skin Appearance.Want to possess that health, water is tender, skin of bullet profit, gloss, the most important condition is exactly that moisture is sufficient and the guarantor of skin in skin It is aqueous good.

Have much with the relevant substance of moisturizing in skin, wherein aquaporin AQP3 is transdermal delivery on a kind of cell membrane The transport protein of the small-molecule substances such as water, glycerine and urea participates in the functions such as skin hydration, skin barrier；Tight junction protein ZO-1 has the function of closing cell gap, is the barrier for hindering substance diffusion, and cell and external body fluid is made to separate；The poly- egg of silk Several GAP-associated protein GAPs that white FLG is related to epidermal differentiation process have important barrier function, can prevent loss of moist, maximum limit Reduce the invasion effect of allergen and microorganism in degree ground；Loricrin Loricrin is the main component of cutin born of the same parents envelope, Ke Yijia Gu cutin born of the same parents seal, enhances permeability barrier, and interact with median fiber silk, increase the flexibility of cutin born of the same parents' seal structure.So And some extraneous factors can cause skin to generate inflammatory reaction, generate excessive inflammatory factor such as TNF-α, can make skin permeation Property enhancing, integrality reduce, influence skin barrier function.

Existing moisturizer is main on the market at present or improves dry skin problem by " water lock+moisturizing " approach.Closely As consumer is to the growing interest of own health and safety over year, relative to the cosmetic industry raw materials of many biochemical synthesis, It is distinguished from natural, effect and can be more favored by daily edible raw material.So exploitation more safely, effectively, tool The product of Saccharideisomerate is very necessary.

Invention content

In view of this, the purpose of the present invention is to provide a kind of composition with moisture-keeping function, active constituent is day Right active constituent is combined and has the function that water lock, moisturizing, and safely and effectively moistening effect is reached.

To achieve the purpose of the present invention, the present invention adopts the following technical scheme that：

A kind of composition with moisture-keeping function, is made of dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis.

Wherein, the perennial herbaceous plant that grows nonparasitically upon another plant of dendrobium candidum (Dendrobium officinale) orchid family Dendrobium, mainly It is distributed in southwest and Jiangnan each province.Early in Qin Han dynasty《Sheng Nong's herbal classic》With regard to claim dendrobium candidum " in main wound, eliminating impediment, lower gas, Tonifying five zang organs consumptive disease is frail, Qiang Yin, long term usage thickness large intestines "；《Compendium of Materia Medica》In, Li Shizhen (1518-1593 A.D.) think the stem of noble dendrobium be " this through top grade, it is sweet, It is light, micro- at, cure mainly in wound, gas under eliminating impediment, mend in never sufficient, flat stomach gas, long muscle, intelligence development is except shying, macrobiosis of making light of one's life by commiting suicide ".Study table Bright, dendrobium candidum is rich in polysaccharide, and dendrobium candidum is better than HERBA DENDROBII in the quantity and content of composition of alkaloids.Iron sheet The amino acid that the stem of noble dendrobium contains is mainly aspartic acid, and glutamic acid, glycine pick propylhomoserin and leucine, and latter two is that human body is " required Amino acid ".Pharmaceutical research shows that Dendrobium officinale polysaccharide has the effects that anti-oxidant, hypoglycemic, enhancing are immune and antitumor.

White fungus (Tremella fuciformis) is high China's economic value, extremely precious a kind of edible mushroom and medicinal Bacterium is traditional product for food and medicine purposes.It is that nourishing is strong containing carbohydrate, protein, vitamin and a variety of amino acid etc. in it The good medicine strengthen, strengthened the body resistance to consolidate the constitution.Polysaccharide is the most important constituent of white fungus, accounts for the 60%~70% of its dry weight.

Dictyophora phalloidea (Dictyophora indusiata), alias " bamboo fungus " are the general names to a major class dictyophora phalloidea category fungi.This Product with the name of bamboo straw mat first recorded in《Dietetic materia medica》, cloud：" moon in cizu woods summer meets rain, and drop juice lands raw straw mat, and like deer horn, white can Food." dictyophora phalloidea has strengthening by means of tonics, QI invigorating cerebrum tonifying, allays excitement and be healthy and strong, boosting qi and nourishing yin, moisten the lung and relieve the cough and clearing heat and promoting diuresis and other effects.Bamboo Sweet-smelling grass can protect liver, reduce the accumulation of Abdominal Wall Fat, therefore be commonly called as " frizing ".Modern study also turns out that dictyophora phalloidea has There is capable of inhibiting tumour, anti-oxidant and antibacterial.

Radix Ophiopogonis (Ophiopogon japonicas (Linn.f.) Ker-Gawl) sweet in flavor, slight bitter, cold nature.With enriching yin Moistening lung, reinforcing stomach reg fluid, relieving restlessness that clears away heart-fire and other effects.Modern pharmacology research shows Radix Ophiopogonis also with anti-oxidant, hypoglycemic, anti-tired The effects that labor, anti-arrhythmia, antibacterial.Ophiopogonpolysaccharide is one of common cosmetic material, and main function is moisturizing and prolongs Slow aging.

Preferably, the dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are (1~4):(1~4):(1~4): (1~4).

In some embodiments, the dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are 4:3:4:3.

In some embodiments, the dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are 3:4:3:4.

In some embodiments, the dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are 1:1:1:1.

The present invention also provides a kind of preparation method of composition, raw material crushes, is sieved, and carries out heat after extracting in water successively Refined filtration and cold refined filtration collect extracting solution.

Preferably, the sieving is to sieve with 100 mesh sieve.

Preferably, the extracting in water is by 1:The solid-liquid ratio of (75~85) adds water, and 55~65 DEG C of extractions 110~ 130min.In some embodiments, the extracting in water is by 1:80 solid-liquid ratio adds water, 60 DEG C of extraction 120min.

The present invention further includes the process impregnated before extraction.It is fully dissolved out immersed with conducive to active ingredient, in can also shortening Medicine decocting time is avoided leading to the consume of drug moiety active ingredient because decocting time is long, be destroyed excessively.

Preferably, the soaking time is 0.5h.

Preferably, the hot refined filtration is to put cardboard H60 in Buchner funnel, the extracting solution for injecting warm soaks, and opens and takes out Filter negative pressure, which is smoked, to be made cardboard be tightly attached to funnel but to ensure that entire cardboard is wetting, and bright extracting solution must be clarified.Wherein hot refined filtration Filter plate aperture be preferably 1 μm.

The preparation method of composition of the present invention further includes the steps that cold refined filtration after hot refined filtration to remove because temperature drops It is low caused to be partly precipitated.

Preferably, the cold refined filtration is to put cardboard H70 in Buchner funnel, the extracting solution of warm is injected, opens to filter and bear Pressure, which is smoked, to be made cardboard be tightly attached to funnel but to ensure that entire cardboard is wetting, and bright extracting solution must be clarified.The filter of wherein cold refined filtration Panel aperture is preferably 0.45 μm.

The preparation method of composition of the present invention controls extracting solution turbidity≤2.0FNU after cold refined filtration.

Composition of the present invention can be applied to the external preparation for skin skin care of the various dosage forms such as gel, mildy wash, face cream, lotion Product.

The present invention also provides a kind of skin care item with moisture-keeping function, including the composition with moisture-keeping function And matrix.

In some embodiments, the dosage of the composition described in the skin care item is preferably 0.25wt%.

Further, in some embodiments the matrix by glycerine, 1,2- pentanediols, hydroxy-ethyl acrylate/propylene Acyl dimethyltaurine sodium copolymer, Phenoxyethanol/Sensiva SC50 and water composition.

The matrix can be prepared as follows：

(1) glycerine, 1,2- pentanediols and water as A phase raw materials be heated to 80 DEG C be uniformly mixed, it is spare

(2) stirring cooling, 60 DEG C of additions are total as hydroxy-ethyl acrylate/sodium acryloyldimethyl taurate of B phase raw materials Polymers (SEPINOV EMT 10), stirs evenly；

(3) stirring cooling, 40 DEG C or less are added Phenoxyethanol/Sensiva SC50 (EUXYL PE as C phase raw materials 9010) it, stirs evenly；

(4) stirring cooling, 38 DEG C of dischargings, obtains matrix.

The extracting solution of composition of the present invention is in after drying pulverulence, is obtained with matrix mixing after being dissolved in water Obtain the skin care item with moisture-keeping function.Preferably, the addition of the composition powder is 0.25wt%.

The present invention also provides a kind of method for moisturizing, skin surface using composition of the present invention or it is above-mentioned containing The skin care item of composition of the present invention.As smeared the gel containing composition of the present invention, face cream, lotion；Or use contains There is the mildy wash of composition of the present invention to carry out the cleaning of face or other area skins；Or use is by above-mentioned matrix and Ben Fa The skin care item of the bright composition composition.

It can be seen that the present invention provides a kind of composition with moisture-keeping function and preparation method thereof and skin care item.This The invention composition with moisture-keeping function is made of dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis, the dendrobium candidum, white fungus, The mass ratio of dictyophora phalloidea and Radix Ophiopogonis are (1~4):(1~4):(1~4):(1~4).The dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis It proportions shared, moistening effect is better than each one-component.Composition of the present invention can be applied to gel, mildy wash, face The external preparation for skin skin care item of the various dosage forms such as frost, lotion.

Description of the drawings

In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technology description to be briefly described.

Fig. 1 shows each component hydroscopicity comparison result；

Fig. 2 shows each component moisturizing rate comparison result；

Fig. 3 shows each embodiment moisture content change amount comparison result；

Fig. 4 shows each embodiment moisture loss variable quantity comparison result；

Fig. 5 shows cell viability curve of the working substance (g) under various concentration in test example 3；

Fig. 6 shows working substance (g) Loricrin Protein Detection fluorograms in test example 3；

Fig. 7 shows working substance (g) Filaggrin Protein Detection fluorograms in test example 3；

Fig. 8 shows working substance (g) Zonula Occludens-1 Protein Detection fluorograms in test example 3；

Fig. 9 shows 3 Protein Detection fluorograms of working substance (g) Aquaporin in test example 3；

Figure 10 shows working substance (g) inflammation-related factor TNF α secretory volume variation tendency in test example 3.

Specific implementation mode

The invention discloses a kind of composition and the preparation method and application thereof with moisture-keeping efficacy.Those skilled in the art Present disclosure can be used for reference, technological parameter realization is suitably modified.In particular, it should be pointed out that all similar substitutions and modifications pair It is it will be apparent that they are considered as being included in the present invention for those skilled in the art.The method and product of the present invention is Be described through passing through preferred embodiment, related personnel obviously can not depart from the content of present invention, in spirit and scope to this Method described in text is modified or suitably changes and combines, to realize and apply the technology of the present invention.

For a further understanding of the present invention, below in conjunction with the embodiment of the present invention, to the technical side in the embodiment of the present invention Case is clearly and completely described, it is clear that and described embodiments are only a part of the embodiments of the present invention, rather than all Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art institute without making creative work The every other embodiment obtained, shall fall within the protection scope of the present invention.

Unless otherwise specified, reagent involved in the embodiment of the present invention is commercial product, can pass through business canal Road purchase obtains.

Embodiment 1

Clean dried beaker is taken, dendrobium candidum is weighed, by 1:80 solid-liquid ratio adds deionized water to impregnate 0.5h in beaker Afterwards, stirring, 60 DEG C of extraction 120min are begun to warm up.Then hot refined filtration is carried out, cardboard H60 (1 μm), injection temperature are put in Buchner funnel Hot water (extracting solution) soaks, and opens and filters negative pressure pumping, so that cardboard is tightly attached to funnel but to ensure that entire cardboard is wetting, obtains Clarify bright extracting solution.Cold refined filtration is carried out later, and cardboard H70 (0.45 μm), injection warm water (extracting solution) are put in Buchner funnel It soaks, opening to filter negative pressure and smoke makes cardboard be tightly attached to funnel but to ensure that entire cardboard is wetting, obtains and clarifies bright extraction Liquid (turbidity control≤2.0FNU).

Embodiment 2

White fungus is weighed, raw material is pre-processed according to the method for embodiment 1, is extracted, hot refined filtration and cold refined filtration, is obtained clear Clear bright extracting solution (turbidity control≤2.0FNU).

Embodiment 3

Dictyophora phalloidea is weighed, raw material is pre-processed according to the method for embodiment 1, is extracted, hot refined filtration and cold refined filtration, is obtained clear Clear bright extracting solution (turbidity control≤2.0FNU).

Embodiment 4

Radix Ophiopogonis is weighed, raw material is pre-processed according to the method for embodiment 1, is extracted, hot refined filtration and cold refined filtration, is obtained clear Clear bright extracting solution (turbidity control≤2.0FNU).

Embodiment 5

Clean dried beaker is taken, by 4:3:4:3 ratio weighs dendrobium candidum, white fungus, dictyophora phalloidea, Radix Ophiopogonis, by 1:80 material After liquor ratio adds deionized water to impregnate 0.5h in beaker, stirring, 60 DEG C of extraction 120min are begun to warm up.Then hot refined filtration is carried out, Cardboard H60 (1 μm) is put in Buchner funnel, injection warm water (extracting solution) soaks, and opens and filters negative pressure pumping, cardboard is made to be tightly attached to leakage It struggles against but to ensure that entire cardboard is wetting, obtain and clarify bright extracting solution.Cold refined filtration is carried out later, and cardboard is put in Buchner funnel H70 (0.45 μm), injection warm water (extracting solution) soak, and opening to filter negative pressure and take out makes cardboard be tightly attached to funnel but to ensure entire Cardboard is wetting, obtains and clarifies bright extracting solution (turbidity control≤2.0FNU).

Embodiment 6

Clean dried beaker is taken, by 3:4:3:4 ratio weighs dendrobium candidum, white fungus, dictyophora phalloidea, Radix Ophiopogonis, by 1:(80±5) Solid-liquid ratio add deionized water to impregnate 0.5h in beaker after, begin to warm up stirring, 60 ± 5 DEG C of 120 ± 10min of extraction.Then Hot refined filtration is carried out, cardboard H60 (1 μm) is put in Buchner funnel, injection warm water soaks, and opens and filters negative pressure pumping, cardboard is made to be close to In funnel but to ensure that entire cardboard is wetting, obtain and clarify bright extracting solution.Cold refined filtration is carried out later, is put in Buchner funnel Cardboard H70 (0.45 μm), injection warm water soak, and opening to filter negative pressure and take out makes cardboard be tightly attached to funnel but to ensure entire cardboard It is wetting, obtains and clarify bright extracting solution (turbidity control≤2.0FNU).

Embodiment 7

Clean dried beaker is taken, by 1:1:1:1 ratio weighs dendrobium candidum, white fungus, dictyophora phalloidea, Radix Ophiopogonis, by 1:(80±5) Solid-liquid ratio add deionized water to impregnate 0.5h in beaker after, begin to warm up stirring, 60 ± 5 DEG C of 120 ± 10min of extraction.Then Hot refined filtration is carried out, cardboard H60 (1 μm) is put in Buchner funnel, injection warm water soaks, and opens and filters negative pressure pumping, cardboard is made to be close to In funnel but to ensure that entire cardboard is wetting, obtain and clarify bright extracting solution.Cold refined filtration is carried out later, is put in Buchner funnel Cardboard H70 (0.45 μm), injection warm water soak, and opening to filter negative pressure and take out makes cardboard be tightly attached to funnel but to ensure entire cardboard It is wetting, obtains and clarify bright extracting solution (turbidity control≤2.0FNU).

Embodiment 8

By 2:1:1 ratio weighs dendrobium candidum, dictyophora phalloidea, Radix Ophiopogonis, is located in advance to raw material according to the method for embodiment 1 Reason, extraction, hot refined filtration and cold refined filtration, obtain and clarify bright extracting solution (turbidity control≤2.0FNU).

Embodiment 9

By 2:1 ratio weighs dendrobium candidum, dictyophora phalloidea, raw material is pre-processed according to the method for embodiment 1, is extracted, Hot refined filtration and cold refined filtration obtain and clarify bright extracting solution (turbidity control≤2.0FNU).

Test example 1, moisture-absorbing moisture-keeping performance measure

1), experiment material

Ammonium sulfate, plate, glass desicator

2), detection method

1. dendrobium candidum that embodiment 1-4 is prepared, white fungus, dictyophora phalloidea, Radix Ophiopogonis extract to be freeze-dried obtain respectively Freeze-dried powder.Freeze-drying condition is：- 76 DEG C of temperature, pressure 60-70mTor, the time is for 24 hours.

2. solid sample directly takes freeze-dried powder to weigh, three, each sample is parallel (sample dry weight is denoted as M1).

3. it is 81% that each plate for filling sample, which is positioned over by the relative humidity that ammonium sulfate saturated solution is reached, first Drier in, place 96h and weigh after sample reaches moisture equilibrium at dry side each sample weight (weight is denoted as M2 behind sample Xishui County), by The weight difference of front and back sample is placed to calculate the hydroscopicity of each sample.

Hydroscopicity calculation formula is as follows：

Hydroscopicity (%)=(M2-M1)/M1*100%

4. by the above-mentioned sample for having reached moisture equilibrium at dry side, it is placed in that reach relative humidity by potassium acetate saturated solution be 46% Drier in.Continue place 96 hours after sample dehydration reach balance after weigh each sample weight (sample water suction after weight It is denoted as M3), calculate the moisturizing rate of each sample.

Moisturizing rate calculation formula is as follows：

Moisturizing rate (%)={ 1-[(M2-M1)-(M3-M1)/(M2-M1)]} × 100%

3), result

Each component hydroscopicity and moisturizing rate the result is shown in Figure 1-2 and table 1-2.

1 each component hydroscopicity result of table

Title	Hydroscopicity
		White fungus	58.89% ± 0.0141
Dictyophora phalloidea	41.84% ± 0.0169
		Radix Ophiopogonis	23.69% ± 0.0142
Dendrobium candidum	23.64% ± 0.00541

2 each component moisturizing rate result of table

Title	Moisturizing rate
		Radix Ophiopogonis	38.76% ± 0.0637
Dendrobium candidum	30.73% ± 0.0117
		Dictyophora phalloidea	24.19% ± 0.0107
White fungus	13.24% ± 0.00684

From the above results：

(1) in hygroscopicity experiment, Yin Er >Zhu Sun >Mai Dong >Dendrobium candidum；

(2) in moisture retention experiment, Mai Dong >Tie Pishihu >Zhu Sun >White fungus.

Test example 2, human body moisture-keeping efficacy measure

1) working substance

(a) 1 dendrobium candidum extracting solution of embodiment is freeze-dried to obtain freeze-dried powder；

(b) 2 tremella extract of embodiment is freeze-dried to obtain freeze-dried powder；

(c) 3 extract from dictyophora of embodiment is freeze-dried to obtain freeze-dried powder；

(d) 4 Radix Ophiopogonis extract of embodiment is freeze-dried to obtain freeze-dried powder；

(e) 5 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder；

(f) 6 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder；

(g) 7 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder；

(h) 8 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder；

(i) 9 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder.

The above freeze-drying condition is：- 76 DEG C of temperature, pressure 60-70mTor, the time is for 24 hours.

2) matrix

In parts by mass, matrix formulations design such as table 3.

3 matrix formulations table of table

Matrix formulation procedure：

(1) A phases raw material be heated to 80 DEG C be uniformly mixed, it is spare；

(2) stirring cooling, 60 DEG C of addition B phase raw materials, stirs evenly；

(3) stirring cooling, 40 DEG C or less addition C phase raw materials, stirs evenly；

(4) stirring cooling, 38 DEG C of dischargings, obtains matrix.

3) test sample

In parts by mass, 0.25 part of each working substance (being freeze-dried to obtain freeze-dried powder) is taken to be added in 9.75 parts of water, and on 90 parts Matrix is stated to be mixed evenly.

4) moisture is tested

Moisture test uses CORNEOMETER- capacitance methods.The humidity measurements that its result passes through setting (Moisture Measurement Value, MMV) change rate indicates.Subject is using product 30min, 1h, 2h and 4h Skin moisture is measured afterwards.Laboratory apparatus is：Corneometer CM825, German CK companies.

The calculation formula of moisture content change amount is：Moisture content change amount=Tn-T0, in formula：T0 is not smear sample When basal data collection value；Tn is each time point data collection value of sample sets.Moisture content change amount is higher, illustrates moisturizing Effect is more apparent.As a result see Fig. 3 and table 4.

4 moisture content change amount result of table

Note：It is analyzed using Duncan ' s multiple range test methods, same row indicates different capital letter matrixs Show group difference extremely significantly (P<0.01)；Indicate different lowercase letter group differences significantly (P<0.05)；Indicate same word Not notable (the P&gt of female expression group difference;0.05).Wherein, the hyaluronic acid that positive control is 0.1%, matrix are as shown in table 3.

Above-mentioned moisture experimental result is shown：

(1) it applies the data measured after sample 5min to show, 11 sample room differences extremely significantly (P<, and f&gt 0.01);g>e>h>d> c>Yang Xing >i>b>a>Matrix, prescription are better than folk prescription；

(2) it applies the data measured after sample 30min to show, 11 sample room differences extremely significantly (P<, and f&gt 0.01);g>e>h>i >d>b>c>a>Yang Xing >Matrix, prescription are better than folk prescription；

(3) it applies the data measured after sample 60min to show, 11 sample room differences extremely significantly (P<, and h&gt 0.01);g>f>e>i >c>d>b>a>Yang Xing >Matrix, prescription are better than folk prescription；

(4) it applies the data measured after sample 120min to show, 11 sample room significant difference (P<, and e&gt 0.05);g>f>d>i> c>Yang Xing >b>h>a>Matrix；

(5) it applies the data measured after sample 240min to show, the not notable (P&gt of difference between c and i and b and e;0.05)；

Based on the above results it is found that the moistening effect of composition of the present invention is better than folk prescription, and deposited between sample and matrix In pole significant difference (P<0.01).

5) water loss amount is tested

Subject and test environment are identical as skin determination of moisture assessment.

The diffusion principle that experimental evidence A.Fick has found measures the variation of neighbouring skin surface moisture vapor pressure, weighs flesh Skin surface moisture wastage, water loss amount TEWL (Trans-Epidermal Water Loss) expression, unit g/ hm², this value is lower, illustrates that sample prevents moisture loss effect more apparent, skin barrier function is more perfect.Subject is using sample Skin water loss amount change rate is measured after product 30min, 1h, 2h and 4h.Laboratory apparatus is：Tewameter TM300, German CK Company.

The calculation formula of skin water loss amount change rate is：Water loss amount change rate %=(Tn-T0)/T0 × 100, In formula：T0 is basal data collection value when not smearing sample；Tn is each time point data collection value of sample sets.Moisture loss Quantitative change rate negative value is lower, illustrates that water lock effect is more apparent.As a result see Fig. 4 and table 5.

5 water loss amount change rate result of table

Note：It is analyzed using Duncan ' s multiple range test methods, same row indicates different capital letter matrixs Show group difference extremely significantly (P<0.01)；Indicate different lowercase letter group differences significantly (P<0.05)；Indicate same word Not notable (the P&gt of female expression group difference;0.05).

Above-mentioned moisture loss experimental result is shown：

(1) after applying sample, water loss amount data are measured in 1h, 2h, 4h, find 11 sample room differences extremely significantly (P< 0.01)；

(2) after 4h stabilizations, it is found that composition of the present invention prevents the effect of moisture loss to be better than folk prescription.

Test example 3, the barrier based on epidermal cell and moisturizing GAP-associated protein GAP content detection

1) working substance

(g) 7 composition of embodiment is made extracting solution and is freeze-dried to obtain freeze-dried powder；Freeze-drying condition is：Temperature -76 DEG C, pressure 60-70mTor, the time is for 24 hours.

2) test sample

By working substance (being freeze-dried to obtain freeze-dried powder) according to each cell experiment sample dilute requirement solvent or water carry out it is dilute It releases.

3) MTT detection methods

Mtt assay is also known as MTT colorimetric methods, is a kind of method of detection cell survival and growth.Its testing principle is living cells Succinate dehydrogenase in mitochondria can make exogenous MTT be reduced to the bluish violet crystallization first a ceremonial jade-ladle, used in libation (Formazan) of water-insoluble simultaneously It is deposited in cell, and dead cell is without this function.Dimethyl sulfoxide (DMSO) (DMSO) can dissolve the first a ceremonial jade-ladle, used in libation in cell, be examined with enzyme linked immunological It surveys instrument and measures its absorbance value at 540 or 720nm wavelength, can reflect living cells quantity indirectly.Therefore, in order to determine moisturizing work( Imitate the maximum safe administration dosage of cell experiment, it is ensured that cell is maintained at higher survival rate and judges without influencing result.Sample 7 administration concentrations are set from high to low, carry out cytotoxicity test experience on keratinocyte, as a result cytotoxicity is shown in Fig. 5 and table 6.

Experimental cell is：Keratinocyte culture solution (Tissue Engineering Study institute).

6 working substance cytotoxicity testing result of table

Sample concentration (ppm)	Survival rate (%)
		25	102.54±6.34
50	101.64±0.58
		100	109.64±8.31
200	101.31±8.68
		500	99.27±0.76
1000	89.99±1.43
		2000	83.25±8.23
Negative control (4%DMSO)	20.67±2.51
		Blank group	100.00±2.40

Above-mentioned cytotoxicity test experience result is shown：

Under 1000ppm dosages, cell survival rate is 90% or more.Therefore, it is below to medicament that 1000ppm may be selected Amount carries out subsequent cell experiment.

4) barrier based on epidermal cell and moisturizing GAP-associated protein GAP content detection

Based on MTT detection methods as a result, determining maximum safe administration dosage of the sample on keratinocyte (SC) 1000ppm.On this basis, it is based on keratinocyte, carries out loricrin (Loricin), mitogen albumen (Filaggrin), tight junction protein (Zonula Occludens-1) and hydrated protein (Aquaporin 3) content detection are real It tests, meanwhile, carry out under UVB incentive conditions, inflammation-related factor (TNF α) secretes Inhibition test.The above experiment uses immunofluorescence Method detects loricrin (Loricin), mitogen albumen (Filaggrin), tight junction protein (Zonula in keratinocyte ) and the expression quantity of hydrated protein (Aquaporin 3) content Occludens-1.

Immunofluorescence technique principle：Combination between antibody and antigen has the specificity of height.Detection tissue is removed with antibody Or the similar antigenic substance in cell.Due to the compound of antigen and antibody be it is colourless, need to use cellular immunofluorescence into Row dyeing is quantitative.In the photo for inspiring green fluorescence under blue light using Image-pro Plus (IPP) software analysis cell Green fluorescence intensity, the fluorescence intensity under unit of account area.Sentence using average optical density value (IOD/Area) as protein expression Disconnected index.IOD/Area is higher, shows that expressing quantity is higher.

This laboratory sample is water-soluble, and positive control vitamin A acid dissolves for DMSO, therefore two groups of this experimental setup is negative Control, respectively SC (cultured epidermal cell liquid) and SC (the cultured epidermal cell liquid for containing 1 ‰ DMSO).

Each Protein Detection fluorogram is shown in Fig. 6 to Fig. 9, data result table 7.

Barrier and moisturizing GAP-associated protein GAP content detection of the table 7 based on epidermal cell

Note：Using with T-Test methods, to for statistical analysis, * is indicated compared with negative control (SC) group, group difference Significantly (P<0.05)；* indicates group difference extremely significantly (P<0.01).

The barrier and moisturizing GAP-associated protein GAP content detection result of above-mentioned epidermal cell are shown：

Compared with working substance negative control (SC) group, working substance is under 500ppm dosages, Human Keratinocytes skin Skin barrier correlation Loricrin albumen, Filaggrin albumen expression quantity have notable (P<0.05) castering action.Cutin is formed Cell skin moisture-keeping correlation Zonula Occludens-1 albumen, 3 albumen of Aquaporin expression quantity have extremely significantly (P< 0.01) castering action.

5) inflammation-related factor TNF-α secretion Inhibition test

Some extraneous factors can cause skin to generate inflammatory reaction, generate excessive inflammatory factor such as TNF-α, can make skin Skin permeability enhances, and integrality reduces, and influences skin barrier function.By detecting working substance to tumor necrosis factor (TNF-α) Influence, come evaluate working substance alleviate inflammation, enhance the effect of skin barrier.It empirically requires to negative control group, sample sets UVB irradiation is carried out with positive controls, establishes the cell model of inflammatory factor generation, using the ELISA detection examinations of Human TNF-α Agent box is tested.

This laboratory sample is water-soluble, and positive control dexamethasone dissolves for DMSO, therefore two groups of this experimental setup is empty White control group, SC (cultured epidermal cell liquid) and SC respectively without ultraviolet light irradiation (contain the cultured epidermal cell of 1 ‰ DMSO Liquid).Cellular damage is caused using ultraviolet irradiation, establishes inflammatory factor hypersecretion model, as the negative control of sample sets, if Two groups of negative control groups are set, the SC (cultured epidermal cell liquid) of ultraviolet light irradiation is respectively applied and apply the SC of ultraviolet light irradiation (the cultured epidermal cell liquid for containing 1 ‰ DMSO).

The result is shown in Figure 10, table 8.

8 inflammation-related factor TNF-α testing result of table

Sample ID	TNF-α concentration (pg/mL)
		Blank control group 1 (SC, 1%DMSO, UVB-)	40.42±7.32Aa
Negative control group 1 (SC, 1%DMSO, UVB+)	84.41±10.17Bb
		Positive control (0.1% dexamethasone)	59.22±17.34Bc
Blank control group 2 (SC, UVB-)	43.69±4.19Aa
		Negative control group 2 (SC, UVB+)	60.73±2.3Ab
500ppm working substances	55.96±8.85Aa

Note：Using with T-Test methods, to for statistical analysis, same row indicates different capitalizations and indicates group difference Extremely significantly (P<0.01)；Indicate different lowercase letter group differences significantly (P<0.05)；Indicate the expression group of same letter Between difference not significantly (P>0.05).

Above-mentioned inflammation-related factor TNF-α testing result is shown：

Compared with blank control group 1 (SC, 1%DMSO, UVB-), (SC, 1%DMSO, the UVB+) inflammation of negative control group 1 because The secretory volume of sub- TNF-α extremely significantly (P<0.01) rise, illustrate that this UVB radiation parameter is effective；With negative control group 1 (SC, 1%DMSO, UVB+) group compares, and positive controls (dexamethasone, 0.1%, UVB+) have the secretory volume of inflammatory factor TNF-α Significantly (P<0.05) inhibiting effect.

Compared to blank control group 2 (SC, UVB-), negative control group 2 (SC, UVB+) inflammatory factor TNF-α secretion amount is aobvious Write (P<0.05) rise, illustrate that this UVB radiation parameter is effective；Compared to negative control group 2 (SC, UVB+), working substance exists Under 500ppm dosages, there is inhibiting effect to the secretory volume of inflammation-related factor TNF-α.

Test example 4, safety evaluatio

1) working substance

Extracting solution (containing polysaccharide, >=3mg/mL) is made in 7 composition of embodiment.

2) test sample

Working substance is diluted according to each safety experiment concentration requirement with solvent or water.

3) erythrocyte hemolysis is tested

Working substance is diluted with PBS (phosphate buffer), is diluted to 40% working substance.According to chemical substance energy loss The principle design for hindering cell membrane passes through hemoglobin in the red blood cell for the fresh separated being incubated with tested material under the conditions of measurement standard The light absorption value of leakage carry out evaluation test result.Hemolysis rate is higher, and irritation is bigger.It can by erythrocyte hemolysis test result Prediction has the stimulus intensity of the sample of minimal irritation, rapidly and efficiently, can be made to plant material irritation quickly, relatively accurately Anticipation.It the results are shown in Table 9.

9 40% working substance erythrocyte hemolysis experimental result of table

Above-mentioned erythrocyte hemolysis experimental result is shown：

40% working substance hemolysis rate is 0%, shows that composition described in embodiment 7 is nonirritant.

4) the multiple Skin Irritation Test of animal

Working substance is diluted with distilled water, is diluted to 60% working substance.Before experiment for 24 hours, by back part of animal backbone two Side back hair is cut, and unhairing range is respectively 3cm × 3cm, takes tested material about 0.5g, is applied on the skin of side, is smeared range and is existed 2.5cm × 2.5cm smears 1 time/d, continuously smears 14d, other side skin is as a control group.Since the 2nd day, smear every time Preceding cropping remains tested material with warm water or nonirritant solvent cleaning, and observation is as a result, press after 1h《Cosmetics safety technical specification (2015 editions)》Skin irritation/corrosion test grade form scores, and every animal average product daily is calculated after experiment Point, judge skin irritatin intensity.Integral is higher, and irritation is bigger.It the results are shown in Table 10.

The multiple Skin Irritation Test result of 10 60% working substance animal of table

Note：One row 0/4 of erythema/oedema indicate that integral is 0 to every animal daily, and daily experimental animal quantity is 4.It calculates Formula is as follows：

The above-mentioned multiple Skin Irritation Test result of animal is shown：

Smear 60% working substance 14 days, every animal integral mean value is 0 point daily, and result judgement is nonirritant.

5) animal eye irritant test

Working substance is diluted with distilled water, is diluted to 80% working substance.80% working substance is instilled into rabbit conjunctival capsule In, eyes are not rinsed in 24 hours, finally carry out clinical examination and scoring.To cornea, iris, conjunctiva carry out observation and it is other Detrimental effect should all record scoring.According to《Cosmetics safety technical specification (2015 editions)》The standards of grading of eye damage/eye thorn Swash property reaction classification and carries out result judgement.Integral is higher, and irritation is bigger.It the results are shown in Table 11.

11 80% working substance animal eye irritant test result of table

Above-mentioned animal eye irritant test result is shown：

By 80% working substance instill rabbit conjunctival capsule in, 1h, for 24 hours, 48h, 72h stimulation is all had no to conjunctiva, iris, cornea Property reaction, result judgement is nonirritant.

6) 3T3 red untakes phototoxicity experiments

3T3 red untake phototoxicity experiments are carried out using working substance stoste.Dimethyl diaminophenazine chloride is a kind of weak dye of positive ion, Easily penetration cell film and assemble in Cytolysosome in such a way that nonionic is spread.In certain chemical substances and external condition Under effect, the change of cell surface or lysosome membrane sensibility can be caused and lysosome brittleness is caused the irreversible changes such as to increase, The ability that dimethyl diaminophenazine chloride is absorbed so as to cause cell declines.It is irradiated through chemical substance and ultraviolet light by measuring 3T3 fibroblasts Cell absorbs the variation of dimethyl diaminophenazine chloride ability to judge whether the chemical substance has phototoxicity after synergy.Experiment parameter PIF (the light stimulus factor) with MPE (average luminous effect) to evaluate phototoxicity, criterion is shown in Table 12, the results are shown in Table 13.

12 external 3T3 red untakes phototoxicity experiments criterion of table

Note：

1, positive control OD540 numerical value is all higher than 0.4 and positive reference substance chlorpromazine hydrochloride meets the following conditions and shows reality Test establishment.

IC50 (+UV) is within the scope of 0.1-3.0 μ g/mL.

IC50 (- UV) is within the scope of 7.0-90.0 μ g/ml.

PIF≥6

2, PIF (the light stimulus factor)=IC50 (- UV)/IC50 (+UV)

3, MPE (average luminous effect) has two kinds of situations of illumination (+UV) and no light (- UV) by comparing on concentration grid The cytotoxic concentration response curve of lower acquisition, concentration of contamination range are having illumination (+UV) and no light (- UV) experiment shared Selection in concentration range.Average luminous effect (MPE) is acquired using special software " PHOTO32 ".

13 working substance 3T3 red untake phototoxicity experiments results of table

Note：*1：Cytotoxicity is not also showed in the maximum concentration value (1000.0 μ g/mL) for reaching permission；

+：It is expected that there are phototoxicities；

—：It is expected that without phototoxicity.

Above-mentioned 3T3 red untakes phototoxicity experiments result is shown：

It is * 1, MPE that IC50 of the working substance stoste under no light and illumination condition, which is all higher than 1000.0 μ g/mL, PIF values, Value is respectively less than 0.1.According to phototoxicity evaluation of result standard scale 9, working substance stoste is without phototoxicity.

In conclusion composition hemolysis rate described in embodiment 7 is 0%, and no haemolysis irritation, no skin and eye irritation, It is safe without phototoxicity.

According to the method described above to described in embodiment 5 and embodiment 6 composition carry out safety detection, as a result with embodiment 7 The composition is similar, and composition is without haemolysis irritation, no skin and eye irritation, unglazed poison described in embodiment 5 and embodiment 6 Property, it is safe.

Claims (11)

1. a kind of composition with moisture-keeping function, which is characterized in that it is made of dendrobium candidum, white fungus, dictyophora phalloidea and Radix Ophiopogonis, it is described Dendrobium candidum, white fungus, dictyophora phalloidea and the mass ratio of Radix Ophiopogonis are (1~4):(1~4):(1~4):(1~4).

2. composition according to claim 1, which is characterized in that the dendrobium candidum, white fungus, dictyophora phalloidea and the quality of Radix Ophiopogonis Than being 4:3:4:3.

3. composition according to claim 1, which is characterized in that the dendrobium candidum, white fungus, dictyophora phalloidea and the quality of Radix Ophiopogonis Than being 3:4:3:4.

4. composition according to claim 1, which is characterized in that the dendrobium candidum, white fungus, dictyophora phalloidea and the quality of Radix Ophiopogonis Than being 1:1:1:1.

5. the preparation method of composition described in claim 1-4 any one, which is characterized in that raw material crushes, is sieved, and water is added to carry Hot refined filtration is carried out after taking successively and cold refined filtration collects extracting solution.

6. preparation method according to claim 5, which is characterized in that the sieving is to sieve with 100 mesh sieve；The extracting in water To press 1:The solid-liquid ratio of (75~85) adds water, after impregnating 0.5h, 55~65 DEG C of 110~130min of extraction.

7. preparation method according to claim 5, which is characterized in that the hot refined filtration is to put cardboard H60 in Buchner funnel, The extracting solution of injection warm soaks, and opening to filter negative pressure and smoke makes cardboard be tightly attached to funnel but to ensure that entire cardboard is wetting, Bright extracting solution must be clarified；The cold refined filtration is to put cardboard H70 in Buchner funnel, injects the extracting solution of warm, opens to filter and bear Pressure, which is smoked, to be made cardboard be tightly attached to funnel but to ensure that entire cardboard is wetting, and bright extracting solution must be clarified.

8. a kind of skin care item with moisture-keeping function, including the composition described in claim 1-4 any one and matrix.

9. skin care item according to claim 8, which is characterized in that the dosage of the composition is 0.25wt%.

10. skin care item according to claim 8 or claim 9, which is characterized in that the matrix is by glycerine, 1,2- pentanediols, propylene Sour hydroxyl ethyl ester/sodium acryloyldimethyl taurate copolymers, Phenoxyethanol/Sensiva SC50 and water composition.

11. a kind of method for moisturizing uses the composition or claim 8- described in claim 1-4 any one in skin surface Skin care item described in 10 any one.