CN108697632A - Including sea cucumber extract improves composition as the skin of active ingredient - Google Patents
Including sea cucumber extract improves composition as the skin of active ingredient Download PDFInfo
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- CN108697632A CN108697632A CN201780011984.5A CN201780011984A CN108697632A CN 108697632 A CN108697632 A CN 108697632A CN 201780011984 A CN201780011984 A CN 201780011984A CN 108697632 A CN108697632 A CN 108697632A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/50—Molluscs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/616—Echinodermata, e.g. starfish, sea cucumbers or sea urchins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
Abstract
The open skin comprising sea cucumber extract as active ingredient improves composition.The sea cucumber extract prepared by being repeated 2 times freeze-drying has effect to skin regeneration activity and skin whitening.
Description
Technical field
The present invention relates to the skins comprising sea cucumber extract as active ingredient to improve composition, is in particular related to comprising logical
Cross the invention for being repeated 2 times freeze-drying the sea cucumber extract for preparing as the skin improvement composition of active ingredient.
Background technology
Sea cucumber is the oceanic invertebrate that one kind belonging to Echinodermata Holothuroidea (holothuroidea), in South Korea
There are stichopus japonicus, Holothuria atra, 14 kinds of 4 sections of Red sea Can Deng.Since ancient times, sea cucumber and ginseng one are all representative nourishing food,
So far also it is the research object of food science.It has hereby been recorded in mountain fish spectrum, Compendium of Materia Medica, Dong-eui-bo-gam etc. for restoring member
Gas, tranquilizing the mind and other effects, it is known that also widely utilized far away from ancient times, and find saponin constituent in a recent study, therefore opened
Have the effect of in food science outstanding.
In recent years, it not only conducts a research in food science, in aspect of the sea cucumber as natural materials newtype drug
Research also actively developing, in particular, demonstrated in nearest result of study sea cucumber for skin regeneration or whitening skin
The effect of skin.But most of research for skin regeneration and skin whitening has used sea cucumber and has been carried by other raw materials simultaneously
The extract taken.Accordingly, it is difficult to determine only have effects that in the case of sea cucumber for skin regeneration and skin whitening.And
And in the case where only studying sea cucumber, does not report and have to skin activity about the type of sea cucumber, position, extracting method etc.
There is the detailed research of which kind of difference, and largely propose the effect according to zoopery, and does not accurately propose there are actual persons
To the effect of skin regeneration and skin whitening in class skin model.
Therefore, it is necessary to study whether sea cucumber has the effect of, to skin regeneration and skin whitening, it is practical there is a need to confirmation
Human skin has effect to the skin regeneration of sea cucumber and whitening.
Invention content
Technical problem
The present invention proposes that the object of the present invention is to provide cold by being repeated 2 times in order to solve the problems, such as described above
The dry skin comprising sea cucumber extract as active ingredient to prepare, which is lyophilized, improves composition.
The means solved the problems, such as
As the technological means for realizing technical problem as described above, according to the skin improvement group of a scheme of the invention
It includes that the sea cucumber extract prepared by being repeated 2 times freeze-drying is used as active ingredient to close object.
Herein, above-mentioned sea cucumber extract can be prepared by the preparation method included the following steps:Sea cucumber is carried out
The step of being freeze-dried for the first time;Freeze-dried sea cucumber is crushed come the step of preparing powder;To being carried out through powder pulverized powder
The step of incubation;The step of concentration is evaporated to the sea cucumber through incubation;Sea cucumber through evaporation and concentration is filtered and is condensed
The step of and the step of carry out second to condensed sea cucumber and be freeze-dried.
Above-mentioned skin improves composition can also be prepared by the preparation method included the following steps:It is cold in above-mentioned first time
It is lyophilized before dry step, superfreeze (deep-freezing) is carried out with -70 DEG C to -100 DEG C of temperature to sea cucumber.
The step of above-mentioned first time freeze-drying, may include:The step of sea cucumber is cooled to -20 DEG C to -100 DEG C of temperature;With
And the step of by being dried 36 hours to 84 hours through cooling sea cucumber.
In the above-mentioned incubation the step of, it can be incubated 1 hour by being put into 55% to 95% ethyl alcohol into sea cucumber powder
To 3 hours.
In the above-mentioned evaporation and concentration the step of, it can be concentrated by evaporation 1.5 hours to 5 hours at a temperature of 55 DEG C to 100 DEG C.
Above-mentioned sea cucumber can be stichopus japonicus and/or Holothuria atra.
Above-mentioned sea cucumber extract can be extracted using only sea cucumber internal organ.
Above-mentioned sea cucumber extract can be prepared by further including the preparation method of following steps:By will be freezed through second
Dry sea cucumber is dissolved in solvent to prepare solution.
Above-mentioned solvent can be selected from by butanediol (butylene glycol), water, ethohexadiol (ethyl
Hexanediol) and 1, one or more of the group of 2- hexylene glycols (1,2-hexanediol) composition.
It can be skin regeneration activity that above-mentioned skin, which improves,.
Above-mentioned skin regeneration activity can be that the expression quantity of type Ⅳ collagen albumen increases, cell active factor (ki-67)
Increase and/or the secretory volume of matrix metalloproteinase is reduced.
Above-mentioned skin improvement can be skin whitening.
Above-mentioned skin whitening can be to inhibit melanin production.
The effect of invention
The skin improvement composition comprising sea cucumber extract as active ingredient of the present invention has the effect for making skin regeneration
Fruit, such as the expression quantity of type Ⅳ collagen albumen increases, the secretory volume of matrix metalloproteinase is reduced etc., and with preventing skin
The effect of melanism, such as inhibit melanin production etc..
Description of the drawings
Fig. 1 is the preparation section shown for extracting sea cucumber extract.
Fig. 2 is the process for showing sea cucumber extract being prepared into solution.
Fig. 3 is to show Comparative Examples 1 and 2 and 4 type Ⅳ collagen albumen and the expression quantity of ki-67.
Fig. 4 is to show embodiment 3 and 4 type Ⅳ collagen albumen and the expression quantity of ki-67.
Fig. 5 is to show that the expression rate of ki-67 compares figure.
Fig. 6 is the standard curve figure for showing MMP-9.
Fig. 7 is to show that the secretory volume of MMP-9 compares figure.
Fig. 8 is to show melanin standard curve.
Fig. 9 is to show that comparative example 2 and the melanin amount of comparative example 3, embodiment 1 to embodiment 6 compare figure.
Specific implementation mode
Hereinafter, illustrating the present invention in further detail.But this is only proposed as example, the present invention is not limited to
This, only range is claimed by invention hereinafter to be defined in the present invention.
The dermato-cosmetic composition that effect is all had to skin regeneration activity and skin whitening is provided.
It includes that the sea cucumber prepared by being repeated 2 times freeze-drying carries to improve composition according to the skin of a scheme of the invention
Take object as active ingredient.
Above-mentioned sea cucumber is Shu Yu Parapet hand mesh (aspiodchirotida), Stichopodidae (stichopodidae), stichopus japonicus category
(stichopus) marine organisms, the whole world have inhabited 1500 kinds.It is stichopus japonicus (stichopus in the scientific name of the sea cucumber of South Korea's survival
Japonicus), and 14 kinds of 4 section has been inhabited.
Preferably, above-mentioned sea cucumber can be stichopus japonicus, Holothuria atra.Stichopus japonicus is for survival in the rock of outer saline waters, rubble, sandy
The sea cucumber in region, Holothuria atra are the sea cucumber for inhabiting the stronger place of influence in eutrophy waters, land.
Above-mentioned sea cucumber can be one kind in the group being made of entire sea cucumber, internal organ and the position other than internal organ
Above position.
As the drying means of seafood, there is natural drying, high-pressure drying, constant pressure and dry and vacuum drying, above-mentioned freezing
Dry is to belong to vacuum drying drying means.Above-mentioned freeze-drying be by freeze and depressurize the material containing large quantity of moisture come
Moisture removal is removed while so that ice is distilled and obtains the drying means of dried object.Freeze-drying has the following advantages, that is, by making
The damage of thermo-responsive substance minimizes to be dried, and dried substance forms the structure for being easy to moisture penetration, from
And it can realize the quick rehydration (rehydration) for freezing substance.
Above-mentioned sea cucumber is easy to cause metaplasia because of heat, since moisture is more, is relatively suitble to dry by freezing
It is dry to dry sea cucumber.Freeze-dried sea cucumber can remain intrinsic tissue to be dried, and excellent to the rehydration of water
It is elegant.
Above-mentioned active ingredient refers to that the principal component that skin improves composition is sea cucumber extract.That is, sea cucumber extract is direct
Or the beautifying skin function of composition is shown indirectly.
Above-mentioned composition, which is skin, improves composition.So-called skin improvement refers to skin whitening, improves around skin, helps
From the function etc. of UV protection skin.
Improve function and dosage form according to required skin, can only be wrapped including the skin of above-mentioned sea cucumber extract improves composition
Be used as active ingredient containing sea cucumber extract, can also include simultaneously sea cucumber extract and other neccessary compositions be used as effectively at
Point.Also, above-mentioned dermato-cosmetic composition can be used as skin preparations for extenal use, also act as the food of such as healthy food or as injected
The drug etc. of dosage form etc..
Above-mentioned sea cucumber extract can be prepared by the preparation method included the following steps:First time freezing is carried out to sea cucumber
Dry (step S110);Freeze-dried sea cucumber is crushed to prepare powder (step S120);To being incubated through powder pulverized powder
It educates (step S130);Concentration (step S140) is evaporated to the sea cucumber through incubation;Sea cucumber through evaporation and concentration is filtered
(step S150) and condensation (condensation) (step S160);And second is carried out to condensed sea cucumber and is freeze-dried
(step S170).
Above-mentioned skin, which improves composition, to be prepared by the preparation method included the following steps:It is freezed in above-mentioned first time
Before dry step, superfreeze (deep-freezing) is carried out with -70 DEG C to -100 DEG C of temperature to sea cucumber.Pass through
Superfreeze, is completely removed the moisture of sea cucumber, and with pole cryogenic quick freezing, therefore the cell of sea cucumber is not impaired and can
Keep complete.Above-mentioned superfreeze can be used as -1 DEG C to -5 DEG C of temperature range of zone of maximum ice crystal formation by running through
(quick freezing) method of fast freezing carries out.
The nutritional ingredient of above-mentioned sea cucumber through first time freeze-drying (step S110) is concentrated.That is, by freeze-drying,
Moisture is removed from 90% or more of weight by the raw sea cucumber that water is formed, compared with raw sea cucumber, under the conditions of with identical weight, through dry
Dry sea cucumber includes highly concentrated nutritional ingredient.Therefore, it is possible to outstanding skin improvement effects can also be presented on a small quantity.
Above-mentioned first time freeze-drying (step S110) may include following preparation method:By sea cucumber freeze for -10 DEG C to -
110℃;And the above-mentioned sea cucumber through freezing is moved and inspired 36 hours to 84 hours.
Preferably, the solidification point of above-mentioned first time freeze-drying (step S110) can be -10 DEG C to -110 DEG C, more excellent
Selection of land can be -20 DEG C to -100 DEG C.According to solidification point, the freeze-off time of freeze-drying changes.Solidification point is lower, freezes
The knot time is then shorter, and the tissue that is big slight and having densification in gap.This is because when this progress quick freezing, quickly
It crosses ice crystal and generates band, therefore form smaller ice pellets.When above-mentioned solidification point is less than -110 DEG C, though sublimation drying
It shortens, but ice crystal formation speed becomes faster and ice crystal becomes smaller, therefore water is penetrated into the inside of the sea cucumber through first time freeze-drying, to
The speed that dried sea cucumber absorbs slows down, to which the presentation of aquation recovery capability is relatively low.Also, when above-mentioned solidification point higher than-
At 10 DEG C, the gap of the inside of sea cucumber becomes larger, therefore aquation recovery capability improves, but sublimation drying is elongated, to prepare
The process of extract is possible to lengthen.
Preferably, the above-mentioned sea cucumber through freezing for the first time can be dried 36 hours to 84 hours, it is highly preferred that it is small to dry 48
Up to 72 hours.Above-mentioned drying refers to that lyophilization and/or second are dry.Lyophilization refers to after freezing by pressure reduction
Below to the three phase point of water and it is directly changed from solid and dries moisture, such as the water of the inside of cell for gas, second
Secondary drying refers to the dehumidification that also remaining moisture after lyophilization is dried.When above-mentioned drying time be shorter than 36 it is small when
When, the moisture in sea cucumber is possible to be unable to fully drying, when being longer than 72 hours, excessive water drying, therefore that there are aquations is extensive
Multiple less able misgivings.
The above-mentioned sea cucumber through first time freeze-drying is crushed to prepare powder (step S120).Preferably, according to using mesh
, the size of powder can be 0.01nm to 1mm.When freeze-dried sea cucumber is prepared into powder, size becomes smaller, therefore
Can be easy to body absorption, and surface area increases, so as to easily with other objects for improving included in skin in composition
Matter is reacted.
(step S130) is incubated to the above-mentioned powder through crushing and may include following preparation method:It is put to sea cucumber powder
Enter 55% to 95% ethyl alcohol and is incubated 1 hour to 3 hours.
Preferably, above-mentioned ethyl alcohol can be 55% to 95% purity, it is highly preferred that can for 65% to 85% it is pure
Degree.And, it is preferable that incubation can carry out 1 hour to 3 hours, it is highly preferred that can carry out 1.5 hours to 2.5 hours.It is above-mentioned to incubate
Educating (step S130) enables the stroma protein of sea cucumber to decompose, and the stroma protein through decomposition can make the skin of sea cucumber extract
Cosmetology function activates.When the purity of above-mentioned ethyl alcohol is less than 55%, because comprising polluter, therefore can be incubated, when big
When 95%, because of the ethyl alcohol of high-purity, sea cucumber protein can be solidified.When above-mentioned incubation time is less than 1 hour, it is possible to nothing
The stroma protein that method fully resolves into sea cucumber can prepare the degree of extract, when more than 3 hours, because of the matrix egg of sea cucumber
The decomposition of white matter and the beautifying skin function of sea cucumber can be made to lose.
Above-mentioned evaporation and concentration (step S140) may include the preparation method of following steps:Extremely at 55 DEG C by the sea cucumber through incubation
It is concentrated by evaporation 1.5 hours to 5 hours at a temperature of 100 DEG C.
Preferably, above-mentioned evaporation and concentration can carry out 1.5 hours to 5 hours at a temperature of 55 DEG C to 100 DEG C, it is highly preferred that
It can be carried out 2 hours to 4.5 hours at a temperature of 65 DEG C to 90 DEG C.The step of by above-mentioned evaporation and concentration, can remove ethyl alcohol, oil at
Point, the pollutant components such as heavy metal and only obtain pure sea cucumber.When be concentrated by evaporation temperature be less than 55 DEG C when, make because of low temperature pollution at
Dividing can not volatilize, and can make the skin of sea cucumber because of high temperature when more than 100 DEG C so as to the residual impurity in sea cucumber extract
It is impaired to improve ingredient.Also, when being concentrated by evaporation the time less than 1.5 hours, the nothings such as ethyl alcohol, pollutant component can be made because the time is short
Method is fully evaporated, and when more than 5 hours, is likely to result in also generating the albumen of sea cucumber ingredient at low temperature because of long-time heating
Qualitative change shape.
The step of (step S150) and condensation (condensation) are filtered to the above-mentioned sea cucumber through incubation (step
S160) for for rejecting impurity from the sea cucumber through evaporation and concentration and the step of concentrate the sea cucumber extract for obtaining high-purity.
The step of to condensed above-mentioned sea cucumber be freeze-dried for second (step S170) is for condensed
Sea cucumber carries out the step of sea cucumber extract of second of freeze-drying to obtain high-purity.Since second of freeze-drying step is
Freeze-drying step for the first time is repeated again, therefore detailed description is omitted below.
Preferably, above-mentioned sea cucumber can be stichopus japonicus and/or Holothuria atra, it is highly preferred that can be inhabit West Coast hundred
The attached inshore stichopus japonicus in clever island and/or Holothuria atra.Stichopus japonicus and Holothuria atra belong to inner bay sea cucumber, and Red sea ginseng belongs to off-lying sea property sea
Ginseng.Not only thick but also short as the stichopus japonicus of inner bay sea cucumber and the Polian vesicle (polian vesicle) of Holothuria atra, front end is in blunt circle
Shape, shrinkage is small, and the form of tentacle osteocomma (ossicle) is simple.In contrast, the Pori as the Red sea of off-lying sea property sea cucumber ginseng
Family name's capsule is elongated, and front end is in the shape of sharp, and shrinkage is strong, the complex shape of tentacle osteocomma.Join phase with the Red sea for only eating red algae
Than above-mentioned stichopus japonicus and Holothuria atra absorb a variety of baits, therefore exist than Red sea ginseng with more kinds of nutritional ingredients.
Above-mentioned sea cucumber extract can be extracted using only sea cucumber internal organ.Because being present in the shadow of the seaweeds in internal organ
It rings, compared with the position other than internal organ, sea cucumber internal organ have high antioxidant.Anti-oxidant refers to the effect for preventing oxidation.Sea
The aging of the cell because of caused by the oxidation of cell can be prevented by removing the active oxygen in human body by joining the inoxidizability of internal organ,
Therefore it can help to beautifying skin.Therefore, include skin of the extract only extracted from the internal organ of sea cucumber as active ingredient
Outstanding function can be played to skin regeneration activity and skin whitening by improving composition.
Above-mentioned sea cucumber extract can be to be prepared by further including the preparation method of following steps:It will be freezed through second
Dry above-mentioned sea cucumber is dissolved in solvent come (step S180) the step of preparing solution.Including sea cucumber extract as effectively at
The skin divided, which improves composition, to be used by a variety of dosage forms.Therefore, according to purpose and method is used, skin improves composition
It also may include the sea cucumber extract of solution.Above-mentioned solvent can be selected from being made of polarity, nonpolarity, organic and inorganic solvent
The solvent of one or more of group.
Preferably, above-mentioned solvent can be selected from by butanediol (butylene glycol), water, ethohexadiol
(ethyl hexanediol) and 1, one or more of the group of 2- hexylene glycols (1,2-hexanediol) composition.
Butanediol, ethohexadiol and 1,2- hexylene glycols are alcohol compound, can dissolve the Multiple components of sea cucumber, therefore can
Preparing can include that the skin improvement composition of sea cucumber extract effectively plays the solution of skin improvement function.
It can be skin regeneration activity that above-mentioned skin, which improves,.Preferably, above-mentioned skin regeneration activity can be type Ⅳ collagen
Expression quantity increase, the increase of cell active factor (ki-67) and/or the matrix metalloprotease of albumen (collagen type IV)
Enzyme (matrix metalloproteinases) secretory volume is reduced.
Above-mentioned skin regeneration activity can be that the expression quantity of type Ⅳ collagen albumen increases.Preferably, with by dimethyl sulfoxide (DMSO)
The negative control group of preparation is compared, and the expression quantity of above-mentioned type Ⅳ collagen albumen can be 200% to 500%, with ascorbic acid phase
Than that can be 103% to 110%.
Collagen (collagen) is the main protein of the skin and flesh layer and connective tissue that constitute mammal, and in structure
At accounting for 25% to 35% in the protein of body.So far, it has been found that the collagen of 29 types, type Ⅳ collagen albumen are substrate
The main composition of film and glomerulus epilepsy matrix.Type Ⅳ collagen albumen is by 7S collagens region, non-collagen 1 (NC1, non
Collagenous 1) region, the region (NC2, non collagenous 2) of non-collagen 2 and three spiral (TH, triple
Helix) 4, region etc. region is constituted.4 molecules of type Ⅳ collagen albumen are connected with NH2 distal portions, wherein disulfide bond
(S-S) more region is 7S collagens region, and the region is not by the multiple proteins of bacillary collagen catabolic enzyme etc.
Catabolic enzyme decomposes.Basilar memebrane refers to the film being present between epidermis and corium, takes on skin regeneration effect.When basilar memebrane by
When damage, the enzyme for decomposing collagen is invaded from epidermis to corium, therefore destroys collagen fabric, to be produced on skin
Raw wrinkle.By making the expression quantity increase of the type Ⅳ collagen albumen for the main composition for being used as basilar memebrane strengthen basilar memebrane
Mechanism, above-mentioned skin, which improves composition, to be had to the active function of skin regeneration.
Above-mentioned skin regeneration activity can be the increase of cell active factor (ki-67).Preferably, with as negative control
0.1% dimethyl sulfoxide (dimethyl sulfoxide) of group is compared, and the expression quantity of above-mentioned cell active factor (ki-67) can
Think 200% to 500%.Cell active factor (ki-67) is the protein of the marker as cell Proliferation, in the cell cycle
In the G1 phases, the G2 phases, the S phases, the interim expression of M.Including sea cucumber extract improves composition as the skin of active ingredient passes through shadow
Skin Cell is rung to make regeneration activity increase.Therefore, improve the cytothesis of cuticula of skin of composition etc. using skin
It becomes active, and the expression quantity of cell active factor (ki-67) increases.
Above-mentioned skin regeneration activity can be that the secretory volume of matrix metalloproteinase is reduced.Preferably, above-mentioned matrix metal
The secretory volume of protease, which is reduced, to be the Expression of Matrix Metalloproteinases amount of the negative control group prepared with dimethyl sulfoxide (DMSO)
50% to 90%, the 60% to 90% of the Expression of Matrix Metalloproteinases amount of ascorbic acid.
Matrix metalloproteinase refers to for decomposing the type Ⅳ collagenase of type Ⅳ collagen albumen (type IV
) and Stromelysin (stromelysin, MMP-3) collagenase.Type Ⅳ collagenase by gelatin enzyme A (gelatinase A,
MMP-2) and Gelatinase B (gelatinase B, MMP-9) is constituted.Gelatin enzyme A is from fibroblast, endothelial cell, macrophage
Etc. being prepared, Gelatinase B is prepared from normal cell and cancer cell, and Stromelysin (MMP-3) is prepared from cancer cell.It is above-mentioned
Sea cucumber extract can by make above-mentioned matrix metalloproteinase secretory volume reduce and inhibit type Ⅳ collagen albumen decomposition machine
System has to the active function of skin regeneration.
Above-mentioned skin improvement can be skin whitening.Skin whitening refers to preventing skin by melanogenesis by melanin
(melanogenesis) and make skin-whitening and beauty.
Above-mentioned dermato-cosmetic composition can before B16 cell, synthesis in or synthesis after play skin whitening function.
It is adjusted by tyrosinase (tyrosinase) transcriptional regulatory and saccharification before B16 cell, passes through junket in B16 cell
Propylhomoserin enzyme hinders, peroxidase hinders, reducing agent and active oxygen are eliminated, decomposed by tyrosinase after B16 cell,
Melanosome movement hinders, stripping promotes to play skin whitening function.
Above-mentioned skin whitening can be to inhibit melanin production.Preferably, with the negative control group that is prepared with 1 × PBS
Melanin secretory volume is compared, including sea cucumber extract improves subtracting for the melanin secretory volume of composition as the skin of active ingredient
It can be to reduce 12% to 15% less, compared with the melanin secretory volume with negative control group prepared by dimethyl sulfoxide (DMSO), can subtract
Few 2% to 6%.
Above-mentioned inhibition melanin production is to be converted to DOPA by tyrosinase in the tyrosine as intracellular matter
(dopa), after DOPA quinone (dopaquinone), melanin is by being used as the dopachrome change of tyrosinase-related protein matter
Isomerase (TRP-2, dopachrome tautomerase) and tyrosinase-related protein 1 (TRP-1,5,6- dihydroxy indoles-
2- carboxylic oxidases (5,6-dihydroxyindole-2-carboxylic acid oxidase)) effect and automation procedure
To generate.Above-mentioned skin, which improves composition, can be reduced by tyrosinase inhibitory activity increase and/or tyrosinase expression to hinder
Tyrosine is hindered to be formed as the mechanism of melanin to inhibit the generation of melanin.
Hereinafter, the present invention is described in detail so that general technical staff of the technical field of the invention can be easy
Implement.But the present invention can be realized by a variety of different forms, and it is not limited to embodiment described herein.
Comparative example 1 is to comparative example 3:Negative control group
Comparative example 1 to comparative example 3 is utilized respectively the maintenance culture medium provided by high (TEGO) scientific & technical corporation (strain) of South Korea's enlightening
(maintenance medium), 0.1% dimethyl sulfoxide (DMSO) provided by high (TEGO) scientific & technical corporation (strain) of South Korea's enlightening
(dimethyl sulfoxide), 1 × PBS (phosphate provided by high (TEGO) scientific & technical corporation (strain) of South Korea's enlightening
Buffer saline) it is prepared for negative control group.
Comparative example 4 and comparative example 5:Positive controls
It is prepared using ascorbic acid (L-Ascorbin acid) in comparative example 4, arbutin is utilized in comparative example 5
(arbutin) positive controls are prepared for.
Embodiment 1:Low concentration sea cucumber extract
By cleaning the stichopus japonicus that acquire in lark island surrounding waters and Holothuria atra and at -30 DEG C at a temperature of is cooling, dries 48
After the first time freeze-drying step of hour, the sea cucumber through freezing is crushed to be prepared into powder.Sea cucumber powder is put into
After being incubated 2 hours in 75% ethyl alcohol, after being concentrated by evaporation 3 hours at a temperature of 70 DEG C, it is run through cotton
(cotton) it is filtered.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-
1112, Tokyo Physico-chemical Apparatus Co., Ltd. (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.With-
30 DEG C, concentrated above-mentioned sea cucumber carried out second and has been freeze-dried by 48 hours conditions.
Embodiment 2:High concentration sea cucumber extract
By cleaning the stichopus japonicus that acquire in lark island surrounding waters and Holothuria atra and at -30 DEG C at a temperature of is cooling, dries 48
After the first time freeze-drying step of hour, the sea cucumber through freezing is crushed to be prepared into powder.Sea cucumber powder is put into
After being incubated 2 hours in 75% ethyl alcohol, after being concentrated by evaporation 3 hours at a temperature of 70 DEG C, cotton is run through to carry out
Filtering.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-1112, Tokyo physics and chemistry instrument
Co., Ltd. (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.With -30 DEG C, 48 hours conditions
Concentrated above-mentioned sea cucumber has been carried out second to be freeze-dried.
Embodiment 3:Low concentration sea cucumber internal organ extract
By cleaning the internal organ of stichopus japonicus and Holothuria atra acquire in lark island surrounding waters and at a temperature of at -80 DEG C using surpassing
Low-temp. refrigerator carries out superfreeze.Sea cucumber internal organ through superfreeze are cooled down at a temperature of -30 DEG C and are dried 48 hours
For the first time after freeze-drying step, the sea cucumber internal organ through freezing are crushed to be prepared into powder.Sea cucumber internal organ powder is put into
After 75% ethyl alcohol is incubated 2 hours, distillation and concentration is run through cotton to carry out after 3 hours at a temperature of 70 DEG C
Filtering.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-1112, Tokyo physics and chemistry instrument strain
Formula commercial firm (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.It will with the condition of -30 DEG C, 48 hours
Concentrated above-mentioned sea cucumber internal organ have carried out second and have been freeze-dried.
Embodiment 4:High concentration sea cucumber internal organ extract
By cleaning the internal organ of stichopus japonicus and Holothuria atra acquire in lark island surrounding waters and at a temperature of at -80 DEG C using surpassing
Low-temp. refrigerator has carried out superfreeze.Sea cucumber internal organ through superfreeze are cooled down at a temperature of -30 DEG C and to dry 48 small
When first time freeze-drying step after, crush sea cucumber internal organ through freezing and be prepared into powder.By sea cucumber internal organ powder
Be put into 75% ethyl alcohol be incubated 2 hours after, after being concentrated by evaporation 3 hours at a temperature of 70 DEG C, be run through cotton come into
Filtering is gone.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-1112, Tokyo physics and chemistry device
Tool Co., Ltd. (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.With -30 DEG C, 48 hours items
Concentrated above-mentioned sea cucumber internal organ have been carried out second and have been freeze-dried by part.
Embodiment 5:Low concentration sea cucumber extract liquid
By cleaning the stichopus japonicus that acquire in lark island surrounding waters and Holothuria atra and at -30 DEG C at a temperature of is cooling, dries
After 48 hours first time freeze-drying steps, the sea cucumber through freezing is crushed to be prepared into powder.Sea cucumber powder is put into
After being incubated 2 hours in 75% ethyl alcohol, after being concentrated by evaporation 3 hours at a temperature of 70 DEG C, cotton is run through to carry out
Filtering.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-1112, Tokyo physics and chemistry instrument
Co., Ltd. (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.With -30 DEG C, 48 hours conditions
Concentrated above-mentioned sea cucumber has been carried out second to be freeze-dried.By will through second freeze-dried sea cucumber extract with
Butanediol (butylene glycol), water, ethohexadiol (ethyl hexanediol) and 1,2- hexylene glycols (1,2-
Hexanediol) solvent mixes to be prepared for the sea cucumber extract solution of 5% concentration.
Embodiment 6:High concentration sea cucumber extract liquid
By cleaning the stichopus japonicus that acquire in lark island surrounding waters and Holothuria atra and at -30 DEG C at a temperature of is cooling, dries
After 48 hours first time freeze-drying steps, the sea cucumber through freezing is crushed to be prepared into powder.Sea cucumber powder is put into
After being incubated 2 hours in 75% ethyl alcohol, after being concentrated by evaporation 3 hours at a temperature of 70 DEG C, cotton is run through to carry out
Filtering.Utilize cold boiler (vacuum evaporator, EYELA N-12, EYEKA CA-1112, Tokyo physics and chemistry instrument
Co., Ltd. (EYELA) produces, Tokyo, Japan) concentrate 5 hours low concentrations to 50mg/ml.With -30 DEG C, 48 hours conditions
Concentrated above-mentioned sea cucumber has been carried out second to be freeze-dried.By by second freeze-dried sea cucumber extract and fourth
Glycol (butylene glycol), water, ethohexadiol (ethyl hexanediol) and 1,2- hexylene glycols (1,2-
Hexanediol) solvent mixes to be prepared for the sea cucumber extract solution of 20% concentration.
Experimental example 1:Wrinkle improves experiment
The substances taken care of at a temperature of -20 DEG C are thawed and are diluted in maintaining culture medium to carry out before treatment
Prepare (with reference to following table 1).By control group and test group per 3D Skins cultures model (High (TEGO) section of South Korea's enlightening
Skill company (strain)) hole (well) respectively handle 3.5ml to culture medium layer, and after respectively handling 30 μ l in cell layer surface, in CO2
Incubator (10%, CO2, 37 DEG C) in cultivated 24 hours.After culture 24 hours, culture medium is replaced, and by control group and examination
Test group per 3D Skins cultures models (High (TEGO) scientific & technical corporation (strain) of South Korea enlightening) hole respectively locates repeatedly to culture medium layer
3.5ml is managed, after respectively handling 30 μ l repeatedly in cell layer surface, CO2Incubator (10%, CO2, 37 DEG C) in additional cultivation 24
Hour, it has always co-cultured 48 hours.After culture 48 hours, all groups of culture solution is collected, and taken care of at a temperature of -70 DEG C,
Block (frozen block) is freezed in tissue preparation.
Table 1
In the case of type Ⅳ collagen albumen, in addition to Triton X-100 (the triton X- using above-mentioned 0.5% are omitted
100) it except the process for carrying out permeability (permeabilization), is tested identically as type Ⅳ collagen albumen, phase
For the total cell number of the basal layer in epidermis, with percentage analysis ki-67 positive cell numbers and in following table 2 and Fig. 5
In show.
Table 2
As shown in above-mentioned table 2, compared with the comparative example 2 as negative control group, type Ⅳ collagen egg in embodiment 3 and 4
White expression significantly increases.Compared with the comparative example 2 as negative control group, the ki-67 tables in the N=3 of embodiment 3 and 4
Up to increase, by the ki-67 expression rates of embodiment 3 in 12.5 (± 3.6) % for the comparative example 2 for being used as negative control group with
49.1 (± 7.8) % increase 3.9 times, and the ki-67 expression rates of embodiment 4 are 12.5 of the comparative example 2 as negative control group
In (± 3.6) % 3.9 times are increased with 48.3 (± 10.2) %.
Experimental example 1-2:MMP-9ELISA
Human body MMP-9ELISA kit (R&D systems/DMP900) kit carry out.It is bent for the standard of MMP-9
Line, using 1 × calibration diluent (calibrator diluent) by the MMP-9 standard reserving solutions of 20ng/ml according to each step
Rapid dilution, and it is prepared for standard solution, it is shown in following table 3.
Table 3
It is put into detection dilution (assay diluent) 100 μ l into each hole of human body MMP-9 microplates, respectively adds 100
Blank solution (blank), standard solution (standard) and the testing liquid of μ l simultaneously reflect 2 hours at normal temperatures.Removal reaction
Liquid is always repeated 3 times the process of removal per hole after the wash buffer (wash buffer) of 400 μ l of addition.It is put into 200 per hole
After the MMP-9 conjugates (conjugate) of μ l, reflected under shading status room temperature 1 hour.Reaction solution is removed, per Kong Tian
After adding 400 μ l wash buffers, it is always repeated 3 times the process of removal.Using matrix solution (substrate solution) it
Before, color developing agent (color reagent) A and B being included in kit is mixed in the same amount, and it is added 200 per hole
μ l are reflected 30 minutes under shading status room temperature.The stop solution (stop solution) of 50 μ l is added per hole and at 30 minutes
Absorbance is determined with inherent 450nm wavelength, the MMP-9 secretory volumes of testing liquid are analyzed using MMP-9 standard curves.
Immunofluorescence decoration method (Immunofluorescence staining) is to utilize to freeze block and tested according to itself
Method carries out type Ⅳ collagen albumen.MMP-9 secretory volumes using collection culture solution and using ELISA kit carried out quantitatively point
Analysis.MMP-9 standard solution absorbance results are shown in following table 4 and Fig. 6, by MMP-9 secretory volumes in following table 5 and Fig. 7
In show.
Table 4
Table 5
As shown in above-mentioned table 4 and table 5, it be comparative example 1 is 14.4 (± 3.1) that result is determined in the average secretion of MMP-9 measurement
Ng/ml, comparative example 2 are 17.9 (± 1.6) ng/ml, and the comparative example 4 as positive controls is measured as 14.8 (± 2.5) ng/
Ml, compared with as the comparative example 1 of negative control group and 2, the secretory volume of MMP-9 is unchanged.The average MMP-9 of embodiment 3 points
The amount of secreting is 12.0 (± 1.4) ng/ml reduces 0.7 times compared with the comparative example 2 as negative control group.
Therefore, as shown in experimental example 1-1 and 1-2, including sea cucumber extract is combined as the beautifying skin of active ingredient
When object is compared with ascorbic acid (L-Ascorbin acid), showing has effects that more than same.
Experimental example 2:Skin whitening is tested
It is thawed with the substances of -20 DEG C of keepings, and is diluted using 1 × PBS (Diluent) before treatment, with as follows
List 6 prepares.
Table 6
By substances on 3D Skins cultures model (Neoderm, high (TEGO) scientific & technical corporation (strain) of South Korea's enlightening) surface
100 μ l are respectively handled per hole.In CO2Incubator (10%, CO2, 37 DEG C) in culture 48 hours after, 3D Skins cultures will be remained on
Residue test matter utilization pipettor removal on model surface.
Then, 3D Skins cultures model is cleaned 2 times with the phosphate buffered saline solution (PBS) of 200 μ l, utilizes blade
(blade) separation is struck off from the insertion edge of 3D Skins culture models.By separated 3D Skins cultures model tissue contain into
In the microcentrifugal tube (e-tube) of 1.5ml, after completely removing phosphate buffered saline solution, adds and split in 3D Skins culture models
Solve liquid (lysis buffer, SolvableTM) each 360 μ l.After being reacted 5 minutes in lysate, 3D Skins culture tissue moulds
Type is removed from UF membrane.It at 95 DEG C, is incubated 45 minutes in micro thermostat (heat block), 10 is centrifuged with 13000rpm
After minute, supernatant is moved into new microcentrifugal tube (eppendorf tube).Utilize the NH of 20mg/ml4OH is prepared for
After the melanin solution of 100 μ l/ml, 100 μ l/ml melanin solution of preparation are carried out as shown in table 7 using lysate
Stage dilutes and prepares standard solution, its result is shown in table 7.Standard solution is respectively moved into 200 μ l into 96 orifice plates
Blank solution, standard solution and substances and the result that absorbance is measured in 450nm is shown in table 8, utilize black
Plain standard curve carries out quantification and is shown in following table 9 and Fig. 9 to analyze the result of melanin amount.
Table 7
Table 8
Table 9
As shown in above-mentioned table, the measurement of the average black element of the N=3 of embodiment 5 is set to 19.3 (± 009) μ g/ml, with than
Reduce 13.6% compared with the comparison of example 3, the average black element measurement of the N=3 of embodiment 6 is set to 19.2 (± 0.4) μ g/ml, with than
Reduce 13.8% compared with the comparison of example 3.The N=3 of embodiment 1 average black element measurement is set to 20.6 (± 1.0) μ g/ml, with than
Reduce 2.2% compared with the comparison of example 2, the average black element measurement of the N=3 of embodiment 2 is set to 19.8 (± 0.7) μ g/ml, compared with
The comparison of example 2 reduces 6.3%.The average black element measurement of the N=3 of embodiment 3 is set to 20.2 (± 1.4) μ g/ml, with comparative example
2 comparisons reduce 4.5%, and the average black element measurement of the N=3 of embodiment 4 is set to 21.0 (± 0.6) μ g/ml, with comparative example 2
Comparison reduces 0.4%.Embodiment 1 to 4 is showed with the white-skinned face function with arbutin (arbutin) similar level.
Explanation present invention as described above is for illustration, and general technical staff of the technical field of the invention answers
It should be appreciated that being also easily deformed as other specific shapes in the case where not changing the technological thought or essential feature of the present invention
State.It is therefore to be understood that upper in all respects in various embodiments described above is illustrative, and limit is not it
Qualitatively.For example, the dispersible implementation of each structural element illustrated in singular form, similarly, what is illustrated in a dispersed form is multiple
Structural element can also be implemented with combining form.Range is claimed to present by invention hereinafter in the scope of the present invention, and
Be not it is above-mentioned illustrate, and should be interpreted that the meaning and range and its equivalents that range is claimed by inventing
Have altered derived from concept or deformation form includes within the scope of this invention.
Claims (14)
1. a kind of skin improves composition, which is characterized in that include the sea cucumber extract prepared by being repeated 2 times freeze-drying
As active ingredient.
2. skin according to claim 1 improves composition, which is characterized in that
Above-mentioned sea cucumber extract is prepared by the preparation method included the following steps:
The step of sea cucumber be freeze-dried for the first time;
Freeze-dried above-mentioned sea cucumber is crushed come the step of preparing powder;
The step of above-mentioned powder through crushing is incubated;
The step of concentration is evaporated to the above-mentioned sea cucumber through incubation;
The step of above-mentioned sea cucumber through evaporation and concentration is filtered and is condensed;And
The step of second of freeze-drying is carried out to condensed above-mentioned sea cucumber.
3. skin according to claim 2 improves composition, which is characterized in that above-mentioned skin improves composition by including
It is prepared by the preparation methods of following steps:Before the step of being freeze-dried above-mentioned first time, to sea cucumber with -70 DEG C to -100 DEG C
Temperature carry out superfreeze.
4. skin according to claim 3 improves composition, which is characterized in that
The step of above-mentioned first time freeze-drying includes:
The step of sea cucumber is cooled to -20 DEG C to -100 DEG C of temperature;And
By the step of being dried 36 hours to 84 hours through cooling above-mentioned sea cucumber.
5. skin according to claim 2 improves composition, which is characterized in that in the above-mentioned incubation the step of, pass through to
55% to 95% ethyl alcohol is added in above-mentioned powder through crushing to be incubated 1 hour to 3 hours.
6. skin according to claim 2 improves composition, which is characterized in that in the above-mentioned evaporation and concentration the step of,
It is concentrated by evaporation 1.5 hours to 5 hours at a temperature of 55 DEG C to 100 DEG C.
7. skin according to claim 1 improves composition, which is characterized in that above-mentioned sea cucumber is stichopus japonicus and/or Holothuria atra.
8. skin according to claim 1 improves composition, which is characterized in that above-mentioned sea cucumber extract is that sea cucumber is used only
Internal organ extraction.
9. skin according to claim 2 improves composition, which is characterized in that above-mentioned sea cucumber extract by further include as
It is prepared by the preparation method of lower step:It is molten to prepare by the way that the above-mentioned sea cucumber through second of freeze-drying to be dissolved in solvent
Liquid.
10. skin according to claim 9 improves composition, which is characterized in that above-mentioned solvent be selected from by butanediol,
Water, ethohexadiol and 1, one or more of the group of 2- hexylene glycols composition.
11. skin according to claim 1 improves composition, which is characterized in that it is that skin regeneration is lived that above-mentioned skin, which improves,
Property.
12. skin according to claim 8 improves composition, which is characterized in that above-mentioned skin regeneration activity is IV type glue
The increase of the expression quantity of former albumen, as cell active factor ki-67 increase and/or matrix metalloproteinase secretory volume
It reduces.
13. skin according to claim 1 improves composition, which is characterized in that it is skin whitening that above-mentioned skin, which improves,.
14. skin according to claim 10 improves composition, which is characterized in that above-mentioned skin whitening is to inhibit melanin
It generates.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020160019462A KR101826296B1 (en) | 2016-02-19 | 2016-02-19 | Skin regenerating and whitening composition containing sea cucumber extract |
| KR10-2016-0019462 | 2016-02-19 | ||
| PCT/KR2017/001690 WO2017142316A1 (en) | 2016-02-19 | 2017-02-16 | Skin improving composition containing sea cucumber extract as active ingredient |
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| CN108697632A true CN108697632A (en) | 2018-10-23 |
| CN108697632B CN108697632B (en) | 2022-03-01 |
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| Country | Link |
|---|---|
| KR (1) | KR101826296B1 (en) |
| CN (1) | CN108697632B (en) |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110538131A (en) * | 2019-10-12 | 2019-12-06 | 烟台华康海洋食品有限公司 | Sea cucumber mask and processing method thereof |
| CN111568799A (en) * | 2020-07-08 | 2020-08-25 | 山东恒鲁生物科技有限公司 | Application of holothurian glycolysis element in whitening products |
| CN115089508A (en) * | 2022-07-08 | 2022-09-23 | 西安惠普生物科技有限公司 | Skin whitening essence containing holothurin and preparation method thereof |
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| JP2006083113A (en) * | 2004-09-17 | 2006-03-30 | Noevir Co Ltd | Skin-beautifying composition |
| CN1872094A (en) * | 2006-04-17 | 2006-12-06 | 山东大学 | Application of active material of sea cucumber as medication or health products |
| CN101385827A (en) * | 2008-10-17 | 2009-03-18 | 大连海晏堂生物有限公司 | Sea cucumber health paste and production method thereof |
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| KR20110084132A (en) * | 2011-02-11 | 2011-07-21 | 재단법인 제주테크노파크 | Compositions for improving skin conditions using an extract of stichopus japonicus red |
| KR20140016461A (en) * | 2012-07-30 | 2014-02-10 | 주식회사 유로코스텍 | Fermented sea cucumber extract and its cosmetic usage |
| KR20150111699A (en) * | 2014-03-26 | 2015-10-06 | (주)신한에코 | Method for Preparing Functional Raw Materials Using Stichopus japonicus |
-
2016
- 2016-02-19 KR KR1020160019462A patent/KR101826296B1/en active IP Right Grant
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2017
- 2017-02-16 CN CN201780011984.5A patent/CN108697632B/en active Active
- 2017-02-16 WO PCT/KR2017/001690 patent/WO2017142316A1/en active Application Filing
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| JP2006083113A (en) * | 2004-09-17 | 2006-03-30 | Noevir Co Ltd | Skin-beautifying composition |
| CN1872094A (en) * | 2006-04-17 | 2006-12-06 | 山东大学 | Application of active material of sea cucumber as medication or health products |
| CN101385827A (en) * | 2008-10-17 | 2009-03-18 | 大连海晏堂生物有限公司 | Sea cucumber health paste and production method thereof |
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| KR20110084132A (en) * | 2011-02-11 | 2011-07-21 | 재단법인 제주테크노파크 | Compositions for improving skin conditions using an extract of stichopus japonicus red |
| KR20140016461A (en) * | 2012-07-30 | 2014-02-10 | 주식회사 유로코스텍 | Fermented sea cucumber extract and its cosmetic usage |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110538131A (en) * | 2019-10-12 | 2019-12-06 | 烟台华康海洋食品有限公司 | Sea cucumber mask and processing method thereof |
| CN110538131B (en) * | 2019-10-12 | 2022-07-01 | 烟台华康海洋食品有限公司 | Sea cucumber mask and processing method thereof |
| CN111568799A (en) * | 2020-07-08 | 2020-08-25 | 山东恒鲁生物科技有限公司 | Application of holothurian glycolysis element in whitening products |
| CN115089508A (en) * | 2022-07-08 | 2022-09-23 | 西安惠普生物科技有限公司 | Skin whitening essence containing holothurin and preparation method thereof |
Also Published As
| Publication number | Publication date |
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| WO2017142316A1 (en) | 2017-08-24 |
| KR20170098342A (en) | 2017-08-30 |
| KR101826296B1 (en) | 2018-02-07 |
| CN108697632B (en) | 2022-03-01 |
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