KR20230065740A - Cosmetic composition comprising snowberry flesh and preparation method thereof - Google Patents

Abstract

A method for separating snowberry pulp and a composition containing snowberry pulp are provided. According to the method according to one aspect, the snowberry pulp can be easily separated. The snowberry pulp isolated by the method does not brown, is non-toxic and non-irritating, has unique appearance and texture and has skin-improving functionality, thereby being used as a cosmetic or food ingredient.

Description

Cosmetic composition comprising snowberry flesh and preparation method thereof {Cosmetic composition comprising snowberry flesh and preparation method thereof}

It relates to a method for separating snowberry pulp and a composition comprising snowberry pulp.

The term "Snowberry" is a plant of the genus Symphoricarpos sp. , and is also called Snowberry, waxberry, ghostberry in English. Patent Document 1 discloses the effects of wrinkle improvement, anti-inflammatory, whitening, moisturizing, antioxidant, and anti-aging effects of a composition containing a snowberry extract. However, snowberry is known to have toxicity, so there is a limit to its application as a cosmetic or food product.

On the other hand, there is almost no research on processing snow berries. Therefore, research on snowberry processing is needed to apply snowberry to cosmetics or food.

KR 10-1601908 B1

A method for separating snowberry pulp is provided.

Snowberry pulp separated by the above method is provided.

A composition comprising snowberry pulp is provided.

One aspect provides a method for isolating snowberry pulp.

The method includes (1) washing snowberry fruits;

(2) immersing the snowberry fruit washed in step (1) in vinegar and peeling to separate the snowberry pulp;

(3) adding fermented alcohol to the snowberry pulp separated in step (2);

(4) using a filter to remove snowberry skin residue and obtain a filtrate;

(5) stirring the filtrate obtained in step (4) at low temperature;

(6) separating the filtrate stirred in step (5) into snowberry flesh and seeds; and

(7) centrifuging the snowberry pulp separated in step (6) to remove the supernatant and obtaining the precipitated snowberry pulp.

The method includes (1) washing the snowberry fruit.

The term "Snowberry" is a plant of the genus Symphoricarpos sp. , and is also called Snowberry, waxberry, ghostberry in English. Snowberry berries come in different colors depending on the variety: white (e.g. Common Snowberry), pink (e.g. Little Leaf Snowberry), red (e.g. Coralberry) or dark purple (e.g. Chinese Coralberry). indicates

Exemplary varieties of snowberry are Common snowberry ( Symphoricarpos albus ), Mexican snowberry (Symphoricarpos mexicanus), Little leaf snowberry ( Symphoricarpos microphyllus ), Creeping snowberry ( Symphoricarpos) mollis ), Western snowberry or Wolfberry ( Symphoricarpos occidentalis ), Coralberry ( Symphoricarpos orbiculatus ), Mountain snowberry ( Symphoricarpos oreophilus ), Roundleaf snowberry ( Symphoricarpos rotundifolius ) , Chinese coralberry ( Symphoricarpos sinensis ), and the like.

In one embodiment, the snowberry may be Common Snowberry, Mexican Snowberry, Little Leaf Snowberry, Creeping Snowberry, Western Snowberry, Coralberry, Mountain Snowberry, Chinese Coralberry, or a combination of two or more thereof. .

In one embodiment, the snow berry may be a common snow berry ( Symphoricarpos albus ).

In the step (1), the washing may be performed according to a conventional washing method. The number of times of washing is not particularly limited, but may be one time or several times, for example, two or more times, two to three times. Any washing solution can be used without limitation as long as it can remove foreign substances attached to the outside of the snowberry fruit. For example, the wash liquid may be water. The washing liquid may be lukewarm water. The washing liquid may be water at 15 to 30 °C.

Foreign substances deposited on the outside of the snowberry fruit can be removed by the above step (1).

The method includes (2) immersing the snowberry fruit washed in step (1) in vinegar and peeling to separate the snowberry pulp.

In the above step (2), the peeling of the snowberry and the separation of the flesh can be smoothly achieved by the addition of vinegar. The vinegar may be conventional vinegar, and the type is not particularly limited.

In the above step (2), some toxic components of snowberries can be removed by adding vinegar.

In the step (2), the vinegar is 0.1 to 10% (w / v), 0.1 to 9% (w / v), 0.1 to 8% (w / v), 0.1 to 7% (w / v), 0.1 to 6% (w/v), 1 to 10% (w/v), 1 to 9% (w/v), 1 to 8% (w/v), 1 to 7% (w/v), 1 to 6% (w/v), 3 to 10% (w/v), 3 to 9% (w/v), 3 to 8% (w/v), 3 to 7% (w/v), 3 to 6% (w/v), 4 to 10% (w/v), 4 to 9% (w/v), 4 to 8% (w/v), 4 to 7% (w/v) or It may contain 4 to 6% (w/v) acetic acid. For example, the vinegar may contain 5% (w/v) acetic acid.

In the step (2), the immersion may be performed for a time sufficient to separate the skin and flesh of the snowberry. For example, the immersion is 1 second to 24 hours, 1 second to 12 hours, 1 second to 6 hours, 1 second to 1 hour, 1 second to 30 minutes, 1 minute to 24 hours, 1 minute to 12 hours, 1 minutes to 6 hours, 1 minute to 1 hour, 1 minute to 30 minutes, 5 minutes to 24 hours, 5 minutes to 12 hours, 5 minutes to 6 hours, 5 minutes to 1 hour, 5 minutes to 30 minutes or 5 minutes to It can be done for 15 minutes.

In step (2), snowberry flesh from which the snowberry skin is removed can be obtained. The snowberry pulp obtained by step (2) may contain seeds therein.

The method includes (3) adding fermented alcohol to the snowberry pulp separated in step (2).

The term "alcohol" means ethyl alcohol that can be diluted and drunk. Specifically, it may mean that a material containing starch or sugar is fermented and distilled to an alcohol content of 85 degrees or more, or a material containing alcohol content is distilled to an alcohol content of 85 degrees or more. Alcohol in a broad sense may include all alcohol obtained by distillation, but alcohol in a general sense may mean 95% pure ethanol obtained through continuous distillation. The alcohol includes fermented alcohol and synthetic alcohol.

The term "fermented alcohol" refers to alcohol produced by fermenting a starch (eg, grain, sweet potato, tapioca) or sugar (eg, candy) raw material by adding a saccharification enzyme and then distilling by a continuous distillation method.

Browning of snowberry flesh can be prevented by the above step (3). Browning occurs when snowberry pulp is exposed to the external environment by peeling. When the snowberry flesh is browned, not only is it not aesthetically pleasing, but the components of the browned part may be transformed. In the step (3), some polyphenols in the snowberry pulp are removed by the addition of fermented alcohol, so that browning of the snowberry pulp can be prevented.

By the above step (3), organic components such as alkaloid components of snowberries are removed, and toxicity of snowberries can be removed.

In the step (3), the fermented alcohol may be diluted fermented alcohol. The diluted fermented alcohol may be diluted fermented alcohol at 1 to 20% (v/v). The dilution solvent may be water. The water may be pure distilled water or ultra-pure distilled water. In general, when preparing an extract of a substance, fermented alcohol or ethanol is used as a stock solution. However, in the step (3), trace amounts of polyphenols, aldehydes, and alkaloids contained in snowberries can be removed by using diluted fermented alcohol rather than the undiluted solution.

After step (3), (3') stirring the mixture of snowberry pulp and fermented alcohol may be further included. The stirring may be performed under mild conditions. The agitation is performed at a speed of 2 to 200 rpm, 2 to 150 rpm, 2 to 100 rpm, 5 to 200 rpm, 5 to 150 rpm, 5 to 100 rpm, 10 to 200 rpm, 10 to 150 rpm, or 10 to 100 rpm. can be performed with The stirring is 1 minute to 300 minutes, 1 minute to 200 minutes, 1 minute to 100 minutes, 1 minute to 90 minutes, 10 minutes to 300 minutes, 10 minutes to 200 minutes, 10 minutes to 100 minutes, 10 minutes to 90 minutes , 30 minutes to 300 minutes, 30 minutes to 200 minutes, 30 minutes to 100 minutes, or 30 minutes to 90 minutes.

The method includes (4) using a filter to remove snowberry skin residue and obtain a filtrate.

In the step (4), the size of the filter may be selected to be suitable for removing snowberry skin residues. The filter may be a 3 to 5 mesh filter. The mesh means the number of nets included in a square having one side of 1 inch (25.4 mm). For example, 3 mesh means a particle diameter of 6730 μm, 4 mesh means a particle diameter of 5000 μm, and 5 mesh means a particle diameter of 4000 μm.

All snowberry peel residues may be removed by the above step (4).

The filtrate obtained in step (4) may include snowberry flesh and seeds.

The method includes (5) stirring the filtrate obtained in step (4) at low temperature.

In the step (5), the low temperature is -20 ℃ to 10 ℃, -20 ℃ to 5 ℃, -20 ℃ to 0 ℃, -20 ℃ to -5 ℃, the low temperature is -15 ℃ to 10 ℃, -15 ℃ °C to 5 °C, -15 °C to 0 °C, -15 °C to -5 °C, low temperature is -10 °C to 10 °C, -10 °C to 5 °C, -10 °C to 0 °C, or -10 °C to -5 °C It may be a temperature of ° C. To achieve the low temperature, the filtrate may be immersed in ice water, but is not limited thereto.

By the above step (5), the softness of the snowberry pulp can be prevented and the hardness can be maintained.

The method includes (6) separating the filtrate stirred in step (5) into snowberry flesh and seeds.

In the step (6), the separation is not limited to a specific method. In one example, the heavier weight of the seeds than the snowberry flesh may be exploited. In another example, the feature that the snowberry seeds are approximately 2 mm in size may be used. In the step (6), the separation may be performed using a separatory funnel, but is not limited thereto.

The method includes (7) centrifuging the snowberry pulp separated in step (6) to remove the supernatant and obtaining the precipitated snowberry pulp.

In the step (7), the centrifugation may be performed at a weak rpm. For example, the centrifugation is 2 to 150 rpm, 2 to 100 rpm, 5 to 150 rpm, 5 to 100 rpm, 10 to 150 rpm, 10 to 100 rpm, 50 to 150 rpm, 50 to 100 rpm, or 100 to 150 rpm. Since the centrifugation is performed at a low rpm, the snowberry pulp is not destroyed and its hardness can be maintained. If the centrifugation is performed at more than 150 rpm, the snowberry flesh may be destroyed.

Only snowberry pulp can be obtained by separating the snowberry pulp from the liquid in step (7). That is, the material obtained in step (7) is not the snowberry oil-soluble extract, but the snowberry pulp itself. Since the snowberry pulp obtained in step (7) has a sherbet texture that melts and permeates into the skin, it can be used as a raw material for natural cosmetic formulations.

The snowberry pulp obtained in step (7) may be used as a cosmetic raw material or an external skin preparation raw material. Snowberries are known to be consumed only by animals because they contain ingredients that cause mild vomiting when ingested by humans. However, since the snowberry pulp obtained through the above method is not cytotoxic, it can be safely used as a cosmetic raw material, a raw material for external skin preparations, or a food raw material. The snowberry pulp obtained in the above step (7) can be used as cosmetics, external preparations for skin, or food itself, and may be included as some components of cosmetic compositions, external preparations for skin, and food compositions.

Therefore, the method of separating the snowberry pulp according to the above aspect may be a method of processing the snowberry pulp. Specifically, the method of separating the snowberry pulp may be a method of processing the snowberry pulp into a cosmetic raw material, an external skin preparation raw material, or a food raw material.

Another aspect provides snowberry pulp separated by the method according to the one aspect.

The snowberry flesh is not cytotoxic. In one embodiment, it was confirmed that the snowberry flesh had no cytotoxicity even at a high concentration of 5 mg/ml. In another example, it was confirmed that the snowberry flesh was non-irritating to the skin when used as a cosmetic raw material. Therefore, the snowberry flesh can be safely used as a cosmetic raw material, an external skin preparation raw material, or a food raw material.

The snowberry pulp may have a sherbet texture. Therefore, when the cosmetic formulation containing the snowberry pulp is applied to the skin, the texture of the sherbet formulation that melts and permeates into the skin can be exhibited.

Since the snowberry pulp has a unique appearance and texture, it can be excellently used as a cosmetic raw material, an external skin preparation raw material, or a food raw material.

The snowberry flesh may have one or more of antioxidant effects, skin aging prevention effects, skin wrinkle improvement effects, skin elasticity enhancement effects, skin whitening effects, and skin moisturizing effects.

Another aspect provides a composition comprising snowberry pulp.

The snowberry pulp may be snowberry pulp separated by the method according to the one aspect.

The composition may be a composition for improving skin conditions, for improving skin beauty, or for preventing or treating skin diseases.

In one embodiment, the composition may be used to improve skin condition.

The skin condition improvement may be antioxidant, skin aging prevention, skin wrinkle improvement, skin elasticity enhancement, skin whitening, skin moisturizing, or a combination of two or more thereof.

The term "antioxidation" means inhibition of oxidation. Free radicals generated in the body attack normal cells in the body and act as causes of aging or various diseases. Therefore, oxidation of cells, that is, aging of cells can be suppressed by removing the active oxygen. The antioxidant may include any action of removing active oxygen, converting active oxygen into a harmless substance, or preventing an increase in active oxygen.

The term "skin aging" refers to both tangible and intangible changes that appear in the skin with age, such as thinning of the epidermis, number of cells or blood vessels in the dermis, ability to repair DNA damage, cell replacement cycle, wound healing, It refers to a decrease in skin barrier function, epidermal moisture retention, sweat secretion, sebum secretion, vitamin D production, physical damage defense, chemical elimination ability, immune response, sensory function, and thermoregulation.

The composition may be used for preventing skin aging caused by extrinsic factors or intrinsic factors, improving skin aging, or anti-aging. The extrinsic factor refers to various external factors, such as ultraviolet rays (light). The endogenous factor is also referred to as a chronological factor and refers to a factor that occurs mainly with the passage of time. That is, the skin aging is not only premature aging symptoms induced by external stimuli such as ultraviolet rays, pollution, cigarette smoke, chemicals, etc., but also a natural aging phenomenon that occurs as the proliferation of skin cells decreases with age. , That is, it includes skin aging due to age, and is a concept including wrinkles, loss of elasticity, skin sagging, and dryness. In addition, wrinkles include stimulation caused by changes in internal and external factors that cause wrinkles by changing components constituting skin tissue.

The aging may be photoaging. The term "photoaging" is a phenomenon caused by external environmental factors, and the most representative factor is ultraviolet rays. Ultraviolet rays cause damage to biocomponents such as activation of proteolytic enzymes, chain scission and abnormal cross-linking of matrix proteins, and repetition of these mechanisms results in apparent skin aging. Accordingly, the composition may be used for preventing, improving, or treating photoaging caused by ultraviolet (UV) rays.

The term “improvement of skin wrinkles” is interpreted to mean not only an action of reducing the number or depth of wrinkles on the skin, but also includes suppression of skin aging through wrinkle improvement.

The term "skin elasticity enhancement" may refer to any action of increasing skin elasticity reduced by causes such as aging or preventing a decrease in skin elasticity.

The term "skin whitening" may mean not only brightening the skin tone by inhibiting the synthesis of melanin pigment, but also improving skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity.

The term “moisturizing the skin” can refer to any action that maintains skin moisture or prevents moisture loss. The skin 　moisturizing effect can help improve skin wrinkles and increase elasticity.

The term "skin disease" may be a disease caused by skin aging, skin hyperpigmentation, or impaired skin barrier function. The term "prevention" includes inhibiting the development of a disease. The term "treatment" includes inhibition of the development of, alleviation of, or elimination of a disease.

The composition may have SOD (Superoxide dismutase)-like activity or DPPH radical scavenging activity. Thus, the composition may exhibit an antioxidant effect.

The composition can inhibit the expression of MMP-1. Accordingly, the composition may exhibit effects of preventing skin aging, improving skin wrinkles, and enhancing skin elasticity.

The composition can inhibit melanin synthesis. Thus, the composition may exhibit a skin whitening effect.

The composition can prevent transepidermal water loss. Thus, the composition may exhibit a skin moisturizing effect.

The snowberry flesh is 0.0001 to 100% by weight, 0.0001 to 90% by weight, 0.0001 to 80% by weight, 0.0001 to 70% by weight, 0.0001 to 60% by weight, 0.0001 to 50% by weight, 0.0001 to 40% by weight based on the total weight of the composition %, 0.0001 to 30% by weight, 0.0001 to 20% by weight, 0.0001 to 10% by weight, 0.01 to 100% by weight, 0.01 to 90% by weight, 0.01 to 80% by weight, 0.01 to 70% by weight, 0.01 to 60% by weight, 0.01 to 50% by weight, 0.01 to 40% by weight, 0.01 to 30% by weight, 0.01 to 20% by weight, 0.01 to 10% by weight, 1 to 100% by weight, 1 to 90% by weight, 1 to 80% by weight, 1 to 70% by weight, 1 to 60% by weight, 1 to 50% by weight, 1 to 40% by weight, 1 to 30% by weight, 1 to 20% by weight, or 1 to 10% by weight, but limited to a specific content It is not.

The snowberry pulp may be included in 100% by weight based on the total weight of the composition. In this case, the composition may consist of snowberry pulp. The snowberry flesh is non-cytotoxic, non-irritating to the skin, and has antioxidant, anti-aging, skin wrinkle improvement, skin elasticity enhancement, skin whitening, and skin moisturizing effects, so it is itself a cosmetic, skin external agent, It can be used as food, etc.

The composition may be a cosmetic composition.

The cosmetic composition may be prepared in any formulation that is conventionally prepared. For example, the cosmetic composition may be cosmetic water, cream, essence, cleansing foam, cleansing water, pack, ampoule, body lotion, body oil, body gel, shampoo, rinse, hair conditioner, hair gel, foundation, lipstick, mascara, makeup It may have a base or a skin-adhesive cosmetic formulation. The skin-adhesive cosmetic formulation may be, for example, a mask pack, but is not limited thereto.

Ingredients included in the cosmetic composition may include ingredients commonly used in cosmetic compositions other than the composition as active ingredients, for example, conventional adjuvants and carriers such as stabilizers, solubilizers, vitamins, pigments and flavors. can include

In addition, the composition may be a composition for external application for skin.

The external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The external skin preparation is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or combinations thereof and may be suitably blended as needed. The external skin preparations include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, and calin. Hot-water extracts of fruits, various herbal medicines, tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts and other drugs, vitamin C, magnesium ascorbate phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars, such as trehalose, etc. can also be mix|blended suitably.

The composition may be a food composition. The composition may be a health functional food composition.

The food composition may be used alone or in combination with other food or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment). There is no particular limitation on the type of health functional food. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The health food composition may also contain nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. carbonation agent used, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.

The composition may be a pharmaceutical composition.

In one embodiment, the pharmaceutical composition may be used for preventing or treating skin aging.

The skin aging may be photoaging or skin aging due to age.

The pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, and magnesium stearate, talc, or a combination thereof as a lubricant. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated for oral or parenteral administration. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrups, or combinations thereof. Parenteral dosage forms may be injections.

Another aspect provides a composition comprising snowberry oil.

The snowberry oil may be extracted from snowberry seeds. Thus, the snowberry oil may be snowberry seed oil.

The composition is as described above.

Redundant content is omitted in consideration of the complexity of the present specification, and terms not defined otherwise in the present specification have meanings commonly used in the technical field to which the present invention belongs.

According to the method according to one aspect, the snowberry pulp can be easily separated. The snowberry pulp separated by the above method does not turn brown, is non-toxic and non-irritant, has a unique appearance and touch, and has skin improving functionality, so it can be used as a cosmetic or food raw material.

1 is a photograph showing a snow berry.
Figure 2 is a photograph showing the finally separated snowberry pulp in Example 1.
3 is a photograph showing the finally separated snowberry pulp in Example 1.
Figure 4 is a photograph showing the essence formulation cosmetic composition of Preparation Example 1.
5 is a photograph showing the essence formulation cosmetic composition of Preparation Example 2.
6 is a photograph showing a creamy cosmetic composition of Preparation Example 3.
7 is a graph showing the results of cytotoxicity evaluation of snowberry flesh.
8 is an evaluation result of antioxidant activity of snowberry pulp.
9 is a graph showing the inhibitory effect of snowberry pulp on MMP-1 activity.
10 is a graph showing the measurement results of transepidermal water loss of the essence formulation cosmetic composition containing 3% by weight of snowberry flesh of Preparation Example 1.
11 is a graph showing the measurement results of transepidermal water loss of the essence formulation cosmetic composition containing 5% by weight of snowberry flesh of Preparation Example 2.
12 is a graph showing the results of measurement of transepidermal water loss of the creamy cosmetic composition containing 3% by weight of snowberry pulp in Preparation Example 3.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1. Separation of snowberry flesh

(1) In order to remove foreign substances, snow berry ( Symphoricarpos albus ) fruit raw material was washed 2-3 times using lukewarm water (15-30 ° C).

(2) To facilitate peeling and pulp separation of snowberries, 100 g of washed snowberries were soaked in vinegar containing 5% (w/v) acetic acid for about 5 to 15 minutes.

(3) After separating only the snowberry pulp, 500 to 1000 ml of fermented alcohol (95% ethanol) diluted to about 1 to 20% (v / v) was added and stirred slowly at 10 to 100 rpm for 30 to 90 minutes. Through the addition of fermented alcohol, browning of snowberry flesh was prevented and toxic substances were removed.

(4) Only snowberry skins were removed using a 3-5 mesh (4000-6730 μm) filter, and the filtrate passed through the filter was taken.

(5) In order to prevent softness of the flesh and maintain hardness, the filtrate was immersed in ice water and stirred slowly.

(6) The flesh and seeds were separated using a separatory funnel.

(7) The separated pulp was centrifuged at a weak rpm (150 rpm or less) to remove the supernatant, and the precipitated pulp was used as a raw material for cosmetics.

1 is a photograph showing a snow berry.

Figure 2 is a photograph showing the finally separated snowberry pulp in Example 1.

3 is a photograph showing the finally separated snowberry pulp in Example 1.

Production Examples 1 to 2. Preparation of Essence Type Cosmetic Composition Containing Snowberry Pulp

According to the ingredients and contents (% by weight) in Table 1 below, cosmetic compositions of essence formulations containing snowberry pulp were prepared.

Specifically, ingredients 1 to 3 were added to mixer tank 1 and then dispersed at room temperature for 30 minutes. Components 4 to 16 were added to mixer tank 2 and completely dissolved at room temperature. Components No. 17 to 19 were added to mixer tank No. 3 and stirred at room temperature. Components No. 20 to 22 were added to mixer tank No. 4, mixed and dissolved by heating to 40 ° C, and then raw material No. 23 was added and stirred. After adding the raw materials of the mixer tank No. 1, the mixer tank No. 2, and the mixer tank No. 3 to the main mixer tank No. 5, they were stirred for 5 minutes at 3500 rpm using a homomixer, and then ingredients No. 24 and 25 were added. Thereafter, all of the raw materials were transferred to a separate weighing container, stirred at 3500 rpm for 5 minutes, and defoamed to finally prepare an essence-type cosmetic composition.

The cosmetic composition of Preparation Example 1 contains 3% by weight of the snowberry pulp of Example 1. The essence-type cosmetic composition of Preparation Example 1 is shown in FIG. 4 .

The cosmetic composition of Preparation Example 2 contains 5% by weight of the snowberry pulp of Example 1. The essence-type cosmetic composition of Preparation Example 2 prepared is shown in FIG. 5.

No. ingredient Preparation Example 1 Preparation Example 2 One Purified water 25.00 25.00 2 Acrylates/C10-30 Alkylacrylate Crosspolymer (product name: Pemulen TR-2) 0.14 0.14 3 Carbomer (product name: Carbopol 940) 0.18 0.18 4 Purified water 38.84 36.84 5 EDTA-3Na 0.02 0.02 6 Betaine (product name: Aminocoat) 2.00 2.00 7 D-panthenol 1.00 1.00 8 Snowberry pulp of Example 1 3.00 5.00 9 Hydrolyzed Acacia Macrostachia Seed Extract/Maltodextrin (Product Name: Aqualicia bio) 0.10 0.10 10 trehalose 0.05 0.05 11 Purslane Extract ( Portulaca Oleracea Extract) 1.00 1.00 12 Saururus Chinensis Extract 1.00 1.00 13 Schisandra chinensis fruit extract/cypress water/Angelica root extract/Green tea extract/Bitter gourd fruit extract (product name: Curerex SSC) 1.00 1.00 14 Olive tree callus culture lysate (product name: OLEA Vitae-C PLF 02) 0.10 0.10 15 Insulin/Glycerin/Phenoxyethanol/Lecithin/Xanthan Gum/Water (Product Name: Polyfructol plus) 0.20 0.20 16 Peruginum Azalea Extract/Glycerin/Water (Product Name: Alpine Rose Active) 0.30 0.30 17 Glycerin 98% 6.00 6.00 18 Dipropylene glycol (product name: DPG-FC) 3.00 3.00 19 Cyclomethicone/Dimethicone (product name: SILICONE Q2-1401) 2.00 2.00 20 Ethanol (product name: S3ETOH) 3.00 3.00 21 spices 0.1 0.1 22 PEG-40 Hydrogenated Castor Oil (Product Name: HCO-40) 0.05 0.05 23 Silica (product name: Sunsil-130) 0.50 0.50 24 Palmitic Acid (Product Name: Unimell-MB) 0.20 0.20 25 Purified water 11.22 11.22 Sum 100.00 100.00

Preparation Example 3. Preparation of cream cosmetic composition containing snowberry pulp

According to the ingredients and contents (% by weight) of Table 2 below, a creamy cosmetic composition containing snowberry pulp was prepared.

Specifically, component No. 19 was placed in a separate weighing container and then completely dissolved. Components No. 6 to 8 were added to mixer tank No. 1 and then dispersed at room temperature for 30 minutes. Components No. 9 to 15 were added to mixer tank No. 2 and heated to 40° C. to completely dissolve them. Components 1 to 5 were added to the mixer tank No. 3, mixed and stirred, and heated to 40 ° C. Components No. 16 to 17 were added to the mixer tank No. 4, and mixed and dissolved by heating to 40 ° C. After adding the raw materials of the mixer tank No. 2 to the main mixer tank No. 5, the raw materials of the mixer tank No. 3 were added while stirring. The first emulsification was carried out at 40 ° C. for 3 minutes with a homomixer at 3000 rpm. Here, the raw material for the mixer tank No. 4 was added to perform secondary emulsification. After adding raw materials No. 20 to 25 here, emulsified for about 3 minutes, de-foamed, and finally prepared a cosmetic composition of cream type.

The cosmetic composition of Preparation Example 3 contains 3% by weight of the snowberry pulp of Example 1. The creamy cosmetic composition of Preparation Example 3 prepared is shown in FIG. 6.

No. ingredient Preparation Example 3 One Dimethicone (product name: DC 200/6CS) 5.00 2 Cyclomethicone/Dimethicone (product name: SILICONE Q2-1401) 5.00 3 Caprylic/capric triglyceride (product name: WAGLINOL 3/920) 1.00 4 Glycerin 98% 6.00 5 Dipropylene glycol (product name: DPG-FC) 8.00 6 Purified water 39.00 7 Acrylates/C10-30 Alkylacrylate Crosspolymer (product name: Pemulen TR-2) 0.10 8 Carbomer (product name: Carbopol 940) 0.48 9 Purified water 9.77 10 EDTA 0.03 11 Hyaluronic Acid (Product Name: HYASOL) 0.50 12 D-panthenol 1.00 13 Snowberry pulp of Example 1 3.00 14 Hydrolyzed Acacia Macrostachia Seed Extract/Maltodextrin (Product Name: Aqualicia bio) 0.10 15 trehalose 0.05 16 Ethanol (product name: S3ETOH) 5.00 17 PEG-40 Hydrogenated Castor Oil (Product Name: HCO-40) 0.50 18 Purified water 11.27 19 Purslane Extract ( Portulaca Oleracea Extract) 1.00 20 Saururus Chinensis Extract 1.00 21 Schisandra chinensis fruit extract/cypress water/Angelica root extract/Green tea extract/Bitter gourd fruit extract (product name: Curerex SSC) 1.50 22 Peruginum Azalea Extract/Glycerin/Water (Product Name: Alpine Rose Active) 0.30 23 Insulin/Glycerin/Phenoxyethanol/Lecithin/Xanthan Gum/Water (Product Name: Polyfructol plus) 0.20 24 Olive tree callus culture lysate (product name: OLEA Vitae-C PLF 02) 0.10 25 Spices 0.10 Sum 100.00

Experimental Example 1. Cytotoxicity evaluation of snowberry flesh

In order to evaluate the cytotoxicity of the snowberry pulp of Example 1, WST-1 assay was performed.

(1) WST-1 assay

first, HaCaT cells, a human keratinocyte cell line, were cultured in DMEM mixed with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin at 37°C in a 5% CO2 incubator. Culture medium was exchanged every 2-3 days.

Experiments for measuring cell viability used WST-1 (water soluble tetrazolium salt-1) assay. Specifically, HaCaT cells were dispensed in 100 μL at a concentration of 4 × 10³ cells/well in a 96-well plate and cultured for 24 hours. Thereafter, the medium was removed, washed with PBS, and starved for 12 hours. The wells were treated with the snowberry pulp of Example 1 at concentrations of 5 mg/ml, 3 mg/ml, or 1 mg/ml, respectively, and cultured for 24 hours. After cell culture, 10 μM of WST-1 solution was added to each well and incubated at 37° C. for 1 hour. Absorbance was measured at 450 nm using a microplate reader.

(2) Experimental results

7 is a graph showing the results of cytotoxicity evaluation of snowberry flesh.

As shown in FIG. 7, even when the snowberry pulp of Example 1 was used at a high concentration of 5 mg/ml, there was no change in cell viability compared to the untreated control group. Therefore, it was confirmed that the snowberry flesh of Example 1 had no cytotoxicity even at high concentrations.

Experimental Example 2. Evaluation of antioxidant activity of snowberry flesh

In order to evaluate the antioxidant activity of the snowberry pulp of Example 1, in vitro SOD (Superoxide dismutase)-like activity measurement experiments and in vitro ( in vitro ) DPPH radical scavenging activity measurement experiments were performed.

(1) In vitro ( in vitro ) SOD-like activity measurement

SOD-like activity evaluation was performed using the SOD assay Kit. Specifically, 20 μl of the snowberry pulp of Example 1 was dispensed into a 96-well plate, and 200 μl of water soluble tetrazolium salt (WST) was added to all wells. Then, 20 μl of xanthine oxidase was added to the sample and reacted at 37° C. for 20 minutes. The absorbance was measured at 450 nm using a microplate reader, and the difference in absorbance between the group with and without the addition of the sample was expressed as a percentage as shown in Equation 1 below. As a positive control, BHT (Butyl hydroxy toluene), which is well known for its antioxidant effect, was used.

[Equation 1]

SOD activity (inhibition rate, %) =

(2) In vitro ( in vitro ) Measurement of DPPH radical scavenging activity

Radical scavenging activity experiments were conducted as follows by applying the DPPH Blois method. After dissolving the snowberry pulp samples of Example 1 for each concentration in a 70% ethanol solution, the samples were dispensed into 96-well plates. DPPH was dissolved in methanol at 0.2 mM and the samples were treated. After stirring, the mixture was allowed to stand at room temperature for 30 minutes, and absorbance was measured at 492 nm. BHT (Butylated Hydroxytoluene) and Ascorbic Acid were used as controls.

(3) Experimental results

8 is an evaluation result of antioxidant activity of snowberry pulp.

As shown in FIG. 8, it was confirmed that the snowberry pulp of Example 1 was excellent in SOD-like activity and DPPH radical scavenging activity at each concentration of 5 mg/ml or 3 mg/ml. Therefore, it was found that the snowberry pulp of Example 1 had excellent antioxidant activity.

Experimental Example 3. Measurement of anti-aging activity of snowberry pulp

In order to measure the anti-aging activity of the snowberry pulp of Example 1, an MMP-1 (Matrix Metalloproteinase-1) measurement experiment was performed.

Among the components constituting the connective tissue of skin cells, collagen is a major constituent protein that accounts for about 90% of the dry weight of the skin. Therefore, the decomposition of collagen has a direct effect on the reduction of elasticity of connective tissue and the formation of wrinkles. Among several types of MMPs produced in the body, MMP-1 is a protease that acts specifically on collagen. By inhibiting the activity of MMP-1 to reduce the degradation of collagen, elasticity of skin tissue can be maintained and wrinkle formation can be prevented.

(1) MMP-1 measurement

Skin fibroblasts were seeded in a 6-well plate at 2 × 10 ⁶ cells/2 ml. The snowberry pulp of Example 1 was added to each well by concentration and cultured for 24 hours in a CO ₂ incubator. At this time, in order to increase the activity of MMP-1, TNF-α was added at a concentration of 10 ng/ml. The cell culture medium was collected and used for experiments, and plate reader (plate reader) using MMP-1 Biotrack Activity Assay Kit (Amersham Bioscience) according to the method of Gross BE (Proc Natl Acad Sci USA. 1962; 48:1014-1022). reader) to measure the absorbance at 450 nm.

MMP-1 activity was measured for snowberry pulp at various concentrations, but among them, 0.1 mg/ml (100 ppm), the lowest concentration without affecting cytotoxicity, was selected as a representative test group. As a comparison group, a group treated with 0.1 ppm of vitamin C, which does not exhibit cytotoxicity, was used.

(2) Experimental results

9 is a graph showing the inhibitory effect of snowberry pulp on MMP-1 activity.

As shown in Figure 9, it was confirmed that the MMP-1 level significantly increased in the group treated with TNF-α alone to increase MMP-1 activity. On the other hand, in the group treated with snowberry pulp of Example 1, the level of MMP-1 was reduced by about 23% or more than in the group treated with TNF-α alone. That is, it was confirmed that the snowberry pulp of Example 1 reduced the MMP-1 level even at a low concentration of 100 ppm. Therefore, it was confirmed that the snowberry pulp of Example 1 can suppress skin aging by inhibiting the activity of MMP-1 that affects collagen decomposition, thereby increasing effects such as improving skin wrinkles and enhancing skin elasticity.

Experimental Example 4. Evaluation of whitening efficacy of snowberry pulp

In order to evaluate the whitening efficacy of the snowberry pulp of Example 1, an experiment was performed to confirm the melanin production inhibition rate.

(One) Confirmation of melanin production inhibition rate

In order to examine the ability of snowberry pulp to inhibit melanin production, an experiment was performed in the following manner using B16/F10 melanoma cells (Korea Cell Line Bank).

DMEM containing 10% FBS was used as a medium. For melanin quantification, B16/F10 melanoma cells were plated in a 6-well plate at 2×10 ⁴ cells/well (final volume 3 ml). In addition, for the MTT assay, B16/F10 melanoma cells were placed in a 96-well plate at 2 X 10 ³ cells/well (final volume: 200 μl). Each was cultured for 24 hours in a 37°C, 5% CO ₂ incubator (MCO-20AIC, Sanyo).

After culturing, the medium was removed, and DMEM medium containing 10% FBS, 2 μM α-MSH (Melanocyte stimulating hormone) and 2 mM theophylline was used as a new medium. The medium was treated daily for 3 days by adding the test substance. The snowberry pulp of Example 1 was used as a test substance. Kojic acid or Melasolv was used as a positive control. As a negative control group, the snowberry pulp-free group of Example 1 was used.

On the 5th day of culture, the culture medium was removed from the 6-well plate and treated with 0.25% trypsin-EDTA (Sigma Chemical Co.) solution. Thereafter, the cell pellet was recovered and transferred to a 1.5 ml tube and centrifuged at 10,000 rpm for 10 minutes to remove the supernatant. The resulting pellet was dried at 60° C., and 100 μl of 1N NaOH was added to dissolve intracellular melanin. After diluting this solution with PBS, the absorbance was measured at 475 nm using an ELISA reader, and the melanin production inhibition rate of the sample was calculated as shown in Equation 2 below.

[Equation 2]

Melanin production inhibition rate (%) = 100 - {(absorbance of each test substance/absorbance of negative control group) × 100}

(2) Experimental results

Table 3 shows the melanin production inhibition rate (%) according to the treatment concentration (0.001% (w/w) or 0.01% (w/w)) of each sample.

0.001% (w/w) 0.01% (w/w) Snowberry pulp of Example 1 19.8 23.6 kojic acid 33.0 50.1 Melasolve 42.2 56.8 negative control group - -

As shown in Table 3, it was confirmed that the snowberry pulp of Example 1 inhibited melanin production in a concentration-dependent manner. Therefore, it was found that the snowberry pulp of Example 1 had excellent whitening efficacy.

Experimental Example 5. Primary skin irritation test of cosmetic composition containing snowberry pulp

For the primary skin irritation test of the cosmetic compositions containing snowberry pulp of Preparation Examples 1 to 3, a patch test on the human body was performed.

(1) Human patch test

Ten healthy adult males and females were tested. A paper disc was attached to the Finn chamber (SmartPractice, Denmark), sufficiently wetted with the test solution, and then adhered to the skin and fixed with Tape (Scanpor tape by Norgesplaster AS). The Finn Chamber with the test substance applied was applied to the test site for 24 hours. After 24 hours, the Finn Chamber was removed, and the degree of skin reaction was evaluated after 30 minutes. The evaluation results were expressed as mean scores according to the standards of the International Contact Dermatitis Research Group (ICDRG), as shown in Tables 4 and 5 below.

mark evaluation score - voice 0 ± Suspicious or mild reactions and erythema 0.5 + erythema + induration One ++ erythema + induration + blisters 2 +++ Erythema + Induration + Bulla 3

Mean score classification 0.00 to 0.75 non-irritating 0.76~1.50 hypoallergenic 1.51 to 2.50 moderate stimulation 2.51~4.00 high stimulation 4.01~ severe high irritation

(2) Experimental results

Table 6 shows the results of the primary skin irritation test of the cosmetic compositions of Preparation Examples 1 to 3.

test group average score judgment result Preparation Example 1 0.065 non-irritating Preparation Example 2 0.08 non-irritating Preparation Example 3 0.035 non-irritating

As shown in Table 6, in the cosmetic compositions containing snowberry pulp of Preparation Examples 1 to 3, no skin abnormalities such as erythema, soreness, and exfoliation were observed. Therefore, it was confirmed that the snowberry pulp of Example 1 was non-irritating to the skin when used as a raw material for cosmetics.

Experimental Example 6. Measurement of skin moisturizing effect of snowberry pulp-containing cosmetic composition

The skin moisturizing effect of the cosmetic compositions containing snowberry pulp of Preparation Examples 1 to 3 was measured using electrical properties.

(1) Measurement of skin moisturizing effect using electrical properties

The skin moisturizing effect of the cosmetic composition containing snowberry pulp was measured using the electrical properties of moisture present in the skin. SKICON-200EX (YAYOI.CO., LTD, Japan), which quantifies the moisture content of the skin, was used as a measuring device. A total of 22 male and female adults with an average age of 46.82±8.02 years were selected as test subjects. The cosmetic composition of each of Preparation Examples 1 to 3 was applied in an amount of 20 µl to an area of 20 mm x 20 mm in diameter on the lower arm once daily for 2 weeks. Before using the product, after 1 day, 7 days, and 14 days after use, the moisture content of the skin was measured over time. The value that appears when the probe is placed on the measurement site and pressed is measured 4 times per area and expressed as an average value, and the measured value is expressed as a change rate (%) of Equation 3 below. The significance probability ( p value) was indicated as *: p <0.05 (Paired sample's T-test), **: p <0.05 (Independent T-test). At the time of measurement, the average temperature was 19.5 ° C (± 1 ° C) and the humidity was 30.0% ( ± 2.0%), and the measurement was performed after the test subject's skin was stabilized in the above conditions for 10 minutes or more.

[Equation 3]

Change rate (%) = {(measured value after product use - measured value before product use) / measured value before product use} × 100

(2) Experimental results

Table 7 shows the measurement results of transepidermal water loss of the essence formulation cosmetic composition containing 3% by weight of snowberry flesh of Preparation Example 1.

10 is a graph showing the measurement results of transepidermal water loss of the essence formulation cosmetic composition containing 3% by weight of snowberry flesh of Preparation Example 1.

Table 8 shows the measurement results of transepidermal water loss of the essence formulation cosmetic composition containing 5% by weight of snowberry pulp of Preparation Example 2.

11 is a graph showing the results of measurement of transepidermal water loss of the essence formulation cosmetic composition containing 5% by weight of snowberry pulp in Preparation Example 2.

Table 9 shows the measurement results of transepidermal water loss of the creamy cosmetic composition containing 3% by weight of snowberry pulp of Preparation Example 3.

12 is a graph showing the results of measurement of transepidermal water loss of the creamy cosmetic composition containing 3% by weight of snowberry pulp in Preparation Example 3.

Transepidermal water loss change (Preparation Example 1) hour Mean±Standard Deviation (℃) Change rate (%) Significance probability ( p value) Comparison within the military comparison between groups Before product use (-1 day) 7.977±1.449 - - - Before product use (day 0) 20.814±4.118 160.912 0.000* 0.495 Before product use (1 day) 18.755±4.460 135.100 0.000* 0.003** Before product use (7 days) 12.068±2.421 51.282 0.000* 0.146 Before product use (14 days) 9.368±1.791 17.436 1.000 0.015**

Transepidermal water loss change (Preparation Example 2) hour Mean±Standard Deviation (℃) Change rate (%) Significance probability ( p value) Comparison within the military comparison between groups Before product use (-1 day) 8.073±1.528 - - - Before product use (day 0) 20.877±4.161 158.615 0.000* 0.483 Before product use (1 day) 18.923±4.458 134.403 0.000* 0.006** Before product use (7 days) 11.918±2.142 47.635 0.001* 0.067 Before product use (14 days) 9.700±1.461 20.158 0.789 0.036**

Transepidermal water loss change (Preparation Example 3) hour Mean±Standard Deviation (℃) Change rate (%) Significance probability ( p value) Comparison within the military comparison between groups Before product use (-1 day) 8.082±1.598 - - - Before product use (day 0) 21.082±4.185 160.855 0.000* 0.510 Before product use (1 day) 19.105±4.535 136.389 0.000* 0.007** Before product use (7 days) 11.736±2.210 45.219 0.002* 0.031 Before product use (14 days) 9.747±1.589 20.607 0.780 0.027**

As shown in Tables 7 to 9 and FIGS. 10 to 12, as a result of using the cosmetic compositions of Preparation Examples 1 to 3 for 2 weeks, it was confirmed that all products reduced transepidermal water loss compared to the uncoated control group. Therefore, it was confirmed that the snowberry pulp of Example 1 had a skin moisturizing effect when used as a cosmetic raw material.

Claims (10)

(1) washing snowberry fruit;
(2) immersing the snowberry fruit washed in step (1) in vinegar and peeling to separate the snowberry pulp;
(3) adding fermented alcohol to the snowberry pulp separated in step (2);
(4) using a filter to remove snowberry skin residue and obtain a filtrate;
(5) stirring the filtrate obtained in step (4) at low temperature;
(6) separating the filtrate stirred in step (5) into snowberry flesh and seeds; and
(7) centrifuging the snowberry pulp separated in step (6) to remove the supernatant and obtaining the precipitated snowberry pulp,
How to separate snowberry pulp. The method according to claim 1, wherein in the step (2), the vinegar contains 0.1 to 10% (w / v) acetic acid. The method according to claim 1, wherein browning of snowberry flesh is prevented by the step (3). The method according to claim 1, wherein in the step (3), the fermented alcohol is diluted to 1 to 20% (v / v). The method according to claim 1, wherein in step (4), the filter is a 3 to 5 mesh filter. The method according to claim 1, wherein in the step (5), the low temperature is a temperature of -20 ° C to 10 ° C. The method according to claim 1, wherein in the step (7), the centrifugation is performed at 100 to 150 rpm. The method according to claim 1, wherein the snowberry pulp obtained in step (7) is used as a cosmetic raw material or an external skin preparation raw material. A cosmetic composition comprising snowberry pulp separated by the method of any one of claims 1 to 8. The cosmetic composition of claim 9, wherein the snowberry pulp is included in an amount of 0.0001 to 100% by weight based on the total weight of the cosmetic composition.

Images