KR102510795B1 - Method for preparing wrinkle improvement composition containing sturgeon water-soluble oil - Google Patents

Abstract

The present invention relates to a skin wrinkle-reducing composition, and more particularly, to a skin wrinkle-reducing composition, which exerts an excellent effect on skin wrinkle reduction by containing Acipenser sinensis ingredients with effects of inhibiting intracellular collagen production and intracellular collagenase activity as active ingredients, and a method for preparing the same.

Description

Method for preparing wrinkle improvement composition containing sturgeon water-soluble oil}

The present invention relates to a composition for improving skin wrinkles, and more particularly, contains a sturgeon component having an effect of inhibiting intracellular collagen production and intracellular collagenase activity as an active ingredient, which is excellent in improving skin wrinkles. It relates to a functional composition for improving wrinkles that exerts an effect and a method for preparing the same.

Sturgeon ( Acipenser sinensis ) is a fish species originating from 450 to 350 million years ago (Devonian Period), belonging to the Acipenseridae family, and is a fish species that is widely distributed in the northern mid-latitude region of the Northern Hemisphere and grows naturally. Currently, 26 species of sturgeon are known, and interest in many countries is increasing due to their economic and ecological value.

In March 1997, Siberian sturgeon, which was first transplanted and bred in Korea at Canakova Farm in Russia, and then produced in 2003 by its own seedlings, has excellent environmental adaptability, grows faster than other breeds farmed in Korea, and has good meat quality. It is gaining popularity as a target fish for aquaculture. The maximum body length is 3 m, the weight can grow up to 200-300 kg, there are species that grow to a maximum length of more than 8 m, and the maximum life span exceeds 100 years. At about 6 to 7 years old, they grow to more than 1 m and produce caviar.

Sturgeon has a chewy taste of flesh, so it is used in raw, smoked, barbecued, steak and skewered dishes, its head is steamed and stewed, its fin is used in shark fin dish, which is one of the three major delicacies in China, and its cartilage is used for special dishes, tonics and industrial ingredients. there is. In addition, sturgeon contains a large amount of active ingredients such as protein, essential amino acids, unsaturated fatty acids, vitamins, EPA, DHA, etc. As the excellent efficacy of is revealed, it is getting more and more attention as a highly nutritious and highly functional material.

In recent years, as sturgeon farming technology has begun to develop, there are more opportunities for common people to come into contact with sturgeon. It became.

However, in the raw material of sturgeon marketed to date, sufficient research has not been conducted on the actual content of sturgeon, the content of active ingredients, processing method, extraction efficiency, etc. In particular, since there is no processing technology for sturgeon and products using it in Korea, sturgeon has not yet been objectively and publicly recognized by consumers as a functional material and product.

On the other hand, one of the fields of interest in the beauty field is aging-related wrinkles, pigmentation, loss of elasticity, skin dryness, hair loss, lack of luster of hair, and the like. The skin undergoes various changes through aging.

The epidermis, dermis, and subcutaneous tissue, which are components of the skin, become thinner and the extracellular matrix (ECM) component that gives skin elasticity changes. As the skin ages, its production rapidly declines and has a close relationship with wrinkle formation, and collagen, elastin, proteoglycans, glucosaminoglycans, Laminin, fibronectin, etc. are oxidized and lose their functions, so the skin loses its elasticity and wrinkles are excessively formed, transforming into aged skin.

With age, the content of collagen and elastin in skin connective tissue decreases, which is due to the promotion of degradation and degradation of collagen and elastin synthesis. Therefore, the decrease in collagen and elastin biosynthesis and promotion of decomposition can be said to be the biggest cause of skin wrinkle formation. In order to solve the problem of skin aging, various ingredients and compositions have recently been studied, and the effect of improving wrinkles on the skin has shown visible results in some cases.

Accordingly, the present applicant has completed this invention after finding out that the sturgeon extract obtained by the method for obtaining raw materials for sturgeon exhibits an excellent effect on skin wrinkle improvement through experiments.

Republic of Korea Patent Publication No. 10-2009-0111066

The present invention is to solve the above problems, and uses a sturgeon component having an effect of inhibiting intracellular collagen production and intracellular collagenase activity as an active ingredient, which exhibits an excellent effect on skin wrinkle improvement, It is intended to provide a functional composition for improving wrinkles and a manufacturing method thereof.

The above and other objects and advantages of the present invention will become apparent from the following description of preferred embodiments.

The above object is to use sturgeon water-soluble oil as an active ingredient, and the sturgeon water-soluble oil has an effect of inhibiting intracellular collagen production and intracellular collagenase activity by a functional composition for improving skin wrinkles. can be achieved

The skin wrinkle improvement functional composition is characterized in that it is made of one or more health functional compositions selected from cosmetic compositions, food compositions, beverage compositions and pharmaceutical compositions.

The sturgeon water-soluble oil is prepared by preparing sturgeon oil according to a method for obtaining raw materials for sturgeon; Performing a first aging of the sturgeon oil to prepare an aged sturgeon oil; A pre-treatment step of foaming the aged sturgeon oil by using a high-pressure processor and a fine nozzle to form changes in physical properties of the oil; Second aging of the pretreated sturgeon oil aging product;

Mixing the aged sturgeon oil and water, in which the second aging has been performed, and introducing the mixture into a distillation extractor and performing a second extraction; After performing the second extraction, water is mixed with the remaining matured sturgeon oil and introduced into a distillation extractor, and third extraction is performed to produce first water-soluble oil; Preparing a first additive composition containing edible snails and a second additive composition containing Puer tea and chrysanthemum, respectively; and distilling and extracting a mixture of the primary water-soluble oil, the primary additive composition, and the secondary additive composition to produce secondary water-soluble oil.

The method for obtaining the raw material of the sturgeon includes a farming step of culturing the sturgeon fry for 3 years or more using natural antibiotics; A pre-treatment step of shipping the farmed sturgeon and pre-processing the whole fish; A cold storage step of rapidly cooling the pretreated sturgeon, wherein the rapid cooling reaches a temperature of -80 to -20 ° C in 5 to 20 minutes; and thawing and washing the sturgeon stored in the cold, then putting the entire sturgeon into a pneumatic extractor and performing a first extraction to obtain sturgeon oil.

According to the present invention, a skin wrinkle-improving functional composition containing a sturgeon component, specifically, a sturgeon extract, and more specifically, a sturgeon water-soluble oil as an active ingredient can be prepared.

More specifically, the water-soluble sturgeon oil is obtained from an extract obtained from whole sturgeon, and has an excellent effect on intracellular collagen production and intracellular collagenase activity inhibition, thereby improving skin wrinkles. can be effective.

In addition, according to the present invention, when preparing sturgeon water-soluble oil, human safety due to non-antibiotics is secured, thereby preparing an eco-friendly, high-quality, and highly functional sturgeon-containing composition compared to conventional sturgeon-containing products. .

However, the effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

1 is a photograph of preparing sturgeon water-soluble oil according to an embodiment of the present invention.
Figure 2 is a table showing the results of the cytotoxicity test of Experimental Example 1.
3 is a graph showing the results of the cytotoxicity test of Experimental Example 1.
Figure 4 is a table showing the results of the intracellular collagenase activity inhibition test of Experimental Example 1.
Figure 5 is a graph showing the results of the intracellular collagenase activity inhibition test of Experimental Example 1.
6 is a table showing the test results of intracellular collagen production in Experimental Example 2;
7 is a graph showing the test results of intracellular collagen production in Experimental Example 2;

Hereinafter, the present invention will be described in detail with reference to embodiments and drawings of the present invention. These examples are only presented as examples to explain the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples. .

In addition, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of skill in the art to which this invention belongs, and in case of conflict, this specification including definitions of will take precedence.

In order to clearly explain the proposed invention in the drawings, parts irrelevant to the description are omitted, and similar reference numerals are attached to similar parts throughout the specification. And, when a certain component is said to "include", this means that it may further include other components without excluding other components unless otherwise stated. Also, a “unit” described in the specification means one unit or block that performs a specific function.

In each step, the identification code (first, second, etc.) is used for convenience of description, and the identification code does not describe the order of each step, and each step does not clearly describe a specific order in context. It may be performed differently from the order specified above. That is, each step may be performed in the same order as specified, may be performed substantially simultaneously, or may be performed in the reverse order.

In the present invention, in the present specification, "sturgeon extract" means sturgeon liquid extract, sturgeon oil or sturgeon solids obtained from whole sturgeon, which will be described later, and is preferably interpreted comprehensively.

Hereinafter, embodiments and embodiments of the present disclosure will be described in detail with reference to the accompanying drawings. However, it may not be limited to these implementations and examples and drawings herein.

One aspect of the present application is characterized by using sturgeon water-soluble oil as an active ingredient, and the sturgeon water-soluble oil has an effect of inhibiting intracellular collagen production and intracellular collagenase activity, skin wrinkle improvement functionality composition is provided.

Preparation of sturgeon water soluble oil composition

<Sturgeon Oil>

First, sturgeon oil obtained according to the method for obtaining raw materials for sturgeon can be obtained.

The method for obtaining the raw material of the sturgeon includes a farming step (S1) of culturing the sturgeon fry for 3 years or more using natural antibiotics; A pre-processing step (S2) of shipping the farmed sturgeon and pre-processing them whole; A cold storage step (S3) of rapidly cooling the pretreated sturgeon, such that the rapid cooling reaches a temperature of -80 to -20 ° C within 5 to 20 minutes; and thawing and washing the sturgeon stored in the cold, then putting the whole sturgeon into a pneumatic extractor and performing a first extraction to obtain sturgeon oil (S4).

More specifically, in the step S1, sturgeon, specifically, sturgeon fry is farmed. The breeds of the sturgeon include Beluga sturgeon (Huso huso), Kaluga sturgeon (Huso dauricus), Siberian sturgeon (Acipenser baerii), Russian sturgeon (Acipenser gueldenstaedti), stellet sturgeon (Acipenser ruthenus), white It may be selected from the group consisting of sturgeon (Acipenser transmontanus), Amur sturgeon (Acipenser schrenckii), Chinese sturgeon (Acipenser sinensis), spatula sturgeon (Polyodon spathula) and hybrids thereof. Sturgeon according to an embodiment of the present invention may be, for example, Siberian sturgeon, but is not limited thereto.

The environment of the aquaculture may be a fresh water farm, and the fresh water source may be river water, ground water, mountain spring water, and tap water, and the fresh water source may be a water source that has undergone purification treatment such as dechlorination treatment with activated carbon.

The farming may be carried out until the sturgeon fry become adult sturgeon weighing at least 1 kg, eg for more than 3 years, eg 3 to 5 years.

The aquaculture is oriented toward an antibiotic-free environment. That is, in the sturgeon farming of the present invention, as in the prior art, chemical components such as various antibiotics and growth promoters administered for weight gain and meat quality improvement of the sturgeon are not used, and natural antibiotics made of natural materials are used. can

For example, bay salt may be used as the natural antibiotic. Specifically, by allowing the sturgeon to consume sea salt while being farmed, impurities including harmful bacteria inside the sturgeon can be effectively removed. Intake of the sun salt is preferably carried out within the range of 1 to 3 times a month.

In the above steps S2 and S3, the cultured sturgeon may be pretreated and then stored in a refrigerator. The pretreatment is performed so that the sturgeon is not cut by part, but in the form of whole fish (separated or uncut whole).

The farmed sturgeon is shipped to perform pretreatment, but first, the sturgeon may be fasted so that it does not consume any food from 10 to 12 days before a predetermined shipment date, for example, 10 days before. By allowing the sturgeon to fast, impurities in the intestines are emptied cleanly, so that the yield of active ingredients of the whole sturgeon can be further increased.

The sturgeon after fasting is shipped out, washed briefly, and then knocked down by hitting the sturgeon head once or twice with a rubber mallet. The stunned sturgeon can then be rapidly cooled directly to carry out storage. The rapid cooling condition may be to reach a temperature of -80 to -20 °C in a short time, for example, between 5 and 20 minutes.

By rapidly cooling the sturgeon after rapidly stunning it in the above-described manner, it is possible to obtain high-quality active ingredients of the whole sturgeon while minimizing the pain of the sturgeon.

In the above step S4, sturgeon raw material, that is, sturgeon oil is obtained.

The cooled sturgeon is thawed for 2 to 6 hours, and then the entire fish body is evenly washed with water or saline to remove various impurities such as residual blood or residual intestines.

The first extraction may be performed with the sturgeon in whole form. The first extraction method may be performed using a pneumatic extractor, specifically, a pneumatic hot water extractor. The whole sturgeon may be put into the pneumatic extractor together with water as a solvent. At this time, the solvent may further include calamansi undiluted solution, bellflower undiluted solution and beet undiluted solution.

In one example, an extraction raw material obtained by mixing 50% by weight of sturgeon, 41% by weight of water, 5% by weight of calamansi stock solution, 2% by weight of balloon flower stock solution and 2% by weight of beet stock solution is introduced into the pneumatic extractor to prepare 1 extraction can be performed.

The pneumatic extractor may be performed under conditions of 100 to 104 ° C, for example, 102 ° C, 2.3 to 2.7 atm, for example, 2.5 atm, and the extraction time is 98 to 102 hours, for example, 100 hours it is desirable

After the first extraction, specific gravity separation may be performed in which upper, middle, and lower layers are separated using the difference in specific gravity. As a specific method for the specific gravity separation, first, the obtained primary extract is directly put into the stirrer tank without removing steam (kim) while the pneumatic extractor is operating. In this case, the primary extract contained in the agitator tank may be a liquid extract of sturgeon, and the liquid extract of sturgeon may account for about 80% by weight of the total weight of the raw material for extraction.

Thereafter, a secondary extract may be obtained by performing compression of the remaining extraction raw materials once or twice or more with a compressor provided in the pneumatic extractor. In this case, the secondary extract may be sturgeon oil, and the sturgeon oil may account for about 5% by weight based on the total weight of the raw material for extraction.

Then, the remaining raw material for extraction, that is, solids of sturgeon can be obtained. The sturgeon solids may include sturgeon meat, cartilage, bone marrow, or a combination thereof. The solid content of the sturgeon may account for about 15% by weight based on the total weight of the raw material for extraction.

The method for obtaining the sturgeon raw material described in step S4 above is to perform extraction and gravity separation at the same time, so that all of the sturgeon liquid extract, sturgeon oil and sturgeon solids obtained from whole sturgeon are obtained in a simpler process. make it possible to obtain

<Sturgeon water-soluble oil composition>

Second, a first aging of the sturgeon oil may be performed to prepare an aged sturgeon oil. Sterilization may be performed before performing the first aging. A method of sterilization may be performed according to known techniques generally used in the art.

The first aging is carried out in a shaded place by putting the sturgeon oil in a container for aging, but may be natural aging performed under conditions similar to natural conditions. Specifically, the aging is preferably performed for 25 days to 35 days, for example, 30 days, for example, at a temperature range of 17 to 21 ° C., for example, 17 ° C., without sunlight and without air. It can be performed under the following conditions.

If the first aging is performed within the above-described condition range, impurities in the sturgeon oil may sink or float and be removed later, thereby increasing the purity of the sturgeon oil and further increasing the content of active ingredients. After the aging, filtration may be performed separately.

Thirdly, the aged sturgeon oil may be pretreated. The pretreatment may be an operation of making the oil state immediately before water solubilization (solubilization) in order to obtain a formulation of water soluble oil later.

A specific method of performing the pretreatment is to cause a change in the physical properties of the sturgeon oil, for example, while passing the sturgeon oil several to tens to hundreds of times through a high-pressure injector and a fine nozzle provided in the high-pressure injector. It could be foaming shark oil. At this time, the efficiency of the foaming operation may be increased while injecting air using a separate injection pump.

This pretreatment process may be repeated a number of times until the oil becomes a formulation like fresh cream. At this time, the point at which the oil becomes a formulation such as whipped cream may be in a state immediately before water-solubilization (solubilization) to obtain water-soluble oil.

This pretreatment process does not use artificial chemical components such as surfactants used in the past, but only physical nanotechnology causes physical property changes in components to form water solubilization (solubilization) to obtain a formulation of water-soluble oil later. make it possible That is, in one embodiment of the present invention, as a method for producing sturgeon water-soluble oil using a natural nanotechnology, in which harmlessness to the human body and environmental friendliness should be given priority, drinking water, food, cosmetics, pharmaceuticals, pet-related fields, and vegetable raw material compositions It can be used in a wide range of fields such as

The pre-treated sturgeon oil-aged product may be stored in a container at room temperature, for example, 1 to 35 ° C, specifically, 18 to 23 ° C, after the second aging. The second aging is natural aging like the first aging, and the specific conditions are the same as the first aging, but the aging period is preferably about 10 days.

Fourth, the first water-soluble oil may be prepared by performing second extraction and third extraction with the pretreated aged sturgeon oil and water. At this time, the method of the second extraction and the third extraction may be distillation extraction.

Specifically, in the second extraction, distillation extraction may be performed after adding the pretreated aged sturgeon oil and water in a 1:1 weight ratio to a distillation extractor. Subsequently, after the second extraction, water vapor is all evaporated and water is added to the remaining sturgeon oil in a weight ratio of 1: 1.2 to 1.5, and then third extraction is performed by distillation extraction. The third extraction is repeated at least 30 times, thereby obtaining primary water-soluble oil.

The primary water-soluble oil may be a formulation in the process of preparing a final water-soluble oil. The primary water-soluble oil may be put into a stirrer and stirred once or twice.

Fifthly, an additive composition may be prepared. The additive composition may be a first additive composition containing edible snails and a second additive composition containing Puer tea and chrysanthemum, respectively.

The primary additive composition may further include at least one selected from the group of primary natural ingredients consisting of Baekbong Ogolgae, white throat mushroom, beet, kelp root, and bellflower. For example, the first additive composition may include a first natural ingredient that is a mixture of edible snails, Baekbong Ogolgae, white throat mushrooms, beets, kelp roots, and bellflowers. At this time, the natural ingredient may be used by washing the raw material cleanly and cutting it into an average size of about 4 to 7 cm.

The first natural ingredient extract and water are mixed at a weight ratio of 1:1 and then put into a pneumatic extractor and extracted for 100 hours at a temperature of 100° C. or higher to prepare a first natural ingredient extract, that is, a first additive composition.

The secondary additive composition is a group of secondary natural ingredients consisting of lotus seeds, sea bamboo shoots, centella asiatica, wormwood, ginseng mugwort, peppermint, yugenpi, hagocho, guccipon, waspong, chaga mushroom, samchae, calamansi and mulberry leaves It may further include one or more selected from. In one example, the secondary additive composition is puerh tea, chrysanthemum, lotus seed, haejuksun, centella asiatica, wormwood, ginseng wormwood, peppermint, yugeunpi, hagocho, guccippong, waspong, chaga mushroom, samchae, calamansi and mulberry leaves It may contain secondary natural ingredients that are mixtures. At this time, the natural ingredient may be used by washing the raw material cleanly and cutting it into an average size of about 4 to 7 cm.

After mixing the secondary natural component and water at a weight ratio of 1:1, it is introduced into a pneumatic extractor and extracted for 100 hours at a temperature of 100° C. or higher to prepare a secondary natural component extract, that is, a secondary additive composition.

Subsequently, the primary natural component extract and the secondary natural component extract were aged at room temperature, 1 to 35 ° C, for example, 19 to 23 ° C, for 10 days, and then put together in a stirrer and stirred, Distillation extraction can be performed. For example, 50 parts by weight of the secondary natural ingredient extract and 50 parts by weight of water may be mixed with 100 parts by weight of the first natural ingredient extract, and then put into a distillation extractor for distillation and extraction.

When the distillation extraction is completed, a mixed additive composition obtained from the mixed extraction of the first natural component extract and the second natural component extract can be obtained.

Sixth, a secondary water-soluble oil may be prepared by distilling and extracting the primary water-soluble oil and the mixed additive composition. The primary water-soluble oil and the mixed additive composition may be mixed in a weight ratio of 1:1, and the distillation extraction may be performed once or two to three times.

The secondary water-soluble oil may be the “sturgeon water-soluble oil composition” in the present invention. That is, the sturgeon water-soluble oil composition may include a mixed additive composition obtained by distilling and extracting a mixture of sturgeon oil and natural additives.

The sturgeon water-soluble oil (composition) is an oil formulation that can be easily solubilized in water without artificial chemicals such as surfactants, and is widely used in the fields of cosmetics, drinking water or food, where harmlessness to the human body and environmental friendliness should be given priority. can be utilized

The water-soluble oil composition of sturgeon, which contains the liquid extract obtained from whole sturgeon, contains a high content of active ingredients in sturgeon and can secure human safety and improved quality due to non-antibiotics.

The sturgeon water-soluble oil composition has an effect of inhibiting intracellular collagen production and intracellular collagenase activity, thereby exhibiting excellent efficacy in improving skin wrinkles. This will be described in more detail through later experimental examples.

The sturgeon water-soluble oil composition is present in an amount of 10 to 100% by weight, specifically, 30 to 100% by weight, more specifically, 50 to 100% by weight, more specifically, 70% by weight, based on the total weight of the skin wrinkle improvement functional composition. to 100% by weight.

The wrinkle-improving functional composition according to an embodiment of the present invention may be applied and commercialized, for example, a cosmetic composition, a food composition, a beverage composition, a pharmaceutical composition, and the like. However, it is not limited thereto, and application to various types of health function compositions is possible.

For example, the composition may be included in a cosmetic composition and made into a skin product. Examples of application of skin products are generally basic cosmetics (toilet, serum, essence, lotion, cream, etc.) for skin care and protection, color cosmetics (make-up base, foundation, powder, eye shadow, lip gloss, lipstick, lip balm, etc.) , makeup for hands and toenails (manicure, nutrients, strengthening agent, top coat, etc.), cosmetics for cleansing (foam cleansing, cleansing oil, cleansing lotion, cleansing cream, cleansing gel, pack, mask, soap, cleansing tissue, etc.), sunscreen Care cosmetics and body cosmetics (body lotion, shower gel, body cream, body oil, etc.) may all be included, but are not limited thereto.

When applied to the skin product, known ingredients commonly used in accordance with the purpose, formulation, and effect of the product to be manufactured may be further included to the extent that the properties of the water-soluble sturgeon oil are not impaired.

As another example, when the composition is prepared as a food composition, it may be prepared in the form of various health supplements provided by conventional methods in the art. For example, the food composition may be liquid such as a suspension or beverage; or solids such as sticks, pills, jellies, chips, etc.; It can be produced in a variety of formulations.

As an example, the food composition may be prepared as a beverage composition, and the shape may be, for example, liquid stick type, liquid pouch type, etc., but is not limited thereto.

When the food composition is prepared as a beverage composition, in addition to the above-described components, for example, acacia honey, pear juice, bellflower juice, apple juice, or a combination thereof may further include natural sweetener components. As an example, based on 100 parts by weight of the sturgeon water-soluble oil, 15 to 20 parts by weight of a natural sweetener obtained by mixing pear juice and acacia honey in a weight ratio of 1: 1 may be further included. By further including such a natural sweetening component, it is possible to improve the sensuality by eliminating the reluctance to eat sturgeon.

According to the present invention, a skin wrinkle-improving functional composition containing a sturgeon component, specifically, a sturgeon extract, and more specifically, a sturgeon water-soluble oil as an active ingredient can be prepared.

More specifically, the sturgeon water-soluble oil is obtained from whole sturgeon, contains an active ingredient having an excellent effect on intracellular collagen production and intracellular collagenase activity inhibition, and is effective in improving skin wrinkles. It can exert excellent effect.

In addition, according to the present invention, when preparing sturgeon water-soluble oil, human safety due to non-antibiotics is ensured, thereby preparing a high-quality and highly functional sturgeon-containing skin wrinkle-improving functional composition compared to existing sturgeon-containing products. .

Hereinafter, the configuration of the present invention and its effects will be described in more detail through specific examples and comparative examples. However, these examples are for explaining the present invention in more detail, and the scope of the present invention is not limited to these examples.

[Example 1]

Manufacture of sturgeon raw materials

Farming with eco-friendly non-antibiotics for more than 3 years, feeding clean sturgeon (Siberian sturgeon) with bay salt at least once a month, fasting for 10 days according to the shipping time, and then hitting the head with a rubber mallet once or twice The samples were stunned without pain and immediately quenched and stored. After washing the rapidly frozen sturgeon, the entire sturgeon is put in a pneumatic hot water extractor with 50% sturgeon, 41% water, 5% calamansi undiluted solution, 2% bellflower seed undiluted solution, and 2% beet undiluted solution at 102℃ and 2.5 atm. After 100 hours of extraction, the mixture was separated at the upper, middle, and lower specific gravity points to obtain 80% sturgeon liquid extract, 15% sturgeon solid content, and 5% sturgeon oil, respectively.

Preparation of sturgeon water soluble oil

The obtained sturgeon oil was aged at 17° C. for more than 30 days and then passed through a fine nozzle using a high-pressure pump (high-pressure sprayer) repeatedly dozens of times to foam like fresh cream. This is an operation that causes a change in physical properties to bring it to a state before water-solubility, and after this is completed, it is stored at room temperature for natural aging for another 10 days.

Aged sturgeon oil and water, which have been subjected to the physical natural separation method described above, are mixed in equal proportions (50% aged sturgeon oil + 50% water), put into a distillation extractor, extracted, and returned to the remaining aged sturgeon oil. Only water was added at 120% and extracted by distillation more than 30 times to prepare a first water-soluble oil.

On the other hand, a primary additive was prepared by extracting 50% + 50% water of a mixture of edible snails, Baekbong Ogolgae, white neck mushrooms, beets, kelp roots, and bellflower seeds in a pneumatic hot water extractor for 100 hours. In addition, 50% of a mixture of puerh tea, chrysanthemum, lotus seed, sea bamboo shoot, centella asiatica, wormwood, wormwood, peppermint, yugenpi, hagocho, guccipon, waspong, chaga mushroom, samchae, calamansi and mulberry leaves and water 50% was put into a pneumatic hot water extractor to prepare a secondary additive extracted for 100 hours, and then stirred in a stirrer together with the primary additive.

Then, based on 100 parts by weight of the first additive, 50 parts by weight of the second additive and 50 parts by weight of the first water-soluble oil were placed in a distillation extractor and distilled and extracted to prepare final sturgeon water-soluble oil.

Referring to Figure 1, it can be seen that the sturgeon water-soluble oil produced by the above-described method has a transparent appearance.

<Experimental Example 1: Intracellular collagenase activity inhibition test>

In order to evaluate the anti-wrinkle activity of the water-soluble sturgeon oil prepared in Example 1, an intracellular collagenase inhibition assay was performed using CCD-986sk cells. The specific test method is as follows.

Preliminary test: cytotoxicity test

Test groups were prepared by diluting the sturgeon water-soluble oil of Example 1 to five concentrations (3.125%, 6.25%, 12.5%, 25%, and 50%).

Then, 200 μl of the cells were dispensed into a 96 well plate at a concentration of 1.5 × 10 ⁵ cell/mL, and cultured for 24 hours in a 37° C., 5% CO ₂ incubator. After 24 hours of cell division, all the medium was removed, and then the serum-free medium, the positive control group, and the test groups by concentration were treated and cultured in a 37°C, 5% CO ₂ incubator for 24 hours.

Then, the WST reagent was treated at a ratio of 10: 1 to the cells and the remaining medium except for 100 μl of the supernatant for confirming the NO production inhibitory activity, and incubated for 2 hours in a 37 ° C., 5% CO ₂ incubator for 2 hours to incubate the cells Toxicity was confirmed. After culturing for 2 hours, absorbance was measured at a wavelength of 450 nm.

When the cell viability was reduced to less than 90% of the sample-free group, it was determined to be potentially cytotoxic, and the cell viability was calculated according to Equation 1 below, and the cytotoxicity test results are shown in FIGS. 2 and 3.

<Equation 1>

Referring to Figures 2 and 3 together, 102.8 ± 1.3%, 108.4 ± 4.4%, 111.8 ± 108.4 ± 4.4%, respectively, for each concentration test group (3.125%, 6.25%, 12.5%, 25%, 50%) in the cytotoxicity test. Cell viability rates were 2.2%, 109.8 ± 0.7%, and 105.6 ± 4.9%.

That is, by showing a cell viability of 105.6 ± 4.9% at a concentration of 50% of sturgeon water-soluble oil, it was judged that cytotoxicity was not shown and used for the collagenase activity inhibition test.

Collagenase activity inhibition test

As shown in Table 1 below, experimental samples including samples having three final concentrations (12.5%, 25%, and 50%) among the test groups in which the preliminary test was performed were prepared.

Cells were dispensed into 48 well plate at a concentration of 5 × 10 ⁴ cell/well by 1 mL, and cultured in a 37°C 5% CO ₂ incubator for 24 hours. After 24 hours of cell dispensing, remove all the medium in the dish, and The medium was completely removed by washing with buffered saline (Phosphate-buffered saline, PBS).

Thereafter, 1 mL of TGF-1β diluted in a serum-free medium and the test groups for each concentration were treated and cultured for 24 hours in a 37°C 5% CO ₂ incubator.

After removing all the culture medium, 200 μL of cell lysis buffer was added to each well and reacted at 150 rpm for 10 minutes. Thereafter, the supernatant was obtained by centrifugation in a microtube. Then, the supernatant was analyzed according to a protein assay kit (BCA protein assay kit (Pierce, USA)) and protein was quantified according to a bovine serum albumin (BSA) standard curve.

Then, the cell lysate was collected and performed according to "Human MMP-1 ELISA kit (Sigma)", and the collagenase content was measured according to a collagenase standard curve.

Intracellular collagenase activity inhibition rate was converted into collagenase content for protein content, and was calculated according to Equation 2 below, and the final results are shown in FIGS. 4 and 5.

<Equation 2>

Referring to Figures 4 and 5 together, with respect to the final concentrations of sturgeon water-soluble oil of 12.5, 25, and 50%, respectively, 51.1 ± 3.5%, 41.5 ± 2.5%, and 22.2 ± 0.9% intracellular collagenase It was confirmed that the activity was inhibited.

That is, the sturgeon water-soluble oil according to one embodiment of the present invention did not show toxicity to CCD-986sk cells at a concentration of 50% or less, and in the intracellular collagenase activity inhibition test (collagen synthesis assay), sturgeon water-soluble oil It was confirmed that the oil exhibits excellent anti-wrinkle activity for inhibition of collagenase activity.

<Experimental Example 2: Intracellular Collagen Production Test>

In order to evaluate the anti-wrinkle activity of the water-soluble sturgeon oil prepared in Example 1, an intracellular collagen synthesis assay was performed using CCD-986sk cells. The specific test method is as follows.

Experimental samples including samples of three final concentrations (12.5%, 25%, and 50%) of the test group having performed the preliminary test (cytotoxicity test) previously conducted in Experimental Example 1 were prepared (Table 2).

Then, the cells were dispensed by 1 mL at a concentration of 5 × 10 ⁴ cell/well in a 48 well plate, and cultured for 24 hours in a 37°C, 5% CO ₂ incubator. After 24 hours of cell division, all the medium in the dish was removed, and then the medium was completely removed by washing with PBS. Thereafter, 1 mL each of L-ascorbic acid diluted with serum-free medium and the test substance for each concentration was treated and incubated for 24 hours in a 37°C, 5% CO ₂ incubator.

After removing all the culture medium, 200 μl of cell lysis buffer was added to each well, followed by reaction at 150 rpm for 10 minutes. Thereafter, the supernatant was obtained by centrifugation in a microtube. This supernatant was performed according to “BCA protein assay kit (Pierce, USA)” and then protein was quantified according to the BSA standard curve. The cell lysate was collected and performed according to “Procollagen type Ⅰ Peptide EIA kit (TaKaRa, Japan)”, and then the content was measured according to a collagen standard curve.

Then, the amount of collagen was converted to the amount of protein, and the intracellular collagen production rate was calculated according to the following equation 3, and the final results are shown in FIGS. 6 and 7.

<Equation 3>

Referring to FIGS. 6 and 7 together, as a result of the collagen production test, the collagen production rate of each test group (12.5, 25, and 50%) of the Siberian sturgeon extract (sturgeon water-soluble oil) was 86.7 ± 4.7%, respectively, 92.2 ± 2.7% and 146.2 ± 14.0% respectively.

In other words, the sturgeon water-soluble oil (Siberian sturgeon extract) of the present invention did not show toxicity to CCD-986sk cells at a concentration of 50% or less, and exhibited a high production rate of collagen, indicating a significant effect on skin wrinkle improvement. It was confirmed that it showed excellent effect.

In this specification, only a few examples of various embodiments performed by the present inventors are described, but the technical spirit of the present invention is not limited or limited thereto, and can be modified and implemented in various ways by those skilled in the art, of course.

Claims (2)

In the skin wrinkle improvement functional composition containing sturgeon water-soluble oil obtained from whole sturgeon as an active ingredient,
The sturgeon water-soluble oil is obtained from whole sturgeon, and the sturgeon water-soluble oil is included in an amount of 10 to 100% by weight based on the total weight of the composition, and the sturgeon water-soluble oil is used for intracellular collagen production and intracellular It is characterized by having a collagenase activity inhibitory effect,
The sturgeon water-soluble oil is prepared by preparing sturgeon oil according to a raw material obtaining method of whole sturgeon; Performing a first aging of the sturgeon oil to prepare an aged sturgeon oil; A pre-treatment step of foaming the aged sturgeon oil by using a high-pressure processor and a fine nozzle to form changes in physical properties of the oil; Second aging of the pretreated sturgeon oil aging product; Mixing the aged sturgeon oil and water, in which the second aging has been performed, and introducing the mixture into a distillation extractor and performing a second extraction; After performing the second extraction, water is mixed with the remaining matured sturgeon oil and introduced into a distillation extractor, and third extraction is performed to produce first water-soluble oil; Preparing a first additive composition containing edible snails and a second additive composition containing Puer tea and chrysanthemum, respectively; and preparing a secondary water-soluble oil by distilling and extracting a mixture of the primary water-soluble oil, the primary additive composition, and the secondary additive composition,
The method for obtaining the raw material of the sturgeon includes a farming step of culturing the sturgeon fry for 3 years or more using natural antibiotics; A pre-treatment step of shipping the farmed sturgeon and pre-processing the whole fish; A cold storage step of rapidly cooling the pretreated sturgeon, wherein the rapid cooling reaches a temperature of -80 to -20 ° C in 5 to 20 minutes; and thawing and washing the cooled sturgeon, then putting the whole sturgeon into a pneumatic extractor and performing a first extraction to obtain sturgeon oil; a functional composition for improving skin wrinkles, characterized in that it comprises a . According to claim 1,
The skin wrinkle-improving functional composition is a cosmetic composition, a food composition, a beverage composition, and a functional composition for improving skin wrinkles, characterized in that prepared from one or more health functional compositions selected from pharmaceutical compositions.

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