KR101339423B1 - cosmetic composition having collagen derived from fishes and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a skin-derived collagen-containing cosmetic composition and a method for manufacturing the same, and more particularly, extracting collagen from skin in a low temperature state to prevent heat denaturation of collagen and to maintain the structure of collagen, thereby providing moisturizing and skin protection effects. The present invention relates to an enhanced fish-derived collagen-containing cosmetic composition and a method for producing the same.
The fish-derived collagen-containing cosmetic composition of the present invention contains collagen isolated from fish, which is the skin of fish. Fish is one or more selected from perch, jeopbolak, flounder, red snapper. And the method of manufacturing a collagen-containing cosmetic composition derived from the fish of the present invention is a drying step of drying the skin while preventing the heat denaturation of collagen contained in the skin of the fish skin, pulverizing the dried skin to form a skin powder Steps; It comprises a separation step of separating the collagen from the skin powder.

Description

Cosmetic composition having collagen derived from fish skin and a method for preparing the same

The present invention relates to a skin-derived collagen-containing cosmetic composition and a method for manufacturing the same, and more particularly, extracting collagen from skin in a low temperature state to prevent heat denaturation of collagen and to maintain the structure of collagen, thereby providing moisturizing and skin protection effects. The present invention relates to an enhanced fish-derived collagen-containing cosmetic composition and a method for producing the same.

Collagen is the most abundant protein in the animal body, accounting for more than 30% of body protein, and has been found to be at least 19 types (Type I-III) (Nakamura, YN et al., Relationship among collagen amount, distribution). and architecture in the M. Longissimus thoracis and M. pectoralis profundus from pigs.Meat Science, 64, pp 43-50, 2003). In addition, collagen is a major protein of animal connective tissue, and supports tissues and organs and surrounds the body surface and plays a role in maintaining body shape. In particular, as a major component of connective tissue such as skin, cartilage and bone in the living body, 40% is widely distributed throughout the body, such as skin, 20% bone and cartilage, and other blood vessels and internal organs.

Unlike myofibrillar proteins, which have a double helix structure, collagen has a triple helix structure, about 14-15 nm in diameter, 280-300 nm in length, and an average molecular weight of about 300 KDa (Lehninger, AL Biochemistry, 2nd ed., Pp. 145, 1975), have a regular sequence of amino acids such as (Gly-XY) n and share within or between tropocollagen molecules, the basic molecules of fibrous proteins. Physically and biologically stable structures are formed by binding crosslinking (McClain, PE et al., Amino acid composition and cross-linking characteristics of collagen intramuscular connective tissue of striated muscle (Bos taurus). of Biochemistry, 2 (7), pp 121-124, 1971).

Collagen has been applied as a raw material of leather or gelatin for a long time, and its field of application has become more diverse in recent years. In the food industry, it is used for meat packaging materials such as sausage and salami as an edible casing raw material. Collagen applied to medicines has been reported to have a healing effect on skin damaged by burns or wounds (Jeyanthi, R. et al., Solid tumour chemotherapy using implantable collagen-poly (HEMA) hydrogel containing 5-fluorouracil. of Pharmacy & Pharmacology, 43, pp. 60-62, 1991). As such, collagen is not only used as a functional material for foods and medicines, but also has a function of enhancing skin moisturizing, and thus is widely used in various fields such as basic materials of cosmetics.

In recent papers, peptides obtained from protein hydrolysates have been used as potential materials to improve wrinkles, improve moisturization, increase elasticity, and exhibit specific skin effects.

Collagen hydrolyzate, called collagen peptide, refers to a low-molecular-weight molecule in the form of peptide by hydrolyzing it through post-treatment process such as enzyme digestion after extraction of polymer collagen from pork skin and fish scales. Collagen products that have been lowered are sold.

Meanwhile, fishery by-products have been classified as food waste so far, and most of them have been used as compost. Dry skin has more than 90% of protein and useful substances, so it can be used in various ways such as collagen extraction, cosmetics, dietary supplements, and adhesives. Recently, various products such as health supplements and cosmetics using collagen are commercialized, but many depend on imports.

Increasing utilization of skin by-products in the context of Korea's per capita aquatic consumption with Japan, the world's number one, is highly suitable for economic synergies and eco-friendly industrial development through the supply of high quality fish protein and collagen.

In addition, it contains a large amount of various polymer proteins such as collagen, albumin, myosin, elastin, and can be utilized as high-quality collagen, and it is easy to collect in domestic where the consumption of sashimi is high. As marine fish farming is consistently in progress, stable yields can be expected. Therefore, it is urgent to evaluate the availability of marine fish by-products.

The present invention was created to improve the above problems, by drying the skin generated during fish processing at a low temperature and then powdered to extract collagen to prevent heat denaturation of collagen and to maintain the collagen structure of the polymer intact gel formation ability The purpose is to provide a cosmetic composition and a method for producing the same having an increased moisturizing effect.

Skin-derived collagen-containing cosmetic composition of the present invention for achieving the above object is characterized in that it contains collagen isolated from the fish skin.

The fish is characterized in that any one or more selected from perch, jeopbolak, flounder, red snapper.

And a method for producing a skin-derived collagen-containing cosmetic composition of the present invention for achieving the above object is a drying step of drying the skin while preventing the heat denaturation of collagen contained in the skin of the fish skin; Grinding the dried fish to form fish powder; It characterized in that it comprises a; separating step of separating the collagen from the skin powder.

The fish is characterized in that any one or more selected from perch, jeopbolak, flounder, red snapper.

The separating step includes adding an alkaline solution to the skin powder to obtain an alkali residue from which the non-collagenic protein has been removed, and adding collagen to the alkali residue to extract collagen.

The present invention can be dried at a low temperature to remove the water from the fish to remove the water and then processed into a powder form to extract the collagen to prevent the deterioration of the skin and at the same time to prevent the heat denaturation of collagen and increase the extraction yield.

In addition, the present invention can provide a cosmetic composition having a gel-forming ability and the moisturizing effect is increased by containing the collagen that is maintained intact the collagen structure of the polymer is not a low molecular weight collagen peptide.

1 is a 100 times optical micrograph of the hot air dried flounder skin powder,
Figure 2 is a 100 times optical micrograph of lyophilized flatfish skin powder,
3 is a photograph showing the dorsal and ventral skin of the perch,
4 is a photograph showing the dorsal skin of the flounder,
5 is a photograph showing the belly skin of the flounder,
Figure 6 is a photograph showing the dorsal and ventral skin of the skin rock rock,
7 is a photograph showing the dorsal and ventral skin of red snapper,
8 is a photograph of SDS-PAGE of perch skin,
9 is a photograph of SDS-PAGE of the flounder skin,
10 is a photograph of SDS-PAGE of skin gel rock
11 is a photograph of SDS-PAGE of red snapper skin.

Hereinafter, the skin-derived collagen-containing cosmetic composition and its preparation method according to a preferred embodiment of the present invention will be described in detail.

The cosmetic composition of the present invention contains collagen isolated from fish skin which is the shell of fish. The collagen may be contained in 0.01 to 20% by weight based on the total weight of the cosmetic composition. If the content of collagen is less than 0.01% by weight, the moisturizing and skin protection effects are insignificant, and if it exceeds 20% by weight, it is difficult to have an elevated effect compared to the content.

Cosmetic composition of the present invention by adding a variety of well-known ingredients and the like using methods known in the art, lotion, lotion, cream, essence, pack, makeup base, foundation, body oil, body lotion, cleansing oil, cleansing foam , Hair oil, hair shampoo and hair rinse, it can be prepared in any one of the formulation selected from soap.

Specifically, when the formulation of the cosmetic composition of the present invention is a paste, cream or gel, pack, as known components, animal fibers, plant fibers, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, Silica, talc or zinc oxide and the like can be used.

When the formulation of the cosmetic composition of the present invention is a powder, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used.

When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylglycol oil, glycerol aliphatic ester, polyethyleneglycol or sorbitan non-dispersion ester and the like can be used.

Water, liquid diluents such as ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester when the formulation of the cosmetic composition of the present invention is a suspension, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tragacanth and the like can be used.

Aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide ether sulfate when the formulation of the cosmetic composition of the present invention is a surfactant-containing detergent , Alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils or ethoxylated glycerol fatty acid esters and the like can be used.

In addition to the cosmetic composition of the present invention can be used as a conventional known ingredients such as stabilizers, solubilizers, vitamins, pigments and fragrances, can be prepared in any formulation commonly prepared in the art, Can be appropriately selected accordingly.

In the present specification, 'fish' means throughout the cartilage fish river, tibia fish river, mudfish river, cephalus river, premature river of vertebrate animals. 'Shell' refers to the outer covering of the fish's body, and the same meaning as the skin. The shell thus contains the epithelium, the corium tissue of fish, and the scales of the epidermis. And 'protein source' means a source capable of supplying a protein, preferably a source having a crude protein content of about 20% or more.

Skin contains some lipids, ash, and pigments, but is composed mainly of collagen. Such collagen is a type of structural protein that denatures when heat is applied above a certain temperature. Therefore, in order to maximize the utilization of collagen, the main component of the fish, it is preferable to process the collagen below the denaturation temperature of the collagen.

Skin applied to the present invention can be applied to the shell of all fish, preferably at least any one selected from the group containing perch, jeopbolak, flounder, red snapper. The four kinds of fish account for about 90% or more (as of 2009) of domestically farmed fish, so it is relatively easy to supply. More preferably the fish is the shell of the flounder. The flounder's skin is more than twice as big as the other three species (perch, zombolak and red snapper), so it is easy to secure raw materials. Flounder is used as a sashimi and bones are used as spicy soup. It is not utilized. In addition, the ash content is low even when the scales are included in comparison with the shells of the three species, and there is no need to remove the scales, and the lipid content is high along with the protein content.

Hereinafter, a method for producing the cosmetic composition of the present invention will be described.

Method for producing a cosmetic composition according to an embodiment of the present invention is a drying step of drying the skin while preventing the heat denaturation of collagen contained in the skin of the fish skin, and to form the skin powder by grinding the dried skin And a grinding step, a separation step of separating collagen from the skin powder, and formulating the separated collagen into a cosmetic composition.

Specifically, first prepare the fish. Fish can be used both wild and farmed. Wash the shell two or three times with clean water to remove any foreign matter or blood from the skin.

After washing the skin, water is removed to increase the extraction yield of collagen, and to prevent deterioration and to dry. In the drying step, the skin is dried to less than 7% by weight, preferably 2 to 5% by weight. Freeze drying and hot air drying may be applied as a drying method. Preferably, freeze drying (FD) is used to prevent denaturation of collagen.

The freeze-drying method rapidly freezes the skin for 10 to 20 hours at a temperature of -50 to -40 ° C, and then for 48 hours at about -40 ° C in a lyophilizer having a vacuum degree of 0.1 to 0.5torr. In addition, it is a matter of course that the conventional freeze-drying method applied in the manufacture of food can be applied.

When lyophilizing the skin, moisture is removed by sublimation in the frozen state, and thus the dried product has a light porous structure, maintains its original shape and size, and is treated at low temperature without applying heat. Transfer of soluble components, non-enzymatic browning and protein denaturation rarely occur in drying.

In the case of hot air drying, the skin may be dried by hot air at 30 to 80 ° C. In order to prevent denaturation of collagen at the time of hot air drying, the temperature of hot air is 65 degrees C or less, Preferably it is 40-60 degreeC. Collagen contained in the skin has a higher denaturation temperature than the separated state.

The dried fish is ground to a suitable size using a grinder to obtain the fish powder. For example, it can be ground to a size of 50 to 150 mesh particle size. Micrographs of the skin powder are shown in Figs. 1 and 12. 1 is a hot air drying method, Figure 2 is a skin powder of the flounder treated by the freeze-drying method.

After the skin powder is obtained, collagen is separated from the skin powder. To this end, an alkaline solution is added to the skin powder. As the alkaline solution, sodium hydroxide or calcium hydroxide solution can be used. The skin powder and the alkaline solution are mixed at a weight ratio of 1: 5 to 10, and then stirred at room temperature (20 to 25 ° C.) for 12 to 24 hours, and an alkali residue from which the non-collagenic protein is removed is separated using a centrifuge. . The obtained alkaline residue was washed with distilled water, and then acid solution was added to extract collagen. As the acid solution, inorganic acids such as hydrochloric acid and sulfuric acid or organic acids such as acetic acid can be used. The alkali residue and the acid solution are mixed at a weight ratio of 1: 5 to 10, and then stirred at room temperature (20 to 25 ° C) for 12 to 24 hours, and the supernatant is separated using a centrifuge to obtain acid soluble collagen.

Various known ingredients are added to the obtained collagen, such as lotion, lotion, cream, essence, pack, makeup base, foundation, body oil, body lotion, cleansing oil, cleansing foam, hair oil, hair shampoo and hair rinse, soap It is prepared in any one of the formulations selected from.

As an example of the formulation, the lotion may contain 0.1 wt% of carbomer, 0.1 wt% of hydroxyethyl cellulose, 2.0 wt% of butylene glycol, 1.0 wt% of glycerin, 0.5 wt% of Fiji-1500, 4.0 wt% of ethanol, poly 0.2% by weight of sorbate 80, 0.1% by weight of triethanolamine, preservative (trace), purified water (remaining amount), collagen 5% by weight.

In addition, as an example of the formulation, the nutrition cream is 1.5% by weight of cetearyl alcohol, 1.0% by weight of glyceryl stearate, 1.0% by weight of polysorbate 60, 0.3% by weight of sorbitan sesquioleate, 6.0% by weight of cetyloctanoate. , 8.0% by weight squalene, 4.0% by weight of apricot kernel oil, 2.0% by weight of dimethicone, butylene glycol 4.0% by weight, glycerin 4.0% by weight, magnesium aluminum silicate 0.4% by weight, xanthan gum 0.03% by weight, preservative ), Purified water (residual amount), collagen may be composed of 5% by weight.

In addition, as an example of the formulation, the lotion may be 5.0% by weight of butylene glycol, 2.0% by weight of oleyl alcohol, 1.0% by weight of polysorbate 20, 6.0% by weight of ethanol, preservative (trace), purified water (remaining amount), 5% by weight of collagen. It can be formulated as.

In addition, as an example of the formulation, the pack is 5.0 wt% butylene glycol, 12.0 wt% polyvinyl alcohol, 2.0 wt% polyvinylpyrrolidone, ethanol 8.0 wt%, polyvinylacetate 1.0 wt%, preservative (trace), purified water (The remaining amount), it may be composed of 5% by weight collagen.

In addition, one example of the formulation is a solid soap. Soaps include a soap base that saponifies fatty acids and collagen. Preferably the soap base is obtained by saponifying fatty acids obtained by hydrolysis of vegetable oils and fats. As a vegetable oil, various kinds of oils such as coconut oil, palm oil, soybean oil, castor oil, olive oil, grape seed oil, apricot seed oil, sunflower seed oil, jojoba oil, etc. may be used alone or in combination.

The soap may further include glycerin and essential oils in addition to the soap base and collagen. In this case, the composition of the soap is 89 to 94% by weight of soap base, 0.5 to 5% by weight of collagen, 0.1 to 5% by weight of glycerin, 1 to 1.5% by weight of essential oil.

Glycerin increases the moisturizing power by forming a protective film on the skin. And as an essential oil, oil extracted from herbs can be used.

 Essential oils can be extracted from the herb according to a conventional essential oil extraction method, the essential oil extraction method can be easily implemented by those skilled in the art. Herbs include juniper berry, chamomile, mindarin, lemon, lavender, rosemary, geranium, peppermint, spearmint, applemint, lime blossom and sage. The essential oils improve the antibacterial and skin regeneration of the soap.

On the other hand, in another embodiment of the cosmetic composition of the present invention, in addition to the collagen isolated from the skin may further contain a bee herb extract.

Gymnaster koraiensis ) is a deciduous perennial grass belonging to the genus Anthoe genus. Hummingbird odor is artificially capable of mass breeding and has excellent economic feasibility, and its roots are strong and its growth is strong. Beetle odorless young sprouts were eaten as herbs or in soups, and the roots were used as herbal medicines because they were used as a herbal medicine. -323) Harmless and safe material

In the present invention, the bee stinger can use the entire plant, such as leaves, roots, stems. Beetle odor extract can be extracted in various ways. For example, after adding the extraction solvent 2 to 20 times by weight with respect to the bee sting odor can be used for hot water extraction, cold needle or warm needle extraction for 10 to 150 ℃ for 1 to 24 hours. Further, a reflux cooling extraction method, an ultrasonic extraction method, or the like can be used. In addition to the above-described extraction method, an extract obtained through a conventional purification process is also included. For example, an active fraction obtained through various purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies, etc., is also included in the extract.

As the extraction solvent, at least one selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof may be used. As the lower alcohol having 1 to 4 carbon atoms, methanol, ethanol and the like can be used. As the polyhydric alcohol, butylene glycol, propylene glycol, pentylene glycol and the like can be used. Mixtures of water and lower alcohols, mixtures of water and polyhydric alcohols, mixtures of lower alcohols and polyhydric alcohols, or mixtures of water and lower alcohols and polyhydric alcohols can be used as the mixture. It goes without saying that the extract obtained by using the above-mentioned extraction solvent can be obtained in the form of powder through filtration, concentration under reduced pressure or freeze-drying, spray drying or the like.

Beetle odor extract is expected to improve the skin's moisturizing effect with collagen. Beetle odor extract may be contained in an amount of 0.01 to 5% by weight based on the total weight of the cosmetic composition.

Hereinafter, the content of the present invention will be described in detail through the following examples and experimental examples. This is for explaining the present invention in more detail, and the scope of the present invention is not limited to the following examples and experimental examples.

<Characteristics of Fish>

Fishes isolated from sea bass, flounder, skinball rock and red snapper were used as the first, second, third and fourth test samples.

1. General composition analysis

The general components were analyzed for four types of fish and are shown in Table 1 below. General component analysis was performed according to the AOAC (2000) method. Moisture was applied at 105 ℃, ash at 550 ℃, direct fat, soxhlet extraction and protein micro-Kjeldahl.

division
moisture(%)
  Dry weight (%) Crude protein Crude fat Ash 1st sample 81.10 ± 0.43 72.81 ± 0.43 25.47 ± 2.59 1.80 ± 0.17 2nd test sample 80.87 ± 1.07 88.91 ± 3.64 17.07 ± 0.45 1.97 ± 0.15 Third test sample 79.37 ± 0.56 80.59 ± 1.68 5.10 ± 0.71 10.90 ± 1.87 Fourth test sample 76.11 ± 0.31 94.11 ± 0.31 4.10 ± 1.36 0.70 ± 0.00

Looking at the Table 1, it was found that the halibut skin of the four species of fish has a high lipid content and a high protein content. This suggests that the flounder may be more advantageous in the amount of collagen extracted than in the other fish.

2.Amino Acid Analysis

In order to analyze the constituent amino acid, 0.5g of the sample was weighed into an 18ml test tube, 3N of 6N HCl was added, and the test tube was sealed using a vacuum pump.

The sealed test tube is hydrolyzed for 24 hours in a heating block set at 121 ℃. After hydrolysis, the sample was removed with a rotary evaporator at 50 ° C and 40 psi. After 10 ml of sodium loading buffer, 1 ml of this sample was filtered through a membrane filter and quantitatively analyzed by an amino acid analyzer. Indicated.

Amino acid
(mg / 100g) 1st sample 2nd test sample Third test sample Fourth test sample Asp 3334 3187 3362 3088 Thr 1579 1391 1358 1424 Ser 2072 2204 2509 2268 Glu 5745 5216 5221 4249 Pro 6940 5737 5271 5744 Gly 11384 10845 10383 11080 Ala 5159 4628 4206 4835 Cys 112 153 195 193 Val 1373 1325 1448 1205 Met 1110 1144 990 1128 Ile 834 763 807 594 Leu 1820 1643 1677 1415 Tyr 479 446 461 441 Phe 1475 1336 1352 1300 His 1086 1010 1099 9248 Lys 2052 1943 1946 1968 Arg 4589 4199 4051 4202 Total 53332 48779 78217 56413 ^* EAA 15918 14754 14727 22484

EEA (essential amino acid): Thr, Val, Met, Ile, Leu, Phe, His, Lys, Arg

Looking at the results of Table 2, the constituent amino acids of collagen, proline, glycine, etc. were found to be high, indicating that the main component of the fish is collagen.

4. Histological observation

Samples from which fish skin was cut for optical microscopy were fixed in aqueous Bouin's solution for 12 hours according to the method of Drury and Wallington (1980), and then washed for 24 hours. Then, after stepwise dehydration using ethanol, it was embedded in Paraplast (McCormick, USA). Embedded samples were serially sectioned to 4-6μm thickness using a microtome (RM2235, Leica, Germany) to make a tissue specimen. The fabricated tissue samples were observed with an optical microscope after Mayer's hematoxylin-eosin (HE) staining, alcian blue-periodic acid and Schiff's solution (AB-PAS, pH 2.5) reaction, Masson's trichrome staining, and the results are shown in FIGS. 5 is shown.

In each photograph, Cc: club cell, Cf: collagen fiber, Dl: dermal layer, El: epidermal layer, Mc: mucous cell, Mf: muscle fiber (muscle fiber).

1 shows dorsal and ventral skin of perch, A is a photograph after AB-PAS reaction of dorsal skin tissue, B is a photograph after Masson's trichrome staining of dorsal skin tissue, and C is an AB-PAS reaction of ventral skin tissue. D is a photograph after D Masson's trichrome staining of the ventral skin tissue. Figure 2 shows the dorsal skin of the flounder, A is a picture after H-E staining, B is a picture after the AB-PAS reaction, C is a picture after Masson's trichrome staining. Figure 3 shows the abdominal skin of the flounder, A is a picture after H-E staining, B is a picture after the AB-PAS reaction, C is a picture after Masson's trichrome staining. Figure 4 shows the dorsal and ventral skin of the skin coat, A and B are photographs after Masson's trichrome staining of the dorsal skin tissue, C and D is a photograph after Masson's trichrome staining of the dorsal skin tissue. 5 shows dorsal and ventral skin of red snapper, A and B are photographs after Masson's trichrome staining of dorsal skin tissue, C is a photograph after H-E staining, and D is a photograph after Masson's trichrome staining.

Referring to Figures 3 to 7, the dome, the dorsal and ventral skin system of the sea bass consisted of the epidermal layer, the dermis layer, the subcutaneous tissue layer and the muscle layer, the dermis layer of the perch consisted of well-developed collagen fiber. The dorsal and ventral skin systems of the flounder consisted of the epidermal and dermal layers of general structure, and the dermal layer was composed of collagen fibers. The dorsal and ventral dermis of the dermis was also composed of the epidermal and dermal layers. The dorsal dermis was composed of collagen fibers and some muscle fibers, and the abdomen was composed of well-developed collagen fibers. The collagen fibers, collagen fibers, were mainly developed in the dorsal or ventral dermis layer of all four species, and the thickness of the dermis layer was found to be the thickest in the epithelium. Because of its large volume, it was confirmed that it is the best material source to utilize collagen fiber protein.

For reference, the skin thickness of the four species of fish is shown in Table 3 below.

division part Epidermal to dermal layer (㎛) Epithelial layer (㎛)
bass Back 485.44 72.380 Abdomen 490.278 75.823
halibut Back 540.237 49.231 Abdomen 736.923 43.077
Zoppybolak Back 367.098 20.127 Abdomen 528.481 23.418
Red snapper Back 287.447 18.515 Abdomen 407.595 67.599

&Lt; Example 1 >

After washing the fish separated from the perch and then rapidly frozen at -45 ℃ for 15 hours, and then dried at -40 ℃ for 48 hours in a freeze dryer having a vacuum degree of 0.5torr and then ground to prepare a fish powder.

The skin powder and 0.1 M sodium hydroxide solution were mixed at a weight ratio of 1:10, stirred at 20 ° C. for 18 minutes, and an alkali residue from which the non-collagenic protein was removed was separated using a centrifuge. The alkaline residue was washed with distilled water, and then the alkali residue and 6M hydrochloric acid solution were mixed at a weight ratio of 1:10, stirred at 20 ° C. for 18 minutes, and the supernatant was separated using a centrifuge to obtain acid-soluble collagen (ASC). Obtained.

Then, the first sample was prepared by mixing 5% by weight of acid-soluble collagen, 94.5% by weight of purified water, and 0.5% by weight of ethanol.

<Examples 2 to 4>

Acid-soluble collagen (ASC) was prepared in the same manner as in Example 1, but fish powders separated from flounder, seaweed rockfish and red snapper were used instead of sea bass.

The second to fourth samples were prepared by mixing 5% by weight of acid-soluble collagen, 94.5% by weight of purified water, and 0.5% by weight of ethanol.

<Example 5>

A fifth sample was prepared in the same manner as in Example 1, but 5% by weight of acid-soluble collagen, 2% by weight of bee odor extract, 92.5% by weight of purified water, and 0.5% by weight of ethanol were mixed.

Beetle odor extract was obtained by drying the outpost containing the leaves, roots, stems and then chopped, and then added 10 times the amount of methanol by weight, repeated extraction four times at 40 ℃ for 6 hours, filtered and concentrated under reduced pressure.

<SDS-PAGE Experiment>

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed using each fish species and acid soluble collagen (ASC) extracted from each fish sample. Protein markers were purchased from Sigma, USA and were sample buffers (60 mM Tris-HCl, 25% glycerol, 2% SDS, 14.4 mM 2-mercaptoethanol, 0.1% bromophenol blue, 10 mL distilled water). )). After adding the same amount of the sample buffer to the protein extracted from each sample, it was heated for 5 minutes in water at 100 ℃ to induce complete denaturation of the protein, followed by electrophoresis (Mini-Protean, BIO-RAD Laboratories, Inc. USA.) . The electrophoretic buffer solution was a 0.025 M-Tris base-0.192 M Glycine (pH8.3) solution containing 0.1% (w / v) SDS. The gels obtained were stained with Coomasie Blue R-250 solution (Coomasie). 1.0 g Coomassie blue R-250, 450 mL methanol, 450 mL H 2 O, 100 mL glacial acetic acid) was shaken at room temperature for 2 hours. Decolorization was performed by shaking the gel with light decolorization reagent (100mL methanol, 100mL glacial acetic acid, 800mL H2O) so that the gel was slightly submerged, and changing the reagent twice over 30 minutes. . The bands stained on the gels obtained at this time are shown in Figs. 8 to 11, respectively, compared to protein markers. 8 to 11, ASC means acid soluble collagen and RS-AL means alkali residues.

8 to 11 all of the acid soluble collagen isolated from the four fish showed the same SDS-PAGE pattern as the skin and alkali residues. The low molecular weight, thermally denatured fragments were not identified, indicating that the extracted acid soluble collagen retained the structure of the collagen without any thermal denaturation.

Moisturizing Experiment

In order to investigate the moisturizing effect on the first to fifth samples prepared in Examples 1 to 5, the moisture content of skin, especially the epidermis, was measured by a biotechnological method. Five women aged 22 to 40 years were examined for changes in the moisture content of the skin before and after sample application. The moisture content of the skin was measured using a skin moisture content meter (MPA5, Corneometer CM 825 probe, Courage & Khazaka electronic, Germany), a sensor device that measures the moisture content of the granular layer on the surface of the stratum corneum by measuring the electrostatic load capacity of the skin.

Subjects who participated in the measurement test acclimated to the temperature and humidity conditions of the evaluation laboratory at least 30 minutes before each measurement was performed. Samples were distributed prior to the evaluation test and trained to apply twice a day to designated skin areas, with the amount applied once, each sample being uniformly applied at a level of 0.5 ml / cm 2. Measure the condition of the skin where nothing is applied as a reference point, and apply twice a day thereafter, and continuously measure the corneometer value after 5 days, 10 days, 15 days, and 20 days. Shown in

division Before application 5 days after application 10 days after application 15 days after application 20 days after application 1st sample 35.23 36.01 39.33 41.74 43.85 2nd sample 34.90 36.18 40.36 42.50 44.51 Third sample 36.83 38.35 39.41 41.83 43.39 Fourth sample 35.49 37.27 38.76 39.23 42.74 Fifth sample 35.47 38.20 40.79 43.54 45.65

Referring to Table 4, by looking at the change in the average value of the skin moisture content of the application site of each sample was able to confirm the excellent moisturizing effect of each sample. In particular, it was found that the moisturizing effect of the second sample was the best among the first to fourth samples containing only collagen. In the second sample, the change in the mean value of skin moisture content was increased about 27.5% after 20 days of application. In addition, the fifth sample containing collagen and bee stinger extract extracted from sea bass was about 4.2% higher than the first sample.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation.

Accordingly, the true scope of protection of the present invention should be determined only by the appended claims.

Claims (5)

delete delete A drying step of drying the fish while preventing heat denaturation of collagen contained in the fish skin;
Grinding the dried fish to form fish powder;
A separation step of separating collagen from the skin powder;
The total weight of the composition is added to contain 0.01 to 10% by weight of the collagen, 0.01 to 5% by weight of honey bee odor extract, lotion, lotion, cream, essence, pack, makeup base, foundation, body oil, body lotion, cleansing oil, It comprises; a formulation step of preparing a formulation selected from any one of cleansing foam, hair oil, hair shampoo, hair rinse, soap,
The separation step is a step of mixing the skin powder and 0.1M sodium hydroxide solution in a weight ratio of 1:10, stirred for 18 hours at 20 ℃ and centrifuged to obtain an alkali residue from which the non-collagenic protein is removed; , Mixing the alkaline residue and 6M hydrochloric acid solution in a weight ratio of 1:10, stirred for 18 hours at 20 ℃ and centrifuged to extract the collagen-derived collagen-containing cosmetic composition comprising the step of extracting Manufacturing method. The method of claim 3, wherein the fish is any one or more selected from sea bass, rockfish, flounder, red sea bream.


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