JP2019014660A - Method for producing proteoglycan - Google Patents

Abstract

To provide a method for producing proteoglycan extracted from cartilage tissue, muscle tissue or skin tissue of fish, mollusks, birds or mammals, with improved efficiency and high purity.SOLUTION: A method for producing proteoglycan includes the step of adsorbing and removing lipid components from proteoglycan extract using a powder cellulose and/or an oil absorption mat, and further the step of separating proteoglycan.SELECTED DRAWING: None

Description

  The present invention relates to a production method for extracting proteoglycan from a biological sample more efficiently and with high purity.

  Proteoglycan is a glycoprotein in which several to dozens of glycosaminoglycans such as chondroitin sulfate and ketalan sulfate are covalently bonded to one core protein, and is widely used in the body such as skin and cartilage as an extracellular matrix. Distributed.

  Glycosaminoglycan has a long straight chain structure without branching, and is negatively charged because it has a large number of sulfate groups and carboxyl groups, and the glycosaminoglycan chain is extended due to its electric repulsive force. Take shape. In addition, due to the water affinity of sugar, it retains a large amount of water and plays a role specific to cartilage such as resilience and resistance to impact. So far, methods for efficiently producing high-purity proteoglycans have been studied.

  For example, a method for extracting proteoglycan from salmon nasal cartilage previously filed by the present applicants using guanidine hydrochloride (Patent Document 1) and a method using acetic acid (Patent Document 2) are known. In addition, a method using an alkali (Patent Document 3), a method using citric acid (Patent Document 4), a method using water (Patent Document 5), a method using an aqueous solution of a surfactant (Patent Document 6), etc. are reported. Has been.

  As described above, various methods for extracting proteoglycans have been studied. However, lipid removal work is not performed, or even if a degreasing operation is performed, the lipid is removed manually and the raw material is washed, or after washing, an organic solvent such as ethanol, acetone, hexane, etc. Or using a hydrophobic carrier or hydrophobic plastic beads. In addition, a degreasing method using activated carbon in the production process of chondroitin sulfate from salmon nasal cartilage has been reported (Patent Document 7). Since proteoglycans are produced using biological samples, residual lipids not only lower the purity as contaminants, but also greatly affect the quality of the obtained proteoglycans such as color and odor. The conventional methods are effective for removing lipids to some extent, but the degreasing effect and work over a long period of time, the high cost due to the use of organic solvents such as alcohol and acetone, beads, danger of ignition, It was not satisfactory from the viewpoints of removal efficiency, time, economy, and safety, such as worker safety. In addition, the use of activated carbon can be expected to have a degreasing effect, but an increase in costs such as a decrease in the yield of the proteoglycan, which is the target substance, and an increase in work time due to the addition of a high-precision filtration process to remove the particulate activated carbon can be avoided. Absent.

JP 2001-172296 A JP 2002-69097 A International Publication No. 2007/094248 JP 2012-236776 A International Publication No. 2012/099216 JP 2014-9164 A JP 2007-252212 A

  Accordingly, an object of the present invention is to provide an efficient method for producing a high-purity proteoglycan that is cheaper from fish, mollusks, birds or mammals.

  The present inventors include a step of adsorbing and removing the lipid component from the extract obtained in the proteoglycan extraction step using powdered cellulose and / or an oil-absorbing mat, so that the proteoglycan, which is a glycoprotein complex, can be efficiently obtained. The present inventors have found that it can be produced at a lower cost from a biological sample containing cartilage tissue and other proteoglycans with high purity.

That is, the present invention includes the following items.
[Item 1] A proteoglycan comprising a step of extracting proteoglycan from a biological sample containing proteoglycan and adsorbing and removing the lipid component from the resulting extract using powdered cellulose and / or an oil-absorbing mat. Manufacturing method.
[Item 2] The production method according to Item 1, comprising a step of separating and purifying proteoglycan after the step of removing the lipid component.
[Item 3] The production method according to any one of Items 1-2, wherein the biological sample containing proteoglycan is fish, mollusk, avian, or mammalian cartilage tissue, muscle tissue, or skin tissue.

  The production method according to the present invention can extract and purify proteoglycan with high efficiency and high purity by a very simple method as compared with the conventional production method. In addition, since a large amount of organic solvent such as alcohol or acetone is not required, there is no danger of flammability and it is suitable for the health of workers, and the proteoglycan that is the target substance is removed together with lipids such as activated carbon. This eliminates the need to add a high-precision filtration process that prevents the collection amount from being reduced and the mixing of fine-particle activated carbon.

3 is an HPLC profile of proteoglycan obtained in Production Example 1. 3 is an HPLC profile of proteoglycan obtained in Comparative Production Example 2. It represents the proteoglycan obtained by the present invention, the proliferative action of cartilage progenitor cells. It represents the cartilage differentiation action of the proteoglycan obtained in the present invention. The calcification inhibitory effect of the proteoglycan obtained by this invention is represented.

  The biological sample containing proteoglycan in the present invention can be applied to, for example, fish, mollusk, avian or mammalian cartilage tissue, muscle tissue, or skin tissue, among which cartilage tissue is preferable. The cartilaginous tissue used in the present invention can be any fish, mollusk, bird or mammal, especially any of their disposal sites. In the present invention, the cartilage tissue means either cartilage alone or a tissue including a cartilage peripheral part, for example, bone, muscle tissue, skin tissue and the like.

  In the present invention, those derived from the nasal cartilage tissue contained in the salmon head are preferably used from the viewpoint of availability and cost, and in particular, salmon ( When salmon (mainly salmon) is processed as various processed products, the head to be discharged can be used.

  Further, in the present invention, in addition to salmon, fish-derived cartilage tissues such as trout, rays, sharks and cod, mollusk-derived epidermis such as squid and octopus, bird-derived cartilage tissues such as chickens, and mammals such as cows and whales Cartilage tissue can also be used.

  A known method can be used for the extraction solvent used in the present invention. For example, acetic acid, water, citric acid, an alkaline solution, a surfactant solution, and the like are appropriately selected. Since the solution after the extraction contains a lot of residues after extracting the proteoglycan, it is preferable to remove these by filtration, centrifugation or other methods.

  Next, by using powdered cellulose and / or an oil-absorbing mat for the proteoglycan extract, it is possible to simply adsorb and remove the lipid components to be mixed. For example, cellulose cellulose (Nippon Paper Co., Ltd .; KC Flock W-50 (S), KC Flock W-50, KC Flock W-100, KC Flock W-100G, KC Flock W-200, KC Flock W-200G, KC Flock W-250, KC Flock W-300G, KC Flock W-400G, NP Fiber W-100F, NP Fiber W-300F, NP Fiber W-10MG2, NP Fiber W-06MG, KC Flock W- 50GK, KC Flock W-100GK, manufactured by Rettenmeier; VITACEL-L10, VITACEL-L20, VITACEL-L600-30, VITACEL-L90, ARBOCEL-BMW40, and VITACEL-L500). As the oil absorbing mat, a fiber nonwoven fabric mainly made of polypropylene is preferable. For example, Maeda Kosen Co., Ltd .; oil adsorption sheet SP-1300N (DX), oil adsorption sheet SP-1100N, Nippon Paper Crecia Co., Ltd .; oil adsorption mat PP-100 series, Tanaka Sangyo Co., Ltd. ); Looklin A-50, Looklin B-50, other non-woven fabrics made from polyolefin and polyester, manufactured by Nippon Paper Crecia Co., Ltd .; Powerful ECO series, oil absorbing mats mainly made from cotton, Daio Paper Co., Ltd .; Prowipe, cotton oil mat and the like.

  The oil absorbing mat from which the lipid has been adsorbed and removed is removed, and the powdered cellulose is separated by filtration. Although any one of them has a degreasing effect, it is more preferable to use them in combination. Proteoglycan obtained from this filtrate can be recovered by solid-liquid separation with a separation membrane having an appropriate fractional molecular weight. In this operation, if a separation membrane having a molecular weight of 50,000 or more is used, low molecular weight impurities (collagen and the like) can be removed from the liquid phase, and the purity of the proteoglycan can be increased. At that time, it is possible to remove a slight odor derived from a biological sample by sufficiently washing with water. It is preferable to use a separation membrane having a molecular weight of 100,000 or more.

  Furthermore, the obtained fraction may be made into a solid using a vacuum freeze dryer. Alternatively, it can be dried with a spray dryer to obtain a powdery solid.

  The proteoglycan obtained by the present invention can be extracted and purified with high efficiency and high purity from a biological sample such as salmon nasal cartilage by a very simple method. The advantages of the present invention are as follows.

  According to the present invention, conventionally, washing of cartilage, use of organic solvents such as ethanol, acetone, hexane, etc., use of hydrophobic carriers, plastic beads, activated carbon, deterioration of quality due to residual lipids, high cost of production, working time However, by using powdered cellulose, oil-absorbing mat, etc. after extraction, it is possible to easily adsorb and remove contaminating lipid components, and not only a high recovery rate of the target product but also time, economy, and environment It is also a great advantage in terms of safety and safety.

  Furthermore, as a result of evaluating the proteoglycan obtained in the present invention in the effect of proliferating cartilage progenitor cells, promoting the differentiation of cartilage and inhibiting the cartilage mineralization, it is equivalent to the proteoglycan extracted using only acetic acid in all tests. The above effects were obtained, and it was confirmed that the product had high quality. EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

(Production Example 1)
400 g of nasal cartilage extracted from the head of chum salmon stored frozen at −30 ° C. was prepared as a starting material. The starting material was added to 2000 g of a 4% by weight acetic acid aqueous solution and extracted at 4 ° C. for 48 hours with stirring.

This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, after the oil in the liquid upper layer is adsorbed with an oil absorbing mat (trade name “Oil Adsorbing Sheet SP-1300N (DX)” manufactured by Maeda Kosen Co., Ltd.), the liquid is sufficiently removed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000. The resulting solution was lyophilized to obtain 7.0 g of proteoglycan, which was measured by Soxhlet extraction method, and the amount of lipid in the obtained proteoglycan was 3.50%.

(Production Example 2)
400 g of nasal cartilage extracted from the head of chum salmon stored frozen at −30 ° C. was prepared as a starting material. The starting material was added to 2000 g of a 4% by weight acetic acid aqueous solution and extracted at 4 ° C. for 48 hours with stirring.

  This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, powdered cellulose (manufactured by Nippon Paper Industries Co., Ltd., trade name “KC Flock W-400G”) was added, and the mixture was stirred for 30 minutes and then filtered. The filtrate was sufficiently dialyzed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000, and the obtained liquid was freeze-dried to obtain 6.2 g of proteoglycan. As a result of measurement by the Soxhlet extraction method, the amount of lipid in the obtained proteoglycan was 2.48%.

(Production Example 3)
400 g of nasal cartilage extracted from the head of chum salmon stored frozen at −30 ° C. was prepared as a starting material. The starting material was added to 2000 g of a 4% by weight acetic acid aqueous solution and extracted at 4 ° C. for 48 hours with stirring.

  This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, the oil in the liquid upper layer was adsorbed with an oil absorbing mat (trade name “Oil Adsorption Sheet SP-1300N (DX)” manufactured by Maeda Kosen Co., Ltd.) and then powdered cellulose (trade name “KC Flock W manufactured by Nippon Paper Industries Co., Ltd. -400G ") was added, and the mixture was stirred for 30 minutes and filtered. The filtrate was sufficiently dialyzed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000, and the resulting solution was freeze-dried to obtain 6.5 g of proteoglycan. As a result of measurement by the extraction method, the lipid content in the obtained proteoglycan was 0.86%, and the proteoglycan content of the powder obtained by HPLC was as shown in FIG.

(Production Example 4)
50 g of cartilage from which meat pieces were manually removed from bean cartilage of domestic chicken was prepared as a starting material. The starting material was added to 250 g of a 4% by weight aqueous acetic acid solution and extracted at 4 ° C. for 48 hours with stirring.

  This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, the oil in the liquid upper layer was adsorbed with an oil absorbing mat (trade name “Oil Adsorption Sheet SP-1300N (DX)” manufactured by Maeda Kosen Co., Ltd.) and then powdered cellulose (trade name “KC Flock W manufactured by Nippon Paper Industries Co., Ltd. -400G ") was added, and the mixture was stirred for 30 minutes and filtered. The filtrate was sufficiently dialyzed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000, and the resulting solution was freeze-dried to obtain 1.5 g of proteoglycan. As a result of measurement by the extraction method, the amount of lipid in the obtained proteoglycan was 1.00%.

(Production Example 5)
50 g of cartilage extracted from the gray shark was prepared as a starting material. The starting material was added to 250 g of a 4 wt% aqueous acetic acid solution and extracted at 4 ° C. for 48 hours with stirring.

This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, the oil in the liquid upper layer was adsorbed with an oil absorbing mat (trade name “Oil Adsorption Sheet SP-1300N (DX)” manufactured by Maeda Kosen Co., Ltd.) and then powdered cellulose (trade name “KC Flock W manufactured by Nippon Paper Industries Co., Ltd. -400G ") was added, and the mixture was stirred for 30 minutes and filtered. The filtrate was sufficiently dialyzed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000, and the resulting solution was freeze-dried to obtain 600 mg of proteoglycan. As a result, the amount of lipid in the obtained proteoglycan was 1.96%.

(Comparative Production Example 1)
100 g of nasal cartilage extracted from the head of a white salmon stored frozen at −30 ° C. was prepared as a starting material. Solid fat was removed from the nasal cartilage as much as possible using tweezers and washed with physiological saline. The salmon nasal cartilage was then finely crushed into mince. Minced salmon nasal cartilage was added to 1000 g of a 4% by weight acetic acid aqueous solution and extracted at 4 ° C. for 48 hours while stirring.

This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, the solution containing the crude proteoglycan was centrifuged with a centrifuge (4 ° C., 10,000 rpm, 20 minutes). After adding 3 times amount of salt saturated ethanol to the obtained supernatant and stirring, the solution was centrifuged again (4 ° C., 10,000 rpm, 20 minutes) to remove the solution, and the cellulose ester membrane having an exclusion limit molecular weight of 1 million. After sufficiently dialyzing with a dialysis tube, the resulting solution was lyophilized to obtain 3 g of proteoglycan. As a result of measurement by the Soxhlet extraction method, the amount of lipid in the obtained proteoglycan was 5.04%.

(Comparative Production Example 2)
40 g of nasal cartilage extracted from the head of white salmon stored frozen at −30 ° C. was prepared as a starting material. A slice of cartilage sliced to about 5 mm with a knife was immersed in acetone and degreased and dehydrated from the nasal cartilage. The treated nasal cartilage was dried under reduced pressure to obtain 12 g of dry cartilage. 80 g of 0.02N sodium hydroxide solution was added to 4 g of this dry cartilage and extracted for 2 hours.

This eluate was filtered through a stainless steel mesh (150 μm) to remove cartilage. Next, the solution containing the crude proteoglycan was centrifuged with a centrifuge (20 ° C., 3,000 rpm, 20 minutes). The obtained supernatant was filtered, and fractionated and washed using VIVA SPIN 100,000 MWCO (fraction molecular weight 100,000) manufactured by sartorius. After adding 3 times the amount of salt saturated ethanol to the passing liquid and stirring, the mixture was allowed to stand overnight and centrifuged again with a centrifuge (4 ° C., 10,000 rpm, 20 minutes), and a precipitated semi-solid proteoglycan was precipitated. Got. This semi-solid proteoglycan was lyophilized to obtain 27 mg of proteoglycan. As a result of measurement by the Soxhlet extraction method, the amount of lipid in the obtained proteoglycan was 18.13%. Moreover, the proteoglycan content of the powder obtained by HPLC was as shown in FIG.

(Comparative Production Example 3)
200 g of nasal cartilage extracted from the head of white salmon stored frozen at −30 ° C. was prepared as a starting material. The starting material is put into 600 g of 0.1% by weight citric acid aqueous solution in an extraction container and stirred at room temperature for 24 hours. The eluate is made of stainless steel mesh (150 μm), filter paper (10 μm), filter paper (1 μm). Then, it was filtered with a glass fiber filter paper (0.4 μm) to remove insoluble matters. Next, 20 ml of polystyrene beads were added to the filtrate, and the mixture was stirred for 30 minutes and filtered. The filtrate was applied to an ultrafilter equipped with a membrane having a molecular weight of 1 million cut, and then applied to an ultrafilter equipped with a membrane having a molecular weight of 100,000 cut to obtain a concentrated solution. After adding 3 times amount of salt saturated ethanol to the concentrated liquid and stirring, it was centrifuged with a centrifuge (15,000 rpm, 5 minutes) to precipitate and separate proteoglycan.

After removing the supernatant, ethanol was added and stirred, and proteoglycan was precipitated with a centrifuge (15,000 rpm, 5 minutes). This washing operation was repeated twice. Distilled water was added to the precipitated proteoglycan, and the solution was lyophilized to obtain 2 g of proteoglycan. As a result of measurement by the Soxhlet extraction method, the amount of lipid in the obtained proteoglycan was 12.50%.

(Comparative Production Example 4)
100 g of nasal cartilage extracted from the head of a white salmon stored frozen at −30 ° C. was prepared as a starting material. Salmon nasal cartilage was put into 500 g of a 4% by weight aqueous acetic acid solution and extracted at 4 ° C. for 48 hours while stirring.

  This eluate was filtered through a stainless steel mesh (150 μm) to remove insoluble matters. Next, activated carbon (trade name “Shirakaba A W-50” manufactured by Osaka Gas Chemical Co., Ltd.) was added, and the mixture was stirred for 30 minutes and then filtered. The filtrate was sufficiently dialyzed with a cellulose ester membrane dialysis tube having a molecular weight of 100,000, and the resulting liquid was freeze-dried to obtain 0.9 g of proteoglycan. As a result of measurement by the Soxhlet extraction method, the amount of lipid in the obtained proteoglycan was 2.50%.

  Table 1 shows a summary of the amount of lipids in proteoglycans by the respective extraction methods in the production examples described above. Furthermore, the color, smell, taste, solubility in water, and the amount of hydroxyproline by amino acid composition analysis were measured and are shown in Table 2.

  As shown in Table 1, the amount of lipid derived from cartilage can be reduced more efficiently by using oil-absorbing mat or powdered cellulose, or using them together, rather than degreasing by washing alone, using acetone, or using polystyrene beads. Was confirmed.

  Furthermore, from Table 2, it was confirmed that the color, smell, taste, and water solubility of the obtained powder were affected as the amount of lipid contained in proteoglycan increased. Similarly, it was confirmed that the content of hydroxyproline decreased by using an oil-absorbing mat or powdered cellulose, or a combination thereof.

  Hydroxyproline is an amino acid characteristic of collagen proteins, and is one of the low molecular weight contaminants contained during proteoglycan extraction. Proteoglycans produced through an oil-absorbing mat or powdered cellulose, or a degreasing process using them in combination, reduce the amount of contaminants such as lipids and collagen mixed in, and can produce high-quality proteoglycans.

(Measurement of proteoglycan)
The HPLC analysis conditions for the proteoglycans obtained in Production Example 1 and Comparative Production Example 2 are as follows.
1) Column: TSKgel G5000-PWXL Φ7.8 mm × 300 mm (manufactured by Tosoh Corporation)
2) Eluent: pH 6.8 Phosphate buffer Flow rate: 0.5 mL / min
Column temperature: 40 ° C
3) Detector: Suggested refractive index detector (RID-10A manufactured by Shimadzu Corporation)
4) Injection volume: 50 μL
5) Molecular weight marker: Shodex STANDARD P-82 (manufactured by Showa Denko)

(Evaluation of proliferative action of cartilage precursor cells)
It is known that the cartilage matrix is denatured by aging, and the number of cartilage cells is reduced in the cartilage tissue that has developed osteoarthritis, and accordingly, the cartilage matrix cannot be produced and the cartilage is worn. The effect on cartilage progenitor cell proliferation was evaluated using ATDC5, a multi-stage cartilage differentiation model.
[sample]
A: The proteoglycan sample obtained in Production Example 3 was prepared and added to the final medium addition concentration. As a comparison,
B: Proteoglycan obtained in Comparative Production Example 2 C: Proteoglycan obtained in Comparative Production Example 3 D: Proteoglycan obtained in Comparative Production Example 4 was subjected to the test. The control was not added.
[Test method]
Seed in a 96-well culture plate at 1200 cells / well in ATDC5 (RIKEN), replaced with 0.5% serum and DMEM / F12 (transferrin, sodium selenite added) after 24 hours, and added the sample to 125 μg / mL. Culturing was performed for 7 days under the conditions of 5% CO ₂ and 37 ° C. After culturing, the number of cells was measured by the MTT method, and the chondrocyte proliferation promoting action of each sample was analyzed based on the relative value when the control was 100.
[result]
The test results are shown in FIG. Compared to the samples (B) to (D), the sample (A) showed a significant effect of promoting the proliferation of cartilage progenitor cells. The results show that the proteoglycan obtained in the present invention is useful as a cartilage progenitor cell proliferation promoter superior to those obtained by other methods.

(Evaluation of cartilage differentiation promoting effect)
Using ATDC5, a multi-stage cartilage differentiation model, the effect on cartilage differentiation promotion was evaluated.
[sample]
A: The proteoglycan sample obtained in Production Example 3 was prepared and added to the final medium addition concentration. As a comparison,
B: Proteoglycan obtained in Comparative Production Example 2 C: Proteoglycan obtained in Comparative Production Example 3 D: Proteoglycan obtained in Comparative Production Example 4 was subjected to the test. In addition, no control was added, and 10 μg / mL insulin was used for the test as a positive control.
[Test method]
ATDC5 (RIKEN) was seeded on a 12-well culture plate at 5 × 10 ⁴ cells / well, replaced with 5% serum and DMEM / F12 (added with transferrin, sodium selenite), and 5% CO ₂ at 37 ° C. The cells were cultured for 7 days to be confluent. Thereafter, each sample was added to 50 μg / mL and cultured for 21 days. After culturing, the images obtained by staining with Alcian blue were quantitatively analyzed with WinROOF to evaluate cartilage differentiation ability.
[result]
The test results are shown in FIG. Compared to the samples (B) to (D), the sample (A) showed a significant cartilage differentiation promoting effect in an insulin-independent manner. From this result, it was obtained that the proteoglycan obtained from the present invention is useful as a cartilage differentiation promoting agent superior to those obtained by other methods.

(Evaluation of inhibitory effect on cartilage calcification)
The cartilage matrix degenerates with aging, and in cartilage tissue that has developed osteoarthritis, the number of chondrocytes is reduced, and as a result, the cartilage matrix cannot be produced and the cartilage is worn. It is also known that bones rub against each other due to ectopic calcification of cartilage, and symptoms progress. Using ATDC5, a multi-stage cartilage differentiation model, the effect on calcification inhibition was evaluated.
[sample]
A: The proteoglycan sample obtained in Production Example 3 was prepared and added to the final medium addition concentration. As a comparison,
B: Proteoglycan obtained in Comparative Production Example 2 C: Proteoglycan obtained in Comparative Production Example 3 D: Proteoglycan obtained in Comparative Production Example 4 was subjected to the test. The control was not added.
[Test method]
ATDC5 (RIKEN) was seeded on a 12-well culture plate at 5 × 10 ⁴ cells / well, and 5% serum and DMEM / F12 (transferrin and sodium selenite added) were used under conditions of 5% CO ₂ and 37 ° C. The cells were cultured for 7 days to be confluent. Thereafter, the cells were further cultured for 21 days under the condition of adding insulin to form cartilage. Thereafter, each sample was added to 50 μg / mL, and further cultured for 24 days under conditions of 3% CO ₂ and 37 ° C. using 5% serum and α-MEM (added with transferrin, sodium selenite, insulin). After culturing, the calcification site was stained with alizarin red, and the acquired image was quantitatively analyzed with WinROOF to evaluate the ability to inhibit cartilage calcification. Furthermore, the cartilage state on the 52nd day of culture was evaluated by Alcian blue staining.
[result]
The test results are shown in FIG. Compared with the samples (B) to (D), the sample (A) showed significant inhibition of cartilage calcification. From these results, it was obtained that the proteoglycan obtained from the present invention is useful as a cartilage mineralization inhibitor superior to those obtained by other methods.

  Specific examples of the health food or health drink and the pharmaceutical composition according to the present invention are not limited to those mainly intended to improve the musculoskeletal system, but include nutritional supplementation, nutrition tonic, fatigue recovery, constitution improvement, beauty and slimming, etc. General-purpose foods or beverages such as sprinkles, herb salt, sauces, dressings and other seasonings, jelly, gum, candy, yogurt, tablets, soft drinks, etc. , Injection drugs, medications to be taken.

  Specific examples of the cosmetic composition according to the present invention are as follows: 1) topical or systemic external pharmaceutical preparations, skin cleansers, skin cosmetics, 2) medicinal or cosmetic preparations applied to the scalp / hair, 3 ) Medicinal or cosmetic bathing agent used in bath water 4) deodorant / deodorant for human body 5) medicinal or cosmetic oral agent such as dentifrice, mouthwash, mouthwash, 6 ) It can be used by appropriately blending in skin patch sheets, cosmetic sheets, cosmetic cotton, sanitary products, sanitary cotton, wet tissue and the like. Ampules, capsules, powders, granules, solids, solutions, gels, bubbles, emulsions, sheets, mists, sprays, etc.

  Such external preparations include basic cosmetics such as creams, gels, ointments, liquid external preparations, lotions, emulsions, creams, ointments, gels, lotions, oils, packs, mists, facial cosmetic sheets, etc. , Shaving agents, facial cleansers, skin cleansers (including soaps), shampoos, rinses, hair treatments, hair styling agents, permanents, hair nickings, hair dyes, hair restorers Makeup cosmetics such as foundation, lipstick, blusher, eye shadow, eyeliner, mascara, perfume, skin deodorant, antiperspirant, bathing agent, mouthwash, toothpaste, mouth freshener, etc. .

  In addition to essential drugs, the components and additives exemplified below can be arbitrarily selected and blended in the health food or health drink, pharmaceutical composition, and cosmetic composition according to the present invention.

(1) Various fats and oils Avocado oil, almond oil, fennel oil, sesame oil, olive oil, orange oil, orange rafa oil, sesame oil, cocoa butter, chamomile oil, carrot oil, cucumber oil, beef tallow fatty acid, cucumber oil, safflower oil , Shea fat, liquid shea fat, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, palm oil, palm kernel oil, owl, palm Oil, beef tallow, pork tallow, squalene, squalane, pristane or hydrogenated products of these oils (hardened oil, etc.).

(2) Waxes Beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, rice wax and the like.

(3) Mineral oils Liquid paraffin, petrolatum, paraffin, ozokelide, ceresin, microcrystalline wax, etc.

(4) Fatty acids Lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, linoleic acid, linolenic acid, docosahexaenoic acid, eicosapentaenoic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, lanolin fatty acid Synthetic fatty acids such as natural fatty acids such as isononanoic acid, caproic acid, 2-ethylbutanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid and isopentanoic acid.

(5) Alcohols: natural alcohols such as ethanol, isopyropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, phenoxyethanol, and synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, 2-octyldodecanol .

(6) Polyhydric alcohols Ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol 1,3-butylene glycol, pentyl glycol, glycerin, pentaerythritol, threitol, arabitol, xylitol, ribitol, galactitol, sorbitol, mannitol, lactitol, maltitol and the like.

(7) Esters Isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyl decyl dimethyloctanoate, cetyl lactate, myristyl lactate, Diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, and the like.

(8) Metal soaps Aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, zinc undecylenate and the like.

(9) gum, sugar or water-soluble polymer compound gum arabic, benzoin rubber, dammar gum, guaiac oil, Irish moss, karaya gum, tragacanth gum, carob gum, quinseed, agar, casein, lactose, fructose, sucrose or ester thereof, Trehalose or derivatives thereof, dextrin, gelatin, pectin, starch, carrageenan, carboxymethyl chitin or chitosan, hydroxyalkyl (C2-C4) chitin or chitosan to which alkylene (C2-C4) oxide such as ethylene oxide is added, low molecular chitin Or chitosan, chitosan salt, sulfated chitin or chitosan, phosphorylated chitin or chitosan, alginic acid or salt thereof, hyaluronic acid or salt thereof, chondroitin sulfate or salt thereof, heparin, ethylcellulose, methylcellulose, carboxy Polyalkylene such as methyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylate, polyethylene oxide and polypropylene oxide Oxide or its crosslinked polymer, carboxyvinyl polymer, polyethyleneimine and the like.

(10) Surfactant Anionic surfactant (alkyl carboxylate, alkyl sulfonate, alkyl sulfate ester salt, alkyl phosphate ester salt), cationic surfactant (alkyl amine salt, alkyl quaternary ammonium salt), amphoteric Surfactant: Carboxylic acid type amphoteric surfactant (amino type, betaine type), sulfate ester type amphoteric surfactant, sulfonic acid type amphoteric surfactant, phosphate ester type amphoteric surfactant, nonionic surfactant ( Ether type nonionic surfactant, ether ester type nonionic surfactant, ester type nonionic surfactant, block polymer type nonionic surfactant, nitrogen-containing type nonionic surfactant), other surfactants ( Natural surfactants, protein hydrolyzate derivatives, polymer surfactants, surfactants containing titanium and silicon, fluorocarbons System surfactant), and the like.

(11) Various vitamins Vitamin A group: retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), vitamin B group: thiamine hydrochloride, thiamine sulfate (vitamin B1), Riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acids, pantothenic acids, biotins, choline, inositols, vitamin C group: vitamin C acid or derivatives thereof, vitamin D group: Ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotaxosterol, vitamin E group: vitamin E or derivatives thereof, ubiquinones, vitamin K group: phytonadione (vitamin K1), menaquinone (vitamin K2), me Dione (vitamin K3), menadiol (vitamin K4), other essential fatty acids (vitamin F), carnitine, ferulic acid, .gamma.-oryzanol, orotic acid, vitamin P such (rutin, eriocitrin, hesperidin), vitamin U and the like.

(12) Various amino acids Valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine , Arginine, ornithine, histidine and the like, and their amino acid derivatives such as sulfate, phosphate, nitrate, citrate or pyrrolidone carboxylic acid.

(13) Various additives derived from plant or animal raw materials These are processed in a conventional manner according to the type and form of the product to be added (for example, grinding, milling, washing, hydrolysis, fermentation, purification) , Compression, extraction, fractionation, filtration, drying, pulverization, granulation, dissolution, sterilization, pH adjustment, deodorization, decolorization, etc., arbitrarily selected and combined), arbitrarily selected from various materials You can use it.

  In obtaining an additive derived from a plant or animal material, the solvent used for extraction may be selected in consideration of the intended purpose, type, or subsequent processing of the product to be provided. Water, methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol, isobutanol and other lower alcohols or hydrous lower alcohols, propylene glycol, 1,3-butylene glycol, glycerin and other polyhydric alcohols or hydrous polyhydric alcohols, acetone, It is desirable to use one or a mixture of two or more selected from various organic solvents such as ethyl acetate. However, when it is not preferable to contain an organic solvent depending on the application, it is sufficient to use only water or ethanol that can be easily removed after extraction, and it may be used alone or in any mixture with water. The thing extracted and extracted may be sufficient.

  When using additives derived from plant or animal-based ingredients for systemic or topical external preparations and cosmetics, it not only protects skin and hair, but also moisturizes, improves touch and texture, imparts flexibility, and reduces irritation , Aroma stress relief, cell activation (cell aging prevention), inflammation suppression, skin and hair quality improvement, rough skin prevention and improvement, hair growth, hair growth, hair loss prevention, glossiness, cleansing effect, fatigue In addition to cosmetic effects such as relaxation, blood flow promotion, and bathing effects, effects such as flavoring, deodorization, thickening, antiseptic, and buffering can also be expected.

  In foods and drinks, nutritional supplementation, fatigue recovery, tonicity, cell activation (cell aging prevention) and other healthy and cosmetic effects, as well as taste improvement, color tone, aroma, gloss, stabilization, thickening, antiseptic, etc. Can be used for the purpose. In addition to this, we expect various cosmetic and pharmaceutical effects of each of the raw materials known so far, and by combining these, we aim to enhance the target effect or to have a multifunctional effect It is also possible to make a product that expects

Specific plants (herbal medicines) as raw materials include, for example, almonds (toen), ai (indigo leaves), aokazura (Keifu Fuji), Aoki (Aoki), Aogiri or Kenashia Ogiri (Ao Tung), Aotsu Samurai (wooden candy) , Aotenma, acacia, red ginseng, red spruce `` fruit '', red clover, red grapes, red ginseng (logwood), red wrinkles (red buds), red crested or red crested or reddish (root), red crested or reddish (ground yellow), Agi (Aki), Aki larch, Akinire (crusted skin), Aguay Guassou, Aguahe (Omiten coconut palm), Akebi (Kitsu), Asa (Asakojin), Maruba morning glory or morning glory (Chicken), Hydrangea (hydrangea), Ashitaba (Tomorrow), Azuki (Red Azuki), Asunaro, Acerola, Asenyaku (Asenyaku), Athira (Shokuyokanna), Anise, Abiyu, A Biurana, oilseed rape, Abemaki `` fruit '', avocado, ama, achacha (Amacha), Achacharu, Amadokoro (Jade bamboo), Amana (a light-hearted), amaranth (Hyuyu, Hagatetou, Himogeito, Senninkoku, Sugimorigateto, Hosoaogeito, Hasoagoito Aozulan, Amaranthus hypochondriacus), Amigasayuri or Baimo (shellfish mother), Aristolomeria (Yurizuisen), Arcana (Alcanna), Algarrobo (Cabe), Altea, Arnica, Alpinia or Katsumadai (Suzukuku), Aloe (蘆薈), Aloe vera, angelica, apricot or honanzu (Anzujin), Ansokkou (benzoic incense), yellow support, igakozolina (geo-biliary head), licorice or gecko licorice (indian oyster), rush (light heart grass), itadori (tiger cane root), yew (one Place), Strawberry, fig (no flower `` fruit, leaf ''), Ichihatsu (first), Ichibi (winter lion), Ichiyakusou (one medicinal herb), ginkgo (Ginkgo `` seed, leaves ''), Japanese yellow peas (distant), carob, Japanese beetle ( Taillexi), Inubiyu (Honagainubiyu), Rice `` Seeds, Seed coat '', Inondo `` Seeds '', Ibuki-so-sou, Nettle, Ylang-ylang, Iwa-tobacco (Iwabuchi), Iwahiba or Iwamatsu (rolling mushroom), kidney beans, oolong tea, fennel (mustard) , Wheat (Chibusanoki), Ukiyara (Sanryo), Uguisukagura `` Fruit '', Himewikyo, Turmeric (Urban), Duckweed, Usubasaishin or Keirinsaishin or Oshusaishin (spicy), Usvenia Oy, Usbeni Hollyhock, phalaenopsis, weevil (summer hay), udo or shishiudo (gung, alone, tangy), unyadegato (unkari) A, Cat's Claw), Uba tea, Ume (Japanese apricot `` seed, pulp ''), Japanese white beetle, Uwaurushi (Kusako-kemomo), Unshuumika (Chen), Unboku, Estragon, Ezoukogi (Tomogoka), Ezo-suzuran, Echinashi (Hosobamurasaki Barengiku) , Enishida, Enokitake (榎 茸), Ebisugusa or Cassia Tora (Keiko), El Campre, Elderberry “fruit”, Elemi, Elingi or Pre-Lotus Elinji, Enju (Safflower, Camellia rice), Pea, Ogi or Kibanaogi (Yellow Twilight), Naruko Yuri or Kagura-mabanaru Koyuri (Yellow), Ohisasa Yuki or Sasano Yuki, Ouren (Yo Ren), Ogata Houken, Okarasuri (Karokon), Oguruma (Civil Works Incense), Otsu Samurai (Prevention), Psyllium , Car forage), toucan thistle (Zhengzhou leakage, bald eagle), toucan Mi (coconut palm), Toucan cactus, Ovana okera or okera (white moth), Osara squirrel (Banaba), Ooyabaya shabushi "fruit", Oohashigusa (cereal sperm), Omia catechu, Oomihasa or hawthorn (Mountain vegetation), Omiyashi, Omiji Barley), Oka, Okazeri (snake bed), Okra `` fruit '', Oguruma (overturn), Oguri higotai (Zhengzhou leakage, indigo head), Panax ginseng or Tochibaninjin (Ginseng), Hypericum pertussis or Pterotus pertussis or Hypericum perforatum (Brother cut grass), Odorikosou (continuous), Onamomi (Circle ear), Onigomi, Onikoro or Tokoro or Nagadokoro (Hikai), Oninoyara (Tempa), Onyuri or Sasayuri or Hakatayuri (Yuri), Onoelan, Ononis, Halibut楡), Ominae (losing sauce), Yuuko, Dutch mustard (watercress), Ora Dazelli, Dutch beech, Dutch bees, olives `` fruit, seeds, leaves '', oregano, orange `` fruits, peels '', silkworm (ripe yellow), cacao `` fruits, peels, seeds '', oysters (bamboo `` leaves ''), Kagi-Kazura (Katsufuji), Kakidooshi or Kantorisou (Lotus), Kakiran, Gaku Hydrangea, Cassia, Casinoki (fruit berries), Gadju (Kashiwa), Kashiwa (Kashiwagi, Kashiwa), Cascarilla, Cascala Sagrada, gypsophila, gypsophila, cynomolgus (Kinshato), kaniwa, carnation, valerian (valerian), hippopotamus, birch or fern birch (white birch), gerbera, pumpkin, kapokoki `` seed '', kahoku salamander (蜀椒), gama Yellow), chamomile or roman chamomile, chamomile (weed), cam cam (camo duck), collar, chrysanthemum, calla or cinnamon , Oat biscak (half summer), oats, guarana `` seed '', carahoo (kokubaku), karaya, galyu, karin (garden), garcinia, kawamidori, kawara-saiko (reeled rape, white millet), kawajisha, kawaratake, kawaranadeshiko ( (Stone bamboo) or Ezokawaranadeshiko (buckwheat noodles, buckwheat), Chinese carrot (blue potatoes), Chinese mugwort (bamboo chin mackerel), kansui (kansui), licorice (licorice), kantarasa, candelilla, kanto, canna, kanran, raspberry (ezo strawberry) , Dutch Strawberry, Shrimp Strawberry, Nawashiro Strawberry, Momiji Strawberry, European Raspberry Strawberry), Kiwi `` Fruits, Leaves '', Kikarasuuri (Gurone), Chrysanthemum (Kikyo, Kikyo-root), Chrysanthemum (Chrysanthemum, Chrysanthemum, Datura), Kikutanigiku, Kisage (Salmon), Gigigishi (sheep hooves), pheasant (salmon), Kizuta, Kidachi aloe, Kidachi mint , Quina, linden (shinkona, red linden), quinoa (quinoa), yellowfin (yellow tuna), bean, gymnema sylvester, kimenkaku, cabbage, cabbage `` unripe fruit '', caraway, cucumber, gyory (Nishikawa willow, tail willow), Kiraja saponaria, Kiraya, Kiranso (golden grass), kumquat "fruit"
, Kinkiju, ginsekai, kimbusen, betel, kim mizuhiki (senzaku), quinlan, gin orchid, kinryougen, guapeba vermelya, guava `` fruit '', guayule, quercus infectria (garlic), kukui nuts, kugenumaran, wolfberry (coconut, coconut , Leaves, ground bones), Kuzusugi Kazura (Tenmon Winter), Kuzu (Kazunori), camphor, gooseberry `` fruit '', carrot carrot (yellow flower moth), cutiti river, gardenia (Yamashiko), Kunugi (Sanso), Kupuas, Kubeba, Kumazasa, Kumatsutsura (horse whip grass), Clara (bitter), cranberry `` fruit '', chestnut `` seed, fruit, astringent skin '', curcumillo latifolia `` fruit '', grapefruit `` fruit and leaf '', buckthorn, kuroganemochi ( Rescue), Kamehiba Kokoshi or Kurobana Hiki Koshi or Hiki Koshi (life extension) ), Clove (chome, cicada), gumbai nazuna (sekimei, sekimeishi), keigai (butterfly, 荊芥 ho), kaito (chicken crown flower, chicken cub), gekkavidin, bay bay (laurel wreath), kenashi saruto raibara (dawn, mountain return), Gentianana, Gennoshouko (Oldweed Grass), Kemponashi (Kugushi), Kokisekoku, Kishumikan (Koji, Tachibana, Obenimikan, Hukurikukan, Sagamikouji, Ponkan, Santara (Tachibana Peel)), Koshinbara (Moonflower), Kouseihakka `` Fruit '', kocha (tea), kohone (river bone), koh hong (Tsubakimoto, Karamotomoto), Koryang, Koryokukyo (Koryo), cilantro `` fruit '', Koouren (Huo Ren), Koganebana (Yellow Gong), cowberry ( Koshibana), coconut `` fruit '', goshuyu (go), gosho strawberry (covered bonsai), pepper (pepper), cosmos, copaiba Roussum, coffee `` seed, leaves '', kobushi or magnolia (hot pepper), burdock (beef bowl, beef eggplant), kobotan lees, sesame (sesame seeds), sesame seeds (genus ginseng), trash beet (pencil), sakanekazura or binankazura or matsubusa, wheat ( Wheat), rice or rice bran `` red cocoon, white birch '', rice oil, cola acminata `` seed '', cola bella `` seed '', colony llya, fenugreek `` fruit '', colombo, condurango, kombu, konjac, comfrey Grass), Sisal (Sisal Asa), Sai Hairan, Sakaneran (Ezosa Kanelan), Sasima button eel or cinnabar eel or Chinese ginseng (weikensen), Sakura (Oshima cherry, Yamazakura, Oyamazakura, Edhigashi, Maizezakura, Miyazakura, Yoshino cherry, Yoshino cherry, Yoshino cherry Kohigan, Satozakura, Kanzakura "Leaf, Flowers, fruit, bark (cherry bark) ''), cherries, pomegranate, Sasa, Sasabagin orchid, sasanqua, Sadamodaka (Sawaka), sweet potato, sugar cane, sugar beet, sunflower jujube, saffron (bancho flower, west safflower), sapodilla , Sapotea mariyo, pomelo `` fruit '', savonso, salmon berry `` fruit '', salamander (sasma), salamander (sand sashimi), salvia (sage), ragweed (bamboo rape), sawagurumi (wild walnuts), sankaku cactus , Sangre de Grado (croton), Sansikuyosou, Sanshichininjin (Ginseng Ginseng), Sanshuyu (Yamamuro), Sansho (Yamamuro), Sandzukon (Yamazuroot), Shia (Kalite), Cyanoki "fruit", Shiitake Shiitake mushrooms, Zion (purple lees), winged bean, jikitarisu, shikunshi (messenger), perilla or aojiso or chirimendiso or catamendizo ( (Soba, Shisoko), Rosewood, Shinanoki, Shirahonoki, Sinalengyo (ream), Shimeji (Hinshimeji, Shakashimeji, Hatakeshimeji, Oshiroi-shimeji, Buna-shimeji, Honjimeshi, Shirotamogitake), Shimokuren (Shigetsu), Shimotsumo Jinja ), Jasmine (Matsurika), Janohige (Wheat Gate Winter), Zodiac Cowpea, Shukukonkassou, Shukushamitsu (Sanjin, Shredded Sandals), Juzudama, Shuro "Fruits", Shunlan, Zhououashi, Ginger (Ginger), Zhou Zan hydrangea (Tsuneyama), Shobu (bamboo shoots, Sone), Shozu `` fruit '', Shirakashi `` Seeds '', Silane (Bikkyu), Shirogori `` Seeds '', White clover (Clover), White clover (Chick-bone grass), White squirrels (Nioiris) ), Shirobanatsuta "flower", Shirobanupin, Shirominan Ten (Minami Tenshin), Shinkonasakushirubura, Jinchoge (Ruika, Ruika, Shencho), Cinnamon, Cinnamomum Cassia (cinnamon), Watermelon (Sairan), Honeysuckle (Gin silver, Shinobi), Sorrel (Acami), Swimberry `` Fruit '', Star Apple, Stevia, Strawberry `` Fruit '', Suzu-Psycho (Xucho), Sugina (Question), Subuhiyu (Horse toothbrush, Horse tooth soap), Plum `` Fruit '', Sulgaran (Oran), Scots pine `` Cone '', Japanese larch, Japanese lizard, Japanese walnut, Japanese hawthorn, Japanese cypress, Japanese dandelion, Japanese horse chestnut (Maronier), Japanese pear `` fruit '', Japanese yellow snow, Japanese elder (Juniper, Japanese pine) , Achillea millefolium (milfoil), rose, pearlfly , Mistletoe, Atlantic mint or Atlantic mint, Horseradish, Berry (Ishizone), Sekkoku (Sakura Sekkoku, Oki Sekkoku, Ohana Sekkoku, Okinawa Sekkoku, Honsekoku, Shiki Sekkokuk, Kibanasekkoku, Shiki) , Cedron (Boushuboku), Zeniaoi, Hirosanega, Senega, Seri, Celery, Senkyu (river cucumber), Senshinren (Pennial ream), Sendang, Sennin cactus, Senna `` Fruit, leaf '', Senninsou (Otsuchi), Senburi (Medicine), senboku, soka (grass), elephant chew, socin waxy, buckwheat `` seed '', some potato, broad bean, daiou (dai yellow), radish, soy, daidai (orange peel, berry), takasagorihigotai (southeast indigo sashimi) , Takasaburo (Shorus), Takatodai (Large)戟), Takara-bi (pig spine), dark sweet cherry `` fruit '', hollyhock, prickly-spotted thyme (thyme, perilla), tachidokoro (hikai), tachiyanagi, tamarind `` seed '', onion, ramshiba (powder), taranoki `` fruit, leaf ``, Root bark '', tarayou (Oba winter blue, Ichiha tea, bitter tea, bitter tea), dahlia (tenjiku button), tarwi, tanukirimame, tanshichiwa, tanjin (dansang), dandelion (Sanko) or Shirobana dandelion or mouko dandelion, Danmaru, cherry “fruit”, chigaya “fruit, root, bud”, chixetsu carrot (bamboo carrot), chicory, chankapiedra
(Kidachi Kumiso), Tulip, Chono Stay, Datura Thistle (Artichoke), Datura (Dark Yellow), Datura (Amber), Datura Worm, Choreimaitake (Amber), Chorogi, Azalea, Azalea, Tsukuritake mushroom, camellia, camellia, camellia, clover (lacquered eel), camellia (duck bean grass), tsuzukizuki (red bean), tsurutsukichibi, tsurudukudami (what neck rice cake), tsuruna (apricot apricot), tsuruminjin (four leaf ginseng), Tsuruma, Tsuwabuki, Deiko, Teuchigurumi, Duberry `` Fruit '', Tengusa, Tencha (bamboo tea), Tendai-yaku (glaze), Toga (winter coconut), Tokagi quail, Capsicum (banban), Toki (toll), Tokinsenka (Tokinsenka) Marigold), red croaker, touna bena (river cut), corn or corn hair (Southern lashes), horned potato (Sadako), Torindo or Ginseng (Dragon gall), Tokusa (wood band), Dokudami (10 medicines), Tokushi (Nippaku), Toshishi or Mamedaoshi or Nekazura, Tochu (bark, bark, Leaves, roots ''), ash (crusts), tomatoes, tragacanth, triacanthos `` seed '', Turkish almonds, tormentilla, dronoki, troro mallow, tonka bean, nigerian berries `` fruit '', Chinese yam or wild yam (wild boar), nagii kada (butcher bloom) ), Naginata Koju, Nazuna, Rapeseed, Natsume or Vertical Spider (Sword Bean), Natsumikan, Jujube (Otsugi), Naniwaibara (Golden Cherry), Navena (Suspended), Nameko, Nankin Bean (Peanut), Nanten (Southern Fruit), Namban Karamushi (Sesame), Niosumire, Nigaki (Nakiki), Nigamugi (Sweetness), Nikuzuku, Kei or Nikkei or Seika Ronnikei or Yabunikei (Cinnamon) or Keishi (Keishi branch), Flying tiger plover, leek (lion), elderberry (bone, `` fruit, flower, stem, leaf ''), garlic (large persimmon), Nurde (penta), leek, nemonoki Or Nebu or Nebi or Nemurino or Digokubana (community), Noazami (Daisetsu), Nobara (fruit), Nozenkazura (Lingen flower), Nogateto, Yarrow, Nodake (former cucumber), Novara, Peach, Pineapple `` fruit '', Hibiscus (Busouge) , Fulinbussouge, Roselle), Hailindo, Hauchiwa, Pau Dorse, Hakama Labosi (osteostomy), Hakukayumatou, Chinese cabbage, Chinese cabbage (white tinea), Hakulberry `` fruit '', Japanese crocodile (breeding), Habami (coconut), Hasidorokoro (Roof root), basil, lotus (lotus, lotus meat, lotus), parsley (holland jellies), hadakamugi, batata, bee or maize Ke (bamboo shoots), patchouli, mint (thin-load, thin-leaves), yellow peas, pearl barley (Yokuinin), Hanasuge (my mother), bananas, hanahakka, Hanabishi (Sitoshiri, Shitsutsuri), vanilla bins, papaya, scallop , Pahalobobo, hub `` whole grass, stem, leaf '', paprika, oysters, hamago or honey bee (Vulgaris), hamasge (Katsukiko), Hamabishi (Shitsuko), Hamanasu (Maikai flower), hamabofu (hamafufu), Hamamelis, palm, rose (rose), balata, balata rubber tree, agaric (agaric, white-faced agaric, quince) (Skin, maple leaves), Barbasco, Harlindow, Handaikai (Taiho Sea), breadfruit, hinoki Sea urchin (hiboshi), lizard carrot (participant), pecan nuts, ganbanana (stone urchin, vine pearl shahua), higurmadaria, higotai, hishi (hishimi), pistachio, beet, human lizard (hisarium), human winged fern, daisies (Daisy), Hinata Taikozuchi (cow knee), chickweed, cypress, hiba, castor, sunflower, bell pepper, Himezu (Tengu), Himamatsu (Kaori), Himematsutake (Kawariharatake, Hiromatsutake), Himemuyoran, Pimenta `` fruit '', Beakshi, beakkyu, jade `` fruit '', chickpea, lentil, belude oyster, hirohaokinagusa (bald eagle), loquat `` fruit, leaf, stem '', betel wax (large clothes skin, coconut), puer tea, futukazura (south rattan), Japanese cypress, Japanese cypress poppy (Fuji winter flower, Japanese cypress winter leaf), Fujibakama (Orchid), Fuji bean (Tsuname), Fujimodoki (Chujizakura, Spike), Bud `` Fruits, pericarp, seeds, leaves '', beech, Fuyumushinatsusatake (corticus), Brazilian licorice, Brazilian carrot, Brazilian palm, Brazilian elephant palm, black currant `` fruit '', blackberry, Branjen, plum `` fruit '' , Frucereria, Blueberry (Prunus japonica), Prunes, Floral Blanca, Blond Psyllium, Bundow (Mung Bean), Hazelnut, Loofah, Hetsukaran, Safflower (Safflower), Heneken, Belladonna, Berry `` Fruits '', Percea, Pervianburg, Pereschia Grandifolia, crassulaceae or nettle (scenic), boysenberry `` fruit '', broomfish or narcissus or nendo or hahagigi kokia (geo-silk), horsiran, garlic (saccharus, acute pupa, parasitoid), pouteria sapota, Powteria Lucuma, cypress, Japanese cypress (windbreak), spinach, physalis (Torone), cypress (Japanese rustic, magnolia), bokeh (wooden moss), hosobaabragiku (thorny horn), squirrels walnuts, sorghum (salmon), cypresses (landscape) Tenseven), Hosobabarengiku, Bodaiju (bamboo tree), buttons (peony `` flowers, leaves, stems, bark ''), buttonpi (peony skin), hops, jojoba, borage (Lulichisha), Bordeaux, white berries `` fruit '' , Hononiku (meat jugo, large art), Honsecek (iron skin stone, sparrow mountain stone wall), Mizuru tennansho (Tennan star), Maitake (maiko), maou (mao), maca, macadamia nut, margaret (mokushungiku) , Makri (marine grass), Magwa (mulberry bark `` bark, leaf ''), Magnolia splengeri, Mazaran Uba, Masalanzuba de Cearer, Masheira de Boi, Mash (Tama no Senhalen, Kyukonkinrenka), Matatabi (Tenten-don), Matsukasa, Pine `` Leaf, Bark, Root '', Matta Orillo, Matsuhodo (茯苓), Matico (Cordon Sillo), Mayoram (Hanahakka), Malvanojajin ( Bitter), Mulberry `` Fruit '', Quince, Malene (Billow Muzu squid), Mango, Mangosteen, Manzanya (Andesuka Mitsuru), Manshugurumi, Mandarin `` Fruit '', Mannentake (Ganoderma biloba), Pheasant (fruit berry `` Fruit ''), Mishimasaiko (Shibako), Mizuoobako or Mizuasagao (Dongweed), Mizokakushi (Half-sided lotus), Misohagi (Chikuma), Mitsuyanagi or Niwayanagi (Henpo), Mitsugasiwa, Mitsuba, Midorisapote, Midorihakka, Mimosa, Miura Fruit "fruit", myrrh, myrobalan, wheat straw, mugweed (mushroom), myrtle, mu Logi (life extension skin), Munya, Murasaki (purple root), Murasakikikibu or Omurasakikibu (purple pearl), Murasaki corn, Murasaki Kinafuji (Kunming chicken blood wisteria), Megakansa Opuntica, Swordfish (beneficial herb), Mebougi, Melaroika, Melissa, Merirot, melon `` fruit '', mouko mugwort, moso chiku, mok kirin, mogi (mushroom), cinnamon, mokko (woody incense), peach leaf, peach (peach `` leaves, seeds, flowers, fruit ''), coconut, morello cherry `` `` Fruit '', Morohaya (burlap), Yakawa Murasakiimo, Yakuchi (Michichi), cornflower (cornflower), cornflower mint, yacon, yashabushi or himeyashabushi `` fruit, pericarp, fruit ear '', yachiyanagi Octagonal metal), mistletoe (willow infestation), willow (kawayanagi, tachiyanagi, weeping) Willow, Red-backed Willow, Pussy Willow, Incolye Nagi, Kinuyanagi, Koryanagi, Eunryu Willow, Miyama Willow, Palm, Willow Willow, Pillus虱, Tenmeisei), pokeweed (commercial land), Yamahanoki (Yamabuchi), bayberry (Plum skin), sagebrush, eucalyptus, saxifrage (tiger ear grass), yucca or flavifolia, yuzu `` fruit '', lily, sagebrush, mugwort (Bamboo leaves), Ligan (Raimaru), Lime `` fruit '', Rye, Lacanca `` Fruit '', Raspberry `` Leaf, fruit '', Latania (Clamelia), Lackey or shallot (palm white), Lavender, Longan (long-eye meat), Agave ( (Aono Agave, Huklin Agave), Ryok (Green tea), apple `` fruit, seeds, leaves, roots '', gentian, lubas or suabishimusu (Suzu Ryo), rulitama thistle (bladder scallop, Xinjiang indigo sashimi), litchi (toothpick, toothpick core), lettuce (chisha) , Red currant `` fruit '', red pitaya, lemon `` fruit '', lemongrass, forsythia or Chinese forsythia (ream), forsythia, wintersweet, wax palm, locon (reed, root), loganberry `` fruit '', rose Examples thereof include marie (mannenrou), rose hips (nobara), wasabi, cotton wisteria (Ming Mongolia), and bituminous (ground).

  As seaweeds, seaweed [Green algae: Chlorella vulgaris, Chlorella pyrenoidosa, Chlorella ellipsoidia, Aonori (Usba aonori, Sugiaonori, Hiraaoori, Boaonori, Hosoeda Aonori), Anaaoosa (Aosa)] (Macombu, Resilibu, Hosocome, Mitsuishi Kombu), Wakame, Hirome, Aowakame, Giant Kelp (Macrocystis Integrifolia, Neocitus Ruetkeana), Hijiki, Hibamata, Umiuchiwa, Usubaumiuchiwa Umiuchiwa, Akaba Umiuchiwa, Konami Uchiwa, Okinawa Uchiwa, Usuki Uchiwa, Etsumi Uchiwa], seaweed [Red algae: Hijirimen, Maca (Tengusa), Hiraksa, Syria, Oaxa Caterpillar, Yatabegusa, Yuukiri, Shimatengusa, Tosakanori, Togekirinsai, Amakusakirinsai, Giraffe, Juniper ginkgo rhinoceros, Tsunomata, Totsuka (Yahazutsu notamata), Ezotsunotamata, Togetu no Takita , Yaleus banori, Kagius banori, Susius banori, Haius banori, Akamo mizunori] and the like.

  Other algae, for example, green algae (Chlamydomonas: Chlamydomonas, Akayukimo, Dunaliella: Dunaliella, Chlorococcus: Chlorococcus, Kuwanomimo: Kuwanomimo, Volbox: Dandelion, Volbox, Palmera, Yotsumeo, Himidori Aoumiduro, Tsurugidomiro, Hibidoro: Hibidoro, Aosa: Anaaaosa, Amiaosa, Nagaaosa, Kawanori: Kawanori, Friccella, Shiogusa : Magatama, Dioscorea: Pheasantata, Surichogita, Heraiwata, Kurokita, Hanemo, Genus: Mill, Chromel, Sakibutimil, Nagamil, Hiramil Casanori: Casanori, Jusmo: Jujusmo, Tamajusmo, Mizojusmo, Mika ヅ kimo, Korekaete, Tsutsumimo, Kikkogusa, Humanexa : Sumiremo, Hosimidro, Fuchinashimidro etc.).

  Cyanobacteria (Suizenjinori: Suizinori, Aoko, Nenju: Kawatake, Ishijelly, Hassai, Yuremo, Rasenmo (Spirulina): Spirulina, Trichodesmium (Iaciasio), etc.

  Brown alga Sanadagusa: Sanadagusa, Fukurimamiji, Commongusa: Commongusa, Yahazugusa: Ezoyahaz, Yahazugusa, Uraboshiyahaz, Jigamusa: Jigmygusa, Umichiwagusa, Umamisakugakua, Hakusou : Navarimo, Chasomen genus: Motsuki Chasomen, Matsumo genus: Matsumo, Nagamatsumo genus: Nagamatsumo, Okinawa mozuku: Okinawa mozuku, fake Tomozuku: Niseft Mozuku, Futomozuku: Futomozuku, Ishimozuku: Ishimozuku, Kuromo: Kuromo, Nisemozuku: Nisemozuku, Mozuku: Mozuku, Ishige: Ishige, Iroro, Ichimegasa: Ichimegasa Umibosu, Urushigusu: Urushigusa, Keurushigusa, Tobaccogusa, Composita: Composita, Haberodomo: Haberodoki, Havanori: Habanori, Habanori: Habanori, Commonbukuro, Ezobukuro, Ezobukuro Chishima fukuronori genus: Chishima fukuronori, Kagomenori genus: Kagomenori, Muraridori genus: Murachidori, shark genus: shark zusa, genus genus: lobster, genus genus: yokojimamanori, Ganoderma genus: cynomolgus, fennel spider: fennel spider, vine genus: turmo, turtle genus: sea turtle, scorpion genus: sujime, misji kumbu genus: misji kumbu, atsuba misji kumbu, kumbu genus: tsutsugara kumbu, oyster kumbu, onikombu, gakonbubu Barracuda, Trollocombu: Trollocombu, Antokume: Antokume, Kajime: Kajime, Tsuruarame, Kurome, Kikuishikombu: Kikuishicombu, Nejikcombu: Nejicombu, Kuroshiome: Kuroshiome, Nekoshikonbu, Aramei Genus: Ainuwakame, Chigaiso, Oniwakame Ezoi Shige: Ezoi Shige, Yabane Moku: Yabane Moku, Rappa Moku: Rappa Moku, Joromoku: Ugano Moku, Jo Lomoku, Hiemoku, Honda Walla: Tamanashimoku, Isomoku, Nagashimamoku, Akamoku, Shidamoku, Honda Walla, Nejimoku, Narasamo, Mametawala, Tatsukuri, Yatsumata Moku, Umitora no Moku, Fushishimoku, Mohmokumoku, Hahamokumoku , Ukimo: Ouokimo, Burukimo: Burukimo, Kayamonori: Kayamonori, etc.).

  Red algae Genus: Sea urchins, Tsukumonori, Anemone crickets, Astragalus, Amaranthus, Amaranthus, Agaricata Genus: Tamaitadaki, Kaikekenori Genus: Kaginori, Kaikekenori, Tengusa: Himetengususa, Hytengususa, Oobusa, Nambugu , Kohira, Yohira, Kinukusa, Hibirodo: Hibirodo, Himebiro, Isomumedo: Isommedo, Aedes: Rhizome, Ryumons: Ryumon, Ranunculaceae, Nisekare Naminohana: Hosobanaminohana, Naminohana, Sangomodoki: Ragaramomodoki, Shiogusagoromo: Shiogusagoromo, Iwanokawa: Etsukiiwanawa, Kinokawa: Kinokawa, Kanonote, Sangori Kyono Shiro, Sakuranori, Nikumukade, Tambanori, Tsurutsuru, Fudarak, Marubahdarak, Isonohana: Nurakusa, Ronulaksa, Oomkadenori, Hirakintoki: Hirakintoki, Matabou: Matabou, Kintoki: Chabokintoki, Kintoki, Matsunori, Rice, Tokamatsu, Hitomatsu, Fushikintoki, Tsunomude, Nagakinito, Crayfish Nagaobane: Naganobane, Funori genus: Hanafonori, Fukurofunori, Mafunori, Karekigusa: Karekigusa, Tosakamodoki: Hosobanatosakamodoki, Hironotosakamodoki, Yatsudegatatosakamodoki, Kurotosakamodokisane, Zotosofomoza Tosakamodoki, Nankaitosakamodoki, Himetosakamodoki, Kinuhada: Kinuhada, Ezotosaka: Ezotosaka, Tsukasanori: Enashikarimenia, Otsukasanori, Hana Tacarimenia, Hoonoo: Honoo, Hikagenoit: Hikagenoit, Usuginu, Nikuhounoo: Nikuhounoo, Venus nago, Venus nago, Susca veni: Suscabeni, Okarugusa: Yamadagusa, Mirin: Mirin, Hosokamiso , Isomocca genus: Isomocca, Yukari genus: Yukari, Hosoyukari, Ibaranori genus: Ibaranori, Saidaibara, Tachiibara, Kagibaranori, Kojinoo: Kizino-o, Isodantsu: Isodantsu, Atsubanori: Atsubanori, Ogonori, Ogonori, Ogori Mizo-okonori, birch, beniogonori, fukureshinori, purple birch, shinkaibanori, spruce birch, kataogonori, ryukyuogono Li, Atlantic scorpion, Ivy genus, Yumigataonori, Kubireogonori, Mosagogonori, Kinubanori, Amanita genus: Hachijotensamodoki, Fushikenori genus: Fushikenori, Namiiwatake, Nymphalanthus Itagususa, Saimi, Hari, Lotus Jigusa: Lotus Jigusa, Sugi nori: Ikanoashi, Hosoibonori, Nobonori, Black Haginansou Genus: gourd, cherries, octopus: octopus, sedge, genus: genus, genus: goat, dals : Dulse, Scorpionus, Anadalus, Weibusa: Uebusca, Benifronori: Benifuronori, Fusunagi: Futsutsunagi, Himefusunagi, Hirohafushitsugi, Otsunagisou, Hirawatagigi, Usuwasugi , Egonori: Egonori, Futogigi, Saeda: Saeda, Chirimomiji: Chirimomiji, Konohori family: Habutaenori, Konohori, Suzuronori, Usubeni: Genus: Kagios banori, Yareus banori, Susius banori, Heius banori, Usubanorimodoki: Usubanorimodoki, Ayanishiki genus: Ayani Genus: Aeginu, Dadia: Enasidadia, Shimadadia: Isohagi, Shimadadia, Dajimodoki: Dajimodoki, Euglena: Morayogusa, Fuyugusa, Makuri: Makri, Yanaginogi, Yuna, Yukinagina , Sasabayanaginori, Sozo genus: Kurosozo, Kobsozo, Hanesozo, Sozonohana, Hanegusa genus: Hananegusa, Hananegusa, Kozanemo genus: Kozanemo, Isomurasaki, Hosozozanemo, Himegoshigusu, Giant genus Isomagusa, Suginasugusa: Suginasusa, Isobashou: Isobasho, Fujimatsumo: Fujimatsumo, Sawweed: Haeshinosakiba, Kawamoku : Kawamoku, Aokawa Mozuku, Himekawa Mozuku, Ideyugogome: Ideyugogome, Okichimozuku: Okimozu, Otosa, Chinomo: Chinomo, Chisujinori, Chisujinori, etc.).

  Axial algae (Shajikumo, Shiratamamo, Hoshitsurimo: Hoshitsurimo, Rikunotumusu, Flaskmo: Himebumo, Chaboflasmo, Tripera, etc.), yellow algae (Hikarimo: Hikarimo, etc.), etc.

  Animal-derived materials include chicken crown extract, cow or pig, human placenta extract, cow or pig stomach, duodenum, intestine, spleen extract or degradation product thereof, cow or pig brain tissue extraction Collagen hydrolysates (acids, alkalis, enzymes, etc.) of products, cattle / pigs or fish, collagen derivatives such as water-soluble collagens or acylated collagens, elastin or elastin hydrolysates of cattle or pigs (acids, alkalis, enzymes, etc.) ) Or water-soluble elastin derivative, keratin and its degradation product or their derivatives, silk protein and its degradation product or their derivatives, pig or cow blood cell protein degradation product (globin peptide), cow or pig hemoglobin degradation product (hemin) Hematin, heme, protoheme, heme iron, etc.), milk, casein and degradation products thereof or derivatives thereof, skim milk powder and degradation products thereof or derivatives thereof Lactoferrin or decomposed product thereof, egg ingredients, fish hydrolyzate, nucleic acid-related substance (ribonucleic acid, deoxyribonucleic acid), or the like.

  As a material derived from these plant or animal-based materials, any site, cell, tissue, organ, metabolite, etc. derived from a gene recombinant or a cell fusion can also be used. Furthermore, for example, an undifferentiated cell group, a cell group in the middle of differentiation, or the like obtained by culturing an arbitrary site, cell, tissue, organ or the like can also be used.

(14) Marine components Seawater such as deep seawater, for example, sea salt, dried seawater, inorganic salt (sodium chloride, magnesium chloride, potassium chloride, etc.), sea mud or mud (from the Dead Sea or Atlantic Ocean) Fango), for example, Italian mud, German fungo, Eifelfango, Freiburg fungo, etc. Sea mud or mud from various locations (containing components: silicon dioxide, titanium dioxide, aluminum oxide, iron oxide, manganese oxide, sodium oxide, potassium oxide, Magnesium oxide, calcium oxide, strontium oxide, sodium, potassium, magnesium, calcium, chromium, iron, copper, nickel, zinc, lead, manganese, arsenic, water), saintoku stone, etc.

(15) Microbial culture metabolite Yeast metabolite, yeast extract, bacterial metabolite, bacterial extract, mold or actinomycete metabolite, mold or actinomycete extract, natto metabolite, natto extract, rice fermentation extract , Fermented rice bran (red rice bran, white birch) extract, Euglena extract or decomposition product thereof or water-soluble derivative thereof, trehalose or derivative thereof, lactic acid fermentation product of raw milk or skim milk powder, fermented lactic acid bacteria of legumes, coconut plant Lactic acid bacteria fermented products.

(16) α-hydroxy acids Glycolic acid, citric acid, malic acid, tartaric acid, lactic acid and the like.

(17) Inorganic pigments Silicic anhydride, magnesium silicate, talc, kaolin, bentonite, mica, titanium mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, Bengala, black iron oxide, gunjo, chromium oxide, chromium hydroxide, carbon black, calamine, etc.

(18) UV absorber / blocker benzophenone derivatives (2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid, sodium 2-hydroxy-4-methoxybenzophenone-5-sulfonate, dihydroxy Dimethoxybenzophenone, dihydroxydimethoxybenzophenone-sodium sulfonate, 2,4-dihydroxybenzophenone, tetrahydroxybenzophenone, etc.), paraaminobenzoic acid derivatives (paraaminobenzoic acid, ethyl paraaminobenzoate, glyceryl paraaminobenzoate, amyl paradimethylaminobenzoate, Octyl paradimethylaminobenzoate), methoxycinnamate derivatives (ethyl paramethoxycinnamate, isopropyl paramethoxycinnamate, octyl paramethoxycinnamate, 2-ethoxycinnamic acid cinnamate) Til, sodium paramethoxycinnamate, potassium paramethoxycinnamate, diparamethoxycinnamate mono-2-ethylhexanoate glyceryl, etc.), salicylic acid derivatives (octyl salicylate, phenyl salicylate, homomenthyl salicylate, dipropylene glycol salicylate, ethylene glycol salicylate, myristyl salicylate, Methyl salicylate, etc.), anthranilic acid derivatives (methyl anthranilate, etc.), urocanic acid derivatives (urocanic acid, ethyl urocanate, etc.), coumarin derivatives, amino acid compounds, benzotriazole derivatives, tetrazole derivatives, imidazoline derivatives, pyrimidine derivatives, dioxane derivatives , Camphor derivative, furan derivative, pyrone derivative, nucleic acid derivative, allantoin derivative, nicotinic acid derivative, vitamin B6 derivative Body, umbelliferone, esculin, benzyl cinnamate, cinoxalate, oxybenzone, dioxybenzone, octabenzone, thrisobenzone, benzoresorcinol, arbutin, guaiazulene, shikonin, baicalin, baicalein, berberine, neoheliopan, escalol, zinc oxide, talc, kaolin etc.

(19) Whitening agent Paraaminobenzoic acid derivative, salicylic acid derivative, anthranilic acid derivative, coumarin derivative, amino acid compound, benzotriazole derivative, tetrazole derivative, imidazoline derivative, pyrimidine derivative, dioxane derivative, camphor derivative, furan derivative, pyrone derivative, Nucleic acid derivatives, allantoin derivatives, nicotinic acid derivatives, vitamin C or derivatives thereof (vitamin C phosphate magnesium salt, vitamin C glucoside, etc.), vitamin E or derivatives thereof, kojic acid or derivatives thereof, oxybenzone, benzophenone, arbutin, guaiazulene, Shikonin, baicalin, baicalein, berberine, placenta extract, ellagic acid, lucinol, etc.

(20) Tyrosinase activity inhibitor Vitamin C or a derivative thereof (vitamin C phosphate magnesium salt, vitamin C glucoside, etc.), hydroquinone or a derivative thereof (hydroquinone benzyl ether, etc.), kojic acid or a derivative thereof, vitamin E or a derivative thereof, N-acetyltyrosine or derivatives thereof, glutathione, hydrogen peroxide, zinc peroxide, placenta extract, ellagic acid, arbutin, lucinol, silk extract, plant extract (camille, mulberry, gardenia, pearl oyster, clam, clam, mugwort, honeysuckle) , Kihada, dokudami, matsuhod, pearl barley, mussel, hops, hawthorn, eucalyptus, yarrow, altea, keihi, mankaishi, hamamelis, yamagwa, lifelong grass, bellflower, toshishi, zetsugo, gyu, mao, senkyu, dod Tsu, Psycho, storm, littoralis, Scutellaria, peony bark, cranesbill, kudzu root, licorice, Fushi aloe, Cimicifuga, safflower, green tea, black tea, Ali elixir), and the like.

(21) Melanin pigment reduction / decomposition substance Phenylmercury hexachlorophene, mercuric oxide, mercuric chloride, hydrogen peroxide, zinc peroxide, hydroquinone or its derivative (hydroquinone benzyl ether).

(22) Turnover promoting action / cell activator hydroquinone, lactic acid bacteria extract, placenta extract, ganoderma extract, vitamin A, vitamin E, allantoin, spleen extract, thymus extract, yeast extract, fermented milk extract, plant extract (aloe, Ogon, Japanese horsetail, Gentian, Burdock, Shikon, Carrot, Camelis, Hop, Yokuinin, Odori, Semburi, Japanese cypress, Japanese red snapper, Achacha, Hypericum, Cucumber, Necklace, Rosemary, Parsley).

(23) Astringents Succinic acid, allantoin, zinc chloride, zinc sulfate, zinc oxide, calamine, zinc paraphenolsulfonate, potassium aluminum sulfate, resorcin, ferric chloride, tannic acid (including catechin compounds) and the like.

(24) Active oxygen scavenger SOD, catalase, glutathione peroxidase and the like.

(25) Antioxidant Vitamin C or a derivative thereof or a salt thereof, stearic acid ester, vitamin E or a derivative thereof, nordihydrogua cetelenic acid, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), hydroxytyrosol , Parahydroxyanisole, propyl gallate, sesamol, sesamorin, gossypol, propolis, etc.

(26) Lipid peroxide production inhibitor β-carotene, plant extract (sesame culture cell, hamcha, hypericum, hamamelis, clove, melissa, birch, birch, salvia, rosemary, minamitenji, jitsu, ginkgo, green tea).

(27) Anti-inflammatory agent Ictamol, indomethacin, kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d-camphor, dl-camphor, hydrocortisone, guaiazulene, camazulene, chlorpheniramine maleate, glycyrrhizic acid or its Salt, glycyrrhetinic acid or a salt thereof, licorice extract, sicon extract, age extract, propolis, etc.

(28) Antibacterial / bactericidal / disinfectant Acrinol, sulfur, calcium gluconate, chlorhexidine gluconate, sulfamine, mercurochrome, lactoferrin or hydrolyzate thereof, alkyldiaminoethylglycine chloride solution, triclosan, sodium hypochlorite, chloramine T, Sarashi powder, iodine compound, iodoform, sorbic acid or its salt, propionic acid or its salt, salicylic acid, dehydroacetic acid, parahydroxybenzoic acid esters, undecylenic acid, thiamine lauryl sulfate, thiamine lauryl nitrate, phenol, cresol, p -Chlorophenol, p-chloro-m-xylenol, p-chloro-m-cresol, thymol, phenethyl alcohol, O-phenylphenol, Irgasan CH3565, halocarban, hexachlorophene, chlorohexidine, ester Tanol, methanol, isopropyl alcohol, benzyl alcohol, ethylene glycol, propylene glycol, 2-phenoxyethanol, 1,2-pentanediol, zinc pyridione, chlorobutanol, isopropylmethylphenol, nonionic surfactant (polyoxyethylene lauryl ether, Polyoxyethylene nonyl phenyl ether, polyoxyethylene octyl phenyl ether, etc.), amphoteric surfactant, anionic surfactant (sodium lauryl sulfate, lauroyl sarcosine potassium, etc.), cationic surfactant (cetyltrimethylammonium bromide, benzalkco chloride) Nium, benzethonium chloride, methyl rosaniline chloride), formaldehyde, hexamine, brilliant green, malachite green, crystal bio Tsu DOO, Jamaru photosensitive No. 101-containing, photosensitizer No. 201, photosensitizer No. 401, N- long chain acyl basic amino acid derivative and its acid shark salts, zinc oxide, hinokitiol, Sophora root, propolis, and the like.

(29) Moisturizer Glycerin, propylene glycol, 1,3-butylene glycol, hyaluronic acid or its salt, polyethylene glycol, chondroitin sulfate or its salt, water-soluble chitin or its derivative or chitosan derivative, pyrrolidone carboxylic acid or its salt, lactic acid Sodium, mini Sasanishiki extract, natto bacteria metabolite, natto extract, loofah water, birch (birch) or red pine sap, plant-derived ceramide, animal-derived ceramide

(30) Elastase activity inhibitor diisopropyl fluorophosphate, plant extract (eg, gongoid, hypericum, clara, mulberry leaves, keihi, genokosho, comfrey, salvia, elderberry, bodaiju, button pie), seaweed extract and the like.

(31) Hair preparations Selenium disulfide, alkylisoquinolinium bromide solution, zinc pyrithione, biphenamine, thianthol, castari tincture, pepper tincture, pepper tincture, quinine hydrochloride, strong ammonia water, potassium bromate, sodium bromate, thio Glycolic acid and the like.

(32) Antiandrogens Follicular hormones (estrone, estradiol, ethinyl estradiol, etc.), isoflavones, oxendrone, etc.

(33) Peripheral vascular blood flow promoter Vitamin E or derivatives thereof, assembly extract, garlic extract, carrot extract, aloe extract, gentian extract, toki extract, cephalanthin, carpronium chloride, minoxidil and the like.

(34) Stimulants Pepper tincture, nonyl acid vanillamide, cantalis tincture, ginger tincture, peppermint oil, l-menthol, camphor, benzyl nicotinate and the like.

(35) Metabolic activator Photosensitive element 301, hinokitiol, pantothenic acid or derivatives thereof, allantoin, placenta extract, biotin, pentadecanoic acid glyceride and the like.

(36) Antiseborrheic agent Pyridoxine or a derivative thereof, sulfur, vitamin B6 and the like.

(37) Keratolytic agent Resorcin, salicylic acid, lactic acid, urea and the like.

(38) Oxidizing agent Hydrogen peroxide water, sodium persulfate, ammonium persulfate, sodium perborate, urea peroxide, sodium percarbonate, sodium tripolyphosphate, sodium bromate, potassium bromate, sodium pyrophosphate, Sodium orthophosphate, sodium silicate hydrogen peroxide adduct, sodium sulfate hydrogen peroxide adduct, sodium chloride hydrogen peroxide adduct, β-tyrosinase enzyme solution, mushroom extract, etc.

(39) Hair remover Inorganic reducing agents such as strontium sulfate, sodium sulfide, barium sulfide, calcium sulfide, thioglycolic acid or its salts (calcium thioglycolate, sodium thioglycolate, lithium thioglycolate, magnesium thioglycolate) Strontium thioglycolate) and the like.

(40) Hair swelling agents such as ethanolamine, urea and guanidine.

(41) Dye agent
5-aminoorthocresol, 2-amino-4-nitrophenol, 2-amino-5-nitrophenol, 1-amino-4-methylaminoanthraquinone, 3,3'-iminodiphenol, 2,4-diaminophenoxyethanol hydrochloride 2,4-diaminophenol hydrochloride, toluene-2,5-diamine hydrochloride, nitroparaphenylenediamine hydrochloride, paraphenylenediamine hydrochloride, N-phenylparaphenylenediamine hydrochloride, metaphenylenediamine hydrochloride, orthoaminophenol, N-phenyl acetate Paraphenylenediamine, 1,4-diaminoanthraquinone, 2,6-diaminopyridine, 1,5-dihydroxynaphthalene, toluene-2,5-diamine, toluene-3,4-diamine, nitroparaphenylenediamine, paraaminophenol, para Nitroorthophenylenediamine, paraphenylenediamine, paramethylamino Phenol, picramic acid, sodium picramate, N, N'-bis (4-aminophenyl) -2,5-diamino-1,4-quinonediimine, 5- (2-hydroxyethylamino) -2-methylphenol, N -Phenylparaphenylenediamine, metaaminophenol, metaphenylenediamine, 5-aminoorthocresol sulfate, 2-amino-5-nitrophenol sulfate, orthoaminophenol sulfate, orthochloroparaphenylenediamine sulfate, 4,4'-diamino sulfate Diphenylamine, 2,4-diaminophenol sulfate, toluene-2,5-diamine sulfate, nitroparaphenylenediamine sulfate, paraaminophenol sulfate, paranitroorthophenylenediamine sulfate, paranitrometaphenylenediamine sulfate, paraphenylenediamine sulfate, parasulfate sulfate Methylaminophenol, metaaminosulfate Diol, metaphenylenediamine sulfate, catechol, diphenylamine, α-naphthol, hydroquinone, pyrogallol, phloroglucin, gallic acid, resorcin, tannic acid, 2-hydroxy-5-nitro-2 ', 4'-diaminoazobenzene-5'-sulfone Sodium acid, hematein, etc.

(42) Fragrance Natural animal fragrance such as musk, civet, castorium, ambergris, anise essential oil, angelica essential oil, ylang ylang essential oil, iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guayakwood essential oil, Cumin essential oil, black letter essential oil, cinnamon essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriandel essential oil, perilla essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, gingergrass essential oil, cedar essential oil, spearmint essential oil, western peppermint essential oil, Otsuka perfume essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, winter green essential oil, trout balsam essential oil, buttery essential oil, rose essential oil, palmarosa essential oil, persimmon essential oil, hiba essential oil, sandalwood essential oil, petit gren essential oil, bay essential oil, vetiva essential oil, bergamot Spirit , Peru balsam essential oil, boad rose essential oil, melamine essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary essential oil, Japanese mint essential oil, and other synthetic flavors Fragrance etc.

(43) Dye and Colorant Red Cabbage Dye, Red Rice Dye, Akane Dye, Anato Dye, Ikumi Dye, Turmeric Dye, Enju Dye, Krill Dye, Amber Dye, Caramel, Gold, Silver, Gardenia Dye, Corn Dye, Onion Dye , Tamarind pigment, spirulina pigment, buckwheat whole plant pigment, cherry pigment, laver pigment, hibiscus pigment, grape juice pigment, marigold pigment, purple potato pigment, purple yam pigment, lac pigment, rutin etc.

(44) Sweeteners Sugar, sweet tea, fructose, arabinose, galactose, xylose, stevia, mannose, maltose, honey, glucose, miraculin, monelin, licorice extract and the like.

(45) Nutrient enhancer Shell calcined calcium, cyanocobalamin, yeast, wheat germ, egg yolk powder, hemicellulose, heme iron and the like.

(46) Dairy products Milk, cheese, fresh cream, butter, margarine, powdered milk, whey, condensed milk, etc.

(47) Others Hormones, sequestering agents, pH adjusters, chelating agents, antiseptic and antifungal agents, cooling agents, stabilizers, emulsifiers, animal and vegetable proteins and their degradation products, animal and vegetable polysaccharides And its degradation products, animal and plant glycoproteins and their degradation products, blood flow promoters, anti-inflammatory agents, anti-inflammatory agents, anti-allergic agents, wound treatment agents, foam increasing agents, thickeners, enzymes, purified water (electronic Water, small clustering, etc.), oral preparations, deodorants / deodorants, bittering agents, seasonings, etc. are raised, and in combination with these, the effects of the present invention or other cosmetic effects are added Or a synergistic improvement can be expected.

[Manufacture of various health foods, health drinks, and cosmetic compositions]
Various cosmetic compositions according to the present invention were prepared. The prescription example is shown below. Ethanol is abbreviated as EtOH, and 1,3-butylene glycol is abbreviated as 1,3-BG.
(Beauty / health tablet)
The composition containing the following additives was compressed into tablets. weight%
1. Proteoglycan powder obtained in Production Example 1 1
2. N-acetylglucosamine 5
3. Chondroitin sodium sulfate 1
4). Collagen powder for food 5
5. Guarana 50% EtOH extraction powder 5
6). Royal jelly powder 5
7). Sodium ascorbate 1
8). Lactose 35
9. Corn starch 25
10. Hydroxypropyl cellulose 4
11. Magnesium stearate 1
12 Dextrin 12

(Cosmetic food for beauty and health)
The following additives were filled into gelatin capsules to produce a capsule-type food (one package).
Weight 1. 0.05 g proteoglycan powder obtained in Production Example 2
2. Collagen powder for food 0.1g
3. Collagen tripeptide 0.05g
4). Pig placental water extraction powder (50% dextrin) 0.01g
5. Soy peptide 0.02g
6). Sodium ascorbate 0.03g

(Natural supplements for beauty and health)
Granules (4 packs) containing the following additives were produced. Weight 1. Collagen powder for food 5g
2. Fat content 0.024g
3. 0.01g sodium aspartate
4). Vitamin A 0.002g
5. Vitamin B1 0.003g
6). Vitamin B2 0.004g
7). Vitamin B6 0.004g
8). Vitamin B12 0.0006g
9. Sodium ascorbate 0.03g
10. Vitamin E 0.002g
11. Niacin 0.004g
12 Pantothenic acid 0.001g
13. Arginine 0.6g
14 Leucine 0.6g
15. Valine 0.4g
16. Eggshell calcium 0.01g
17. 0.01 g of proteoglycan powder obtained in Production Example 1
18. Hydroxypropylcellulose 2g
19. N-acetylglucosamine 5g
20. 1g sodium chondroitin sulfate
21. Dextrin 2g

(Beauty and health drinks)
weight%
1. Pomegranate water extract 20
2. Rose Hot Water Extract 20
3. Sodium ascorbate 0.1
4). Proteoglycan powder obtained in Production Example 2 0.01
5. Porcine placental water extract 0.02
6). Honey 5
7). Citric acid 0.1
8). Water-soluble collagen for food 2
9. dl-malic acid 0.1
10. D-sorbitol solution (70%) 10
11. Sodium benzoate 0.03
12 Perfume appropriate amount13. Residue with 100 purified water

(Lactic acid bacteria fermented beverage for beauty and health)
weight%
1. Proteoglycan solution obtained in Production Example 1 (solid content: 1.0%) 2
2. Carrot juice lactic acid bacteria fermentation 40
3. Orange concentrated reduced juice 10
4). Sodium ascorbate 0.2
5. Lactulose syrup 1
6). Citric acid 0.1
7). Water-soluble collagen for food 2
8). Perfume appropriate amount 9. Residue with 100 purified water

(candy)
Weight 1. Minamata 500g
2. 500g sugar
Is mixed and heated at 120 ° C. under reduced pressure (−500 mmHg), and then cooled to 90 ° C. to this,
3. 5 g proteoglycan concentrate obtained in Production Example 2
4). 30g orange fruit ground
5. Stevia extract 20g
6). Citric acid 10g
7). Fragrance 1.5mL
Was added and kneaded sufficiently, and this was punched and molded into a circle having a diameter of 15 mm and a thickness of 8 mm to produce a bag.

(lotion)
weight%
1. Proteoglycan solution obtained in Production Example 3 (solid content: 1.0%) 1
2. Sorbit 2
3.1,3-BG 2
4). Polyoxyethylene oleyl ether (25 EO adduct) 2
5. EtOH 20
6). Ascorbic acid 2-glucoside 0.2
7). Green tea 30% EtOH extract 0.01
8). Chamomile 30% EtOH extract 1.0
9. pH adjuster appropriate amount10. Phenoxyethanol appropriate amount11. Residue with 100 purified water

(Milky lotion)
weight%
1. Proteoglycan solution obtained in Production Example 4 (solid content: 1.0%) 1
2. Squalane 3
3. Vaseline 1
4). Stearyl alcohol 0.3
5. Sorbitan monostearate 1.5
6). Polyoxyethylene (20) sorbitan monooleate 3
7.1,3-BG 10
8). Ascorbic acid phosphate magnesium salt 0.1
9. Acerola 30% 1,3-BG extract 1.0
10. Scarlet root 30% 1,3-BG extract 1.0
11. Sodium hyaluronate 0.005
12 Preservative (paraoxybenzoic acid ester) Residue with 100 purified water

(Milky lotion)
weight%
1. Proteoglycan solution obtained in Production Example 5 (solid content: 1.0%) 2.0
2. Squalane 8.0
3. Jojoba oil 7.0
4). Maltose 3.0
5. Ascorbic acid phosphate magnesium salt 0.1
6). Cetyl alcohol 1.5
7). Glycerol monostearate 2.0
8). Polyoxyethylene (20) cetyl ether 3.0
9. Polyoxyethylene (20) sorbitan monooleate 2.0
10. Yuzu 50% 1,3-BG extract (containing Yuzuceramide) 1.0
11. Porcine placenta extract 0.1
12.1, 3-BG 1.0
13. Glycerin 2.0
14 Sodium hyaluronate 0.001
15. Preservative (paraoxybenzoic acid ester) Residue with 100 purified water

(Cosmetic oil)
mass%
1. Proteoglycan solution obtained in Production Example 3 (solid content: 1.0%) 1.0
2. Liquid paraffin 30.0
3. Squalane 20.0
4). Olive oil 20.0
5. Isopropyl palmitate 10.0
6). Olive oil 1.0
7). Shea fat 1.0
8). Butylhydroxyanisole 0.1
9. DL-α-tocopherol derivative 0.1
10. Ascorbic acid phosphate magnesium salt 0.1
11. Bilberry 30% 1,3-BG extract 1.0
12 Indigo Luros 30% 1,3-BG extract 1.0
13. Preservative (paraoxybenzoic acid ester) Perfume appropriate amount 15. Residue with 100 purified water

(cream)
mass%
1. Proteoglycan solution obtained in Production Example 4 (solid content: 1.0%) 1.0
2. Sara honey bees 11.0
3. Liquid paraffin 22.0
4). Lanolin 10.0
5. Olive oil 5.0
6). Chamomile oil 5.0
7). Palm oil 5.0
8). Oat bark 30% 1,3-BG extract 2.0
9. Water-soluble collagen solution 3.0
10. Porcine placenta extract 0.1
11. Ascorbic acid phosphate magnesium salt 0.1
12 Liquid shea fat 1.0
13. Hydroxypropyl cellulose 2.0
14 Residue with 100 purified water

(Face washing cream)
mass%
1. Proteoglycan solution obtained in Production Example 5 (solid content: 1.0%) 2.0
2. Stearic acid 10.0
3. Palmitic acid 10.0
4). Myristic acid 12.0
5. Lauric acid 4.0
6). Oleyl alcohol 1.5
7). Lanolin derivative (EO adduct) 1.0
8). Glycerin 18.0
9. Potassium hydroxide 6.0
10. Glycyrrhizic acid 2-potassium 1.0
11. Soap nut (Rita) extracted saponin 0.1
12 Perfume appropriate amount13. Residue with 100 purified water

(Hair shampoo)
weight%
1. Lauryl sulfate triethanolamine 5
2. Sodium polyoxyethylene lauryl ether sulfate 12
3. 1,3-butylene glycol 4
4). Lauric acid diethanolamide 2
5. 2-Phenylchromone 0.01
6). Proteoglycan solution obtained in Production Example 3 (solid content: 1.0%) 1.0
7). Arginine or its salt 0.2
8). Preservative (ethyl paraben) Appropriate amount 9. Residue with 100 purified water

  The production method according to the present invention can extract and purify proteoglycan with high efficiency and high purity by a very simple method as compared with the conventional production method. In addition, since a large amount of organic solvent such as alcohol or acetone is not required, there is no danger of flammability and it is suitable for the health of workers, and the proteoglycan that is the target substance is removed together with lipids such as activated carbon. This eliminates the need to add a high-precision filtration process that prevents the collection amount from being reduced and the mixing of fine-particle activated carbon.

The proteoglycan produced by the method according to the present invention has higher quality than the proteoglycan obtained by the conventional method, such as color, odor, taste and solubility, because there are few hybrids such as lipid and collagen. The effects of promoting the proliferation of cartilage progenitor cells, the promotion of cartilage differentiation, and the suppression of cartilage calcification are also high, and the utilization as a raw material for health foods, health drinks, pharmaceutical compositions and cosmetic compositions is remarkably high.

Claims (3)

A method for producing a proteoglycan comprising the steps of extracting proteoglycan from a biological sample containing proteoglycan and adsorbing and removing the lipid component from the resulting extract using powdered cellulose and / or an oil absorbing mat. The production method according to claim 1, comprising a step of separating and purifying proteoglycan after the step of removing the lipid component. The method according to any one of claims 1-2, wherein the biological sample containing proteoglycan is fish, mollusk, avian, or mammalian cartilage tissue, muscle tissue, or skin tissue.