JP5538675B2 - Pelago algae extract-containing composition, food and cosmetics containing the Pelago algae extract-containing composition - Google Patents

Description

  The present invention is an extract obtained from the algal body of the sarcinolysis of Pelago algae which is a kind of microalgae, and contains, for example, a Pelago algae extract having actions such as hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability and antibacterial activity The present invention relates to a composition and a food and a cosmetic containing the Pelago algae extract-containing composition.

  In recent years, the number of allergic patients has been increasing rapidly, and not only the constitution but also the influence of living environment and disorder of eating habits can be considered. As tea ingredients and perilla leaf extracts are already widely recognized as anti-allergic food materials that are safe for the human body and have no side effects, clinical studies and animal experiments have been confirmed to alleviate and prevent allergic symptoms. (Otolaryngology perspective; Vol. 38, No. 4, 519 to 532 1995, inflammation; Vol. 15, No. 2, 167 to 173 1995) (Fragrance journal; Vol. 23, No. 5, 43 to 48 1995) Vol. 23, No. 7, 90-94 1995).

  Plant-derived hyaluronidase inhibitors can be taken daily to prevent or improve inflammation, throat swelling, hay fever, cough, etc. associated with colds. At the same time, cosmetics containing these hyaluronidase inhibitors It is possible to improve the itching of the skin, and as a more potent hyaluronic acid degradation inhibitor, it suppresses the decrease in the hyaluronic acid content contained in the skin, arterial wall, joint space, etc. There are known inventions that increase the moisture retention and flexibility of the skin, prevent arterial effects associated with aging, and contribute to the improvement of arthritis. (See Patent Document 1)

  On the other hand, stress, ultraviolet rays, air pollution, and the like cause active oxygen, and when oxidation progresses in the body, it appears as a lifestyle-related disease or aging. In the modern social environment, stress is not an exaggeration to say that it is the cause of all illness, and it is difficult to deal with it. In addition, protecting the body from various active oxygens such as an increase in the amount of ultraviolet rays due to ozone layer destruction and exhaust gas from automobiles is important for maintaining health. In recent years, for the purpose of reducing stress, it is also recommended to use vitamin C and vitamin B group supplements, which are water-soluble antioxidants, and vitamin C, a water-soluble antioxidant, is also used to prevent and improve stains. ing.

  Hyaluronic acid, which is widely recognized as a cosmetic material, is fermented using animal origin and microorganisms and has a high moisturizing effect, so it is used in many products as a moisturizing agent for cosmetics. Hyaluronic acid is not only a skin moisturizing effect but also a synovial fluid and cartilage component and is used as a food to suppress inflammation.

  Furthermore, most antibacterial agents currently used in cosmetics are synthetic raw materials, and parabens are considered to have a high possibility of causing causative dermatitis and allergic dermatitis. Furthermore, its chemical structure is similar to the environmental hormone nonylphenol, and it is said to cause abnormalities in the normal hormonal action of humans and to accumulate in the human body and adversely affect the fetus during pregnancy.

  By the way, in recent years, cosmetics and foods made from natural algae, which are natural materials, have attracted attention. As a technique for using this microalgae, for example, a polysaccharide constituting a unicellular algae, which is a polysaccharide constituting an envelope peeled from a unicellular algae belonging to the Pelagophyceae There is a polysaccharide obtained from the outer coat of algae (see Patent Document 2).

  The known technique of Patent Document 2 does not use the algal body of Pelago algae itself, but relates to a technique that uses a polysaccharide constituting the outer shell peeled from the single-celled algae of Pelago algae.

Japanese Patent Laid-Open No. 7-10768 Japanese Patent Laid-Open No. 2005-28197

  Conventionally, sarcinolysis, which is a yellow plant gate Pelagophyceae sarcinolysis, has a polysaccharide around the alga body and has the property of being easily detached by heat. So far, biodegradable plastics and water purification agents have been developed using the polysaccharide, but there are no reports on structural analysis or physiological activity.

  Therefore, as an object of the present invention, there is provided a material or antibacterial agent having both hyaluronidase inhibitory activity, antioxidant activity and moisturizing ability, which contains an extract from a kind of microalgae not reported in the past as an active ingredient. It is an object of the present invention to provide foods and cosmetics containing the agent.

  As a result of intensive studies on microalgae in order to solve the above-mentioned problems and achieve the intended purpose, the present inventors have found that the extract of microalgae belonging to sarcinolysis, which is the yellow plant Pelagophyceae sarcinolysis Was found to have hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability or antibacterial activity, and the present invention was completed.

Therefore, in the present invention, as a first means according to the present invention as a specific means for solving the problems of the above-described conventional example, the algal body of the sarcinolysis that is a yellow plant gate Pelagophyceae Salsinolysis is frozen after desalting. After drying, water was added to the dried alga body and the mixture was stirred and extracted at 90 ° C. or higher for 3 hours , and then the hot water extract obtained by filtering the supernatant by centrifugation was concentrated to 1/5 to 1/6, It can be obtained as a precipitate after adding ethanol to it overnight, or after adding 50% ethanol to the dried alga and performing reflux extraction at 90 ° C. or more for 3 hours , and then supernatant by centrifugation Pelago alga extract-containing composition characterized by containing an extract having hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability and antibacterial activity obtained by filtering and completely removing ethanol You It is intended.

Further, as a second invention, the composition contains 0.1% of a Pelago algae extract-containing composition having the hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability and antibacterial activity of the first invention. As a third invention, there is provided a cosmetic comprising 0.1 to 0.3% of the Pelago algae extract-containing composition of the first invention.

The Pelago algae extract-containing composition according to the first aspect of the present invention has hyaluronidase inhibitory activity, antioxidative activity, moisturizing ability and antibacterial activity extracted from the algal body of Salsinoclisis, which is a yellow plant Pelagophyceae Salsinolytica. Since it contains an extract with a combination, it can be used as a material that takes advantage of these characteristics, and it can be used as a product that takes advantage of these characteristics , such as a hyaluronidase inhibitor or an antibacterial agent. In addition , foods and cosmetics that make use of characteristics such as hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability, and antibacterial activity can be obtained by incorporating the above-mentioned composition of Pelago algae extract into foods or cosmetics. There is an excellent effect that it can be provided.

Next, the present invention will be described in detail based on specific embodiments.
Perago alga extract containing composition according to the embodiment of the present invention are those containing an extract extracted from algae of a yellow plant Gate Perago Momo (Pelagophyceae) Sarushinokurishisu th (Sarcinochrysidales) Sarushinokurishisu (Sarcinochrysis) Moreover, it is the foodstuffs and cosmetics which mix | blended the said Pelago algae extract containing composition.

  As the sarcinolysis of Pelago algae, which is a kind of microalgae used as a raw material, natural ones can be used, but it is desirable to grow them by culturing from the viewpoint of stable supply. Since algae perform photosynthesis and generate their own energy, the culture can be performed by a normal culture method using a culture medium for algae under light irradiation. However, since the culture method differs slightly depending on the algae, it is necessary to set optimal culture conditions. The specific culture conditions are as follows.

[Culture of sarcinolysis]
If it is used when cultivating general marine algae, there will be no special restriction | limiting, For example, f / 2 medium can be used. That is, a solution prepared by adding 75 mg of NaNO ₃ , 10 mg of Na ₂ SiO ₃ .9H ₂ O, 0.5 μg of Thiamine HCl, 0.5 μg of Biotin, and 100 μg of Cyanocobalamin to 999 mL of fresh filtered seawater was prepared, and EDTA 2Na · 2H ₂ O 4.4 g / L, FeCl ₃ · 6H ₂ O 3.16 g / L, CoSO ₄ · 7H ₂ O 12 mg / L, ZnSO ₄ · 7H ₂ O 21 mg / L, MnSO ₄ · 4H ₂ O 180 mg / L , CuSO ₄ · 5H ₂ O 7 mg / L, Na ₂ MoO ₄ · 2H ₂ O 7 mg / L containing 1 mL of a trace element mixture was added, then autoclaved at 121 ° C. for 20 minutes, and NaH ₂ PO ₄ · The medium was prepared by adding 6 mg of H ₂ O. The medium was inoculated with Sarcinochrysis sp. Cells and cultured in a 2-liter glass flat flask. The illuminance at this time is preferably set to 10-30 μEinsteins / m ² / sec, and it is preferably performed under continuous light irradiation. The culture temperature is 20-30 ° C, preferably around 25 ° C.

  A method for producing an extract-containing composition containing Pelago algae by extracting an extract from the algal bodies of the sarcinolysis obtained in this way, a hyaluronidase inhibitory activity test, an antioxidant test, and a moisturizing composition of the Pelago algae extract-containing composition The present invention will be described in more detail by giving more specific examples regarding tests, safety tests, antibacterial tests, and methods for producing anti-inflammatory agents, antioxidants, humectants or antibacterial agents.

[Example 1]
In this Example 1, it manufactured as follows.
(Extraction method of extract with hot water)
The harvested salsinolysis algal bodies were desalted and then freeze-dried. To 5 g of the obtained dried algal bodies, 1000 mL of water was added, and the mixture was extracted by stirring at 90 ° C. or more for 3 hours. Thereafter, centrifugation was performed, and the supernatant was filtered through a glass filter to obtain a hot water extract. This hot water extract was concentrated to 1/5 to 1/6, 4 volumes of ethanol was added and allowed to stand overnight, and then the precipitate was recovered and freeze-dried to obtain a powder. It was set to 1.

[Confirmation of acid sugar]
Since some algae having a physiological activity are known for algae, in order to investigate the acidic sugar contained in Example 1, purification with an ion exchange resin was performed. DEAE-TOYOPAR (650M) was used as the ion exchange resin, and the elution solvent was gradient elution with water and 1M sodium chloride solution. The results are shown in Table 1.

(Table 1)

[Confirmation of component sugar]
Next, in order to investigate the constituent sugars constituting Example 1, analysis by gas chromatography was performed. That is, 10 mg of sodium borohydride and 0.1 mL of 1M aqueous ammonia were added to the hydrolyzed Example 1, and maintained at 40 ° C. for 90 minutes, and then the reduction reaction was stopped with 0.1 mL of acetic anhydride. Next, 0.2 mL of 1-methylimidazole and 2 mL of acetic anhydride were added in this order, and acetylation was performed by applying ultrasonic waves at room temperature for 40 minutes. Further, after stopping the reaction with about 2 mL of distilled water, 2 mL of chloroform was added while cooling and extraction was performed. The chloroform layer was washed with distilled water, dehydrated with anhydrous sodium sulfate, and concentrated, and then a small amount of acetone was added. The results are shown in Table 2.

(Table 2)

  From the results of Tables 1 and 2, the polysaccharide of sarcinolysis is 95.8% neutral sugar and 3.69% acidic sugar, and the composition ratio is arabinose: xylose: mannose: glucose = 1.7: It was found to be 1.2: 0.4: 0.04.

[Test Example 1]
In Test Example 1, the hyaluronidase inhibitory activity of Example 1 was measured by the following method. The results are shown in Table 3.
(Test method for hyaluronidase inhibitory activity)
A test sample dissolved in 0.1 M acetate buffer (pH 4.0) and the same buffer are combined, taken into a 0.2 mL test tube, and a hyaluronidase solution (type IV-S final enzyme activity: 400 NF units) dissolved in the same buffer. / ML, 0.1 mL of Sigma), and incubated at 37 ° C. for 20 minutes. Further, final concentrations of activator solution (Compound 48/80 (Sigma), CaCl ₂ , and NaCl are 0.1 mg each. / ML, 2.5 mM, 0.15 M dissolved in the same buffer) was added and incubated at 37 ° C. for 20 minutes.

To this, 0.5 mL of a sodium hyaluronate solution (final concentration: 0.4 mg / mL) dissolved in the same buffer was added, incubated at 37 ° C. for 40 minutes, and then 0.2 mL of 0.4N sodium hydroxide solution was added. It was. After cooling with ice, 0.2 mL of boric acid solution (pH 9.1) was added, boiled for 3 minutes, cooled on ice, added with 6 mL of p-dimethylbenzaldehyde reagent, incubated at 37 ° C. for 20 minutes, and then measured after absorption at 585 nm. . As an index, DSCG (sodium cromoglycate) (Comparative Example 1) was used. In addition, the same buffer was used instead of the test sample as a control, and the same buffer was used instead of the enzyme solution as each blank. The inhibitory activity was expressed as an inhibition rate obtained from the following formula.
Inhibition rate (%) = [(A−B) − (C−D)] / (A−B) × 100
A: A _{585 of the} control solution
B: Control solution blank A ₅₈₅
C: A _{585 of the} sample solution
D: Sample solution blank A ₅₈₅

(Table 3)

  From the results of Table 3, the hyaluronidase inhibitory activity in the composition containing the extract of Pelago algae according to the present invention (Example 1) is DSCG (sodium cromoglycate) which is a histamine release inhibitor used as an index (Comparative Example 1). Of 2.4 times as much as the inhibitory activity.

  This test method is widely used as a primary screening for suppression of type I allergy such as hay fever. By the way, according to Japanese Patent Laid-Open No. 9-118627, this test method can prevent and improve inflammation, hay fever, cough, etc. by daily ingestion of a hot tea hot water extract that has the same activity as DSCG. It has also been reported that itching can be improved or skin itchiness can be improved by using cosmetics.

  In recent years, hay fever is also said to be a national disease, and about 20% of the people are diagnosed with hay fever, and it is thought that there are about 20% of reserves. In addition, the side effects of arrhythmia have occurred due to the use of antihistamines that are used as therapeutic agents for allergic rhinitis, bronchial asthma, dermatitis, etc., and development of antiallergic agents that are safe and free of side effects is being promoted. Since the tea is one of them, it is considered that the composition containing the Pelago algae extract according to the present invention has sufficient potential.

[Test Example 2]
In Test Example 2, the antioxidant activity of Example 1 was measured by the following method. The results are shown in Table 4.
(Antioxidation test method by β-carotene fading method)
Take 7.5 mL of β-carotene solution (100 mg / 100 mL, chloroform), 1.0 mL of linoleic acid solution (10 g / 100 mL, chloroform), and 5.0 mL of Tween 40 solution (20 g / 100 mL, chloroform) in an eggplant-shaped flask. After completely skipping chloroform, 500 mL of water was added and dissolved to prepare a linoleic acid-β-carotene solution.

  To 45 mL of this solution, 4 mL of 0.2 M phosphate buffer (pH 6.8) was added and gently stirred. 1 mL of the test sample was added thereto, and the mixture was heated at 55 ° C., and the absorbance at 470 nm was measured every 20 minutes. BHA (butylhydroxyanisole) (Comparative Example 2) was used as an index. Moreover, it carried out similarly about what did not add (beta) -carotene solution as each blank. For the antioxidant activity, the absorbance value immediately after addition of the test sample was taken as 100.

(Table 4)

  From the results of Table 4, the antioxidant activity in the composition containing Pelago algae extract (Example 1) according to the present invention is that of BHA (butylhydroxyanisole) (Comparative Example 2), which is an antioxidant used as an index. Similar antioxidant activity was observed at 1000 times the concentration.

[Test Example 3]
In Test Example 3, the moisture retention ability of Example 1 was measured by the following method. The results are shown in FIGS.
(Moisturizing test method)
Wash the inner forearm portion in advance measurement site with soap, temperature 22 ° C., after resting for 20 minutes or more at a measuring chamber kept at a humidity of 44%, 0.2% to inner forearm part test sample 10 [mu] L / cm ² The skin moisture content was measured using a skin moisture meter (SKIN DIAGNOSTIC SD 27). Sodium hyaluronate (Comparative Example 3) was used as an index. As a control, water was used in place of the test sample, and a blank area was measured as a blank. The amount of moisture increase was defined as 1 for the amount of skin moisture before application.

  From these results of FIG. 1 and FIG. 2, the moisturizing effect in the composition containing Pelago algae extract (Example 1) according to the present invention is 7 times the amount of water before coating after 1 minute of coating, and until 120 minutes thereafter. The water content of 3 to 5 times or more before coating was maintained. Sodium hyaluronate having a high moisturizing effect used as an index was 4 times the amount of water before coating 1 minute after coating, and about 3 times the amount of water before coating until 120 minutes thereafter. Furthermore, as a result of continuing to apply Example 1 and sodium hyaluronate for 7 days, Example 1 maintained a higher water content than sodium hyaluronate. From this, the moisturizing effect superior to the sodium hyaluronate with a high moisturizing effect was recognized by the composition containing the extract of Pelago algae (Example 1).

[Test Example 4]
In Test Example 4, the safety test of Example 1 was performed by the following method. The results are shown in Table 5.
(Safety test method by patch test)
Apply 0.3 mL of a 0.3% test sample (Example 1) and water (Comparative Example 4) to a patch test adhesive bandage (manufactured by Riba Tape Pharmaceutical Co., Ltd.) and apply it to the inside of the upper arm. The skin reaction was examined.

(Table 5)

  From the results of Table 5, the results of the patch test (safety test) in the composition containing the extract of Pelago algae according to the present invention (Example 1) showed no positive reaction in all cases of 6 subjects. The safety of the extract-containing composition was confirmed.

[Test Example 5]
In Test Example 5, the pH stability of Example 1 was examined by the following method. The results are shown in Table 6.
(PH stability test method)
0.1 mol / L HCl or 0.2 mol / L NaOH was added to a 0.3% test sample, adjusted to each pH, allowed to stand at the pH and appearance immediately after that, and at each temperature for one week, and the appearance was observed.

○：変化なし、△：濁り発生、×：沈殿発生
(Table 6)

○: No change, △: Turbidity, ×: Precipitation

  From the results of Table 6, the stability of the solution prepared by adjusting the composition containing Pelago algae extract (Example 1) according to the present invention to pH 3 to 11 shows a change in the appearance after standing at each temperature for 1 week. Therefore, the property stability was confirmed as a cosmetic raw material.

[Test Example 6]
In Test Example 6, the compatibility with the cosmetic raw material was examined for Example 1 by the following method. The results are shown in Table 7.
(Test method for compatibility with cosmetic ingredients)
A 0.3% test sample and the cosmetic raw materials described in Table 7 were mixed at a ratio of 1: 1, and left at the pH and appearance immediately after that and at each temperature for one week, and the appearance was observed.

○：変化なし、△：濁り発生、×：沈殿発生
(Table 7)

○: No change, △: Turbidity, ×: Precipitation

  From the results in Table 7, the stability of the solution obtained by mixing the composition containing the extract of Pelago algae (Example 1) according to the present invention with a cosmetic raw material does not change in appearance after standing at each temperature for one week. The property stability was confirmed as a cosmetic raw material.

[Example 2]
In Example 2, it was manufactured as follows.
(Extraction method of extract with alcohol)
The harvested algal bodies were desalted and then lyophilized. To 10 g of the obtained dried algal bodies, 100 mL of 50% ethanol was added, and reflux extraction was performed at 90 ° C. or higher for 3 hours. Thereafter, centrifugation was performed, and the supernatant was filtered through a glass filter. Then, ethanol was completely removed and freeze-dried to obtain a powder, which was designated as Example 2.

[Test Example 7]
In Test Example 7, the antibacterial activity of Example 1 and Example 2 was measured by the following method. The results are shown in Table 8.
(Antimicrobial test method by challenge test)
10 ⁶ S. aureus ( Staphylococcus aureus subsp. Aureus Rosenback 1884 IAM1011) pre-cultured per 1 mL of the test sample prepared at each concentration was inoculated and stored at 25 ° C. The number of viable cells was measured immediately after inoculation, 1 day, 3 days, and 7 days after using SCDLP medium and culturing at 30 ° C. for 48 hours. As an index (Comparative Example 5), paraben which is usually used as a preservative for cosmetics was used.

(Table 8)




(Unit: cfu / mL)

  From the results of Table 8, the antibacterial activity of the Pelago algae extract-containing composition of Example 1 according to the present invention was measured. As a result, antibacterial activity against Staphylococcus aureus was not observed. The antibacterial activity of the composition containing Pelago algae extracted with% ethanol against Staphylococcus aureus was 2000 times that of parabens, which are usually used as preservatives in cosmetics.

  With the increasing number of patients with sensitive skin due to atopic dermatitis, cosmetic allergic diseases, and recent lifestyles, the selection of raw materials used in cosmetics is important. The Pelago algae extract-containing composition extracted from 50% ethanol in Example 2 is a cosmetic raw material that can be expected as a preservative that is extracted from natural raw materials and uses a small amount.

  The action of the Pelago algae extract-containing composition extracted with hot water (Example 1) and the Pelago algae extract-containing composition extracted with alcohol (Example 2) are combined with food or cosmetics to achieve the action. It can be applied as a product having Hereinafter, the case where the composition containing Pelago algae extract is mixed with food or cosmetics will be described.

[Example 3]
In Example 3, a cookie containing 0.1% of Example 1 was produced with the composition shown in Table 9 as an example of a food containing the Pelago algae extract-containing composition. The food is not limited to the above-mentioned cookies. For example, functional foods, strawberries, troches, gums, yogurts, ice creams, puddings, jellies, mizuyokan, sprinkles, coffee drinks, juices, carbonated drinks, soft drinks Any food containing water, milk, whey drink, lactic acid bacteria drink, etc. may be used, and it can also be added to food additives or feed / food.

(Table 9)

[Example 4]
In Example 4, a lotion containing 0.3% of Example 1 was prepared in a conventional manner with the composition shown in Table 10 as an example of a cosmetic containing the Pelago algae extract-containing composition.

(Table 10)

[Example 5]
In Example 5, a lotion containing 0.1% of Example 2 was prepared in a conventional manner with the composition shown in Table 11 as an example of a cosmetic containing the Pelago algae extract-containing composition.

(Table 11)

  The cosmetic is not limited to the lotion, and may be any cosmetic including, for example, lotion, milky lotion, cream and the like, and a pharmaceutical containing shampoo, body shampoo, rinse, soap, and the like. It can also be incorporated into quasi-drugs.

It is a graph showing the result of Test Example 3 (humidity retention test) for confirming the effect in the Pelago algae extract-containing composition according to Example 1 of the present invention, and representing the immediate moisture retention effect. It is a graph showing the result of the same test example 3 (humidity retention test) and showing the effect on the stratum corneum moisture content.

Claims (3)

Freeze-dry the algal bodies of sarcinolysis, which is a yellow plant gate Pelagophyceae sarcinolysis, after desalting,
After adding water to the dried alga body and stirring and extracting at 90 ° C. or more for 3 hours , the hot water extract obtained by filtering the supernatant by centrifugation is concentrated to 1/5 to 1/6, and ethanol is added to this. Obtained as a precipitate after being added and left overnight, or
It was obtained by adding 50% ethanol to the dried alga body and performing reflux extraction at 90 ° C. or higher for 3 hours , and then filtering the supernatant by centrifugation to completely remove ethanol.
A composition containing an extract of Pelago algae, comprising an extract having hyaluronidase inhibitory activity, antioxidant activity, moisturizing ability and antibacterial activity . A food comprising 0.1% of the Pelago algae extract-containing composition according to claim 1. A cosmetic comprising 0.1 to 0.3% of the Pelago algae extract-containing composition according to claim 1.

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