KR101459591B1 - Anti inflammantory composition comprising anthocyanin - Google Patents

Abstract

An anti-inflammatory composition comprising anthocyanin extracted from sweet potato is provided. The composition for anti-inflammation according to an embodiment of the present invention may include anthocyanin having anti-inflammatory activity extracted from sweet potato.

Description

An anti-inflammatory composition comprising anthocyanin,

The present invention relates to a composition for anti-inflammation, and more particularly to a composition for anti-inflammation containing anthocyanin extracted from sweet potato and having anti-inflammatory activity.

Anthocyanin is a water-soluble flavonoid pigment that exhibits red, purple, and blue colors widely found in flowers, fruits, stems, and leaves of plants. The term anthocyanin is derived from the word anthos, which means flower in Greek, and kyanos, which means blue, and was first used by Marquart in 1835 to name a blue pigment in petals.

Recently, anthocyanin pigments have been used as additives for processed foods such as soft drinks, jams and candies. Processed foods have distinctive colors, but their color and nutritional value change during processing or storage. The addition of anthocyanin has the effect of preventing discoloration or discoloration of the inherent color of the food itself, or enhancing the value of the food by complementing the original color. Anthocyanin has also been reported to function as a food preservative to enhance the storage stability of preserved and processed foods. In addition to food additives, anthocyanins are also actively utilized in coloring raw materials and pharmaceuticals of the coconut industry.

Anthocyanin pigment is mainly used as a substitute for synthetic colorants (red No. 2, No. 4, No. 40), which is toxic to red and can be produced as a by-product of vegetables and fruits, Products.

At present, anthocyanins contained in grape, strawberry, red cabbage and red pepper are extracted and used globally, and the demand for domestic anthocyanins is not reported accurately. However, in 1998, It is estimated that the total domestic demand is about 150 tons and the market of about 80 billion won is formed. On the other hand, in Japan, demand for 385 tons was created in '94, and it is expected that the demand of natural pigment will continue to increase due to the increase of national income and the advancement of dietary life.

On the other hand, the inventors of the present invention have learned through the researches that the anthocyanin exhibits various physiological activities in addition to the function as a natural pigment. Typically, anthocyanins are used for antioxidant, cholesterol lowering, vision improvement effect, vascular protection, arteriosclerosis, Anti-ulcer function, anti-cancer, anti-inflammatory, anti-diabetic, and protection from ultraviolet rays. Therefore, anthocyanin has been shown to be a new target for the creation of health foods and new medicines since it exhibits various physiological activities and is actively utilized in pharmaceuticals.

Accordingly, the present invention is intended to protect the matters obtained by the above-mentioned study,

A problem to be solved by the present invention is to provide a composition for anti-inflammation containing anthocyanin having an anti-inflammatory activity extracted from sweet potato.

The problems to be solved by the present invention are not limited to the above-mentioned technical problems and other technical problems which are not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the object, the anti-inflammatory composition according to an embodiment of the present invention may include an anthocyanin having anti-inflammatory activity extracted from sweet potato.

On the other hand, the anthocyanin is a cyanidin-based anthocyanin or a peonidin-based anthocyanin.

On the other hand, the cyanidin-based anthocyanin is cyanidin 3-sophoroside-5-glucoside.

On the other hand, the cyanide-based anthocyanin is a cyanidin-3-p-hydroxycarboxylic acid having mono-acylation of p-hydroxybenzoic acid, caffeic acid and ferulic acid Cyanidin 3-p-hydroxybenzoylsophoroside-5-glucoside, cyanidin 3- (6 "-caffeoylsophoroside-5-glucoside) -5-glucoside or cyanidin 3- (6 "-feruloylsophoroside) -5-glucoside.

On the other hand, the cyanide-based anthocyanin may be cyanidin-3-caffeoyl-p-hydroxybenzoic acid, diacylated p-hydroxybenzoic acid, caffeic acid, ferulic acid, p-hydroxybenzoyl-sophoroside-5-glucoside, cyanidin 3- (6 ", 6" -dicapheylene microsoposide) 5-glucoside) or cyanidin 3- (6 "-caffeoyl-6" -feruloyl-5-glucoside - (6 "-caffeoyl-6" '- feruloylsophoroside) -5-glucoside.

On the other hand, the phenionine-based anthocyanin is cyanidin 3-sophoroside-5-glucoside.

On the other hand, the phenionine-based anthocyanins include p-hydroxybenzoic acid, caffeic acid, and mononoacylated aconylated phenonidine 3-p- Peonidin 3-c-hydroxybenzoylsophoroside-5-glucoside, peonidin 3-caffeoylsophoroside-5-glucoside, (6 "-feruloylsophoroside) -5-glucoside. ≪ / RTI >

On the other hand, the phenonidine-based anthocyanin may be a p-hydroxybenzoic acid, a caffeic acid, a di-acylated phenonidine-3-caffe oil of ferulic acid p-hydroxybenzoyl-sophoroside-5-glucoside, peonidin 3- (6 ", 6" -dicapheyloic microside- -5-glucoside, peonidin 3- (6 "- caffeoyl-6"'- ferulic oil microsoposide) -5- Feruloyl-5-glucoside and peonidin 3-feruloyl-5-glucoside and peonidin 3-feruloyl-6-feruloylsophoroside- -hydroxybenzoylsophoroside-5-glucoside).

According to the present invention, there is provided a composition for anti-inflammation comprising anthocyanin having an anti-inflammatory activity extracted from sweet potato.

FIG. 1 is a graph showing an analysis of anthocyanin obtained by the examples by autopurification prep LC-DAD-ESI / MS chromatogram. FIG.
FIG. 2 is a graph showing the protective effect evaluation from oxidative stress induced by hydrogen peroxide. FIG.
Figure 3 is a graph showing the results of cytotoxicity assay of anthocyanin on NO production induced by LPS in RAW 264.7.
FIG. 4 is a graph showing the anti-inflammatory effect of peak 2 anthocyanin, Peonidin-3-sophoroside-5-glucoside.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

Unless defined otherwise, all terms (including technical and scientific terms) used herein may be used in a sense commonly understood by one of ordinary skill in the art to which this invention belongs. Also, commonly used predefined terms are not ideally or excessively interpreted unless explicitly defined otherwise.

Hereinafter, a composition having anti-inflammatory activity according to one embodiment of the present invention will be described.

The composition having anti-inflammatory activity according to an embodiment of the present invention may include anthocyanin extracted from sweet potato.

First, the description of sweet potato is as follows. Also known as potatoes and sweet potatoes. It grows widely throughout Korea. The length is about 3m. The stem is routed along the ground for a long time. The leaves are alternate, the leaves are shallowly divided into a heart shape, and the leaves and stems are juiced. Roots are formed at the base of petiole at the bottom of the stem, and part of it grows in the ground to become sweet potato root. The shape varies from pointed cylindrical to spherical on both sides, and its color varies from white to yellow to light red to red to light purple. In particular, the composition having anti-inflammatory activity according to an embodiment of the present invention may include anthocyanin extracted from purple sweet potato.

On the other hand, the flowers hang on a peduncle from July to August, which are from the axilla of the leaves, in the shape of light purple morning glory. Calyx is divided into 5 pieces. The corolla is funnel-shaped and has 5 stamens and 1 pistil. The fruit is a ball-shaped seed vessel with two to four black-brown seeds.

When the frost falls in autumn, the leaves and stems are sown. At this time, the sweet potatoes are kept warm and stored, and the seeds are planted in the seedlings in the spring of next year. Cut the buds, plant them in the field and lower their roots. In the subtropical and tropical regions, they do not die throughout the year, so they cut and breed the stem at the right time. Do not use seeds when growing.

There are many varieties around the world. Varieties vary in appearance, taste, speed, and color of white, yellow, red, purple, and viscosity. For industrial use, it is good to have high starch content and high yield, and for feed, pick sweet potatoes and vines. In Korea, varieties such as Chungsung 100, Suwon 147, Shinmi, and Hwangmi are cultivated.

Purple sweet potato can be partially purified by a sequential solvent fractionation method to extract anthocyanin from purple sweet potato. More specifically:

First, the raw sweet potatoes of purple sweet potatoes are lyophilized and powdered. Thereafter, the powder is put into a water tank containing a formic acid solution and stirred at 20 to 50 ° C for at least one day to extract a first extract containing anthocyanin from the powder. The first extract contains fat-soluble and other water-soluble compounds in addition to the flavonoid compound (anthocyanin) with sugar.

If the stirring temperature is less than 20 캜, extraction of the first extract from the powder is not easy and it may be difficult to obtain anthocyanin. On the other hand, if the stirring temperature exceeds 50 캜, the active ingredient of the first extract may be damaged, which is not preferable. That is, when the temperature is higher than 50 ° C, the active ingredient in the first extract may be damaged and it may become difficult to obtain a good anthocyanin.

On the other hand, the mixing ratio of the powder: formic acid solution is preferably 1: 2 to 1: 5. If the mixing ratio is less than 1: 2, the extraction of the first extract from the powder may not smoothly take place. If the mixing ratio exceeds 1: 5, the solution becomes unnecessarily large, and the first extract is dissolved in the solution, so that it may take more time to separate the first extract from the solution in a subsequent step.

Subsequently, the first extract extracted from the cinnabar is filtered from the solution under reduced pressure. Vacuum filtration can proceed with a pressure of 5 to 20% relative to atmospheric pressure. By the filtration under reduced pressure, the damage of the first extract is minimized and the first extract can be filtered from the solution. Thereby, a first extract of good quality can be obtained.

Subsequently, anthocyanin is fractionated from the first extract. As described above, since the first extract contains fat-soluble and other water-soluble compounds in addition to anthocyanin, anthocyanin is fractionated therefrom.

At this time, an organic solvent may be used. Examples of the organic solvent include hexane, ethyl acetate, and butanol. More specifically, it is as follows.

First, add the extracted first extract to a container containing hexane. Thereafter, after stirring at room temperature, anthocyanin is separated from the first extract. The anthocyanin of the first extract moves to the hexane layer, which is an organic solvent, and the water-soluble substance of the first extract does not move to the hexane layer. Thereby, the anthocyanin and the water-soluble substance can be fractionated. On the other hand, in order to increase the yield of anthocyanin by fractionation, the first fractionation step with hexane can be repeated several times. In the following description, the first extract obtained through fractionation with hexane is referred to as a second extract for convenience of explanation.

Subsequently, a fractionation step for the second extract is carried out. The second extract has been removed in anthocyanin, a lipid soluble compound, and a nucleic acid fractionation process, but contains a minor amount of water soluble compounds. At this time, ethyl acetate can be used in the second fractionation process.

More specifically, the second extract is placed in a container containing ethyl acetate. Thereafter, after stirring at room temperature, anthocyanin is separated from the second extract. That is, the anthocyanin of the second extract moves to the ethyl acetate layer and the lipophilic substance of the second extract does not migrate to the ethyl acetate layer. Thereby, the anthocyanin and the lipid-soluble substance can be fractionated. On the other hand, in order to increase the yield of anthocyanin by the fraction, the second fractionation step with ethyl acetate can be repeated several times. In the following, the second extract obtained through fractionation with ethyl acetate is referred to as a third extract for convenience of explanation.

Subsequently, a fractionation process for the third extract is carried out. The third extract contains anthocyanin and a small amount of a fat-soluble compound and a water-soluble compound. At this time, butanol can be used in the third fractionation process.

More specifically, the third extract is placed in a container containing butanol. Then, after stirring at room temperature, anthocyanin is separated from the third extract. That is, the anthocyanin of the third extract is transferred to the butanol layer, and the liposoluble and water soluble substance of the third extract is unable to migrate to the butanol layer. Thereby, the anthocyanin and the oil-soluble and water-soluble substance can be fractionated. On the other hand, in order to increase the yield of anthocyanin by fractionation, the tertiary fractionation step with butanol can be repeated several times. On the other hand, the third extract obtained by fractionation with ethyl acetate is hereinafter referred to as the fourth extract for convenience of explanation.

Subsequently, the butanol layer containing the fourth extract is depressurized to concentrate the anthocyanin contained in the fourth extract. At this time, the butanol layer can be depressurized with a rotary vacuum evaporator, for example, to concentrate the anthocyanin contained in the fourth extract.

After the concentration under reduced pressure, the solution, water-soluble and lipid-soluble substances remaining in the fourth extract are finally removed to finally obtain anthocyanin.

On the other hand, the anthocyanin can be purified rapidly and in a large amount from the saccharide powder at a high purity. That is, a high resolution large prep. An autopurification prep (LC-PDA-ESI / MS) automated system equipped with a column allows rapid and large-scale purification of anthocyanin from high-purity powders.

The anthocyanin may have the following formula (1).

Referring to Formula 1, anthocyanins include cyanidin 3-sophoroside-5-glucoside and peonidin 3-sophoroside-5-glucoside, glucoside. That is, the anthocyanin may include a cyanidin system and a peonidin system. Herein, the cyanide group is defined as an anthocyanin compound wherein R ₁ is H in the above formula (1). On the other hand, the phenionine system is defined as an anthocyanin compound wherein R ₁ is CH ₃ in the above formula (1).

That is, the anthocyanin is a basic compound of cyanidin 3-sophoroside-5-glucoside and peonidin 3-sophoroside-5-glucoside Structure. On the other hand, when cyanidin 3-sophoroside-5-glucoside and peonidin 3-sophoroside-5-glucoside were substituted with p P-hydroxybenzoic acid, caffeic acid, and ferulic acid may be mono- or di-acylated to form additional anthocyanins.

Specifically, there are the following kinds of cyanide-based anthocyanins.

The basic form of cyanidin 3-sophoroside-5-glucoside is cyanidin 3-sophoroside-5-glucoside.

In addition, the compound of the above formula (2), p-hydroxybenzoic acid, caffeic acid, ferulic acid, mono-acylated cyanidin 3-p- 5-glucoside, cyanidin 3-p-hydroxybenzoylsophoroside-5-glucoside, cyanidin 3- (6 "- caffeoylsophoroside) -5- glucoside and cyanidin 3- (6 "-feruloylsophoroside) -5-glucoside.

Further, the compound of the above formula (2), p-hydroxybenzoic acid, caffeic acid, ferulic acid, di-acylated cyanidin 3-caffeoyl- 5-glucoside, cyanidin 3-caffeoyl-p-hydroxybenzoyl-sophoroside-5-glucoside, cyanidin 3- (6 " cyanidin 3- (6 ", 6" -dicaffeoylsophoroside) -5-glucoside and cyanidin 3- (6 "-caffeoyl-6" "-caffeoyl-6" '-feruloylsophoroside) -5-glucoside.

Next, the phenonidine anthocyanins are as follows.

There is a basic type of cyanidin 3-sophoroside-5-glucoside.

Also, p-hydroxybenzoic acid, caffeic acid, and ferulic acid, which are compounds of Formula 2, are mono-acylated phenonidine 3-p-hydroxybenzoyl Peonidin 3-p-hydroxybenzoylsophoroside-5-glucoside, peonidin 3-caffeoylsophoroside-5-glucoside and phenonidine 3- (6 "-feruloylsophoroside) -5-glucoside. ≪ / RTI >

In addition, p-hydroxybenzoic acid, caffeic acid, and ferulic acid, which are compounds of Formula 2, are di-acylated phenonidine 3-caffeoyl-p- 5-glucoside, peonidin 3-caffeoyl-p-hydroxybenzoyl-sophoroside-5-glucoside and phenonidine 3- (6 " Glucoside, peonidin 3- (6 ", 6" '- dicaffeoylsophoroside) -5-glucoside, 3-feruloyl-p-hydroxybenzoylsophoroside-5-glucoside) and 3-feruloyl-6-feruloylsophoroside -glucoside.

The composition having anti-inflammatory activity according to an embodiment of the present invention includes the anthocyanin component extracted from sweet potato, and the contained anthocyanin component has anti-inflammatory activity. The composition according to an embodiment of the present invention can be used wherever anti-inflammatory activity is required. That is, the composition according to one embodiment of the present invention can be used as an additive for cosmetics, foods, medicines, detergents and the like.

Hereinafter, the contents of the present invention will be described in detail by way of examples. However, the scope of the present invention is not limited to these embodiments, but the present invention is not limited thereto.

[ Example ] - From Shinzami  Anthocyanin extraction

50 g of the lyophilized powders of microcrystalline cellulose were placed in a water bath containing 1% formic acid solution. At this time, the temperature of the water tank was 40 占 폚, and the solution of the syneresis powder and the solution were stirred in the water tank. Thereafter, the mixture was stirred for 3 days, and the extraction operation was carried out three times per day. Thereafter, a vacuum filtration operation was performed on the extract in a chamber depressurized to 0.9 atm. Thereafter, the extract under reduced pressure was placed in hexane at a concentration of 0.1 M, and the first fractionation work was carried out. The first fractionation work was performed three times. Subsequently, the second fractionation work was performed three times using ethyl acetate, and the third fractionation work was performed three times using butanol.

Thereafter, the extract was subjected to a third fractionation work at 0.9 atm with a rotary vacuum evaporator and concentrated under reduced pressure to obtain about 3 g of a concentrate. Subsequently, a concentrate was prepared at 500,000 ppm, and an individual anthocyanin was obtained by autopurification prep LC-MS.

[ Experimental Example  1] - Anthocyanin component analysis

The anthocyanin obtained by the above examples was analyzed by autopurification prep LC-DAD-ESI / MS chromatogram. The results are shown in Fig. Referring to Figure 1, approximately 15 anthocyanins were detected. The detected results are summarized in Table 1. As can be seen from the following Table 1, it can be seen that high purity anthocyanin can be obtained as a whole.

Compound name Vial (g) After concentration (g) Species (mg) Purity (%) Peak 1
(Cyanidin-3-sophoroside-5-glucoside) 6.1929 6.1929 6.1972 6.1971 4.30 100.00 6.1929 6.1970 6.1929 6.1971 Peak 2
(Peonidin-3-sophoroside-5-glucoside) 6.2080 6.2080 6.2190 6.2190 11.00 99.40 6.2080 6.2191 6.2080 6.2190 Peak 3
(Cyanidin 3- p- hydrocyvenzoyl sophoroside-5-glucoside) 6.1694 6.1694 6.1890 6.1890 19.60 98.26 6.1694 6.1889 6.1695 6.1890 Peak 4
(Peonidine 3- p -hydroxybenzoyl sophoroside-5-glucoside) 6.2044 6.2044 6.2272 6.2271 22.80 100.00 6.2045 6.2272 6.2044 6.2270 Peak 6
(Cyanidin 3- (6 "-feruloyl sophoroside) -5-glucoside) 6.2016 6.2015 6.2045 6.2045 2.90 96.31 6.2015 6.2044 6.2015 6.2045 Peak 7
(Cyanidin 3- (6 "-cyclohexylsophoroside) -5-glucoside) 6.1501 6.1502 6.1555 6.1555 5.40 98.02 6.1502 6.1555 6.1502 6.1555 Peak 9
(Cyanidin 3-caffeoyl- p -hydroxybenzoyl sophoroside-5-glucoside) 6.1388 6.1388 6.1568 6.1568 18.00 82.43 6.1387 6.1569 6.1388 6.1568 Peak 10
(Peonidine 3-caffeoyl sophoroside-5-glucoside) 6.1972 6.1972 6.2005 6.2006 3.30 98.48 6.1972 6.2007 6.1972 6.2006 Peak 11
(Cyanidin 3- (6 "-cyclohexyl-6" -feruloyl sophoroside) -5-glucoside) 6.1243 6.1242 6.1311 6.1311 6.80 96.41 6.1242 6.1311 6.1242 6.1310 Peak 12
(Peonidine 3- (6 ", 6 " -dicaffeoyl sophoroside) -5-glucoside) 6.2166 6.2167 6.2193 6.2193 2.70 99.27 6.2167 6.2193 6.2168 6.2193 Peak 13
(Peonidine 3-caffeoyl- p -hydroxybenzoyl sophoroside-5-glucoside) 6.2229 6.2228 6.2403 6.2403 17.40 94.67 6.2228 6.2403 6.2228 6.2404 Peak 14
(Peonidine 3- (6 "-cyclohexyl-6" -feruloyl sophoroside) -5-glucoside) 6.2220 6.2220 6.2611 6.2611 39.10 90.78 6.2219 6.2611 6.2220 6.2610

[ Experimental Example  2] - Hydrogen Peroxide Induced Oxidative  Evaluation of protective effects from stress

The protective effect evaluation from oxidative stress induced by hydrogen peroxide is shown in Fig. Specifically, the protective effect of the individual anthocyanins on the oxidative stress induced by H ₂ O ₂ (0.9 mM) in HepG2 was evaluated, and as a result, the protective effect was excellent overall. Among them, peak 12 (Peonidin 3- (6 ", 6" -dicaffeoyl sophoroside) -5-glucoside) showed significant cytoprotective effect ( p <0.05).

[ Experimental Example  3] - RAW  At 264.7 LPS Induced NO  Assessment of cytotoxicity of anthocyanin for production

The results of the cytotoxicity evaluation of anthocyanin on NO production induced by LPS in RAW 264.7 are shown in FIG. Individual anthocyanins showed low toxicity to NO production induced by LPS at RAW 264.7. On the other hand, individual anthocyanin significantly inhibited the production of NO induced by LPS 2 and 4 in RAW 264.7 ( p <0.05).

[ Experimental Example  4] - peak  2 anthocyanin, Peonidine -3- sophoroside -5- glucoside  Anti-inflammatory effect test

peak 2 anthocyanin, and peonidin-3-sophoroside-5-glucoside were tested. The inhibition of the production of NO by LPS of anthocyanin of Peak 2 inhibited NO production by 15% and 8%, respectively, at concentrations of 10 μM and 1 μM (p <0.05). In other words, Peonidin-3-sophoroside-5-glucoside of peak 2 inhibited the production of NO, indicating that the anti-inflammatory effect was excellent.

While the present invention has been described in connection with what is presently considered to be practical exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, It will be understood that the invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (8)

A composition for anti-inflammation comprising peonidin-3-sophoroside 5-glucoside having anti-inflammatory activity extracted from sweet potato. delete delete delete delete delete delete delete

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