KR101640217B1 - Antioxidative Composition Contianing Enzymatic Extract of Clove - Google Patents

Abstract

The present invention relates to an antioxidant composition containing an enzyme-treated extract of clove and a preparation method thereof, and more particularly to a composition for antioxidation wherein an extract of clove is treated with an enzyme to improve antioxidation ability and a method for producing the composition.
The clove extract having improved antioxidative function by the enzyme treatment according to the present invention can be used for the production of cosmetics and foods which are derived from natural products and which require a substance having excellent antioxidant ability.

Description

[0001] The present invention relates to an antioxidative composition containing an enzyme-treated extract of cloves,

The present invention relates to an antioxidant composition containing an enzyme-treated extract of clove and a preparation method thereof, and more particularly to a composition for antioxidation wherein an extract of clove is treated with an enzyme to improve antioxidation ability and a method for producing the composition.

Clove is an evergreen small arboreal tree with a dicotyledonous plant plating thick sheep plating, and is the origin of the Moluka Islands. The stem is 4 ~ 7m in height, the leaves are opposite, fragrant and bushy. The flower is white, with four petals. The shape of the bud is nail-like and fragrant, so it is called clove. The English clove is also derived from the French clou (nail). It was already known in China around the third century BC and was known in Rome and Europe in the Middle Ages. Because it is very fragrant, it is used as it is or as powder, and it uses clove oil extracted with water or steam. Used in foods, medicines, preservatives, or as an analgesic in dentistry, including seizures. Cloves are used not only for buds but also for flowers and berries because their yields are small.

Free radicals, on the other hand, are chemically reactive chemicals that can be generated when the skin is exposed to ultraviolet light or when the skin cells are inhaling. Most of the free radicals are reactive oxygen species (ROS), such as superoxide radicals, hydrogen peroxide (H2O2), hydroxy radicals, singlet oxygen have. Since free radicals are related to skin aging, it has been common to provide antioxidative effects by adding superoxide dismutase (SOD), vitamin-E derivatives, vitamin C and derivatives thereof, and flavonoid to cosmetics in cosmetics . However, the above-mentioned materials are used as synthetic raw materials, and in recent years, compositions having antioxidative effects derived from natural products have been spotlighted.

It is known that the antioxidant effect of cloves is known, and antioxidant compositions containing clove oil and other plant oils (Korean Patent Laid-Open No. 10-2011-0060602), cosmetic compositions having antioxidant ability containing a solvent extract of cloves (Korean Patent Laid- 2004-0076103).

However, the clove extracted with the well-known solvent extraction method has insufficient antioxidative effect, and a method for producing a natural extract having an improved antioxidative effect is required.

Accordingly, the present inventors have made intensive efforts to develop a plant extract having an excellent antioxidative effect. As a result, it has been confirmed that the antioxidative ability is improved when the enzyme is treated with the solvent extract of clove, and the present invention has been completed.

It is an object of the present invention to provide a method for producing a clove extract having improved antioxidative ability and a use of clove extract prepared by the method.

In order to accomplish the above object, the present invention provides a method for producing an extract, comprising: (a) extracting a clove with a solvent to obtain an extract of clove; (b) removing the solvent from the clove solvent extract; And (c) treating the extract from which the solvent has been removed with a hydrolytic enzyme to obtain an enzyme-treated clove extract, as an active ingredient.

The present invention also provides a composition for antioxidation, which comprises the extract prepared by the above method and hydrolyzed with the clove solvent extract as an active ingredient.

The present invention also provides a cosmetic composition comprising an extract prepared by the above method and hydrolyzed with the clove solvent extract as an active ingredient.

The clove extract having improved antioxidative function by the enzyme treatment according to the present invention can be used for the production of cosmetics and foods which are derived from natural products and which require a substance having excellent antioxidant ability.

FIG. 1 is a graph showing DPPH radical scavenging activity of an enzyme-treated extract of clove according to the present invention.
FIG. 2 is a graph showing the ABTS radical scavenging activity of an enzyme-treated extract of clove according to the present invention.
FIG. 3 is a graph showing SOD activity of an enzyme-treated extract of clove according to the present invention.

In one aspect, the present invention relates to a process for the preparation of (a) an extract of cloves with a solvent to obtain an extract of cloves; (b) removing the solvent from the clove solvent extract; And (c) treating the extract from which the solvent has been removed with a hydrolytic enzyme to obtain an enzyme-treated clove extract, as an active ingredient.

In the present invention, the hydrolytic enzyme may be selected from the group consisting of? -Amylase,? -Glucosidase, pectinase and peptidase, preferably? -Glucosidase, But is not limited thereto.

In the present invention, the solvent may be any solvent capable of extracting the antioxidant active ingredient in the clove, and may be selected from the group consisting of water, ethanol, dichloromethane and ethyl acetate, Is more preferable.

In the present invention, a step of solvent extraction of the enzyme-treated clove extract may be further included.

In one embodiment of the present invention, the clove is extracted with water at 50 캜 and then concentrated, then treated with? -Glucosidase, further extracted with 50% ethanol, and then filtered to obtain an antioxidant (DPPH radical scavenging activity, ABTS radical Scavenging activity and SOD activity) were measured.

(2, 2-diphenyl-1-picrylhydrazyl) radical scavenging activity, and ABTS (2,2'-azino-bis (3-ethyl benzothiazoline-6-sulphonic acid)) radicals of the ethanol extract of clove and the enzyme- As a result of measuring the scavenging activity and SOD (superoxide dismutase) activity, it was confirmed that the enzyme-treated extract of clove had a much higher antioxidative capacity than the ethanol extract of clove (Figs. 1, 2 and 3)

In another aspect, the present invention relates to an antioxidative composition prepared by the above method and containing an extract obtained by hydrolyzing a clove solvent extract as an active ingredient.

The composition of the present invention may comprise a functional food. The functional food of the present invention can be used in the form of powder, granule, tablet, capsule or beverage, for example, various foods, candy, chocolate, beverage, gum, tea, vitamin complex and health supplement.

The extract of the present invention may be added to foods or drinks for the purpose of antioxidation. At this time, the amount of the above extract in the food or beverage may generally be 0.01 to 50% by weight, preferably 0.1 to 20% by weight, of the total food weight of the health functional food composition of the present invention, In a proportion of 0.02 to 10 g, preferably 0.3 to 1 g.

The health beverage composition of the present invention has no particular limitation on the liquid ingredient other than the above-mentioned extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose, polysaccharides such as maltose, sucrose and the like, such as dextrin, cyclodextrin and the like And sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

In another aspect, the present invention relates to a cosmetic composition prepared by a method and containing an extract obtained by hydrolyzing an extract of clove solvent as an active ingredient.

The composition of the present invention may contain a cosmetic composition that requires an antioxidant effect. Useful substances which can be further used in the cosmetic composition of the present invention include antioxidants, binders, bulking agents, chelating agents, pigments, softeners, emulsion stabilizers, film formers, fillers, directional ingredients, gelling agents, But are not limited to, hair fixatives, wetting agents, plasticizers, preservatives, skin conditioners, solvents, sunscreens, surfactants, ultraviolet absorbers, viscosity modifiers, waxes and the like.

The cosmetic composition of the present invention may contain a cosmetically acceptable carrier and may be an aqueous solution such as water or a polar organic solvent, alcohol such as ethanol or other polar solvent, natural or synthetic oil; Oil-in-water emulsion; Water-in-oil emulsion; Or wax. The carrier is clearly non-toxic. A preferred carrier for the composition of the present invention is an aqueous solution.

Depending on the function, the cosmetic composition of the present invention can be applied to a variety of cosmetic compositions such as a solution (lotion-like composition), a concentrated solution, a gel, ointment, emulsion (cream, emulsion), vesicle dispersion, powder, dense powder, And the like. More specifically, the cosmetic composition of the present invention can be used as a cosmetic composition for oral use such as ball puff, cream (facial cream, hand cream, moisturizing cream, sunscreen cream), cream powder, eyeliner, eye shade, eyebrow pencil, foundation, lotion, Emulsions, ointments, pomades, lozes, and the like, but are not limited thereto. They may be packed in a squeeze pack containing a filler that can be applied in the form of a foam or spray.

The cosmetic composition may contain natural or artificial wax. Natural waxes include animal and carnauba such as lanolin, beeswax, spermaceti or lanolin alcohol, hardened or acetylated lanolin, lanolin derivatives such as lanolin fatty acids or acetylated lanolin alcohols, carnauba, candelilla, There are vegetable such as kapok, rice, hardened jojoba, alfa or japanese wax or cork fiber, sugar cane wax, cocoa butter and the like. Alternatively, the inorganic wax may include paraffin, montan, lignite, petrolatum, petrolatum wax or microcrystalline wax, ceresin, or ozokerite. Examples of synthetic waxes used include linear esters such as myristyl myristate, a reaction product of a polyethylene wax obtained by Fischer-Tropsch synthesis and a saturated C10-C40 carboxylic acid and a saturated C10-C40 alcohol. Other waxes used include calcium lanolate or stearate or hardened coconut oil and the like.

The cosmetic composition may include modified or unmodified oils of vegetable or animal oils. For example, sweet almond oil, avocado oil, castor oil olive oil, jojoba oil, sunflower oil, turmeric oil, sesame oil, peanut oil, grape seed oil, soy milk, safflower oil, coconut oil, maze oil, hazelnut oil, Butter, palm oil, apricot seed oil, calf pill oil or perhydro-squalene. Furthermore, the oil phase may be an inorganic oil such as liquid paraffin, liquid petrolatum, and the like. The oil may be selected from the group consisting of fatty acid esters such as isopropyl myristate, isopropyl palmitate, 2-ethylhexyl palmitate, penicillin oil (stearyloctonate), unsaturated fatty acids such as oleate, palmitate, stearate, behen, linoleic, Volatile or non-volatile isoparaffins such as fatty acids or C8-C16 isoparaffins.

Moreover, the oil may be C12-C18 fatty alcohols such as oleyl alcohol, cetyl alcohol and stearyl alcohol.

If provided as an emulsion, the emulsified composition of the present invention comprises an oil phase and an aqueous phase. The oil phase is preferably provided in a range of about 1 to about 75 wt%, more preferably about 5 to about 60 wt%, and most preferably about 40 to about 60 wt%, based on the total weight of the composition desirable.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1: Preparation of Clove Extract

Cling (origin: China) was purchased from Kyungdong market and crushed to a size of 1 mm or less. The ground cloves were extracted with methanol and ethanol solvent. The clove used was 200 g per each solvent. At the time of extraction, 2,300 ml of the solvent was divided into four portions in 800 ml, 500 ml, 500 ml and 500 ml, and they were mixed together. A water bath at 50 ° C was used for the extraction. After extraction, the solid content was removed by using filter paper.

In order to remove impurities such as flavonoid contained in the clove in the production of the clove extract, β-glucosidase enzyme (Vision Biochem, Korea), peptiaase (available as a source), pectinase (available as a source) The α-amylase (available) was treated with 0.03% by weight of the extract to hydrolyze the flavonoids and the like so that they did not dissolve in water.

During the enzyme treatment, the concentrate is warmed to a temperature suitable for the enzyme reaction. The temperature of the concentrate was adjusted to 52 占 폚, and? -Glucosidase was added to the concentrate so as to be 0.1% (w / w) by mass. The hydrolysis reaction was carried out for 12 hours while stirring the concentrate with the enzyme.

The extracts were extracted by adding 2000 mL of 50% ethanol to the enzyme - treated clove extract and extracted with water bath at 80 ℃ for 4 hours.

The ethanol extract was concentrated using a rotary evaporator to remove ethanol. Concentration was performed to 1 to 2-fold concentration, and the concentrated extract was concentrated until it was confirmed that ethanol remained and remained.

Example 2: Analysis of DPPH activity of an extract of Clostridial enzyme

2-1 Analysis of DPPH activity by enzyme

200 μl of the enzyme-treated extract obtained in Example 1 and 800 μl of a DPPH solution (Aldrich, USA) were added, and the mixture was shaken with a vortex mixer for 10 minutes, and the absorbance at 517 nm was measured.

After the DPPH standard UV value was adjusted to 1, the UV value of each sample was measured.

The DPPH radical scavenging activity (%) was calculated by the following formula.

DPPH radical scavenging activity (%) = (absorbance of sample / absorbance of control) x 100

DPPH radical scavenging activity of enzyme-treated extract DPPH activity by enzyme 0 min 5 min 10 min Pepdidase 50.295 56.011 60.717 50.315 56.042 60.800 50.893 56.032 60.872 Pectinase 49.803 50.126 53.267 49.833 50.115 53.308 49.864 50.177 53.277 β - glucosidase 63.226 63.401 63.463 63.257 63.411 63.442 63.268 63.391 63.473 α-amylase 34.670 35.280 35.722 34.988 35.260 35.804 35.245 35.280 35.804

As a result, as shown in Table 1, the activity of DPPH was the best after 10 minutes of treatment, and the DPPH activity of the extract treated with? -Glucosidase was the highest, and the analysis was carried out with? -Glucosidase- Were used.

2-2 Comparison of DPPH activity between enzyme-treated extract and ethanol extract

The extracts with and without enzymatic treatment of clove extracts were compared with those of clove extracts with only 50% ethanol extraction.

As a result, as shown in Table 2 and Fig. 1, it was confirmed that the DPPH radical scavenging activity of the extract added with the enzyme treatment was nearly three times higher than that of the ethanol extract.

Comparison of DPPH Radical Scavenging Activity of Clove Extracts Clove extract enzyme treatment supernatant Clove extract enzyme treated precipitate 50% ethanol extract 1 50% ethanol extract 2 DPPH scavenging activity (%) 63.70 13.04 23.81 25.30

Example 3: Analysis of ABTS activity of the extract of Clostridial enzyme

ABTS (Sigma, USA) 7.4 mM and potassium persulphate 2.6 mM were left in a dark place for 24 hours to form ABTS cations. 900 쨉 l of ABTS solution was added to 100 쨉 l of the enzyme-treated extract prepared in Example 1, and the mixture was shaken with a vortex mixer for 10 minutes, and the absorbance was measured at UV 732 nm.

After setting the ABTS Standard UV value close to 1, the UV value of each sample was measured (0.1 mM).

The ABTS radical scavenging activity (%) was calculated by the following formula.

ABTS radical scavenging activity (%) = (absorbance of sample / absorbance of control) x 100

As a result, as shown in Table 3 and FIG. 2, it was confirmed that the ABTS radical scavenging activity of the extract added with the enzyme treatment was nearly double that of the ethanol extract.

Clove extract enzyme treatment supernatant Clove extract enzyme treated precipitate 50% ethanol extract 1 50% ethanol extract 2 ABTS scavenging activity (%) 83.1 1.2 47.8 34.1

Example 4: Analysis of SOD activity of the extract of Clostridial enzyme

SOD activity of clove extract was measured by SOD assay kit (Cayman chemical, USA). The working solution containing SOD was prepared according to the manufacturer's method of the kit, and SOD activity was analyzed by the manufacturer's method.

The SOD activity (%) was calculated by the following equation through the SOD assay kit.

SOD activity (%) = {[(S1-S3) - (SS-S2)] /

S1: Slope of blank 1

S2: Slope of blank 2

S3: Slope of blank 3

SS: Slope of Sample

As a result, as shown in Table 4 and FIG. 3, it was confirmed that the ABTS radical scavenging activity of the extract added with the enzyme treatment was significantly higher than that of the ethanol extract.

Clove extract enzyme treatment supernatant Clove extract enzyme treated precipitate 50% ethanol extract 1 50% ethanol extract 2 SOD scavenging activity (%) 62.89 24.22 45.33 44.44

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereto will be. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (6)

A method for producing an antioxidative composition containing an extract of cloves as an active ingredient comprising the steps of:
(a) extracting the clove with a solvent to obtain an extract of clove solvent;
(b) removing the solvent from the clove solvent extract; And
(c) treating the extract from which the solvent has been removed with? -glucosidase hydrolyzing enzyme to obtain an enzyme-treated clove extract.
delete The method of claim 1, wherein the solvent is selected from the group consisting of water, ethanol, dichloromethane and ethyl acetate.
The method of claim 1, further comprising the step of solvent extraction of the enzyme-treated clove extract.
A composition for antioxidation, which is prepared by the method of any one of claims 1, 3, and 4, and contains an extract obtained by hydrolysis of the clove solvent extract as an active ingredient.
A cosmetic composition comprising an extract obtained by the method of any one of claims 1, 3, and 4 and hydrolyzed with the clove solvent extract as an active ingredient.

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