KR102017572B1

KR102017572B1 - Manufacturing method of sasa quelpaertensis nakai extract using high temperature-alkali extraction technique and cosmetic compostion comprising the extract

Info

Publication number: KR102017572B1
Application number: KR1020160037135A
Authority: KR
Inventors: 구창섭; 서수경; 장동일; 김대현
Original assignee: 주식회사 콧데
Priority date: 2016-03-28
Filing date: 2016-03-28
Publication date: 2019-09-03
Also published as: KR20170111528A

Abstract

The present invention relates to a preparation method of Jeju Joridae extract using the High Temperature-Alkali Extraction Technique (HTAE Technique) and to a cosmetic composition containing the Jeju Joridae extract prepared by the hot-alkali extraction method as an active ingredient. . Jeju Joridae extract prepared by the high temperature-alkali extraction method developed in the present invention contains not only high concentration of p-coumaric acid (p-coumaric acid) but also various kinds as compared to the extract prepared using other extraction methods. And it contains a large amount of potency ingredient can maximize the effect of the potency ingredient.

Description

Manufacturing method of sasa quelpaertensis nakai extract using high temperature-alkali extraction technique and cosmetic compostion comprising the extract}

The present invention not only contains a high concentration of p-coumaric acid (p-coumaric acid) more than 10% compared to the extract prepared using other extraction methods, and contains various types and large amounts of efficacy ingredients Regarding the cosmetic composition containing the preparation method of Jeju chopsticks using the High Temperature-Alkali Extraction Technique (HTAE Technique) and the Jeju chopstick extract prepared by the hot-alkali extracting method as an active ingredient will be.

Jeju jeretdae is one of the most abundant plant species in the forests of Jeju Island. Jeju choridae (rice family) is an edible bamboo leaf, dried leaves have been used in the manufacture of leaf tea for the treatment of diabetes and gastritis. Among the compound components constituting the plant, p-coumaric acid has already been identified.

Republic of Korea Patent Application No. 2005-59961 'Jeju jeokdae leaf extract with whitening activity' is a extract obtained from the sake extract, jeju jeokdae leaves showing the antioxidant activity, anti-inflammatory activity, tyrosinase inhibitory activity, melanin formation inhibitory properties The extract is described.

Republic of Korea Patent Application No. 2007-134908 'method of purifying para-coumarin acid from jeju choridae and functional cosmetics and pharmaceutical compositions using the para-coumarin acid' describes a method of purifying p-coumaric acid from jeju chori .

Such prior art documents include extracting the active ingredient from jeju jeoldae, hot water extraction method and 30% to 80% of ethanol (ethanol) by adding, extracting, filtering and concentrating the methylene chloride (chloroethylene), chloroform ( After fractionation with chloroform and normal hexane (n-hexane), the fractions were concentrated again ethyl acetate, methyl acetate, n-butanol and isopropanol, etc. Disclosed is a solvent extraction-fractionation method to extract the active ingredient by fractionation.

However, the hot water extraction method and the solvent extraction-fractionation method have a need for a new extraction method because it is difficult to realize the desired efficacy effect with the practically applicable amount due to the problem that the extraction yield and the content of the active ingredient are very low.

1. Republic of Korea Patent Application No. 2005-59961 2. Korean Patent Application No. 2007-134908

Accordingly, the present invention is repeated to search for an extraction method that can produce a jeju jeoldae extract containing not only a high concentration of p-coumaric acid (p-coumaric acid) more than 10%, but also contains a variety of types and a large amount of potent ingredients. As a result, Jeju Joridae extract prepared using high temperature-alkali extraction method not only contains high concentration of p-coumaric acid at more than 10%, but also contains various kinds and large amounts of potent ingredients. It was found that the activity, anti-inflammatory activity and the whitening effect is maximized and completed the present invention.

Accordingly, an object of the present invention is to provide a high concentration of p-coumaric acid (p-coumaric acid) by using a high-alkali extraction technique, as well as containing various types and large amounts of potent ingredients It is to provide a manufacturing method and a cosmetic composition containing the extract as an active ingredient.

In order to achieve the above technical problem, the present invention provides a method for producing Jeju boiled extract comprising a high temperature alkali extraction step of treating the Jeju boiled with alkali at a high temperature of 80 ~ 130 ℃.

In another method for preparing a sachet extract according to the present invention, the alkali is preferably at least one selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium bicarbonate and sodium carbonate.

In another manufacturing method of the present invention,

The jeju jeoldae and alkali may be mixed in a weight ratio of 1: 0.01 to 1: 100.

The preparation method according to another embodiment of the present invention may further include neutralizing, filtering, concentrating, and solvent extracting the hot alkali extracted extract.

In order to achieve another technical problem, the present invention comprises the Jeju chopsticks extract obtained by the preparation method of Jeju chopsticks extract including the high temperature alkali extraction step of treating the Jeju chopsticks with alkali at a high temperature of 80 ~ 130 ℃ as an active ingredient It provides a cosmetic composition.

In the cosmetic composition according to the present invention, the Jeju jejudae extract is contained in an amount of 0.001 to 20% by weight based on the total weight of the composition shows excellent antioxidant activity, anti-inflammatory activity, whitening effect.

Jeju Joridae extract prepared by the hot-alkali extraction method according to the present invention contains not only a high concentration of p-coumaric acid (p-coumaric acid) but also various kinds and Contains a large amount of potency ingredients can maximize the effect of the potency ingredients.

Jeju choridae extract provided by the present invention is excellent in whitening effect by inhibiting DPPH radical removal effect, NO production inhibitory activity and melanin synthesis can be used as an antioxidant, anti-inflammatory and whitening cosmetic composition.

The present invention provides a method for producing a Jeju chopsticks extract comprising a high temperature alkali extraction step of treating the Jeju chopsticks with an alkali at a high temperature of 80 ~ 130 ℃.

In addition, the present invention provides a cosmetic composition containing the jeju jeoldae extract prepared by the hot-alkali extraction method as an active ingredient.

Hereinafter will be described in detail with reference to Examples and Test Examples for carrying out the present invention.

The present invention is prepared by using the high temperature and alkali decomposition in combination with Jeju jinjadae extract lignin-based polyphenol components and high content of p-coumaric acid (p-coumaric acid) and ferulic acid ( Efficacy ingredients such as ferulic acid can be extracted. In addition, it is possible to maximize the effect of the potent ingredients because it can extract a large amount of potent ingredients.

The preparation method of Jeju choridae extract according to the present invention includes a high temperature alkali extraction step of treating the Jeju choridae with alkali at a high temperature of 80 ~ 130 ℃. In addition, the method may further include neutralizing, filtering, concentrating, and solvent extracting the hot alkali extracted extract.

The step-by-step description of the preparation method of jeju jeoldae extract using the hot-alkali extraction method of the present invention.

1) a high temperature alkali extraction step of treating Jeju jejudae with alkali at a high temperature of 80 ~ 130 ℃;

Decompose Jeju jeori stand by mixing with alkali at 80 ~ 130 ℃. At temperatures below 80 ° C., active ingredients are not sufficiently extracted, and at temperatures above 130 ° C., alkali hydrolysis is performed at a temperature in the above range because the increase in effective ingredient extraction is insignificant and inefficient. The most preferable temperature is 110 ~ 125 ℃, the most efficient considering the amount of extraction of the active ingredient and the amount of destruction in the extraction process at this temperature.

The alkali that can be used in the present invention is at least one member selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate.

It is preferable that the jeju choridae and alkali are mixed in a weight ratio of 1: 0.01 to 1: 100. This is because if the amount of alkali is less than 1: 0.01, the extraction of the active ingredient is not performed properly, and if the amount is greater than 1: 100, the amount of increase in the amount of the extracted component is small and inefficient.

2) neutralizing the alkali decomposed solution;

It is possible to prevent the deterioration of the active ingredient by neutralizing the alkali hydrolysis solution in step 1). As the acid usable in the present invention, hydrochloric acid, sulfuric acid, acetic acid, and the like, which are commonly known in the art, may be used. For example, hydrochloric acid may be added to adjust pH to 2.0 to 7.0 to stably store the active ingredient for a long time. The stock solution of alkaline hydrolyzate extract of Jeju jerk can be obtained.

3) filtering the neutralized solution;

By filtering the stock solution of the jeju jeoldae neutralized in step 2) to remove impurities, it is possible to obtain a desired alkali-decomposed jeju jeoldae extract. As a filtration method usable in the present invention, all methods conventional in the art may be used, and for example, an alkaline hydrolysis extract stock solution of Jeju chopsticks from which impurities are removed may be obtained by passing through a fine filter paper.

4) concentrating the filtered solution;

The jeju jeoldae extract stock solution filtered in step 3) is concentrated to reduce the amount of the solvent used in step 5). At this time, a vacuum concentration method commonly used in the art may be used.

5) solvent extraction of the concentrated solution;

Extracted from the jeju jeoldae extract concentrated in step 4) with a suitable solvent to enhance the content of the active ingredient. At this time, all solvents commonly used in the art may be used, but are preferably extracted using ethanol, acetonitrile, ether, methanol, butanol, ethyl acetate or isopropanol, or a mixed solvent thereof.

In another aspect, the present invention provides a cosmetic composition containing the extract jeju jeoldae prepared by the above-mentioned high-alkali extraction method.

The cosmetic composition containing the jeju jeoldae extract prepared by the high temperature-alkali extraction method of the present invention is excellent in whitening effect by inhibiting DPPH radical removal effect, NO production inhibitory activity and melanin synthesis, and thus as an antioxidant, anti-inflammatory and whitening cosmetic composition. Can be used.

Jeju rind extract of the present invention is contained in an amount of 0.001 to 20% by weight, more preferably 0.1 to 20% by weight relative to the total weight of the composition. If the content is less than 0.001% by weight, the effect is insignificant, and if it exceeds 20% by weight, the increase in efficacy is not large compared to the increase in content because it is rather inefficient.

The cosmetic composition of the present invention is a flexible cosmetics, astringent cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, shampoo, conditioner, body It may be formulated as a detergent, toothpaste or mouthwash, but is not limited thereto.

Hereinafter, although an Example and a test example are given and this invention is demonstrated in detail, this invention is not limited only to these examples.

[ Example 1] Preparation of Jeju Joridae Extract Using Hot-Alkali Extraction Method

2 liters of 2% sodium hydroxide solution was added to 100 g of Jeju chopsticks at a temperature of 121 ° C., and reacted for 5 hours to obtain a hydrolyzed Jeju chopstick extract extract. Subsequently, hydrochloric acid was added to this stock solution to have a pH of 2.0, and then filtered with a filter paper to remove debris and concentrated under reduced pressure. The concentrate was extracted three times with 1 liter of acetonitrile, and the filtrate was concentrated to dryness to prepare Jeju extract.

[ Comparative example 1] Preparation of Jeju Joridae Extract Using High Temperature Extraction Technique

After extracting 100 g of Jeju chopsticks with 2 liters of purified water at a temperature of 121 ℃, the solution filtered with filter paper was concentrated to dryness to prepare the Jeju chopsticks extract.

[ Comparative example 2] Preparation of jeju jeoldae extract using alkaline solution

2 liters of 2% alkali solution was added to 100 g of Jeju cooking liquor at 25 ° C, and the mixture was reacted. Then, hydrochloric acid was added to the stock solution so that the pH was 2.0. It was. The concentrate was extracted three times with 1 liter of acetonitrile, and the filtrate was concentrated to dryness to prepare Jeju extract.

[ Comparative example 3] Preparation of Jeju Litter Extract Using Ethanol Extraction Method

After extracting 100 g of jeju bar with ethanol 2 liters at a temperature of 25 ℃, the solution filtered with filter paper was concentrated to dryness to prepare a jeju bar extract.

[ Comparative example 4] Preparation of jejudae extract using enzymatic digestion technique

After treating 100 g of Jeju boiled crab with glycosidase at a temperature of 25 ° C. and extracting it with 2 liters of purified water, the solution filtered with filter paper was concentrated to dryness to prepare Jeju boiled extract.

[ Test Example 1] Comparison of Components Contents of Jeju Joridae Extract Using High-alkali Extraction Method and Conventional Extraction Method of the Present Invention

The content of P-cumarin was analyzed in Jeju jejudae extract of Example 1 and Comparative Examples 1 to 4 using the following HPLC analysis. The results are shown in Table 1 below.

1) HPLC conditions

Column: Atlantis dC18 (4.6 × 250 mm) inverted column (Waters)

HPLC: Young Lin (Pump, SP930D; Detector, UV730D; Vacuum degasser & mixer, Acme 9000)

-Analysis temperature: 30 ℃

Detection wavelength: 280 nm

Flow rate: 1 ml / min

Eluent: 2% acetic acid: acetonitrile = 80:20 (v / v)

ingredient Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 p-coumarin acid 12.09% 0.04% 0.04% 0.03% 0.02% Ferulic acid 4.70% - - - - Lignin Polyphenols 83.21% - - - -

As can be seen in Table 1, when comparing the total polyphenol content of the jeju jeoldae extract obtained by using the conventional extraction method (Comparative Examples 1-4) and jeju jeoldae extract prepared by the hot-alkali extraction method, the hot-alkali extraction technique The p-coumarinic acid and total polyphenol contents of Jeju Joridae extract were 300 ~ 600 times and 2,500 ~ 5,000 times higher than those of conventional extraction methods. It can be confirmed that the extraction in the extraction method according to the invention.

Therefore, Jeju Joridae extract extracted using the high temperature-alkali extraction method of the present invention could contain a large amount of potency components than other conventional extraction methods.

[ Test Example 2] DPPH ( Diphenylpicryl Hyrazyl ) Radical Removal effect

In order to measure the antioxidant effect of the Jeju boreal extract using the hot-alkali extraction method, the removal effect of DPPH radicals of the extract of the Jeju bori extract using the hot-alkali extraction method of the present invention and the extract of the comparative example was compared to the conventional method.

Experiment of the removal effect of the DPPH radical is a method commonly used as a measure of antioxidant activity, through a change in absorbance generated by reduction of free radical 1,1-diphenyl-2-picryl hydrazyl (Diphenylpicryl Hyrazyl, DPPH) It is a method of evaluating antioxidant activity. The degree to which the oxidation of DPPH was inhibited and the absorbance was reduced compared to the control group was measured, and the concentration (IC50) showing an absorbance of 50% or less compared to the absorbance of the control group was evaluated as the effective antioxidant concentration. Lower IC50 means higher radical scavenging effect, which means higher antioxidant capacity.

In detail, the experimental procedure, 190 μl of 100 μM (in ethanol) DPPH solution and 10 μl of Example 1 and Comparative Examples 1 to 4 were prepared for each concentration, and the reaction solution was made for 30 minutes at 37 ° C., followed by absorbance at 540 nm. Was measured. The results are shown in Table 2 below.

division IC ₅₀ (μg / ml) Example 1 150 Comparative Example 1 500 Comparative Example 2 480 Comparative Example 3 510 Comparative Example 4 500

As can be seen in Table 2, Example 1 using the high temperature-alkali extraction method of the present invention was confirmed that the antioxidant capacity is 3.3 times superior to Comparative Examples 1 to 4.

[ Test Example 3] Nitric oxide (NO) production inhibitory activity

Cell culture was obtained by using a Murine macrophage cell line RAW 264.7 cells from the Korean Cell Line Bank, 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA) ) Was cultured in a 5% CO ₂ incubator at 37 ° C using Dulbecco's modified Eagle's medium (DMEM) medium, and passaged once every two days.

To evaluate the inhibition of nitric oxide (NO) production, RAW 264.7 cells were placed in a 24 well plate at 2.0 × 10 ⁵ cells / ml using DMEM medium containing 10% FBS and incubated for 18 hours. After preparing the samples by concentration, the prepared samples were mixed with LPS at a concentration of 1µg / ml and treated at the same time in a cell at 37 ℃, 5% CO ₂ incubator for 24 hours. 100 μl of the cell culture supernatant and 100 μl of Griess reagent were mixed and reacted in a 96 well plate for 10 minutes, and the absorbance was measured at 540 nm. The amount of NO produced was in the form of NO ₂ ^- present in the cell culture solution using Griess reagent [1% (w / v) sulfanilamide, 0.1% (w / v) naphylethylenediamine in 2.5% (v / v) phosphoric acid]. Sodium nitrite (NaNO ₂ ) was used as standard.

division IC ₅₀ (μg / ml) Example 1 125 Comparative Example 1 1,003 Comparative Example 2 980 Comparative Example 3 1,005 Comparative Example 4 1,012

As can be seen in Table 3, Example 1 using the high temperature-alkali extraction method of the present invention was confirmed that the anti-inflammatory activity is about 8 times superior to Comparative Examples 1 to 4.

[ Test Example 4] Measurement of melanin biosynthesis inhibitory activity

Cell culture was obtained by using B16F10 cell, a melanoma cell line, from Korean Cell Line Bank, and 1% antibiotic (Gibco, USA) and 10% fetal bovine serum (FBS; Gibco, Grand Island, USA). Dulbecco's modified Eagle's medium (DMEM) medium was used to incubate in 37 ° C., 5% CO 2 incubator and passaged once every two days.

To evaluate the melanin biosynthesis inhibitory activity, B16F10 cells cultured in DMEM medium were inoculated at 2.0x10 ⁴ cells / well in 24 well plates. Samples were prepared for each concentration after 24 hours incubation in 37 ℃, 5% CO ₂ incubator. In order to stimulate each sample and cell at each concentration, α-MSH, a stimulant, was added at 200nM. After incubation for 48 hours at 37 ℃, 5% CO ₂ incubator, the culture medium was removed and each well was washed with phosphate buffer (pH 7.4). 100 μl of 1N NaOH was added to remove the cells and then transferred to a 1.5ml tube. After reacting at 80 ° C. for 1 hour, the absorbance was measured at 405 nm. Inhibition of melanin biosynthesis was shown by the decrease in absorbance of the addition and the addition of the sample solution.

division Concentration (μg / ml) Melanin production inhibition rate (%) Example 1 250 80 Comparative Example 1 250 12 Comparative Example 2 250 10 Comparative Example 3 250 15 Comparative Example 4 250 8

As can be seen from Table 4, Example 1 jeju jeoldae extract using the hot-alkali extraction method of the present invention showed a melanin production inhibition rate of about 80%, but Comparative Examples 1-4 is about 8-15% low melanin production Inhibition rate was shown.

Claims

(a) adding 2 liters of 2% sodium hydroxide solution to 100 g of sasa quelpaertensis at a temperature of 121 ° C. and reacting for 5 hours to obtain a hydrolyzed Jeju sachet extract;
(b) adding hydrochloric acid to the stock solution of step (a) so that the pH is 2.0 and then filtering to remove debris and concentrating under reduced pressure; And
(c) extracting the concentrate of step (b) three times with 1 liter of acetonitrile and then concentrated to dryness;
Preparation method of Jeju choridae extract comprising a.

(a) adding 2 liters of 2% sodium hydroxide solution to 100 g of sasa quelpaertensis at a temperature of 121 ° C. and reacting for 5 hours to obtain a hydrolyzed Jeju sachet extract;
(b) adding hydrochloric acid to the stock solution of step (a) so that the pH is 2.0 and then filtering to remove debris and concentrating under reduced pressure; And
(c) extracting the concentrate of step (b) three times with 1 liter of acetonitrile and then concentrated to dryness;
Cosmetic composition characterized in that it contains an antioxidant activity and whitening effect by containing the extract jeju jeoldae prepared by the manufacturing method comprising an.

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