KR20130071918A - Method for extracting bambusoideae's leaves using subcritical solvents - Google Patents

Abstract

PURPOSE: A method for preparing a bamboo leaf extract is provided to quickly extract and to selectively obtain useful substances according to an extraction condition. CONSTITUTION: A method for preparing a bamboo leaf extract comprises a step of treating bamboo leaves with a subcritical solvent. The subcritical solvent is prepared by treating with a mixture of water and ethanol at 120-220 deg. C and 100-2500 PSI. The volume ratio of water and ethanol is 0.1-10:1. The extract contains one or more compounds selected from the group consisting of coumaric acid, tricin, p-coumaryl alcohol, 3- (4-hydroxyphenyl)-2-propenoic acid ethyl ester, and ferulic acid ethyl ester.

Description

[0001] METHOD FOR EXTRACTING BAMBUSOIDEAE'S LEAVES USING SUBCRITICAL SOLVENTS [0002]

The present invention relates to a bamboo leaf extract extraction method.

Bamboo (bamboo) is a perennial plant belonging to Gramineae and Bambusoidae. There are about 280 kinds of bamboo in the world and 70 kinds of bamboo in Korea. Phyllostachys nigra , Phyllostachys pubescens ), the throne ( Phyllostachys bambusoides ), cotton ( Phyllostachys nigra there is . henonis ), Sasa ( Sasa borealis ) and Jeju borealis ( Sasa quelpaertensis ) is known as a representative bamboo.

Phyllostachys nigra is also known as 紫 竹, 水 竹子, and 胡 竹, and is used as a medicinal product for vomiting, inflammation, span, stomach, heritage, warts, pelvic torture, tetanus, sweating, It is used. Despite the various efficacies of the leaves, it is not easy to obtain the leaf extracts which can be obtained with high efficiency and low cost.

Conventional organic solvent extraction methods are detrimental to human health, safety and the environment through the use of excessive organic solvents. Microwave and ultrasonic extraction methods have been developed to overcome the limitations of the solvent extraction method. However, the extraction efficiency is low and the cost is high and it is not widely used.

Accordingly, the present inventors have found that when extracting bamboo leaves using an accelerated solvent extractor (ASE) or a pressurized liquid extraction (PLE) And that the content of the physiologically active ingredient contained in the extract is increased. Thus, the present invention has been completed.

Korean Patent Publication No. 10-2006-0129607

It is an object of the present invention to provide a bamboo leaf extract extraction method.

In order to achieve the above object, the present invention provides a bamboo leaf extract extraction method comprising treating a bamboo leaf with a subcritical solvent.

In addition, the present invention provides a composition comprising the bamboo leaf extract extracted by the above extraction method.

The present invention also relates to a process for the preparation of the compound of formula (I), which comprises reacting coumaric acid, tricin, p-coumaryl alcohol, 3- (4- hydroxyphenyl) -2- propanoic acid- (4-hydroxyphenyl) -2-propenoic acid ethyl ester, and ferulic acid ethyl ester.

The extraction method of the present invention has a faster extraction speed than an organic solvent extraction method commonly used in the art and is harmless to the environment and can selectively obtain a useful substance according to extraction conditions such as temperature, pressure, and addition amount of an organic solvent But it is useful because it has a high extraction rate and contains a large amount of a physiologically active substance containing an antioxidant substance.

FIG. 1 is an experimental condition for measuring the total polyphenol content in the extract.
2 is a graph showing changes in the content of total phenolic compounds in an accelerated solvent extractor (ASE) according to respective temperatures and solvent conditions (ethanol / water mixing ratio) using Response Surface Method (RSM) The results obtained.
Figure 3 shows the ABTS radical scavenging ability at each temperature condition.
4 is an online antioxidant HPLC chromatogram at extraction conditions of 40 and 160 degrees. A negative peak in the chromatogram indicates the degree of antioxidant activity.
5 is the NMR data of the compound.
Figure 6 is the LC / MS spectrum of the compound.

In one aspect, the present invention relates to a method for extracting bamboo leaf extract, which comprises treating a bamboo leaf with a subcritical solvent.

In one aspect of the present invention, the bamboo may be a Bambusoidae plant belonging to the family Graminea, which is distributed mainly in Southeast Asia. As used herein, the term "subcritical solvent" means a solvent that maintains a liquid phase at a temperature above the boiling point, and can be obtained by subcritical water extraction (SWE). The term " subcritical water extraction method "as used herein means a method of extracting a specific organic material in a short time using a change in dielectric constant (e) of water according to temperature and pressure in a solid or semi-solid sample. Specifically, in the present specification, the subcritical water extraction method may include a technique of extracting water and ethanol as a solvent under high pressure and high temperature conditions.

In an extraction method of bamboo leaf extract according to one aspect of the present invention, the subcritical solvent can be obtained by treating a mixture of water and ethanol at a temperature of 120-220 DEG C and a pressure of 100-2500 PSI. In one aspect of the present invention, the subcritical solvent may be obtained by using an accelerated solvent extractor (ASE) or a pressurized liquid extraction (PLE), but the present invention is not limited thereto, Devices commonly known in the art may be used.

In the above extraction method, when the extraction temperature is lower than 120 ° C, the extraction yield of the physiologically active substance with low polarity is lowered because the dielectric constant of water is increased. When the temperature is higher than 220 ° C, It was confirmed to be degraded. In view of the above, the extraction temperature of the extraction method may be 130-210 ℃, 140-200 ℃ or 150-190 ℃.

In addition, in the above extraction method, when the extraction pressure was less than 100 PSI, it was observed that the solvent could not maintain the liquid phase at a temperature higher than the boiling point, and when it exceeded 2500 PSI, it was confirmed that the subcritical extraction device could not withstand pressure. In such a viewpoint, the extraction pressure of the extraction method may be 300-2300 PSI, 500-2100 PSI, 700-1900 PSI, 900-1700 PSI or 1100-1500 PSI.

In the method for extracting bamboo leaf extract according to one aspect of the present invention, the volume ratio of the mixture of water and ethanol may be 0.1-10: 1. When the volume ratio of water to ethanol is out of the above range, it has been observed that it is not easy to extract low polarity substances from the extract. In the above-mentioned aspect, the volume ratio of the mixture of water and ethanol in the bamboo leaf extract extraction method of the present invention may be 0.3-8: 1, 0.5-6: 1 or 0.7-4: 1, 1, but is not limited thereto.

In one aspect of the present invention, bamboo leaf extract is selected from the group consisting of Phyllostachys nigra , Phyllostachys Pubescens , Phyllostachys bambusoides , Phyllostachys nigra there is . henonis), bamboo (Sasa borealis) and Jeju Sasa (Sasa quelpaertensis ), and in particular, the bamboo may be disrupted.

In one aspect of the present invention, the bamboo leaf extract is selected from the group consisting of coumaric acid (Formula 1), tricin (Formula 2), p-coumaryl alcohol 4-hydroxyphenyl) -2-propenoic acid ethyl ester (formula 4) and ferulic acid ethyl ester (formula 4) ester (Formula 5). < / RTI >

The present invention relates to a composition comprising bamboo leaf extract extracted from the bamboo leaf extract according to another aspect. The composition comprising bamboo leaf extract which is one aspect of the present invention may be a composition for antioxidation.

In a composition comprising bamboo leaf extract which is an aspect of the present invention, the composition may be a cosmetic composition, and may further comprise a cosmetic composition for antioxidation. The cosmetic composition is not particularly limited in formulation, and may be appropriately selected as desired. For example, skin lotion, skin softener, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing But the present invention is not limited thereto, and may be manufactured by any one or more formulations selected from the group consisting of a foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, and especially in the case of spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The content of the active ingredient is not particularly limited, but may be included as 0.0001 to 10% by weight based on the total weight of the composition. When the active ingredient satisfies the content, it may exhibit excellent efficacy without side effects.

In addition to the bamboo extract prepared above, the cosmetic composition may further contain ingredients included in a functional additive and a general cosmetic composition. The functional additives may include water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

In addition to the above-mentioned functional additive, in addition to the cosmetic composition of the present invention, components contained in a general cosmetic composition may be blended if necessary. In addition to the other components included, oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, blood circulation And accelerators, cooling agents, limiting agents, purified water, and the like.

In another aspect, the present invention relates to a process for the preparation of a compound of formula (I), which comprises reacting a compound selected from the group consisting of coumaric acid, tricin, p-coumaryl alcohol, 3- (4-hydroxyphenyl) A bamboo leaf extract comprising at least one compound selected from the group consisting of 3- (4-hydroxyphenyl) -2-propenoic acid ethyl ester and ferulic acid ethyl ester.

In one aspect of the present invention, the bamboo leaf extract comprises coumaric acid in an amount of 2.5% by weight or more based on the total weight of the extract, and the tricin is present in an amount of 3% Or the p-coumaryl alcohol comprises 1% by weight or more based on the total weight of the extract, or the 3- (4-hydroxyphenyl) -2-propenoic acid ethyl ester (3 - (4-hydroxyphenyl) -2-propenoic acid ethyl ester is contained in an amount of not less than 1.3 wt% based on the total weight of the extract, or the ferulic acid ethyl ester is contained in an amount of not more than 1.2 wt% Or more.

Specifically, the coumaric acid may be contained in an amount of 2.5 wt% or more and 99.9 wt% or less based on the total weight of the extract. When the extract contained less than 2.5% by weight of cumaric acid, the antioxidant activity was insignificant, and it was confirmed that the extract had not exceeded 99.9% by weight due to the presence of impurities. In the above-mentioned aspect, in the extract, the cumaric acid is contained in an amount of 3 to 80 wt%, 3.5 to 70 wt%, 4 to 60 wt%, 4.5 to 50 wt% By weight or more and 40% by weight or less, or 5.5% by weight or more and 30% by weight or less.

The tricin may be contained in an amount of 3 wt% or more and 99.9 wt% or less based on the total weight of the extract. When the extract contained less than 3% by weight of cumaric acid, the antioxidant activity was insignificant and it was confirmed that the extract had not exceeded 99.9% by weight due to the presence of impurities. In the above-mentioned aspect, in the extract, tricine is contained in an amount of 3.5 to 80 wt%, 4 to 70 wt%, 4.5 to 60 wt%, 5 to 50 wt%, and 5.5 wt% % Or more and 40 wt% or less, or 6 wt% or more and 30 wt% or less.

The p-coumaryl alcohol may be contained in an amount of 1 wt% or more and 99.9 wt% or less based on the total weight of the extract. When the extract contained less than 1% by weight of cumaric acid, the antioxidant activity was observed to be insignificant, and it was confirmed that the extract had not exceeded 99.9% by weight due to the presence of impurities. In the above-mentioned aspect, the p-coumaryl alcohol in the extract is present in an amount of 1 to 80 wt%, 3 to 70 wt%, 5 to 60 wt%, 7 to 50 wt% By weight or less, 9% by weight or more and 40% by weight or less, or 11% by weight or more and 30% by weight or less.

The 3- (4-hydroxyphenyl) -2-propenoic acid ethyl ester may be present in an amount of 1.3 to 99.9% by weight based on the total weight of the extract. ≪ / RTI > When the extract contained less than 1.3% by weight of cumaric acid, the antioxidant activity was insignificant, and it was confirmed that the extract had not exceeded 99.9% by weight due to the presence of impurities. In the above-mentioned aspect, the extract of the present invention contains 3- (4-hydroxyphenyl) -2-propenoic acid-ethyl ester in an amount of 3 to 80% by weight, 3.5 to 70% Or more and 60 wt% or less, 4.5 wt% or more and 50 wt% or less, 5 wt% or more and 40 wt% or less, or 5.5 wt% or more and 30 wt% or less.

The ferulic acid ethyl ester may be contained in an amount of 1.2 to 99.9% by weight based on the total weight of the extract. When the extract contained less than 1.2% by weight of cumaric acid, the antioxidant activity was insignificant, and it was confirmed that the extract had not exceeded 99.9% by weight due to the presence of impurities. In the above-mentioned aspect, in the extract, the ferulic acid ethyl ester is contained in an amount of 3 to 80 wt%, 3.5 to 70 wt%, 4 to 60 wt%, 4.5 to 50 wt% , 5 wt% or more and 40 wt% or less, or 5.5 wt% or more and 30 wt% or less.

The bamboo extract which is one aspect of the present invention may have antioxidant ability.

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.

Accelerated solvent extraction system ( accelerated solvent extracter , ASE ) Was used to determine total polyphenol content

Dry oak leaves were filled into an accelerated solvent extractor (ASE; Dionex, Sunnyvale, Calif., USA) with different extraction temperature and solvent conditions to extract ocher extract. Thereafter, the total polyphenol content in the extract was measured. The respective experimental conditions and the total polyphenol content contained in each extract are shown in Fig.

Reaction surface method ( Response Surface Method , RSM Analysis of polyphenol content optimization conditions

Using the data in Table 1 above, conditions for optimizing the total polyphenol content by Response Surface Method (RSM) were analyzed (FIG. 2). As a result, it was expected that the total polyphenol content of the extract would be optimized when the extraction temperature was 160 ° C or higher and 50% ethanol was used.

Determination of total polyphenol content in Ojuk extract

In order to confirm the result expected by Example 2, the polyphenol content of the oyster extract obtained in Example 1 was measured. The content of phenol was colorimetrically determined according to the total phenol content analysis (Folin-Denis method). To 20 uL of the sample solution (1,000 ppm), 60 uL of distilled water and 20 uL of phenol reagent (2N) of Folin-Ciocalteu were added. After 5 minutes, 100 uL of 20% sodium carbonate solution was added and mixed. After 30 minutes, absorbance was measured at 730 nm. The total phenolic content was expressed as catechine equivalents (mg catechin / g dry sample) after catechin (catechine) was prepared as a standard product at a concentration of 10, 50, 100 and 200 ppm respectively (Table 1).

Temperature (℃) EE (%) ^a TP (mg / g DW) ^b 40 5 1.8 160 21 15 190 28 28

* ^a EE: extraction efficiency for dry weight of leaves.

** ^b TP: total phenolic acid expressed as catechine equivalent value in dry weight (g) of leaves

As can be seen from the above Table 2, at a high temperature of 160 ° C or higher, the extracting ratio of Ojuk extract was high and the total polyphenol content contained in the extract was high.

Measurement of antioxidative activity by changing temperature condition

The antioxidant activity of the extracts was investigated using an online antioxidant system and the components having radical scavenging activity of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)

The ABTS method is a colorimetric method for measuring the reduction of ABTS free radicals by measuring the reducing power of ABTS free radicals.

A mixture of 3.5 mM potassium persulfate and 2 mM ABT was prepared, diluted 8-fold and reacted for 14 hours under dark conditions. 95 uL of ethanol and 100 uL of ABTS reaction solution were added to 5 uL of the extract (1,000 ppm) obtained in Example 1. After 5 minutes, the absorbance was measured at 723 nm. The 50% ABTS radical scavenging value of the extract was determined at 40 ° C and 160 ° C, respectively, using trolox as a positive control. As a result (FIG. 3), it was confirmed that ABTS radical scavenging ability was high at a high temperature of 160 ° C.

Identification and purification of compounds having antioxidative activity

On-line antioxidant HPLC chromatograms at extraction conditions of 40 ° C and 160 ° C were compared to identify antioxidant components extracted only at high temperature of 160 ° C or higher (FIG. 4, peak 1 to 5).

In order to identify the structures of peaks 1 to 5 in FIG. 4, separation and purification experiments were conducted. The dried oak leaves which had been dried at room temperature were cut into a size of 2 cm, and 43 g (dry weight) of the oak leaves were extracted under the condition of 50% ethanol extraction at 160 캜 to obtain 13 g of oak leaf extract. The oak leaf extract was subjected to solvent fractionation successively with nucleic acid, ethyl acetate and butanol to obtain 3.5 g of the nucleic acid fraction and 3.5 g of the ethyl acetate fraction.

The ethyl acetate fraction was subjected to preparative HPLC (Jasco prep-HPLC, YMC-PACKC18RP 250 * 20 mm, ACCN-water 10-90% in 60 min) using silica gel open column chromatography (ethyl acetate: hexane = 8: To obtain 2.5 mg of Compound 1, 3 mg of Compound 2, 1 mg of Compound 3 and 1.3 mg of Compound 4, respectively. The nucleic acid fractions were purified by preparative HPLC (Waters prep-HPLC4000, YMC-PACKC18RP 250 * 20 mm, MeOH-water 60 (0) -60 (5) -80 (40) -100% (50 min) mg.

NMR and LC-MS of the separated compound proceeded to identify the structure. NMR analysis was performed to obtain the results shown in Fig. 5, and as a result of LC / MS analysis, Fig. 6 was obtained. NMR and LC / MS data analysis showed that the bamboo extract of the present invention contained coumaric acid (Compound 1), tricin (Compound 2), p-coumaryl alcohol (Compound 3) , 4-hydroxyphenyl) -2-propenoic acid ethyl ester (Compound 4) and ferulic acid ethyl ester (compound 5). ≪ / RTI >

Hereinafter, formulation examples of the cosmetic composition will be described, but the present invention is not intended to be limited thereto but is specifically described.

Formulation Example 1 Lotion Compounding ingredient weight% Omega extract 3.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Water soluble collagen (1% aqueous solution) 1.00 Sodium citrate 0.10 Citric acid 0.05 White pickle extract 0.20 1,3-butylene glycol 3.00 Purified water to 100

Formulation Example 2 Cream Compounding ingredient weight% Omega extract 1.00 Polyethylene glycol monostearate 2.00 Self emulsifying monostearic acid glycerin 5.00 Cetyl alcohol 4.00 Squalene 6.00 Tri-2-ethylhexane glyceryl 6.00 Sphingolipid 1.00 1,3-butylene glycol 7.00 Purified water to 100

Formulation Example 3 Pack Compounding ingredient weight% Omega extract 5.00 Polyvinyl alcohol 13.00 L-ascorbic acid-2-phosphate magnesium salt 1.00 Lauroylhydroxyproline 1.00 Water soluble collagen (1% aqueous solution) 2.00 1,3-butylene glycol 3.00 ethanol 5.00 Purified water to 100

Formulation Example 4 Serum Compounding ingredient weight% Omega extract 2.00 Hydroxyethylene cellulose (2% aqueous solution) 12.00 Xanthan gum (2% aqueous solution) 2.00 1,3-butylene glycol 6.00 Concentrated glycerin 4.00 Sodium hyaluronate (1% aqueous solution) 5.00 Purified water to 100

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

As a method of extracting bamboo leaf extract,
Comprising treating subcritical solvent to bamboo leaves,
Extraction method of bamboo leaf extract. The method of claim 1, wherein the subcritical solvent
A mixture of water and ethanol
A method of extracting bamboo leaf extract, which is a solvent obtained by treatment under a temperature of 120-220 ° C. and a pressure of 100-2500 PSI. The method of extracting bamboo leaf extract according to claim 2, wherein the volume ratio of the mixture of water and ethanol is 0.1-10: 1. The method of extracting bamboo leaf extract of claim 1, wherein the bamboo comprises at least one selected from the group consisting of bamboo porridge, bamboo shoot, kingdae, cotton swab, bamboo shoot, garrison, and jeju jangri. The method of extracting bamboo leaf extract of claim 4, wherein the bamboo comprises bamboo porridge. The method of claim 1, wherein the bamboo leaf extract is coumaric acid (coumaric acid), tricin (tricin), p- coumaryl alcohol (p-coumaryl alcohol), 3- (4-hydroxyphenyl) -2-propeno A method for extracting bamboo leaf extract, comprising one or more compounds selected from the group consisting of 3- (4-hydroxyphenyl) -2-propenoic acid ethyl ester and ferulic acid ethyl ester. A composition comprising the bamboo leaf extract extracted by the extraction method of any one of claims 1 to 6. The composition of claim 7, wherein the composition comprises a cosmetic composition. Coumaric acid, tricin, p-coumaryl alcohol, 3- (4-hydroxyphenyl) -2-propenoic acid-ethyl ester (3- (4-hydroxyphenyl) Bamboo leaf extract containing one or more compounds selected from the group consisting of) -2-propenoic acid ethyl ester) and perulic acid ethyl ester. 10. The method of claim 9,
The coumaric acid (coumaric acid) is included 2.5% by weight or more based on the total weight of the extract,
The tricin is included at least 3% by weight based on the total weight of the extract,
The p-coumaryl alcohol (p-coumaryl alcohol) is included by weight of 1% by weight or more based on the total weight of the extract,
The 3- (4-hydroxyphenyl) -2-propenoic acid-ethyl ester (3- (4-hydroxyphenyl) -2-propenoic acid ethyl ester) is included 1.3 wt% or more based on the total weight of the extract,
Or the perulic acid ethyl ester (ferulic acid ethyl ester) is contained 1.2% by weight or more based on the total weight of the extract, bamboo leaf extract.

Images