KR101838266B1 - Method for Preparation of polymethoxyflavones from citrus peel by using supercritical fluid extraction - Google Patents

Abstract

The present invention relates to an extract of citrus peel having an increased content of polymethoxyflavone which can be utilized in fields of bioengineering, pharmacy and food, and a method for manufacturing the same. The present invention provides a method for manufacturing an extract of citrus peel having an increase content of polymethoxyflavone, comprising the following steps: 1) a step for pretreating by cutting, washing and drying citrus peel; 2) a step for obtaining an alcohol extract by soaking the peel pretreated in step 1 in alcohol, filtering and enriching; and 3) a step for obtaining a supercritical fluid extract from the alcohol extract obtained in step 2 by using supercritical carbon dioxide as a solvent. According to the present invention, a yield of nobiletin and tangeretin is increased by performing alcohol extraction and supercritical carbon dioxide extraction, thereby increasing a yield of polymethoxyflavone.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a method for preparing citrus peel extract, which comprises increasing the content of polymethoxy flavone using a supercritical fluid extraction method,

The present invention relates to a citrus peel extract having an increased amount of polymethoxy flavone which can be utilized in biotechnology, medicine, and food field, and a method for producing the same. More particularly, the present invention relates to a citrus peel extract having a content of novoletine and anthraquinone, Citrate peel extract using supercritical extraction method and a method for preparing the same.

Recently, as the number of consumers who prefer foods and drugs using natural products with little side effects or excessive stimulation has been increasing, the development value of natural raw materials is increasing.

Citrus fruit of various varieties including citrus fruit (C.sunki hort. Ex Tanaka) which is native citrus cultivated in Jeju Island has diverse functional materials which are beneficial to the body as well as vitamin C, and polyphenols Flavonoids < / RTI > There are about 60 functional substances in citrus fruit. The flavonoid is a generic name of a yellow pigment compound consisting of the basic skeleton of C6-C3-C6. In addition to common flavonoids such as rutin and theosmin in citrus fruits, flavonoids specific to citrus such as naringin, hesperidin, and poly It contains methoxy flavone-based nobiletin and tangeretin. Most flavonoid substances have a hydroxyl group (-OH) and seem to be related to functions such as antioxidation. Nobiletin belongs to a flavone group having a methoxyl group (CH3O-) among flavonoids, It is known that the biological activity is also high with a smaller content. In addition to nobiletin, there are tangeretin and sinensitin, especially those found only in citrus fruits. Citrus, especially, C.sunki hort (ex Tanaka) is one of the 12 indigenous native orchids native to Jeju. The dermis, which has dried the peel of citrus fruits, has been used as herb medicine, and C. hybrid Shiranuhi Jeju is a high-grade varieties of citrus citrus fruit, Jean-ri and Halabong have been reported to extract nobilisetin and tangerine. In particular, it has been reported that nobiletin and tangeretin are contained in a large amount in the pulp and are contained in a large amount in the remnant of the juice, but it is known that a sufficient amount can not be obtained by a simple extraction method. Therefore, there is a need for a method of effectively recycling a large amount of citrus residues after the fruit juice is produced when producing confectionery and beverages.

Korean Patent No. 10-0892180 Korean Patent Publication No. 10-2016-0097771

It is an object of the present invention to provide a process for producing a citrus peel extract in which the content of polymethoxy flavone is increased by immersing the citrus peel in the alcohol and extracting supercritical carbon dioxide with a solvent.

In order to accomplish the above object, there is provided a method for preparing a citrus fruit, comprising the steps of: (1) a pretreatment step of cutting, washing and drying the citrus peel; (2) dipping the pre-treated peel in the step 1, filtering and concentrating the same in step 1 to obtain the alcoholic extract; (3) obtaining the alcohol extract obtained in step 2 as a supercritical fluid extract using supercritical carbon dioxide as a solvent; The present invention also provides a method for producing citrus peel extract having an increased content of polymethoxy flavone.

The above-mentioned two-step alcohol may contain 10 to 20 times the dry weight of the perilla.

The step of preparing the supercritical extract of step (3) comprises: (a) preparing a first supercritical fluid extract using supernatant carbon dioxide as a solvent; (b) preparing the first supercritical fluid extract as a second supercritical fluid extract using supercritical carbon dioxide as a solvent; . ≪ / RTI >

Wherein the first supercritical fluid extract is extracted at a temperature of 30 to 70 DEG C under a pressure of 100 to 400 bar and the second supercritical fluid extract is pressurized at a temperature of 30 to 80 DEG C to a pressure of 100 to 600 bar Lt; RTI ID = 0.0 > conditions. ≪ / RTI >

The polymethoxy flavone may include at least one member selected from the group consisting of noviracin and anthraquinone.

The citrus peel extract prepared by the above-described method may contain 13.8 to 74.0% by weight of the total amount of polymethoxy flavone.

According to the present invention as described above, by performing alcohol soaking and supercritical carbon dioxide extraction, the yield of the Novartin and anthraquinone is increased, and the yield of the polymethoxy flavone is increased.

FIG. 1 is a diagram illustrating a method for preparing citrus peel extract having an increased content of polymethoxy flavone according to an embodiment of the present invention.
FIG. 2 shows the results of an analysis of polymethoxy flavone in alcohol extract of the present invention.
FIG. 3 shows the results of an analysis of polymethoxy flavone in the first supercritical fluid extract according to an embodiment of the present invention.
Figure 4 shows the results of an analysis of polymethoxy flavone HPLC in a second supercritical fluid extract according to an embodiment of the present invention.
FIG. 5 shows the result of repeated analysis of the content of polymethoxy flavone in the second supercritical fluid extract according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

According to the practice of the present invention, (1) a pretreatment step of trimming, washing and drying the citrus peels; (2) dipping the pre-treated peel in the step 1, filtering and concentrating the same in step 1 to obtain the alcoholic extract; (3) obtaining the alcohol extract obtained in step 2 above as a supercritical fluid using supercritical carbon dioxide as a solvent; The present invention also provides a method for producing citrus peel extract having an increased content of polymethoxy flavone.

Citrus fruits are a lot of fruits produced in Korea, and vitamin and polyphenols have a high content of antioxidants, so they are well known for their effects such as skin fatigue recovery and immunity enhancement. In addition, fruit shells belonging to the citrus fruit contain various flavonoids such as anthrax, novirretin, hesperidin, and naringin, and exhibit anti-inflammatory and anti-cancer functions. The citrus fruits include citrus depressa, C. tachibana, C. leiocarpa, C. tardiva, C. succosa, C. kinokuni, C.erythrosa, C.deliciosa, C. nobilis, C.retuculata, C. tangerina, C.hanaya, And one or more citrus fruits selected from the group consisting of C. nippokoreana, C.sunki hort. Ex Tanaka and C. hybrid Shiranuhi.

In the method for preparing the citrus peel extract having increased content of polymethoxy flavone according to the present embodiment, the polymethoxy flavone is a mixture of Nobiletin (2- (3,4-Dimethoxyphenyl) - 5,6,7,8-tetramethoxychromen4-one, Hexametho xyflavone) and tangeretin (5,6,7,8-tetramethox y-2- (4-methoxyphenyl) -4H- 1-benzopyran-4-one), wherein the content of the polymethoxy flavone is increased.

[Chemical Formula 1]

(2)

The supercritical fluid extraction method sets the pressure and temperature above the vapor phase of the substance and the liquid phase to set the dissolution power in the liquid state and the diffusion coefficient and the viscosity in the gaseous state, As a method enabling extraction, the extraction solvent uses supercritical fluid having a critical temperature and a critical pressure or higher. Supercritical carbon dioxide is used in the extraction process because it is mainly used in the vicinity of room temperature and is less susceptible to thermal denaturation. Supercritical carbon dioxide can make supercritical fluid compared to other gases, and it can be used for colorless, odorless, non-toxic, non- And non-reactivity. In addition, supercritical fluids such as liquefied carbon dioxide, liquefied propane, liquefied butane, etc. can be used for low-temperature extraction, so that components that are weak to heat can be extracted without being destroyed, and essential components can be obtained. Therefore, this Example uses a supercritical fluid extraction method to obtain a biodegradable polymethoxy compound having a hydroxyl group (-OH) in a flavonoid group having a methoxyl group (CO 3 O-) and having a smaller capacity than most flavonoids The present invention also provides a method for preparing citrus peel extract having increased flavon content.

The pretreatment step of step (1) of the present invention may be a step of preparing the extractive raw material by pretreating the citrus fruits by cutting, washing, washing and drying the citrus fruits.

In step (2), the step (2) is carried out by immersing the extract in the step (1) for 10 to 20 times the weight of the extracted raw material for 24 hours, filtering the impurities, Lt; / RTI >

The step of obtaining the supercritical fluid extract of step (3) comprises the step of preparing a supercritical fluid extract using supercritical carbon dioxide as a solvent in the alcohol extract obtained in step (2), wherein the supercritical fluid extract of step Producing a fluid extract, and (b) preparing a second supercritical fluid extract.

(A) is a step of obtaining a first supercritical fluid extract using supernatant carbon dioxide as a solvent of the alcohol extract obtained in step (2), (b) The supercritical fluid extract is prepared by using the supercritical carbon dioxide as a solvent. Preferably, the first supercritical fluid extract of step (a) may be extracted under a pressure of 100 to 400 bar at a temperature of 30 ° C to 70 ° C, and the second supercritical fluid extract of step (b) More preferably, the first supercritical fluid extract at the step (a) may be extracted at a temperature of 30 to 80 ° C under a pressure of 100 to 600 bar, bar, and the second supercritical fluid extract of step (b) may be extracted under a pressure of 300 to 500 bar at a temperature of 50 ° C to 80 ° C.

The citrus peel extract prepared according to the method of manufacturing the citrus peel extract having an increased content of polymethoxy flavone according to the present invention may have a total content of novile retin and anthraquinone of 13.8 to 74.0% by weight.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings. It will be apparent to those skilled in the art that this embodiment is for illustrative purposes only and that the scope of the present invention is not construed as being limited by these embodiments.

Examples. Preparation of Citrus Peel Extract with Increased Polymethoxy Flavone Content.

In the example of the present invention, the extraction raw material was prepared by pretreating the citrus peel, which was then washed, washed, and then dried. The dried citrus peel was dipped in a 10 to 20 times dry weight alcohol for 24 hours, filtered to remove impurities, concentrated under reduced pressure, and dried. The first supercritical fluid extract was obtained under the conditions of a pressure of CO ₂ of 300 bar and a temperature of 60 ° C by using a diaphragm type compressor, 200 g of the dried extract of the alcohol was put into a supercritical CO ₂ extractor (RTI, Korea) _{A second} supercritical fluid extract was obtained under the conditions of a pressure of 500 bar and a temperature of 70 캜. The alcohol extract, the first supercritical fluid extract and the second supercritical fluid extract were collected and dried under reduced pressure.

Experimental example. HPLC  Analysis of alcohol extracts Supercritical  Fluid extract Novyletin , Anthrax Retin  And Polymethoxy flavone  Comparison of yield.

The supercritical fluid extract and the alcoholic extract were dissolved in methanol according to the operating conditions shown in Table 1 and Table 2, and filtered through a syringe filter to obtain a high-speed liquid Chromatography (HPLC) and ultraviolet absorption spectrophotometer were used for the analysis. The maximum absorption wavelength was quantitatively analyzed at 254 nm. The analytical time was 60 minutes. The contents of the Novartin and anthraquinone were calculated by plotting a calibration curve using a standard reagent (Wako). The concentration of each extract was fixed at 10 mg / ml and analyzed by high performance liquid chromatography. The resultant value was calculated according to the following formula. The content of polymethoxy flavone was calculated by taking the sum of novirretin and anthraquinone as molecules in terms of the denominator of the total weight of the extract.

 Item  Condition  Dose  20 μl  Detector wavelength  254 nm  Column temperature  25 ℃  Mobile phase A: B MeOH: H ₂ O  Flow rate  1.0 ml / min

 Time (minutes) Solvent A (%) Solvent B (%)  0  50  50  70  100  0

[formula]

Yield of polymethoxy flavone of extract = (novilletin content in extract / dried citrus peel weight * 100) + (anthragelin content in extract / dried citrus peel weight * 100)

As a result of comparing the extracts obtained by the above examples, it was found that the orange extract of citrus peel, which is a citrus fruit, was a yellowish and viscous extract, the first supercritical fluid extract was a yellow viscous extract but the second supercritical fluid extract Was a pale yellow viscous extract. After the extraction, the results of measuring the yields of the nobiletin, anthraquinone and polymethoxy flavone of the respective extracts were calculated as shown in the above formula, and the results are shown in Table 3 below.

Novyletin
(%) Anthrax Retin
(%) Polymethoxy flavone
(%)  Alcohol extract 9.4 4.4 13.8  First supercritical fluid extract 33.6 23.9 57.5  Second supercritical fluid extract 50.0 24.0 74.0

According to the above Table 3, it was confirmed that the yield of the polymethoxypropane was increased as it went through each extraction step. The polymethoxy flavone of the alcohol extract was 13.8%, the polymethoxy flavone of the first supercritical fluid extract was 57.5%, and the polymethoxy flavone of the second supercritical fluid extract was 74.0% Which was about 5 times higher than that of the alcohol extract.

In addition, three second supercritical fluid extracts were repeatedly extracted, and the respective extracts, novirretin and anthraquinone, were confirmed by HPLC analysis, and the yield of the polymethoxy flavone was measured, ].

Novyletin
(%) Anthrax Retin
(%) Polymethoxy flavone
(%) (a)     42.73     23.18         65.91 (b)     40.81     22.95         63.76 (c)     39.37     21.92         61.29

According to the results of the HPLC chromatogram analysis shown in Fig. 5 and the results of Table 4, it was confirmed that there was no significant difference in the yield of the polymethoxy flavone of the second supercritical fluid extract repeatedly conducted. The polymethoxy flavone content of the second supercritical fluid extract of Table 4 (a) and 5 (a) was 65.91%, the polymethoxy flavone content of Table 4 (b) and 5 (b) was 63.76% The content of the polymethoxy flavone of 4 (c) and 5 (c) was 61.29%.

Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (7)

1) a pretreatment step in which citrus fruit is cut, washed and dried;
2) dipping the pre-treated rind in the step 1, filtering and concentrating the rind in the step 1 to obtain a mussels extract;
3) extracting the alcohol extract obtained in step 2 above with supercritical carbon dioxide as a supercritical fluid extract; And
4) drying the fluid extract obtained in the above step 3 under reduced pressure,
The alcohol in the step 2) is 10 to 20 times the dry weight of the perilla,
Wherein the step 3) comprises: preparing a first supercritical fluid extract using supernatant carbon dioxide as the solvent of the alcohol extract of step 2) and a second supercritical fluid extract using supercritical carbon dioxide as a solvent, Preparing a critical fluid extract,
Characterized in that the first supercritical fluid extract is extracted under a pressure of 300 bar at a temperature of 60 ° C,
Wherein the second supercritical fluid extract is extracted under a pressure of 500 bar at a temperature of 70 ° C.
delete delete delete The method according to claim 1,
Wherein the polymethoxy flavone comprises at least one member selected from the group consisting of noviracin and anthraquinone, wherein the content of the polymethoxy flavone is increased.
A citrus peel extract having increased content of polymethoxy flavone prepared by the method of claim 1. delete

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