KR20210145543A - Hot water extract of Suaeda japonica including Narcissin and Rutin, and isolation method for Narcissin and Rutin from the same - Google Patents

Abstract

The present invention relates to a method for isolating and purifying antioxidant compounds from Suaeda japonica. More specifically, the present invention relates to a hot water extract of Suaeda japonica including narcissin and rutin, and a purification method which can purify narcissin and rutin from the hot water extract of Suaeda japonica.

Description

Hot water extract of Suaeda japonica including Narcissin and Rutin, and isolation method for Narcissin and Rutin from the same}

The present invention relates to a method for separating and purifying useful components from turkey, and more specifically, a turkey extract containing narcissin and rutin, and a method for purifying narcissin and rutin from the turkey extract It relates to a purification method.

Chickweed ( Suaeda japonica ) is an annual halophyte of the Amaranthaceae family distributed mainly in the west coast of Korea. Turkey vinegar is green when young, but changes to various colors as it grows up and turns red in autumn.

As such, turkey vinegar has a red color and is used as a natural dye and red salt, a vegetable salt. In oriental medicine, the above-ground part is used as an antipyretic. Turkey vinegar is known to have a high concentration of NaCl content and rich in minerals such as magnesium, calcium, and potassium. Recently, it has been reported that it inhibits antioxidant, tyrosinase activity, and melanin production, and also has a preventive effect on diabetes.

However, research on various physiological activities and useful ingredients is still very insufficient, so the indicator ingredients for quality control of turkey vinegar and its applied products are not yet known.

On the other hand, rutin is known to exert anti-inflammatory, antioxidant, and neuroprotective effects. In particular, antidiabetic effects such as inhibiting carbohydrate absorption in the small intestine or inhibiting intracellular gluconeogenesis, promoting glucose absorption, and stimulating insulin secretion by stimulating pancreatic beta cells have been reported. ), the existence of natural products containing a large amount of it is well known.

On the other hand, narcissin (Isorhamnetin 3-rutinoside) is a type of flavonoid and has been reported to have physiological activities such as antioxidant and antibacterial activity, but it contains 0.06% in the hatchery of Narcissus tazetta var. chinensis ROEM flower, It is known that it is contained in pollen of the wild lily flower (Lilium auratum Lindley). As described above, since narcisin is contained in a very small amount in natural products and the price of a commercial product is 600,000 won per 2mg, more specific and systematic research on various physiological activities of narcissin is not being conducted, of course, with other types of flavonoids. Comparisons have also not been made.

Therefore, for more diverse research and practical use of narcissin, a technology capable of securing a natural product containing a large amount of narcissin and purifying narcissin in a simple and large amount from the natural product will be developed. There is a need.

Republic of Korea Patent Registration No. 10-1261029

The present inventors completed the present invention by confirming that turkey contains a large amount of narcissin and rutin, particularly narcissin, as a result of numerous studies.

Therefore, it is an object of the present invention to confirm that turkey contains a large amount of narcisin and rutin, particularly narcisin, which can act as useful ingredients such as antioxidants, and turkey vinegar containing a significant amount of narcisin and rutin that can be used for purifying narcisin and rutin It is to provide hot water extract.

Another object of the present invention is to provide a method for purifying narcisin and rutin, which can easily separate narcisin and rutin from turkey extract.

Another object of the present invention is to increase the utility of narcisin and rutin by securing narcisin and rutin in high yield as well as high purity, and a method for purifying narcisin and rutin, which can contribute to the activation of research using narcisin and rutin is to provide

The object of the present invention is not limited to the object mentioned above, and other objects not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above-described object of the present invention, first, the present invention provides a hot water extract of turkey containing narcissin and rutin.

In a preferred embodiment, the hot water extract for turkey is obtained through hot water extraction by mixing after drying and pulverizing turkey vinegar powder and water in a weight ratio of 1:10 to 1:15.

In a preferred embodiment, the turmeric extract contains 0.15 parts by weight or more of narcisin, and 0.05 parts by weight or more of rutin per 100 parts by weight of the turmeric powder.

In a preferred embodiment, the hot water extraction is made by heating at 80 to 95° C. for 10 to 30 minutes based on 600 to 800 ml of the water.

In addition, the present invention comprises the steps of preparing a hot water extract of turkey; a solvent fractionation step of fractionating the turkey extract with a solvent to obtain a solvent fractionation layer; a concentration step of concentrating the solvent fraction layer under reduced pressure to obtain a concentrated fraction; and a purification step of purifying the concentrated fraction.

In a preferred embodiment, the turmeric vinegar extract is obtained by mixing turkey vinegar powder and water obtained by drying and pulverizing turkey vinegar in a weight ratio of 1:10 to 1:15, followed by heating.

In a preferred embodiment, the solvent used in the solvent fractionation step is butanol.

In a preferred embodiment, the solvent fractionation step is performed twice by treating 4 to 2 volume ratios of butanol per 6 to 8 volume ratios of the turmeric extract.

In a preferred embodiment, the purification step comprises the steps of dissolving the concentrated fraction in distilled water to obtain a fraction solution; and high-performance liquid chromatography using distilled water (solvent A) and acetonitrile (solvent B) containing trifloroacetic acid (TFA) as an elution solvent for the fraction solution using an octadecylsilane (ODS) column. step; includes.

In a preferred embodiment, the pH of the solvent A and the solvent B is adjusted to 2.5.

In a preferred embodiment, after the fraction solution is injected into the high-performance liquid chromatography, the elution solvent is maintained at A/B = 9:1 (v/v) for the initial 20 minutes under the condition of a flow rate of 50 mL/min. Then, when eluted with a linear gradient elution method such that A/B = 7:3 (v/v) ratio until 80 min, rutin, a peak fraction eluted at t _{R 31 min at a detection wavelength of 280 nm, is obtained,} The peak eluted at t _R 39 min, narciscin, is obtained.

In a preferred embodiment, the obtained narcisin and rutin have a purity of 95% or more.

The present invention has the following excellent effects.

First, according to the present invention, it is confirmed that turkey contains narcisin and rutin, which can act as useful antioxidants, and using this, it is possible to provide a hot water extract of turkey containing a significant amount of narcisin and rutin.

In addition, according to the present invention, it is possible to easily separate narcisin and rutin from the turmeric extract.

In addition, according to the present invention, by securing narcisin and rutin in a high yield as well as in high purity, it is possible to increase the utility of narcisin and rutin, and to contribute to the activation of research using narcisin and rutin.

These technical effects of the present invention are not limited only to the above-mentioned range, and even if not explicitly mentioned, the effect of the invention that can be recognized by a person of ordinary skill in the art from the description of the specific content for the implementation of the invention to be described later is also of course included.

1 is a flowchart schematically showing a method for simultaneous separation of flavonoids rutin and narcissin from prepared turkey according to an embodiment of the present invention.
2 is an ODS-MPLC chromatogram of a butanol fraction of turkey herb prepared according to an embodiment of the present invention.
3 is an HPLC chromatogram and UV-spectra of indicator substances in a butanol fraction of turkey herb prepared according to an embodiment of the present invention.
^{4 is a 1} H-NMR spectrum of rutin obtained according to an embodiment of the present invention.
^{5 is a 1} H-NMR spectrum of narcissin obtained according to an embodiment of the present invention.
6 is a graph showing the DPPH free radical scavenging activity of rutin and narcissin obtained according to an embodiment of the present invention.
7 is a graph showing the cell free radical scavenging activity of rutin and narcissin obtained according to an embodiment of the present invention.
8A and 8B are structural formulas of the compounds rutin and narcisin finally isolated and identified by performing purification and isolation by the method of FIG. 1, respectively.

The terms used in the present invention are only used to describe specific embodiments, and are not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly dictates otherwise. In the present application, terms such as “comprise” or “have” are intended to designate that a feature, number, step, operation, component, part, or combination thereof described in the description of the invention exists, but is not limited to one or more other It should be understood that it does not preclude the possibility of addition or presence of features or numbers, steps, operations, components, parts, or combinations thereof.

Terms such as first, second, etc. may be used to describe various elements, but the elements should not be limited by the terms. The above terms are used only for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, a first component may be referred to as a second component, and similarly, the second component may also be referred to as a first component.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and should not be interpreted in an ideal or excessively formal meaning unless explicitly defined in the present invention. does not

In interpreting the components, it is interpreted as including an error range even if there is no separate explicit description.

In the case of a description of a temporal relationship, for example, if the temporal relationship is described as 'after', 'following', 'after', 'before', etc., 'immediately' or 'directly' This includes cases that are not continuous unless ' is used.

Hereinafter, the technical configuration of the present invention will be described in detail with reference to the accompanying drawings and preferred embodiments.

However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Like reference numbers used to describe the invention throughout the specification refer to like elements.

First, the technical feature of the present invention lies in confirming that narcisin and rutin are contained in turmeric. It is also present in turkey hot water extract, which contains significant amounts of narcisin and rutin. In other words, turkey grass is widely distributed in intertidal zones such as tidal flats and salt marshes, unlike other halophytes, including green tea plants, which have a low yield, and thus has the characteristic of being able to secure a large amount of raw materials when used as a biological resource. This is because the research on this topic is very insufficient and its utilization is still very low.

Accordingly, the present invention provides a turkey extract containing narcissin and rutin (Narcissin and Rutin).

In the present invention, the turkey vinegar extract is obtained through hot water extraction by heating after mixing turkey vinegar powder obtained by drying and pulverizing turkey vinegar and water in a weight ratio of 1:10 to 1:15, and the narcisin per 100 parts by weight of turkey vinegar powder 0.15 parts by weight or more, and rutin may be included in 0.05 parts by weight or more. In one embodiment, narcisin may be contained in an amount of 0.15-0.20% by weight based on the weight of turmeric powder, and rutin may be included in an amount of 0.05-0.08% by weight. At this time, hot water extraction may be performed by heating at 85 to 95° C. for 10 to 30 minutes based on 600 to 800 ml of water.

In addition, a technical feature of the present invention is a purification method for isolating and purifying high-purity narcissin and rutin from a turmeric extract.

Therefore, the purification method of narcisin and rutin of the present invention comprises the steps of preparing an extract of turkey hot water; a solvent fractionation step of fractionating the turkey extract with a solvent to obtain a solvent fractionation layer; a concentration step of concentrating the solvent fraction layer under reduced pressure to obtain a concentrated fraction; and purification for purifying the concentrated fraction.

Here, the turkey vinegar extract is obtained by mixing turkey vinegar powder and water obtained by drying and pulverizing turkey vinegar as described above in a weight ratio of 1:10 to 1:15, followed by heating.

The solvent used in the solvent fractionation step may be butanol, and the solvent fractionation may be performed twice by treating 4 to 2 volume ratios of butanol per 6 to 8 volume ratios of the turmeric extract.

In the purification step, high-performance liquid chromatography may be further performed on the fraction solution after dissolving the concentrated fraction in distilled water to obtain a fraction solution. For example, high-performance liquid chromatography using an octadecylsilane (ODS) column and distilled water (solvent A) and acetonitrile (solvent B) containing trifloroacetic acid (TFA) as an elution solvent can be performed. , as an embodiment, it can be carried out under the conditions of a flow rate of 50 mL/min and a detection wavelength of 280 nm. At this time, the solvent conditions are A/B = 9:1 (v/v) for the initial 20 minutes at a pH of 2.5 for solvent A and solvent B, and then A/B = 7:3 (v/v) until 80 minutes. ), it may be eluted using a linear gradient elution method. In this way, while performing high-performance liquid chromatography, in particular , the peak fraction eluted at t _R 31 min and t _R 39 min can be isolated, respectively, and the peak fraction eluted at t _R 31 min is rutin purified to a high purity of 95% or more, The peak fraction eluted at t _R 39 min is narcisin purified to a high purity of 95% or more.

On the other hand, in the embodiment to be described later, only the above-ground portion of turmeric is illustrated, but the root is not excluded.

Example 1. Preparation of turkey hot water extract containing narcisin and rutin

Chilmyeon grass ( Suaeda japonica ) was collected from the tidal flat in Sinan-gun, Jeollanam-do in August 2017, and washed with water to remove foreign substances such as the tidal flat. The above-ground portion of the washed turkey vinegar was steamed for 3 minutes and then dried with hot air at 50° C. for 48 hours, and turkey vinegar powder pulverized with a grinder was used.

After adding 700 mL of distilled water to the dried turkey powder (50 g), extraction was performed at 90 ° C. for 20 minutes, and filtered through filter paper (Whatman No. 2 filter paper, Maidstone, UK) to obtain a turkey extract containing narcisin and rutin. .

Example 2. Purification of high purity narcisin and rutin from turkey superhot water extract

The procedure shown in FIG. 1 was performed as follows to obtain a turmeric extract as in Example 1, and high-purity natural narciscin and rutin were obtained from the turmeric extract.

1. Solvent fractionation of turkey superhot water extract

The solvent was fractionated with water-saturated n -butanol (300 mL) in 700 mL prepared in Example 1, and this was repeated twice to obtain a butanol fractionation layer.

2. Concentrating the butanol fraction layer under reduced pressure

The butanol fraction layer obtained was concentrated under reduced pressure at 40° C. or lower using a vacuum evaporator (N-2N, Eyela, Tokyo, Japan) equipped with a cooling aspirator (CA-111, Eyela, Tokyo, Japan), followed by a butanol fraction (1.05 g) ) was obtained. The yield of the butanol fraction of turkey vinegar was about 2.1% based on the weight of dry turkey powder.

3. Purifying the concentrated butanol fraction

The water-saturated butanol fraction (1.05 g) obtained by solvent fractionation of the hot water extract of turkey herb was purified by MPLC equipped with an ODS reversed-phase column as follows. That is, the saturated butanol fraction was dissolved in distilled water and then Isolera one (Biotage) equipped with SNAP Ultra C18 120 g (Biotage) was injected. The mobile phase solvent was distilled water (H ₂ O, pH 2.5, solvent A) containing trifloroacetic acid (TFA) and acetonitrile (MeCN, pH 2.5, solvent B), A/B = 9:1 for the first 20 minutes (v/v) was maintained, and then eluted with a linear gradient system to a ratio of A/B = 7:3 (v/v) until 80 min. At this time, the flow rate was set to 50 mL/min, and the wavelength of the detection (PDA detector) was set to 280 nm.

As shown in FIG. 2 , ODS-MPLC was performed on the saturated butanol fraction (1.05 g) obtained by solvent fractionation of hot water extract of turkey grass, and rutin was eluted at 31 minutes and narcissin was eluted at 39 minutes. These fractions were concentrated under reduced pressure to obtain 27.6 mg and 77.6 mg of rutin and narcissin, respectively.

Experimental Example 1

To confirm the purity of the rutin and narcissin separated in Example 2, ODS-HPLC was analyzed as follows, and the results are shown in FIG. 3 . HPLC (LC-20AD, Shimadzu, Kyoto, Japan) equipped with Capcell pak C18 AQ (4.6 × 250 nm, Shiseido, Kyoto, Japan) was used, and the mobile phase was H ₂ O (pH 2.5) and TFA contained in TFA. MeCN contained in (pH 2.5) was maintained at a ratio of 85:15 (v/v) for 10 minutes, and then H ₂ O (pH 2.5) contained in TFA and MeCN contained in TFA (pH 2.5) until 30 min. was eluted with a linear gradient system to a ratio of 72:28 (v/v). At this time, the flow rate was 1.0 mL/min, and the UV wavelength was 254 nm.

As shown in FIG. 3 , rutin was detected at 22.2 min and narcissin was detected at 26.5 min on the ODS-HPLC chromatogram. It was confirmed that the two compounds correspond to the retention time of the standard, respectively. In addition, it was confirmed that the UV spectra of rutin and narcissin isolated from turkey were also consistent with each other. And from the results of ODS-HPLC analysis, it was found that the purity of rutin and narcissin isolated from turkey was more than 95%.

Experimental Example 2

To reconfirm the purity of rutin and narcissin isolated in Example 2, these compounds _{were dissolved in CD 3} OD and then ¹ H-NMR analysis ( ^unity INOVA 500 spectrometer; Varian, Walnut Creek, CA, USA) was performed, and the result is shown in Figures 4 and 5.

As shown in FIG. 4 ^{, protons corresponding to the aglycone quercetin in 1} H-NMR spectrum of rutin isolated from turkey grass [δ 6.14 (1H, d, J = 2.5 Hz, H-6), 6.33 (1H, d) , J = 2.0 Hz, H-8), 7.59 (1H, d, J = 2.0 Hz, H-2'), 6.80 (1H, d, J = 8.5 Hz, H-5'), 7.56 (1H, dd , J = 8.5, 2.0 Hz, H-6')] and protons corresponding to rutinose [δ 5.04 (1H, d, J = 8.0 Hz, H-1''), 3.25-3.82 (10H, m, H) -2''~H-6'' and H-2'''~H-5'''),4.44 (1H, d, J = 1.5 Hz, H-1'''), 1.05 (3H, d , J = 6.0 Hz, H-6''')] was observed. ^{The 1} H-NMR spectrum of rutin isolated from turkey was consistent with that of rutin, a standard material.

As shown in FIG. 5 ^{, protons corresponding to isorhamnetin, an aglycone, in 1} H-NMR spectrum of narcissin isolated from turkey grass [δ 6δ 6.13 (1H, d, J = 2.0 Hz, H-6), 6.33 (1H, d, J = 2.0 Hz, H-8), 7.86 (1H, d, J = 2.0 Hz, H-2'), 6.83 (1H, d, J = 8.5 Hz, H-5'), 7.54 (1H, dd, J = 8.5, 2.0 Hz, H-6'), 3.87 (3H, s, _{-OCH 3} )] and protons corresponding to rutinose [δ 5.15 (1H, d, J = 7.5 Hz, H-1') '), 3.25-3.83 (10H, m, H-2''~H-6'' and H-2'''~H-5'''), 4.44 (1H, d, J = 1.5 Hz, H -1'''), 1.01 (3H, d, J = 6.5 Hz, H-6''')] was observed. ^{The 1} H-NMR spectrum of narcissin isolated from turkey grass was consistent with that of the standard material.

These results show that the two compounds isolated from turkey in Example 2 are rutin and narcissin. In particular, from their ¹ H-NMR spectrum, the purity of rutin and narcssin isolated from turkey is 95% or more once again. could check

Experimental Example 3

The antioxidant activity of rutin and narcissin obtained in Example 2 was measured as follows using 1,1-diphenyl-2-picrylhydrazyl (DPPH, Sigma-Aldrich, St. Louis, MO, USA) free radical scavenging activity method. and the results are shown in FIG. 6 . A solution (final concentration of 50 μM) prepared from the compounds isolated from turkey grass and 990 μL of DPPH ethanol solution (final concentration of 100 μM DPPH) were mixed with a vortex mixer. After reacting the mixed solution in the dark at room temperature for 30 minutes, absorbance was measured at 517 nm (UV/VIS Spectrophotometer, V-550, Jasco, Tokyo, Japan). In addition, ferulic acid was used as a positive control. The antioxidant activity of the compounds isolated from turkey grass was expressed as a percentage of the absorbance decrease compared to the absorbance of the DPPH ethanol solution to which the sample solution was not added.

As shown in FIG. 6 , both rutin and narcissin isolated from turkey grass exhibited DPPH free radical scavenging activity. Narcissin exhibited relatively low DPPH free radical scavenging activity compared to ferulic acid, a positive control, but rutin exhibited superior activity compared to ferulic acid, a positive control.

Experimental Example 4

The oxidative stress inhibitory effect of rutin and narcissin obtained from turkey in Example 2 was tested as follows, and the results are shown in FIG. 7 .

Huh7 cells (American Type Culture Collection, USA) were cultured in Dulbecco's modified Eagle's medium/high glucose (DMEM) containing 10% Fetal Bocine Serum (Gibco) and 1% Pen Strep (Gibco) in a 5% CO ₂ incubator at 37°C. cultured. Huh7 cells were seeded in a 96-well plate at a ^{level of 0.7 × 10 4} per well, and 100 μL of DMEM was seeded into each well and cultured for 24 hours. Here, rutin and narcissin (20 μM) obtained from turkey and quercetin (10 μM) as a positive control were treated, and then cultured in _{a 5% CO 2 incubator at 37° C. for 24 hours.} After DMEM was removed, it was washed with Phosphate Buffer Saline solution. Thereafter, 100 μL of Hanks' Balanced Salt Solution (HBSS) containing 15 M 2',7'-dichlorofluorescin diacetate (H _{2 DCFDA) was treated and incubated in a 5% CO 2} incubator at 37° C. for 30 minutes. After H ₂ DCFDA was removed and _{100 μL of HBSS containing 1 mM H 2} O ₂ was treated, the fluorescence was measured at 485 nm and 520 nm using a SYNERGY HTX multi-modereader (BioTek). was evaluated.

As shown in FIG. 7, both rutin and narcissin isolated from turkey showed an effect of reducing reactive oxygen species (ROS) generated from hydrogen peroxide. Rutin and narcissin had lower activity than quercetin, a positive control, but it was found that they exerted an effect of reducing oxidative stress in cells.

Based on the above-mentioned results, rutin and narcissin isolated from turkey according to the present invention were identified as rutin and narcissin having the structural formulas shown in FIGS. 8A and 8B, respectively. became In particular, narcisin is contained in a small amount in natural products, so it was not easy to extract, but through the present invention, not only is it contained in a large amount in turkey, but also a method for easily purifying it is provided. It is expected to make a significant contribution to material development.

Although the present invention has been illustrated and described with reference to preferred embodiments as described above, it is not limited to the above-described embodiments, and those of ordinary skill in the art to which the present invention pertains within the scope not departing from the spirit of the present invention Various changes and modifications will be possible.

Claims (12)

Herbal turkey extract containing narcissin and rutin.
The method of claim 1,
The turkey super hot water extract is a turkey super hot water extract, characterized in that it is obtained through hot water extraction by heating after mixing the turkey vinegar powder and water obtained by drying and pulverizing the turkey vinegar in a weight ratio of 1:10 to 1:15.
3. The method of claim 2,
The turkey super hot water extract, per 100 parts by weight of the turmeric powder, the narcisin is 0.15 parts by weight or more, and the rutin is 0.05 parts by weight or more.
3. The method of claim 2,
The hot water extraction is a hot water extract of turkey, characterized in that it is heated for 10 to 30 minutes at 80 to 95 ℃ based on 600 to 800 ml of the water. Preparing a turkey hot water extract;
a solvent fractionation step of fractionating the turkey extract with a solvent to obtain a solvent fractionation layer;
a concentration step of concentrating the solvent fraction layer under reduced pressure to obtain a concentrated fraction; and
A purification step of purifying the concentrated fraction; narcissin and rutin purification method using turkey, including a.
6. The method of claim 5,
The turkey vinegar extract is narcisin and rutin purification method using turkey vinegar, characterized in that it is obtained by mixing turkey vinegar powder obtained by drying and pulverizing turkey vinegar and water in a weight ratio of 1:10 to 1:15, and then heating.
6. The method of claim 5,
The solvent used in the solvent fractionation step is narcisin and rutin purification method using turmeric, characterized in that butanol.
8. The method of claim 7,
The solvent fractionation step is a method for purifying narcisin and rutin using turkey vinegar, characterized in that it is performed twice by treating 4 to 2 volume ratios of butanol per 6 to 8 volume ratios of the turkey extract.
6. The method of claim 5,
The purification step includes dissolving the concentrated fraction in distilled water to obtain a fraction solution; and high-performance liquid chromatography using distilled water (solvent A) and acetonitrile (solvent B) containing trifloroacetic acid (TFA) as an elution solvent for the fraction solution using an octadecylsilane (ODS) column. narcissin and rutin purification method using turkey vinegar, comprising:
10. The method of claim 9,
The solvent A and solvent B are narcissin and rutin purification method using turmeric, characterized in that the pH is adjusted to 2.5.
10. The method of claim 9,
After the fraction solution is injected into the high-performance liquid chromatography, the elution solvent is maintained at A/B = 9:1 (v/v) for the initial 20 minutes under the condition of a flow rate of 50 mL/min, and then until 80 minutes When eluted with a linear gradient elution method maintaining a ratio of A/B = 7:3 (v/v), rutin, a peak eluted at t _R 31 min at a detection wavelength of 280 nm, was obtained, and at t _R 39 min. A method for purifying narcisin and rutin using turmeric, characterized in that the eluted peak powder, narcissin, is obtained.
12. The method of claim 11,
The method for purifying narcisin and rutin using turkey vinegar, characterized in that the obtained narcisin and rutin have a purity of 95% or more.

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