KR19990048809A - Natural Antioxidant Components Extracted from RUMEX CRISPUS - Google Patents

Abstract

The present invention relates to a natural antioxidant component extracted from the extract, and more particularly, extract the underground part of Rumex crispus with a mixed solution of methanol and water, and then washed and fractionated using several organic solvents to prevent antioxidant effects. Relates to a method of separating high natural product components.

Description

Natural Antioxidant Components Extracted and Extracted from the Root Jelly

The present invention relates to a natural antioxidant component extracted from the extract, and more particularly, extract the underground part of Rumex crispus with a mixed solution of methanol and water, and then washed and fractionated using several organic solvents to prevent antioxidant effects. Relates to a method of separating high natural product components.

In general, antioxidants are known to be very beneficial to the human body. However, conventional synthetic antioxidants are toxic and unlike natural products, they are regulated by the US FDA or regulated. Efforts are being made to gain.

For example, the extract obtained by extracting degreasing rice bran with a mixture of water and ethanol is filtered and concentrated, and then an antioxidant is prepared [Domestic Patent Publication No. 95-10536] and green tea leaves are extracted with calcium carbonate-added water. After adsorbing to an adsorption resin, eluting with an organic solvent, converting it into ethyl acetate, and concentrating and drying to obtain a catechin powder, a method of preparing a natural antioxidant (Korean Patent Publication No. 93-5694) and the like are known. However, these methods are difficult to industrialize because the use of relatively rare materials such as rice bran and green tea has a problem in supply and demand of raw materials in mass production.

In addition, there is also a method of extracting and concentrating ethanol from turmeric, hawthorn, wormwood, donkey, larvae, gojija, and sanjoin to obtain an antioxidant substance [Korean Patent Publication No. 96-3620]. Although it is possible to obtain a good natural antioxidant, in this case, too, since the material used is a relatively expensive herbal medicine, there is a problem in mass production as in the above case.

In addition, there are known methods for separating natural antioxidants from various plant materials in foreign countries. For example, a method of separating antioxidant components from a spermatozoon plant using a water extraction method [US Pat. No. 4,923,679], and a method of separating components having an excellent antioxidant effect according to the polarity of a solvent from a perfume plant [US Pat. ], A method of using rosemary extract as an antioxidant component [US Patent No. 5,102,659] and the like. In addition, a method of preparing an antioxidant component from Lamiaceae plants using an alkaline solution to exclude lipid components [US Patent No. 5,209,870] and a method of extracting an antioxidant component using an acidic aqueous solution from green tea [US Patent No. 5,427,806. However, these methods have problems of supply and demand of raw materials in domestic production because the plant materials are foreign varieties, and the separation methods are also concentrated on the separation of pure materials, which has a fundamental problem of costly and time-consuming separation.

However, there have been no studies on or antioxidation of Rumex crispus extracts at home and abroad.

Therefore, the present invention, in order to improve the various problems appearing in the extraction and separation method of the conventional antioxidant component, in the present invention using a rumex crispus (material), unlike the prior art by applying a simple extraction and separation method The purpose of the present invention is to produce a natural antioxidant component with excellent anti-oxidant effect while having low manufacturing cost compared to the existing antioxidant.

1 is a process diagram schematically illustrating a process of extracting an antioxidant component from Rumex crispu s according to the present invention,

2a and 2b is a process diagram schematically showing a method for separating the antioxidant component through a solvent fractionation process from the extract obtained through the process of Figure 1,

3 shows the IR spectrum for the diethyl ether fraction,

Figure 4 shows the IR spectrum for the ethyl acetate fractions.

The present invention is a natural antioxidant obtained by rinsing the subterranean Rumex crispus subterranean , extracted with a mixed solution of water and methanol, concentrated, and then sequentially washing and fractionating the concentrated solution using different organic solvents for each polarity. The component is characterized by that.

Referring to the present invention in more detail as follows.

The present invention extracts antioxidant components from Rumex crispus with a mixture of water and an organic solvent, and simply washes and fractionates them by polarity using several organic solvents to obtain natural products having excellent antioxidant effects.

Rumex crispus used in the present invention is a perennial genus belonging to the genus Madipulaceae, and from olden times the leaves of the larva were used for food, and the root was used as a dry agent. Roots of Roots are straight and thick, with long stems growing straight, often purple with green background. The leaves from the roots are long, lanceolate or long, oval-shaped, and edged. It lives mainly near wetlands and is distributed in Korea, Manchuria, China, Japan, Taiwan, Europe, and North Africa.

According to the extraction separation method according to the present invention, there was no sulfate effect at the ground portion, but only at the underground portion showed an antioxidant effect.

In addition, in order to make it suitable at the time of extraction, such as to facilitate the penetration of the solvent, the groundwood part is used after shaving to an appropriate size and drying it.

In the present invention, the organic solvent used in the initial solvent extraction and mixed with water is suitable for methanol, the mixing ratio of methanol: water mixed with a volume ratio of 5: 1 to 1: 5 is used to complete the active ingredient Good in terms of extraction,

On the other hand, the present invention is an important feature to greatly improve the extraction effect by the method of washing the extract sequentially with different organic solvents for the separation of effective antioxidants, solvent extraction is degreased with, for example, heptane It can then be washed with ethyl acetate to effectively separate the antioxidant components. At this time, degreasing with heptane is for removing fat-soluble components, and for effective degreasing, it is preferable to use after dissolving the extract before use of heptane in methanol.

In addition, to obtain a better antioxidant component, the solvent extraction is degreased with heptane, and then washed sequentially with diethyl ether and ethyl acetate having a similar polarity.

The extraction and separation method according to the present invention will be described in more detail in a series of processes with reference to FIGS. 1 and 2 as follows.

The process of obtaining the extract from the brood is as shown in FIG. The dried raw material is chopped to a size of 10 mm or less for easy penetration of the solvent, immersed in a mixed solution in which methanol: water is mixed in a volume ratio of 5: 1 to 1: 5, left for several days, and then filtered to obtain an extract. Extraction at this time can be carried out repeatedly several times to increase the yield of the extract. The extracts thus obtained are combined and concentrated to about 1/10 of the original volume using a reduced pressure concentrator. In this state, mostly water is present.

The concentrated extract is washed with a process as shown in FIG. 2A. First, methanol is added to completely dissolve all the components, followed by degreasing through layer separation with heptane, which is repeated several times. Then, ethyl acetate is added to the concentrated solution which has been washed with heptane, followed by washing several times with fractions to remove nonpolar components. The ethyl acetate fraction thus obtained is concentrated and spray-dried to complete the commercialization of the natural antioxidant component.

In another method, the washing fraction is subjected to a process as shown in FIG. 2B. First, methanol is added to completely dissolve all the components, and then heptane is added to perform degreasing through layer separation. Diethyl ether was added to the concentrated solution which had been washed with heptane, and the mixture was washed several times to remove nonpolar components, and then washed several times using ethyl acetate in the same manner. This continuous washing and fractionation of diethyl ether and ethyl acetate will increase the antioxidant capacity compared to the method using only one of the two solvents. The fraction thus obtained is concentrated and spray dried to complete the commercialization of the natural antioxidant component.

Thus, as a result of the antioxidant activity measurement using DPPH for the antioxidant natural extract prepared according to the present invention, it was found that the ED50% value was 3 μl / ml or less, and the total phenol content was obtained at 60% or more.

The present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1 by the method of FIG. 2A

After grinding the dried and dried Rumex crispus basement to a size of 10 mm or less, this pulverized sample was immersed in 10 times of methanol: water (3: 1, v / v) mixed solution for 2-3 days. It was left to stand and filtered to collect only the extract. In order to increase the yield of the extract was again replaced with a new methanol: water mixture solution, and the extraction was left for 2 to 3 more days. The obtained extracts were combined and concentrated to about 1/10 of the original volume using a vacuum concentrator. In this state, mostly water is present.

By the solvent fractionation method shown in FIG. 2A, the same amount of methanol was added to completely dissolve all the components, and then, the same amount of heptane was added thereto, followed by vigorous shaking. Thereafter, when the layer was formed, degreasing was performed by removing the heptane layer, and the above procedure was repeated twice (preparation of heptane fraction). Ethyl acetate (EA) was poured into the concentrated solution after the heptane washing was completed, and washing fractions were carried out three times in the same manner as in the case of heptane (EA fraction preparation). Each fraction thus obtained and the methanol / aqueous layer remaining after fractionation were concentrated and dried using a spray dryer to powder.

Example 2 by the method of FIG. 2b

After grinding the dried and dried Rumex crispus basement to a size of 10 mm or less, this pulverized sample was immersed in 10 times of methanol: water (3: 1, v / v) mixed solution for 2-3 days. It was left to stand and filtered to collect only the extract. In order to increase the yield of the extract was again replaced with a new methanol: water mixture solution, and the extraction was left for 2 to 3 more days. The obtained extracts were combined and concentrated to about 1/10 of the original volume using a vacuum concentrator. In this state, mostly water is present.

By the solvent fractionation method shown in Fig. 2b, the same amount of methanol was added to completely dissolve all the components, and then, the same amount of heptane was added thereto, followed by vigorous shaking. Thereafter, when the layer was formed, degreasing was performed by removing the heptane layer, and the above procedure was repeated twice (preparation of heptane fraction). An equal amount of diethyl ether (Et ₂ O) was poured into the concentrated solution of heptane washing, and washing fractions were carried out three times in the same manner as in the case of heptane to remove nonpolar components (Et ₂ O fraction preparation). For a more thorough wash fraction, the same amount of ethyl acetate (EA) having a polarity similar to diethyl ether was poured into the concentrate and washed three times in the same manner as above (EA fraction preparation). Each fraction thus obtained and the methanol / aqueous layer remaining after fractionation were concentrated and dried using a spray dryer to powder.

Experimental Example

The extracts of the extracts prepared in Examples 1 and 2 were measured in the following method to compare the antioxidant capacity with the existing antioxidants, the results are shown in Table 1 below.

-How to measure antioxidant activity-

The antioxidant activity of the extract was measured by the following method using a spectrophotometer.

First, 2.96 mg of DPPH (1,1-diphenyl-2-picrylhydrazine) was dissolved in 50 ml of methanol to make a standard DPPH solution. Samples of fractions for which antioxidant activity was to be measured were dissolved in methanol, and seven samples were prepared such that the concentrations were 1.25, 2.5, 5.0, 10.0, 20.0, 40.0, and 80.0 μg / ml, respectively. Then, 1 ml of standard DPPH solution was added thereto, shaken and left for 30 minutes, and then absorbance was measured at 520 nm using a spectrophotometer. The result was obtained from the regression equation of absorbance at each concentration, and the concentration having the absorbance corresponding to half of the standard DPPH solution (hereinafter referred to as "ED50% value") was taken as the antioxidant capacity of the sample. ].

In general, the lower the value of ED50% is evaluated to have excellent antioxidant capacity.

division Antioxidant ED50% value (μg / ml) Existing Antioxidants α-tocopherol 6.53 Butylhydroxyanisole (BHA) 5.15 BHT (dibutylhydroxytoluene) 10.82 Roots underground extract (Example 1) Heptane fraction 316.32 EA fraction 2.20 Methanol / water layer 56.13 Roots underground extract (Example 2) Heptane fraction 320.21 Et ₂ O Fraction 2.30 EA fraction 2.06 Methanol / water layer 52.40 Mix Et ₂ O Fraction with EA Fraction 2.14

According to Table 1, the antioxidant effect is different for each organic solvent used at the time of solvent fractionation, it is possible to use different antioxidant components depending on the method of use.

When the step of washing with diethyl ether was omitted by the fractionation method of Example 1 and washed with ethyl acetate immediately after washing with heptane, an antioxidant effect of 2.45 times was obtained compared to tocopherol, a known natural antioxidant. Compared to Example 2, the antioxidant effect is somewhat lower, but in terms of industrial use, the simplicity of the process may be an important factor, and the yield is also higher than that of Example 2. Therefore, it is preferable to apply the fractionation method of Example 1 in a process in which yield is considered an important factor, and it is preferable to apply the fractionation method of Example 2 in a process in which an antioxidant effect is considered first.

The Et ₂ O fraction and the EA fraction according to Example 2 showed an improved antioxidant capacity of 2.84 times and 3.17 times, respectively, compared to the known natural antioxidant tocopherol. In addition, when the Et ₂ O fraction and the EA fraction were mixed, they also showed about 3.05 times higher antioxidant activity compared to the tocopherol. When the yield of the extract was considered to be higher than the effect, the Et ₂ O fraction and the EA fraction were When used in combination, sufficient antioxidant effect can be obtained. However, methanol / aqueous layer showed 0.12 times lower antioxidant activity than tocopherol. In addition, IR spectra of the Et ₂ O fraction and the EA fraction obtained in Example 2 are shown in FIGS. 3 and 4, respectively.

-Bark Extract Characteristics and Antioxidant Activity-

Some characteristics of Rumex crispus extracts were analyzed in terms of antioxidant activity, and the correlation coefficient was 0.93, indicating that the total phenol content was closely related to antioxidant activity. there was. In other words, the higher the antioxidant activity fraction, the higher the total phenolic content. This result is consistent with the known fact that phenolic substances have an antioxidant effect.

Total phenolic content (%) by fraction of the extract from the extract of Example 2 division Heptane fraction Et ₂ O Fraction EA fraction Methanol / water layer Total phenolic content 2 ± 1% 70 ± 2% 74 ± 3% 11 ± 4%

As shown in Table 2, the total phenolic content of the Et ₂ O fraction and EA fraction having better antioxidant capacity than tocopherol was 70 ± 2% and 74 ± 3%, respectively. This is consistent with a known fact that a substance has an antioxidant effect.

As described above, the present invention can be produced very economically more than two times better than conventional natural antioxidants by using a cheap insulator as a raw material and different extraction and separation process, unlike the conventional.

Claims (8)

After cutting the underground part of the rumex crispus , extracting and concentrating with a mixed solution of water and methanol, the concentrated solution is sequentially washed and fractionated using different organic solvents for each polarity to obtain a natural extract. A method for extracting and isolating an antioxidant component from a bullock. The method of claim 1, wherein the mixed solution of water and methanol is water: methanol in a 5: 1 to 1: 5 volume ratio. The method of claim 1, wherein the sequential washing using the organic solvent for each polarity is performed by degreasing with heptane and sequentially using ethyl acetate. The method of claim 1, wherein the sequential washing using the organic solvent for each polarity is performed by using diethyl ether and ethyl acetate sequentially after degreasing with heptane. 2. The method of claim 1 wherein the concentrate is sufficiently dissolved in methanol prior to degreasing to heptane. Antioxidant extract from Rumex crispu s, characterized in that the extract is isolated according to claim 1. The method of claim 6, wherein the extract of Rumex Crispus antioxidant extract from (Rumex crispu s) to the antioxidant activity measurement results, wherein the ED50 value% 3 ㎍ / ㎖ or less using DPPH. 7. The antioxidant extract from Rumex crispu s according to claim 6, wherein the extract has an average total phenol content of at least 60%.