KR100650016B1 - Separation method of antioxidant active substance from bokbunja - Google Patents

Abstract

The present invention relates to a method for separating an antioxidant active substance from bokbunja, and more particularly, to a method for separating a substance having strong antioxidant activity from a residue obtained after juice from bokbunja.

Separation method of the antioxidant active material from the bokbunja of the present invention comprises the step of extracting the bokbunja juice extract, extracting the bokbunja residue, separating the antioxidant active material from the bokbunja residue extract using an aromatic sorbent.

Description

Isolation method of anti-oxidation active material from Rubus coreanus Miquel}

The present invention relates to a method for separating an antioxidant active substance from bokbunja, and more particularly, to a method for separating a substance having strong antioxidant activity from a residue obtained after juice from bokbunja.

Bokbunja ( Rubus coreanus Miquel, Korean black raspberry) is a deciduous shrub belonging to the family Rosaceae, native to Korea, China, and Japan. The fruit of Bokbunja is round and turns black when ripe, and grows well in both dry and wetland. Bokbunja fruit has been used for food and medicinal use since ancient times, and has a tonic, tonic effect, and is also used for diabetes, hemostasis, and hemostasis. Recently, the immune enhancing action, superoxide anion radial scavenging activity, xanthine oxidase inhibitory activity, anticancer activity, and antibacterial activity of bokbunja fruit extract and polyphenol have been reported (Lee Kyung-kyung et al. 6, Korean Journal of Medicinal Plants, 11 (1), 5-12 (2003); Sungwoo Woo and 5 others, Korean Journal of Food Science and Nutrition, 33 (5), 769-777 (2004); ), 96-101 (2002)).

The production of liquor such as fermented liquor has been made a lot by using bokbunja berries having various pharmacological effects, and recently, various products are being developed for various uses of bokbunja berries, and some beverages, health supplements, etc. It is manufactured in the form and is also commercially available. However, despite many studies related to bokbunja, the development of products and the separation process of specific ingredients have not been developed. Recently, Yunin et al. Reported quercetin, five phenolic acids, and organic acids as antioxidants contained in bokbunja fruits to utilize bokbunja fruit as a functional food material (Korean Journal of Food Science, 35 (3), 499-). 502 (2003)), yet no functional food using these ingredients has been released.

Free radicals or free radicals promote oxidation of lipids, protein denaturation, damage to biological membranes and genes, and eventually cause adult disease, carcinogenesis, and aging. In order to artificially inhibit oxidation by such free radicals or active oxygen, research and development of various natural antioxidants are actively underway, and many patents have been applied (Japanese Patent No. Hei 11-80148; International Patent, WO 2000/64883; International Patent, WO 2004/080993 A1). On the other hand, alcohol extract of bokbunja fruit is known to have a very high antioxidant activity, and some varieties are known to have a strong antioxidant effect comparable to vitamin C (Korean Journal of Forestry, 94 (1), 11-15 (2005)).

However, the juice obtained from juice of the bokbunja fruit is very low in antioxidant activity as shown in Table 1. In other words, the juice residue is mostly extracted in the flesh or seeds without extracting the ingredient that leads to the antioxidant activity, it can be seen that in the case of beverages using the juice of bokbunja juice, the excellent effect of the bokmo molecule is not sufficiently exhibited.

Currently, residues after extracting bokbunja juice from farms are mostly discarded or supplied for feed to livestock farms. That is, most of the components having excellent physiological activity effects of bokbunja fruit are not used well.

The present invention was completed by using polystyrene-based adsorbents, which are synthetic resin adsorbents, to extract components having strong antioxidant activity from bokbunja residues left after juice of bokbunja juice using environmentally friendly solvents.

In the present invention, simply extracting the bokbunja fruit is not enough to extract an effective ingredient having a strong antioxidant capacity, and by examining a variety of extraction conditions, having excellent physiological activities such as hydrolyzed tannins and anthocyanins, as well as having a strong antioxidant capacity It is an object of the present invention to establish extraction conditions for obtaining a large amount of active ingredients and to provide a method for separating antioxidant active substances from bokbunja which can increase the active ingredient content of antioxidant active substances by adsorbing and eluting these extracts to an aromatic adsorbent.

The present invention for achieving the above-mentioned object comprises extracting the bokbunja residue obtained by juice of bokbunja, separating the antioxidant active substance using an adsorbent to the bokbunja residue extract obtained by extracting the bokbunja residue.

The antioxidant active substance of the present invention can be isolated from the remaining bokbunja residues after extracting the bokbunja.

In the present invention, the bokbunja juice is obtained and the remaining residue is extracted and the extract is obtained by extracting the bokbunja residue.

The bokbunja residue extract can be obtained by using any one method selected from the supercritical extraction method using hot water extraction, organic solvent extraction, supercritical fluid.

The hot water extraction of the bokbunja residue is the extract of bokbunja residue by extracting the bokbunja residue in hot water of 60 ~ 100 ℃, preferably hot water of 80 ~ 90 ℃.

The organic solvent extraction of the bokbunja residue in the extract can be obtained by extracting the bokbunja residue in the organic solvent. In this case, ethanol may be used as the organic solvent.

Supercritical extraction of the bokbunja residue in the extract can be obtained by extracting the bokbunja residue using a supercritical fluid. In this case, carbon dioxide (CO ₂ ) may be used as an example of the supercritical fluid.

In the present invention, an aromatic adsorbent is used for the extract of Bokbunja residue, and antioxidant compounds such as anthocyanins and hydrolyzed tannins having strong antioxidant activity can be separated from the extract of Bokbunja residue.

In the present invention, any adsorbent may be used as long as it can separate the substance having antioxidant activity from the bokbunja residue extract.

The adsorbent may be an aromatic adsorbent, a polystyrene adsorbent may be used as such an aromatic adsorbent, and one commercially available as a commercial product may be used as an example of the polystyrene adsorbent. Examples of polystyrene-based adsorbents that are commercially available as above are DIAION SP-205, DIAION SP-206, DIAION SP-207, DIAION HP-2MG (Mitsubishi Chemical Corporation), Amberite XAD-7, Amberite XAD-8 ( One or more selected from Organo Corporation), DIAION, HP-10, DIAION HP-20, DIAION HP-30, DIAION HP-40, and DIAION HP-50 (Mitsubishi Chemical Corporation) can be used.

On the other hand, the present invention includes an antioxidant active substance isolated from the bokbunja residue by the above-mentioned method.

In addition, the present invention obtains an antioxidant active material separated from the bokbunja residue by the above-mentioned method, food containing such antioxidant active substances, health supplements containing antioxidant active substances, cosmetics containing antioxidant active substances, antioxidant Includes medicines containing active substances.

Hereinafter, the content of the present invention will be described in detail through examples. However, these are intended to explain the present invention in more detail, and the scope of the present invention is not limited thereto.

Example 1 Hydrothermal Extraction and DPPH Free Radical Scavenging Activity of Bokbunja Fruit Juice Extract

The bokbunja residue after bokbunja fruit juice was frozen at -45 ℃ and dried by lyophilizer, and 5g of lyophilized bokbunja residue was placed in 50ml of distilled ion-exchanged water at 60 ℃, 70 ℃, 80 ℃, 90 ℃ and 100 ℃, respectively. Extraction for 10 minutes. After extraction, the bokbunja residue extract was filtered and washed with a 1G4 glass filter, and the filtrate was concentrated and lyophilized. The bokbunja fruit lyophilized juice was used as a comparative sample.

The yield of the bokbunja residue extract obtained above was determined by dividing the total dry weight of each hot water extract by the total dry weight of the sample. DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging ability was analyzed using Munevver et al. (Munevver et al., Life Sciences, 76, 2981-2993 (2005)).

Extracting the bokbunja fruit juice residue at 60 ~ 100 ℃, the extraction yield increased with the increase of the extraction temperature, especially as shown in Table 1, the extract yield was constant at a temperature of 80 ℃ or more. The DPPH free radical scavenging ability of the extract increased with increasing extraction temperature, but dropped sharply from 100 ° C. That is, it turns out that 80-90 degreeC of the residue extraction temperature which shows high antioxidant capacity is suitable. In addition, DPPH free radical scavenging ability of bokbunja extract extracted from bokbunja residue in the above temperature range was 2.5 to 2.9 times better than bokbunja juice shown as a comparison. It can be confirmed that it can be extracted.

Table 1. Yield and DPPH Free Radical Scavenging Capacity of Bokbunja Fruit Juice Extract

sample Extract yield (%) DPPH free radical scavenging activity (%) Bokbunja Juicer - 10.1 60 ℃ -10 minutes 7.6 17.0 70 ℃ -10 minutes 8.3 21.6 80 ℃ -10 minutes 8.7 25.2 90 ℃ -10 minutes 8.7 29.1 100 ℃ -10 minutes 8.8 23.0

Example 2 Isolation of Antioxidant Activity and Its Characteristics

100 g (whole dry weight basis) of bokbunja fruit juice residue was put in 1 liter (L) of distilled ion-exchanged water and extracted at 80 ° C for 10 minutes. After extraction, the mixture was filtered through a large Buchner filter, and the residue was washed with 2 liters of distilled ion-exchanged water at 80 ° C. The filtrate was concentrated under reduced pressure at 60 ° C. so that the total liquid was 600 ml. 300 ml of the solution was added and 35 ml of an adsorbent DIAION HP-50 was added thereto, followed by stirring at room temperature for 18 hours.

After stirring, DIAION HP-50 was transferred to a 17G3 glass filter, filtered, and washed three times with ion-exchanged water. After desorbing the components adsorbed on the adsorbent with anhydrous ethanol and concentrated under reduced pressure at 50 ℃. In order to completely remove moisture, the desorbed and concentrated sample was vacuum dried again for 12 hours, and used as an analytical sample such as DPPH free radical scavenging ability.

In Table 2, the DPPH free radical scavenging ability of the purified sample after adsorbing the antioxidant component of the bokbunja residue extract with the adsorbent was measured by the method of Munevver et al. Mentioned in Example 1, and the results are shown. Purification of the hot water extract of the bokbunja residue with an adsorbent showed a 13.6-fold improvement in DPPH free radical scavenging ability compared to the bokbunja juice, as shown in Table 2. Therefore, it can be seen that the adsorbent is very effective for separating the antioxidant component contained in the bokbunja residue.

Table 2. DPPH free radical scavenging ability of bokbunja residues purified after adsorption with adsorbent

sample DPPH free radical scavenging activity (%) Bokbunja Juicer 4.1 Bokbunja residue 80 ℃ -10 minutes hot water extraction 28.0 After Purifying Adsorbent 55.7

Table 3 shows the chemical composition after the adsorbent purification of the bokbunja residue extract.

As a result of analyzing the freeze-dried extracts of bokbunja juice juice, as shown in Table 3, more than 60% was composed of pectin, which is a neutral sugar, and 17% was an acidic sugar. When the juice residue was hydrothermally extracted and purified with an adsorbent, no free neutral sugars and pectin were detected. However, hydrolysis of this resulted in 30% of the neutral sugars containing glucose as a main component, and it can be seen that a large part of the sugars exist in the bound form. It can be seen that such bound sugars are due to hydrolysis of hydrolyzed tannins and anthocyanates in the sample after the adsorbent purification.

From the above results, it can be seen that the components having strong antioxidant activity in the bokbunja can be separated and purified by extracting the hot water extract of the bokbunja juice residue and separating and purifying it with an adsorbent. The components separated by these adsorbents were found to be mainly hydrolyzable tannins and anthocyanins, and these components are considered to be the cause of strong DPPH radical scavenging ability.

Table 3. Chemical Composition of Samples after Purification of Adsorbents from Bokbunja Fruit Juice and Bokbunja Residue Extracts

sample Bokbunja Juicer After Purifying Adsorbent Neutral sugars (%) Before hydrolysis 61.1 - After hydrolysis N.D. 30.3 Organic acids (%) Aliphatic 18.77 - Aromatic 0.1 10.4 * Anthocyanates (%) 4.7 34.4 pectin(%) 16.9 - Total phenols (%) 5.5 62.6

N.D .: Not analyzed,-: Not detected.

*: Aromatic organic acids are mainly composed of gallic acid and ellagic acid, which are derived from hydrolyzed tannins contained in bokbunja.

As described above, although described with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and modified within the scope of the present invention without departing from the spirit and scope of the present invention described in the claims below. It will be appreciated that it can be changed.

As can be seen from the results of the above example, it can be seen that the antioxidant active substance can be easily separated from the bokbunja residue using the aromatic sorbent extract from the extract of the bokbunja residue.

According to the present invention, antioxidant active substances can be separated from bokbunja residues that have been conventionally discarded or mainly used in feed, thereby contributing to increasing farm income.

The antioxidant active substance isolated from the bokbunja residue by this invention can be used for food, a dietary supplement, cosmetics, a pharmaceutical, etc.

Claims (4)

Extracting bokbunja residue obtained by extracting bokbunja residue from hot water at 80-90 ° C. to obtain bokbunja residue hot water extract, Separation method of antioxidant active substance from bokbunja using an adsorbent on the bokbunja residue hot water extract obtained above. delete The method of claim 1, wherein the adsorbent is a polystyrene-based adsorbent. The method of claim 3, wherein the polystyrene-based adsorbent DIAION SP-205, DIAION SP-206, DIAION SP-207, DIAION HP-2MG, Amberite XAD-7, Amberite XAD-8, DIAION HP-10, DIAION HP-20, Separation method of antioxidant active material from bokbunja, characterized in that at least one selected from DIAION HP-30, DIAION HP-40, DIAION HP-50.