KR20140144429A - Composition for antioxidant from main component using extract of stem, leaf & seed of rubus coreanus mique - Google Patents

Abstract

The present invention relates to an antioxidant composition including extracts of a stem, a leaf, and a seed of Rubus coreanus as active ingredients. More specifically, the extracts obtained by extracting the stem, the leaf, and the seed of Rubus coreanus with ethanol are phenolic compounds and flavonoid compounds and have activities of removing DPPH radicals and ABTS radicals of the phenolic compounds. When the antioxidant composition of the present invention is used for antioxidant medicines, health-promotional food, food or the like, the antioxidant composition can contribute to promoting health of people.

Description

TECHNICAL FIELD The present invention relates to an antioxidant composition comprising a bacterium stem and an extract of a leaf and a seed as main components.

The present invention provides an antioxidant composition comprising as an essential component an extract of bokbunja extracted with hot water or ethanol on a bokbunja stem. More specifically, the method comprises the steps of extracting stem, leaves and seeds of bokbunja by drying and pulverization with hot water or ethanol, filtering and storing the freeze-dried extract, and testing the antioxidant efficacy. Especially, these extracts obtained by reflux extraction with hot water into bamboo stalks, leaves or seeds, or by ultrasonic extraction at different temperature and ethanol concentrations have high total phenol and total flavonoid contents and thus have excellent antioxidative effect.

Rubus occidentalis ) is a deciduous shrub of Rosaceae and is grown in the southern and central parts of Korea. It ripens in mid-June to July-August, ripens in round and red color and then matures into reddish-brown color with sweetness, sourness and unique fragrance It contains phenol compounds such as kaemferol, sanguinin H-5, 3-Ο-β-D-glucuronide, quercetin, ferulic acid, coumaric acid, caffeic acid, rutin , luteolin, etc. have been reported. Studies on bokbunja have been reported on tannin and flavonoids compounds from leaves and stems.

Bokbunja is used for edible and medicinal purposes. Fruits are mainly used in food, beverage and alcoholic products. In Oriental medicine and medicinal berry, fruit protects the liver, clears the eyes, darkens the hair and treats infertility. It is recorded. There are various studies on the components and physiological activity of the bokbunja fruit. The functionalities of the bokbunja fruit known to date include antioxidant, antibacterial, antiallergic, anticancer, antiinflammatory, hormone control, skin whitening, obesity, Blood sugar, blood pressure, cholesterol).

Antioxidants are antioxidants. Antioxidants are polyphenol compounds, carotenoids (β-carotene, lycopene, lutein), flavonoids (eg, Anthocyanins, catechins, resveratrol, proanthocyanidins), isoflavones (genistein, daidzein), vitamin C, vitamin E and minerals.

Reactive oxygen species (ROS) generated by free radical reaction in vivo causes inactivation and peroxidation of DNA segments and proteins, which causes various diseases by lowering biological function. The active oxygen species produced in the body are known to be cleared by intracellular antioxidant enzymes or by antioxidant vitamins and polyphenol components taken from foods. Synthetic antioxidants such as buthylated hydroxy anisole (BHA) and buthylated hydroxy toluene (BHT), which have been used to inhibit damage caused by these active oxygen species, have been limited in their use as safety concerns despite their excellent antioxidative and economic properties Development of natural antioxidants capable of replacing these synthetic antioxidants has been demanded.

Korean Patent Registration No. 10-0650016 (a method for separating an antioxidative active substance from a bokbunja) of the present invention relates to a method for extracting an antioxidative active substance from a bokbunja by extracting a bokbunja to obtain a residue, extracting a bokbunja residue, using an aromatic adsorbent And separating the antioxidant active substance. Korean Patent Laid-open Publication No. 10-2012-0119265 (Antioxidant Functional Composition and Functional Food Containing the Functional Food Containing the Antioxidant Functional Component) The present invention relates to an antioxidant functional ingredient and a functional food containing the same, Functional compositions and foods. However, these prior arts have different technical constructions from the present invention.

Although bokbunjae have various efficacies, it is difficult to industrialize as a functional food because the berry itself is high in price. Therefore, in order to replace the bokbunjae, it is necessary to use alternative materials such as stems, leaves and seeds The research on this is still not very clear. In addition, after the bokbunja was cultivated, it was disposed of, causing environmental pollution.

In addition to berries Leaves, stems and seeds contain various polyphenols. However, the bokbunja leaves are registered as a food ingredient in Korea, but stem and seeds are not registered because of lack of grounds for domestic ingestion. Accordingly, the present invention will provide basic data on the functional properties of the leaves, stem and seeds of the bokbunja, and further develop various processed products using the bokbunja leaf and stem through comparative study on the functionality of the bokbunja leaf, stem and seed. In addition to the berries, Since leaves, stems and seeds also contain various polyphenols, studies on their application are needed.

The present invention comprises a step of extracting the stem, leaves and seeds of the bokbunja strawberry with hot water or ethanol, filtering the extract, freeze drying and storing, and testing the antioxidant efficacy.

The present invention provides basic data according to functional analysis of bramble leaves, stems and seeds, and develops them as a variety of medicines, health foods or foods using bramble leaves and stems according to their functionalities, have.

Figure 1 is the total phenolic content for the bract stalks, leaves and seed extracts according to the extraction method and EtOH concentration.
Figure 2 is the total flavonoid content for the bract stalks, leaves and seed extracts according to the extraction method and EtOH concentration.
FIG. 3 shows the results of measurement of DPPH radical scavenging activity at 100 μg / mL concentration of bacterium stem, leaf and seed extracts according to the extraction method and EtOH concentration.
FIG. 4 shows the ABTS radical scavenging activity at 100 μg / mL concentration of extracts of Rubus trunks, leaves and seeds according to extraction method and EtOH concentration.
Figure 5 is the reduction power at 100 μg / mL concentration of bract stalk, leaf, and seed extract according to extraction method and EtOH concentration.
FIG. 6 shows DPPH radical scavenging activities of phenolic compounds detected in berry stalks, leaves and seeds.
FIG. 7 shows the results of measurement of ABTS radical scavenging activity of phenolic compounds detected in berry stalks, leaves, and seeds.

The present invention comprises a step of extracting the stem, leaves and seeds of the bokbunja strawberry with hot water or ethanol, filtering the extract, freeze drying and storing, and testing the antioxidant efficacy.

≪ Materials and methods >

The bacterium stem and leaf used in the present invention were collected from Bukbun Bokbunja Test Center in Gochang-gun, Jeollabuk-do, Korea at the beginning of May, 2012. After washing with water, they were dried in hot air for 3 days at 60 ° C, In June, 2012, bokbunjae used berries obtained from Seonwon Nonghyup, Kochang-gun, Jeollabuk-do using seaweeds as a sample. The extraction solvent is ethanol (manufactured by Samcheon Chemical Co.), which is a folin-Ciocalteau reagent, butylated hydroxyanisole (BHA), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Diethylene glycol, 2,2'-Azino- 6-sulfonic acid, trichloroacetic acid, rutin, gallic acid, ellagic acid, caffeic acid, luteolin, quercetin, resvertrol, ρ-coumaric acid and tannin.

100 g of the dried stem or leaf of the bokbunja were weighed and 1000 mL of water, 25, 50, 75, and 100% EtOH, respectively, were added to a round flask, followed by reflux extraction at 80 ° C for 2 hours. The filtrate was filtered through a filter paper (Whatman NO. 2, Whatman International Ltd., Maidstone, England), and the resulting filtrate was concentrated using a vacuum concentrator (Buchi R210, Switzerland), lyophilized and dissolved in 70 ° C deep freezer (CLN-71UWM, Nihon , Japan) and used as a sample. As another method, ultrasonic extraction by ethanol concentration was repeated twice at 70 ° C for 2 hours with an ultrasonic wave extender (Sonic Medical Plus up, MIRCO, Korea) in the same manner as above. To understand the effect of the present invention, the analysis of the experimental results (total phenol content, total flavonoid content, scavenging activity by DPPH radical assay, scavenging activity by ABTS radical assay, reduction power, phenolic compound analysis, DPPH radical scavenging activity, ABTS radical scavenging activity of phenolic compounds) were performed according to known methods.

Example 1 Extraction Yield by Ethanol Concentration

1-1. Reflux Extraction Yield by Ethanol Concentration

Table 1 shows the yields of reflux extracts (water, 25, 50, 75, 100%) of EtOH concentration of berry strawberry (stem, leaf, seed) The yields of 25, 50% EtOH extracts showed the highest yields of 14.07 and 14.75%, respectively. The yield of water extracts of leaves was the highest at 20.30%. In particular, 100% EtOH extract showed a high yield of 20.85%, indicating that the fats present in the seeds were extracted by the nonpolar solvent EtOH and showed a high yield. The highest yields were obtained in the order of leaves, seeds and stems, except for the extracts obtained by extracting 100% EtOH with solvent.

1-2. Ultrasonic extraction yield by ethanol concentration

Table 1 shows the yield of ultrasonic extracts according to the EtOH concentration in the berries of the bokbunja. In the case of stem, the yield of 25% EtOH extract was 13.3% and the yield of 100% EtOH extract was the lowest. The yields of 25%, 25%, 20%, 20%, 20%, 20%, 20%, 20%, 20%, 20%, 20% and 20% Respectively. Seed extract also showed the highest yield (23.05%) as in reflux extraction. As a result of comparing the yield of ultrasonic extracts of Rubus coreanus, the yields of leaves were higher than those of 100% EtOH solvent.

Example 2: Antioxidative experiments according to extraction conditions

2-1. Total phenol content

Phenolic compounds present in the plant are secondary metabolites that have at least one hydroxyl group in the molecule and therefore have the property of binding to proteins and macromolecules and have excellent binding power with divalent metal ions. (33, 34). In addition,

The total phenol content of the extract of Rubus coreanus according to the extraction method and EtOH concentration is shown in FIG. The total phenol contents of the extracts were 40.74 ± 5.92 and 400.62 ± 12.12 mg / g, respectively. The contents of total phenol contents in the extracts were 50 ~ The content of the sex compound was found to be higher.

2-2. Total flavonoid content

The total flavonoid contents of the extracts of the bokbullus strawberry according to the extraction method and EtOH concentration are shown in Fig. 100% EtOH extract showed the highest contents of 143.99 ± 1.08 ㎎ / g in refluxed leaves, and water, 25%, 50% and 75% EtOH extract showed lower contents than stem. The total flavonoid contents of water, 25%, 50%, 75% and 100% EtOH extracts were 95.4 ± 2.49, 104.39 ± 9.07, 110.30 ± 13.93, 109.11 ± 4.80 and 70.93 ± 7.10 mg / g, respectively The content of total flavonoids in the extracts according to the extraction method and EtOH concentration was higher than that of the extracts by the reflux extraction.

2-3. Scavenging activity by DPPH radical assay

DPPH uses antioxidative substances from various natural materials by using principle of decolorization of dark purple color due to reduction and formation of irreversibly stable molecules by receiving electrons or hydrogen by using ascorbic acid and tocopherol, polyhydroxy aromatic compounds and aromatic amines It is widely used for searching.

The results of measuring DPPH radical scavenging activity at 100 μg / mL concentration of the extract of Rubus coreanus according to the extraction method and EtOH concentration are shown in FIG. In both reflux and ultrasonic extraction methods, the radical scavenging ability of the stem was highest in the order of stem>seed> leaf. The positive control, BHA, showed the activity of 78.06 ± 4.69% at the same concentration. The 50% EtOH extract of the stem was similar to 73.87 ± 0.332%. These results are similar to those reported by Choi et al., And the results of the RC ₅₀ values of extracts according to the extraction method and EtOH concentration are shown in Tables 2, 3 and 4. The extracts of EtOH concentration of 60.81 ~ 118.8 μg / mL showed that the higher the content of total phenol, the higher the DPPH radical scavenging ability. Choi et al.

2-4. Scavenging activity by ABTS radical assay

In the plasma ABTS, metmyoglobin is activated by hydrogen peroxide to form ABTS ⁺ . The ABTS assay is a method of analyzing the extent to which ABTS ⁺ is eliminated in the presence of antioxidants based on the inhibition of ABTS by antioxidants in the presence of antioxidants.

The ABTS radical scavenging activity at 100 μg / mL extract of Rubus coreanus according to extraction method and EtOH concentration is shown in FIG. In both reflux and ultrasonic extraction, stem was most abraded in order of stem>seed> leaf. It appeared to reflux extraction and ultrasonic extraction are both 50% EtOH extract showed the highest activity, reflux extraction RC ₅₀ value of 50% EtOH extract of the excellent scavenging activity to 82.57 μg / mL of the stem (see Table 2). Similar to DPPH radical scavenging activity, the highest radical scavenging activity was observed in stem extracts with high total phenol content.

2-5. Reducing power

The absorbance value in the reducing power itself indicates the reducing power of the sample, and the absorbance value of the substance having a high antioxidant activity is high. The reducing power at 100 μg / mL concentration of the extract of Rubus coreanus according to the extraction method and EtOH concentration is shown in FIG. Both the reflux and ultrasonic extraction showed the highest reducing power of the stem. Compared with the control, BHA, the reducing power of BHA was lower than that of BHA but showed a high level of reducing power. Therefore, it is considered that the reflux extraction method is suitable for the extraction of strawberry parts of bokbunja, and to develop a functional tea using the stem with the highest antioxidant activity as a raw material when manufacturing tea using bokbunja strawberry part.

2-6. Phenolic compound analysis

The phenolic compounds of Rubus coreanus extract according to extraction method and EtOH concentration were analyzed as gallic acid and ellagic acid as the highest contents of compounds as shown in Tables 5, 6 and 7. Gallic acid was found to be more than 4 times higher than water extract in reflux and ultrasound extracts, and water extract in reflux extract (245.00 μg / g) 261.88 μg / g), respectively. In addition, ellagic acid showed higher contents in reflux extract than gallic acid, and the extracts (145.70 μg / g) and antioxidant activity of leaves (23.44 μg / g) The extracts (136.94 μg / g) were analyzed to be higher. Quercetin, ferulic acid, coumaric acid, caffeic acid, rutin and luteolin were analyzed in all extracts. As a result, the content of phenolic compounds was higher than that of the extraction method in the reflux extraction method. The main phenolic compounds were gallic acid and ellagic acid regardless of extraction method and EtOH concentration.

2-7. DPPH radical scavenging activity of phenolic compounds

The DPPH radical scavenging activity of the phenolic compounds detected in the bokbunja strawberry stem, leaf and seed were measured as shown in FIG. gallic acid> ellagic acid>luteolin> quercetin, gallic aicd and ellagic acid showed the highest scavenging activity.

2-8. ABTS radical scavenging activity of phenolic compounds

The results of ABTS radical scavenging activity of the phenolic compounds detected in the bokbule strawberry stem, leaf, and seed were as shown in FIG. Gallic acid> ellagic acid>quervetin> resveratrol showed higher antioxidative activity than gallic acid and ellagic aicd as DPPH radical scavenging activity.

<Application>; Formulation

1) Manufacture of powders

2 g of the bacterium stem extract of the present invention was mixed with 1 g of lactose and filled in an airtight container to prepare a powder.

2) Preparation of tablets

200 mg of the bacterium stem extract of the present invention, 150 mg of corn starch, 150 mg of lactose, 2 mg of magnesium stearate and the above components were mixed and then tableted according to a conventional method for preparing tablets.

3) Preparation of capsules

200 mg of the bacterium stem extract of the present invention, 150 mg of corn starch, 150 mg of lactose, and 2 mg of magnesium stearate were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules .

4) Manufacture of rings

1.5 g of lactose, 1 g of glycerin and 0.5 g of sorbitol were mixed with 2 g of the bacterium stem extract of the present invention, and the mixture was prepared to be 5 g per one ring according to a conventional method.

5) Preparation of granules

After mixing 300 mg of the bacterium stem extract of the present invention, 50 mg of soybean extract, 100 mg of isomaltooligosaccharide, 600 mg of starch and the above components, 120 mg of 25% ethanol was added and dried at 60 ° C to form granules And filled in foams.

6) Manufacture of beverages

300 mg of the bacterium stem extract of the present invention, 100 mg of sugar, 10 mg of citric acid, 50 mg of sorbitol and 1,000 mg of purified water were mixed and filtered to prepare a drink.

By using stem extract, which is a byproduct of bokbunja, we can confirm the improvement effect of antioxidant and develop various products using it. Because it utilizes waste resources, income is guaranteed and waste disposal costs can be reduced.

Claims (5)

An extract of bokbunja stem, leaf and seed as an active ingredient. The antioxidative composition according to claim 1, wherein the composition is a medicament mixed with a pharmaceutically acceptable excipient. The antioxidative composition according to claim 1, wherein the composition is a health functional food mixed with an acceptable excipient in the Health Function Food Act. The antioxidative composition according to claim 1, wherein the composition is a foodstuff to be mixed with an acceptable excipient according to the Food Sanitation Act. The antioxidative composition according to claim 1, wherein the composition is selected from the group consisting of liquid, powder, tablet, capsule, ring and granule.

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