KR101646516B1 - Food composition comprising extract of nelumbo nucifera leaf, seed, and olive leaf - Google Patents

Abstract

The present invention relates to an antioxidant-functional food composition containing an olive leaf, a lotus seed and a lotus leaf extract as an active ingredient. More specifically, the present invention relates to an antioxidant-functional food composition comprising an olive leaf extracted with alcohol and other organic solvent, Which is excellent in antioxidant power.

Description

TECHNICAL FIELD [0001] The present invention relates to an antioxidant-functional food composition comprising a mixed extract of olive leaf, lotus seed and lotus leaf. [0002] FOOD COMPOSITION COMPRISING EXTRACT OF NELUMBO NUCIFERA LEAF, SEED, AND OLIVE LEAF [

The present invention relates to an antioxidant-functional food composition containing an olive leaf, a lotus seed and a lotus leaf extract as an active ingredient. More specifically, the present invention relates to an antioxidant-functional food composition comprising an olive leaf extracted with alcohol and other organic solvent, Which is excellent in antioxidant power.

Nelumbo nucifera is a paleontologic plant widely distributed in tropical and temperate East Asia, Korea and Japan, mainly in India and China. It is a sacred plant in Buddhism, flowers have been used as ornamental plants and a tea plant, It has been edible. It is used for the treatment of diarrhea, headache and dizziness, hemorrhage, postpartum hemorrhage, enuresis, and detoxification. The components are analgesic, sedative roemerine, nuciferin, armepavine, n-nornuciferine, (Yuk CS 1990). It has been reported that the metabolites of lysine, lysine, and lysine are known to be involved.

Among them, roots and leaves of Nelumbo nucifera Gaertn have been used in folk remedies for metabolic diseases such as diabetes, hyperlipemia and hypertension, and are known to be effective for diarrhea, headache, dizziness, bleeding and detoxification (Xiao et al 2005, Ling et al 2005). Generally, root and leaves of kite are widely used for medicinal and edible purposes. Especially, lotus root has a lot of water-soluble fibers and it is known to have constipation-relieving effect and also to lower blood pressure. In addition, it has been found that the glycine contained in the glycoprotein, mucin, has a hypotensive effect of lowering cholesterol. Tannin contained in the lotus root is known to have a strong convergence action, and thus has excellent hemostatic effect and antioxidant activity (Ling et al 2005).

Olive is a common name for olive trees, not only about one plant and its representative genus, but about 900 species in 24 genera, Italy, and Greece are the major olive producing countries. Due to the growing interest in the health benefits of olives, they are also grown in many countries and regions of the world (USA, Canada, Japan, etc.). Fruits are used as raw materials, as raw materials for olive oil, It is currently used as a medicinal food.

As medicinal food, olive leaf has been used as a private drug for the treatment of malaria fever, etc. It is known to be effective for symptoms such as hypertension, atherosclerosis, colon cancer, inflammation and food poisoning. In particular, olive leaf extract is known to have the ability to lower blood pressure, increase blood flow velocity in coronary arteries, alleviate arrhythmias, prevent seizure of small intestine muscles, etc. Recently, it has been shown that AIDS (Acquired Immune Deficiency Syndrome) (Campeol et al ., 2003; Flamini et al ., 2003; Garcia-Gomez et al ., 2003; Ryan et al ., 2003).

The major physiologically active substances of olive leaves are phenolic compounds such as hydroxytyrosol, tyrosol, catechin, caffeic acid, vanillic acid, p- P-coumaric acid, diosmetin, vanillin, rutin, oleuropein, demethyloleuropein, oleuroside, verbascoside, verbascoside, ligstroside, and luteolin 7-rutinoside, luteolin 4-glucoside, apigenin 7-glucoside, glucoside, and flavonoid glycosides such as apigenin 7-rutinoside. Among them, oleuropein is a phenolic substance most abundant in olive leaves. Recently, studies on the bioactivity of oleuropain have been actively carried out Physiologically active substances are high-value-added substances that exhibit remarkable activity in the human body even in a very small amount (Bianco et al ., 2000; Ryan et al ., 2002).

Oleuropein is a major phenolic compound contained in olive fruit and leaves. It enhances resistance to microorganisms such as bacteria, viruses and fungi which are harmful to human body. It also helps to relieve hypertension, antioxidant effect, (Raffaella and Patumi, 2002). In addition, the phenolic compounds extracted from olive leaves lead to a study of these diseases (Raffaella and Patumi, 2002).

On the other hand, Korean Patent Registration No. 10-0386640 (registered on May 23, 2003) discloses a washing step of washing a lotus leaf with clean water; A first room-temperature drying step of drying the washed lotus leaves in a well-ventilated shade at normal temperature; A cutting step of chopping the dried lotus leaves at a temperature of 0.5 to 1.0 cm; A second room-temperature drying step in which the finely chopped leaves are dried again at a room temperature so that the moisture content is 30% or less; Heating and drying the secondarily dried lotus leaf at a temperature of 90 to 110 캜 for 3 to 8 minutes and drying; And a cooling step of cooling the heated and dried lotus leaf to complete a fused green tea leaf.

In addition, Korean Patent Registration No. 10-0386640 (registered on May 23, 2003) discloses a method for manufacturing a lotus leaf tea beverage, which comprises cutting and washing washed leaves with steam at a pressure of 90 to 100 kgf / cm 2 for 40 to 50 seconds A cooling step for cooling for 1 minute by cold air drying; A refluxing step of refluxing the heated lotus leaf for 15 to 25 minutes with hot air at 75 to 110 DEG C; The process of keeping the above-mentioned dried long leaves at room temperature for 5 to 10 minutes; A heavy oil process in which the above-mentioned lotus leaf is subjected to heavy oil treatment for 20-30 minutes with hot air at 30-40 ° C; The heavy oiled leaves are rebuilt by blowing off foreign matter and powder in hot air at 40 to 50 ° C for 15 to 25 minutes to rebuild; Drying the reconstituted lotus leaf with hot air in a dryer at a temperature of 60 to 70 DEG C for 30 to 40 minutes; The dried leaf is dried at 80 to 90 DEG C for 20 to 40 minutes to remove the lion leaves. A water-soluble 5-20% oligosaccharide is added to the lecheal tea finished with the above-mentioned inclusion, and the mixture is cooled at 70-100 ° C for a short time and then cooled; Wherein the liquid is filtered and sealed in a container, and then sealed and sterilized at a temperature of 80-90 DEG C for 30-60 seconds for sterilization, cooling and packaging.

Korean Registered Patent No. 10-0659483 (registered on Dec. 13, 2006) discloses a plant having a rootstock; Yan of fruit; With lotus leaves; A rice bowl; A stigma; After the lotion was washed with water, these were put in a vacuum decompression concentrator together with water and concentrated under reduced pressure at 110 ° C for 2 hours to obtain an extract. Oligosaccharide was added to the extract to dilute it, Which is boiled for 30 to 40 minutes, filtered through a filter to remove contaminants therein, and then stored in a refrigerator at 0 to 5 ° C.

However, the above-mentioned inventions employ only a method of simply drying a leaf of a green tea to prepare tea or putting it in water and heating it, so that not only useful components of the yeast are sufficiently exposed but also there is a limit to use as various kinds of food materials. Also, there is no known method for using the lotus seed and the olive leaf together with the lotus leaf as in the present invention.

A problem to be solved by the present invention is to provide a functional food composition having excellent antioxidant ability comprising an extract of olive leaf, lotus seed and lotus leaf as an active ingredient.

In order to solve the above problems, the present invention provides an antioxidant food composition comprising 2.0 to 50.0% by weight of a mixed extract of olive leaf, lotus seed and long leaf, wherein the mixing ratio of olive leaf, lotus seed, 3: 2. ≪ / RTI >

According to an embodiment of the present invention, the mixed extract may be prepared by pulverizing dry olive leaves, lotus seeds, or lotus leaves and adding 20 to 70 wt% of alcohol; Repeatedly extracting the solution twice or more while refluxing it in the range of 60 to 80 캜; Filtrating the extracted mixed solution, concentrating it under reduced pressure, and freeze-drying it; And optionally water and an organic solvent, followed by fractionation at a reduced pressure in the range of 0 to 80 ° C followed by lyophilization.

According to an embodiment of the present invention, the reflux cooling step is preferably performed in a temperature range of 60 to 80 ° C, and the decompression concentration step is preferably performed in a temperature range of 30 to 80 ° C, more preferably 40 ° C.

According to another embodiment of the present invention, the alcohol is preferably 40 to 90% ethanol, and the organic solvent is preferably at least one selected from the group consisting of hexane, chloroform, ethyl acetate and butanol.

According to another embodiment of the present invention, the food comprising the mixed extract of olive leaf, lotus seed and lotus leaf according to the present invention may be any one selected from the group consisting of powders, tablets and capsules, The content of the mixed extract of olive leaf, lotus seed and long leaf is preferably 2.0 to 50.0% by weight.

According to another embodiment of the present invention, the food containing the mixed extract of olive leaf, lotus seed and long leaf according to the present invention is selected from the group consisting of bread, cake, cookie, cracker, noodle, dairy product, And the beverage may be any one selected from the group consisting of carbonated or non-carbonated drinks, vegetable juice, and fruit juice. At this time, the content of olive leaf, lotus seed, and lotus leaf extract in the food is preferably 2.0 to 30.0% by weight.

Since the mixed extract of olive leaf, lotus seed and long leaf according to the present invention has a high total phenol content and excellent SOD-like activity and nitrite scavenging ability, it is possible to provide various functional preparations including olive leaf, lotus seed, Foods are expected to contribute to the promotion of public health through excellent antioxidant power.

Hereinafter, the present invention will be described in more detail with reference to examples and drawings.

The antioxidant food composition according to an embodiment of the present invention is an antioxidant food composition comprising 2.0 to 50.0% by weight of a mixed extract of olive leaf, lotus seed and long leaf, wherein the mixing ratio of olive leaf, lotus seed, 5: 3: 2.

According to an embodiment of the present invention, the mixed extract may be prepared by pulverizing dry olive leaves, lotus seeds, or lotus leaves and adding 20 to 70 wt% of alcohol; Repeatedly extracting the solution twice or more while refluxing it in the range of 60 to 80 캜; Filtrating the extracted mixed solution, concentrating it under reduced pressure, and freeze-drying it; And optionally water and an organic solvent, followed by fractionation at a reduced pressure in the range of 0 to 80 ° C followed by lyophilization.

According to an embodiment of the present invention, the reflux cooling step is preferably performed in a temperature range of 60 to 80 ° C, and the decompression concentration step is preferably performed in a temperature range of 30 to 80 ° C, more preferably 40 ° C.

According to another embodiment of the present invention, the alcohol is preferably 40 to 90% ethanol, and the organic solvent is preferably at least one selected from the group consisting of hexane, chloroform, ethyl acetate and butanol.

According to another embodiment of the present invention, the food comprising the mixed extract of olive leaf, lotus seed and lotus leaf according to the present invention may be any one selected from the group consisting of powders, tablets and capsules, The content of the mixed extract of olive leaf, lotus seed and long leaf is preferably 2.0 to 50.0% by weight.

According to another embodiment of the present invention, the food containing the mixed extract of olive leaf, lotus seed and long leaf according to the present invention is selected from the group consisting of bread, cake, cookie, cracker, noodle, dairy product, And the beverage may be any one selected from the group consisting of carbonated or non-carbonated drinks, vegetable juice, and fruit juice. At this time, the content of olive leaf, lotus seed, and lotus leaf extract in the food is preferably 2.0 to 30.0% by weight.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to examples. However, the scope of the present invention should not be construed as being limited to the examples, and should be construed to facilitate understanding of the present invention.

Example 1: Preparation of methanol extract of olive leaf, lotus seed and lotus leaf

The olive leaves ( Olea europaea L. var. Kalamata) from Greece, the domestic lotus seeds and the lotus leaves were mixed at 50:30:20, washed well, removed foreign matter, dried naturally and then used as a sample.

10% of 60% methanol was added to 25 g of each of the crude olive leaves, lotus seeds and lotus leaves, respectively, before and after 30 mesh, and the mixture was refluxed with a soxhlet apparatus and repeatedly extracted three times at 80 ° C in a water bath for 3 hours. After filtration, it was left at 4 캜 for 24 hours and re-filtered. This solution was concentrated under reduced pressure using a rotary vacuum evaporator (R-124, BUCHI, Switzerland) at 70 ° C and freeze-dried to prepare an extract. The extract was washed with different solvents Followed by stepwise addition. Specifically, the extract was dissolved in water, placed in a fractionation funnel, and hexane was added thereto to separate the hexane layer and water layer. The filtrate was concentrated under reduced pressure to obtain hexane fraction. Through the same procedure, chloroform, ethyl acetate and butanol were added sequentially to obtain respective fractions. The remaining solution was called a water fraction. These fractions were depressurized, concentrated and lyophilized to remove the solvent and used in the experiment.

Example 2: Preparation of ethanol-mixed extract of olive leaf, lotus seed and lotus leaf

Ethanol extracts of olive leaf, lotus seed, and lotus leaf and respective organic solvent fractions were prepared in the same manner as in Example 1, except that 10 g of 80% ethanol was added to 25 g of each of olive leaf, lotus seed and lotus leaf Were used in the following evaluation experiments.

Comparative Example 1: Preparation of extract of olive leaf

Extraction and fractionation experiments were conducted according to the method described in Example 1 for olive leaves from Greece.

Evaluation Example 1: Evaluation of total flavonoid and phenol content

Total flavonoid content was determined by taking a certain amount of olive leaf, lotus seed, lotus seed, 80% ethanol extract and solvent fraction of the lotus leaf and massaging it with 5 mL of 50% methanol solution. Then, add 1 mL of this sample solution, 10 mL of diethylene glycol, The solution was incubated at 37 ° C for 1 hour and absorbance was measured at 420 nm using a UV / VIS spectrophotometer (Optizen 2120UV, Mecasys, Korea) naringin (Sigma, Saint Louis, USA).

The phenolic substances for the extracts obtained in the above Examples and Comparative Examples were measured using the Folin-Dennis method (Teresa-Sartue et al ., 1995) using a substance showing blue color by reacting with phosphomolybdic acid .

Add 1 mL of sample dissolved in a certain amount of dimethyl sulfoxide (DMSO), 7 mL of distilled water, and 0.5 mL of Folin-Dennis reagent. After exactly 3 minutes, add 1 mL of anhydrous sodium carbonate saturated solution and 0.5 mL of distilled water. Absorbance was measured using a spectrophotometer. At this time, the standard calibration curve was prepared using tannic acid (Fluka, Buchs, Switzerland). The Folin-Dennis reagent was prepared by adding distilled water to 10 g of sodium tungstate, 2 g of molybdophosphoric acid and 5 mL of phosphoric acid (enough to melt the sample) and then refluxing and cooling on a 80 ° C water bath.

Table 1 below shows the total phenolic content of the mixed extract of olive leaf, lotus seed and long leaf according to the present invention.

Sample Total phenol content Methanol extract 32 ± 2 Chloroform fraction 17 ± 3 Hexane fraction 12 ± 2 Ethyl acetate fraction 22 ± 4 Butanol fraction 19 ± 3 Water fraction 10 ± 2

On the other hand, the total flavonoids and the total phenol contents of the 80% ethanol extract of olive leaf, lotus seed and long leaf of Example 2 and hexane, chloroform, ethyl acetate, butanol and water fractions were compared with each other, .

Total flavoid contents were highest in ethyl acetate fraction (24.9%) and low in hexane and water fractions. The total phenol content of ethyl acetate fraction was the highest at 18.2%, and hexane and water fractions were similar to flavonoid due to lower phenol content.

The content of flavonoids and phenols in olive leaves, lotus seeds and lotus leaf fractions were 1.57.13% and 3.4-17.5%, respectively. . However, the content of flavonoids and phenols in butanol fraction, ethylacetate fraction and olive leaf 80% ethanol extracts was relatively high, and the content of water and hexane fractions was relatively low.

Evaluation Example 2: Measurement of SOD-like activity

SOD is a physiologically active enzyme in human and animal organs and blood. It plays a role to remove harmful oxygen. SOD-like active substance plays a similar role to SOD in phytochemicals, so that the reactivity of superoxide radical (Murakami et al ., 2003).

In this experiment, SOD-like activity (Superoxide dismutase-like activity) was measured by Marklund et al. (1975). Add 0.2 mL of a certain concentration of sample, 3 mL of Tris-HCl buffer (50 mM Tris (hydroxymethyl) aminomethane + 10 mM EDTA, pH 8.5) and 0.2 mL of 7.2 mM pyrogallol. And the reaction was stopped by adding 1 mL of 1N HCl. The amount of oxidized pyrogallol in the reaction solution was measured by using a UV / VIS spectrophotometer at 420 nm. The SOD-like activity was expressed as a percentage of absorbance difference between the addition of the sample solution and the addition of no sample.

SOD-like activity (%) = (1-absorbance of sample added / absorbance of no additive) × 100

The following table shows the ethanol extracts (Table 3) of olive leaves, lotus seeds, lotus seeds, methanol blended extracts (Table 2), olive leaves, lotus seeds, (Table 4). According to the results, it can be confirmed that the SOD-like activity of the mixed extract according to the present invention is remarkably improved as compared to the SOP-like activity for the olive leaf alone.

Samples (1000 ppm) SOD-like activity (%) Chloroform fraction 14.7 ± 2.2 Butanol fraction 91.7 ± 6.6 Ethyl acetate fraction 90.3 ± 4.9 60% methanol extract 82.2 ± 5.4 Hexane fraction 14.6 ± 4.4 Water fraction 62.3 ± 6.1

sample 1000ppm Chloroform fraction 5.82 + - 0.46 Butanol fraction 5.29 ± 0.41 Ethyl acetate fraction 4.67 ± 0.68 80% ethanol extract 29.54 + - 4.62 Hexane fraction none Water fraction 1.58 ± 0.29

Samples (1000 ppm) SOD-like activity (%) Chloroform fraction 4.99 ± 0.88 Butanol fraction 4.31 ± 0.61 Ethyl acetate fraction 3.33 ± 0.17 60% methanol extract 24.71 ± 0.59 Hexane fraction None Water fraction 0.78 ± 0.17

The SOD-like activity of the olive leaf, lotus seed and lotus leaf fractions according to Example 2 ranged from 29.54% to 80.5%, and the highest activity was observed in the concentration dependent manner. The SOD-like activities of 82 Korean medicinal plants including Gugija (21.27%), Chunmun-dong (11.7%), Bokbunja (13.2%), Puerariae (17.13%), Ginger (9.03% Compared with the results, SOD - like activity was higher or similar at 1,000 ppm concentration, except 3-4 kinds of medicinal plants.

It has been confirmed that SOD-like activity is caused by the phytochemical components of phenolic compounds in olive leaf, lotus seed and lotus leaf fractions. Taking such SOD-like activity inhibits senescence inhibition and oxidation It can be said that this method can prevent.

Evaluation Example 3: Measurement of nitrite (Nitrite) resolution

According to the method of Gray and Dungan (1975), 1 mL of 1 mM NaNO ₂ solution was added to 1 mL of a certain concentration of sample, and the pH of the reaction solution was adjusted to 1.2 with 0.1 N HCl, and the total amount was adjusted to 10 mL. Take 1 mL of this solution at 37 ° C for 1 hour, add 5 mL of 2% acetic acid, and use a Griess reagent (1: 1 mixture of 1% sulfanilic acid and 1% naphthylamine in 30% acetic acid) 0.4 mL) was added and mixed well. The mixture was allowed to react at room temperature for 15 minutes and absorbance was measured at 520 nm using a UV / VIS spectrophotometer to determine the amount of remaining nitrite. The control was prepared by adding the same amount of distilled water instead of the Griess reagent.

N (%) = (1-A-C / B) 100

N: nitrite scavenging ability, A: sample loading absorbance

B: absorbance of 1 mM NaNO ₂ , C: absorbance of control

Table 5 and Table 6 show the results of confirming the nitrite removal activity, which is a causative substance of nitroamine production, which is a carcinogen substance for Examples 1 and 2. With the exception of hexane and water fractions, the nitrite scavenging ability of olive and leafy leaf fractions tended to increase in a concentration-dependent manner. A study of nitrite scavenging activity of medicinal plant extracts used as herbal medicines showed that 35% of Angelicae gigas, 42% of corn germs, 33% of gingerbreads and 64% of bryophytes were found under the same conditions. In addition, the extracts of pine needles and mugwort extracts, which have antioxidant ability and prevent aging disease, showed nitrite scavenging ability of 77.85% and 37%, respectively, suggesting that the removal activity of olive leaf fractions used in this experiment shows high level of activity.

Nitrite itself contained in food is not harmful, but it is toxic if it is reduced by nitrite reductase. The term "phytochemical" refers to all the chemical substances produced in the roots and leaves of a plant. Among these phytochemicals, flavonoids and phenolic compounds have strong conjugated double bond structure as a radical acceptor and have a strong reducing ability, so they serve as nitrite scavengers by donating electrons to free radicals.

As a result, it was confirmed that olive leaf, lotus seed and lotus leaf fractions have the ability to inhibit the carcinogenic substance nitroamine by donating electrons to free radicals by the influence of phytochemical components.

The sample (1000 ppm) Nitrite Resolution (%) Chloroform fraction 48.7 ± 5.6 Butanol fraction 93.5 ± 7.2 Ethyl acetate fraction 95.3 ± 5.4 80% ethanol extract 91.6 ± 7.1 Hexane fraction 54.8 ± 7.0 Water fraction 64.7 ± 6.3 Vitamin C 94.76 ± 3.28 Vitamin E 86.48 + - 2.46 BHT 82.53 + - 4.16

sample 100ppm 500ppm 1000ppm Chloroform fraction none 11.25 + 1.24 71.37 + - 8.26 Butanol fraction none 28.26 ± 3.26 77.22 + - 4.28 Ethyl acetate fraction 29.95 + - 4.23 69.16 + - 5.26 86.57 ± 3.23 80% ethanol extract none 11.56 ± 1.36 75.73 + - 2.37 Hexane fraction none none none Water fraction none none none Vitamin C 38.62 ± 1.93 66.14 + - 5.57 94.76 ± 3.28 Vitamin E 28.58 ± 1.82 58.58 + - 4.36 86.48 + - 2.46 BHT 28.54 + - 2.24 49.53 + - 4.52 82.53 + - 4.16

Evaluation Example 4: Measurement of DPPH radical scavenging activity

Chu et al . (2000), 0.8 mL of 4 × 10 ^-4 M DPPH (1, 1-diphenyl-2-picrylhydrazyl) solution was added to 0.2 mL of the extract at a constant concentration, mixed for 10 seconds, left at room temperature for 10 minutes, The absorbance was measured using a UV / VIS spectrophotometer. The DPPH radical scavenging ability was expressed as a percentage by the difference of the absorbance between the addition of the sample solution and the no addition.

DPPH radical scavenging activity (%) =

(Absorbance of 1-sample added / absorbance of no added) x 100

Tables 7 and 8 below show the DPPH measurement results of Example 1 and Comparative Example.

Sample DPPH scavenging ability (%) Methanol extract 83.2 + -5.4 Chloroform fraction 60.4 ± 3.9 Hexane fraction 57.4 ± 4.1 Ethyl acetate fraction 74.6 ± 6.3 Butanol fraction 66.3 ± 6.1 Water fraction 53.8 ± 4.2

sample 100ppm 500ppm 1000ppm Chloroform fraction 20.24 0.88 29.23 + - 0.42 53.52 + - 2.59 Butanol fraction 10.49 + - 0.72 18.21 + - 0.14 29.19 ± 0.15 Ethyl acetate fraction 7.74 ± 0.65 17.24 0.31 31.47 ± 0.63 80% ethanol extract 15.64 + 1.59 40.86 ± 1.59 59.35 ± 3.17 Hexane fraction 17.89 ± 1.92 33.62 ± 0.37 43.48 ± 0.67 Water fraction 12.94 + 0.36 18.45 + 0.21 23.46 ± 1.12 Vitamin C 29.14 + - 0.93 50.43 + - 0.10 74.24 + - 5.23 Vitamin E 27.26 + 0.07 48.37 ± 1.58 71.53 + - 4.12 BHT 29.07 ± 0.77 43.27 + - 0.49 68.83 + - 4.50

The ethanol extract and chloroform fractions of olive leaf, lotus seed, and lotus leaves showed relatively high activity, and the DPPH radical scavenging activity of olive leaf fractions increased to 5.9056.86% as the treatment concentration increased Table 8)

The DPPH radical scavenging activity of olive leaf, lotus seed, and lotus leaf fractions was similar to the radical scavenging activity of 43.0% and 60.0% of medicinal plants, Korean medicinal herbs, Angelica gigas Nakai, and catechin, , DPPH radical scavenging activity was reported to be 50.56%, and it was confirmed that the antioxidative effect of olive leaf, lotus seed, and lotus leaf fractions is not inferior to that of well known medicinal plants.

The cells are oxidized and damaged by the free radicals generated by the growth and growth of the cells. Phenolic compounds are known to have antioxidant ability to inhibit oxidation by donating electrons to free radicals because of their strong reducing ability. DPPH is a compound that has stable free radicals and is widely used for confirming antioxidant ability by being reduced by antioxidant. As a result, it was confirmed that olive leaf, lotus seed, and lotus leaf fractions have the ability to inhibit oxidation by donating electrons to free radicals, which is judged by the action of phenolic compounds in olive leaves.

Evaluation Example 5: Automatic oxidation inhibitory activity of linolenic acid

Kiharu et al . (1990). Add 2 mL of 99.5% ethanol, 2.05 mL of 2.5% linoleic acid diluted with ethanol, and 4 mL of 0.05 M phosphate buffer (pH 7.0) aeration at least 1 hour to a given amount of sample at each treatment concentration. Add distilled water After adjusting the total volume to 10 mL, take 0.1 mL of the reaction solution at a rate of 3 hours while reacting the solution at 70 ° C for 24 hours, add 9.7 mL of 75% ethanol, 30% ammonium thiocyanate, , 0.1 mL of a mixed solution of 3.5% HCl and 0.02 M ferric chloride were added in this order, followed by reaction at room temperature for 3 minutes, and then the absorbance at 500 nm was measured using a UV / VIS spectrophotometer.

In the autoxidation process, the reaction is terminated by generating a stable compound of two radicals. In this process, a strong reducing substance which inhibits oxidation by donating electrons to free radicals is required. Antioxidants play a role in eliminating these free radicals. Mainly phenolic compounds have this function and oleuropein and hydroxytyrosol, the main phenolic compounds of olive leaves, (Park et al., 2005).

The degree of inhibition of lipid peroxidation by treating samples with linolenic acid is shown in FIG. The antioxidant activity was relatively constant in a concentration dependent manner over time. In the case of 80% ethanol extract of olive leaf, lotus seed and lotus leaf, the antioxidant activity was similar to that of vitamin C, non-time E and BHT according to the elapsed time. The antioxidant activity of chloroform fraction and water fraction was confirmed in the order of It is thought that the activity of linolenic acid is also inhibited by the action of phenols present in the extract.

Preparation Example 1: Preparation of powder

4 g of the mixed extract or fraction of olive leaf, lotus seed, or lotus leaf prepared in the above example was mixed with 1 g of lactose and filled in an airtight container to prepare a powder.

Production Example 2: Preparation of tablets

The tablets were prepared by using the olive leaf, lotus seed, lotus seed, mixed extract or fraction 200 mg, corn starch 100 mg, lactose 100 mg, and magnesium stearate 2 mg prepared in the above Example according to a conventional preparation method.

Preparation Example 3: Preparation of capsules

The capsules were prepared according to the conventional preparation method of capsules using 200 mg of olive leaf, lotus seed, lotus seed extract, or fraction 200 mg, corn starch 100 mg, lactose 100 mg, and magnesium stearate 2 mg prepared in the above Examples.

Production Example 4: Production of flour food

A bread, a cake, a cookie, a cracker or a noodle is prepared by adding 2.0 to 7.0% by weight of a mixed extract of olive leaf, lotus seed and lotus leaf or fractions thereof according to the present invention to wheat flour, .

Production Example 5: Production of dairy products

5-10 wt% of olive leaf, lotus seed, lotus seed extract or fraction thereof according to the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

Production Example 6: Preparation of wire

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and then the mixture was ground to a powder having a particle size of 50 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a known method, and then they were made into powder having a particle size of 50 mesh by a grinder. The olive leaf, lotus seed, lotus seed extract or fraction thereof according to the present invention was concentrated under reduced pressure in a vacuum concentrator, dried by spraying, and dried with a hot air drier, and the dried material was pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder. 5% by weight of the dry powder of the olive leaf, lotus seed and lotus leaf extract or fractions thereof according to the present invention, 30% by weight of brown rice, 15% by weight of yamu, 20% by weight of barley, (7% by weight of perilla, 8% by weight of black soybeans, 5% by weight of black sesame), 0.5% by weight of manure, and 0.5% by weight of zirconium.

Production Example 7: Production of carbonated beverage

The syrup is prepared by mixing additives of 5 to 10% of oligosaccharide, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel and 0.1 to 1% of vitamin C and 79 to 94% of purified water thereto, ° C for 20 to 180 seconds and mixed with cooling water at a ratio of 1: 5, then 0.5 to 0.8% of carbon dioxide gas was injected to prepare a mixed extract of olive leaf, lotus seed, Carbonated beverages were prepared.

Production Example 8: Preparation of health drinks

A mixture of an ingredient such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%) and water (75%) and olive leaf, lotus seed, His fractions were uniformly blended and instant sterilized and packaged in small containers such as glass bottles and plastic bottles to produce health drinks.

Production Example 9: Preparation of vegetable juice

The vegetable juice for health promotion was prepared by adding 5 g of the mixed extract of olive leaf, lotus seed and long leaf or its fraction according to the above example to 1,000 ml of tomato, carrot or celery juice.

Production Example 10: Preparation of fruit juice

The fruit juice for health promotion was prepared by adding 1 g of the mixed extract of olive leaf, lotus seed, or lotus leaf or its fraction according to the above example to 1,000 ml of orange, mandarin or grape juice.

FIG. 1 is a flowchart illustrating a process for producing an ethanol extract of olive leaf, lotus seed, and lotus leaf according to an embodiment of the present invention.

FIG. 2 is a flowchart illustrating a process of fractionating an ethanol extract of olive leaf, lotus seed, and lotus leaf using an organic solvent according to an embodiment of the present invention.

FIG. 3 is a graph showing the total flavonoid contents of the ethanol extract of the olive leaf, lotus seed, the lotus leaf, and the organic solvent fraction according to an embodiment of the present invention.

FIG. 4 is a graph showing the total phenol contents of the ethanol extract of olive leaf, lotus seed, long leaf, and organic solvent fraction according to an embodiment of the present invention.

FIG. 5 is a graph showing the automatic oxidation inhibitory activity of the ethanol extract of olive leaf, lotus seed, or lotus leaf and the organic solvent fraction according to an embodiment of the present invention.

Claims (8)

Wherein the mixture ratio of olive leaf, lotus seed and long leaf of the mixed methanol extract is in the range of 5: 3: 1 to 5: 3, 2. ≪ / RTI > The method according to claim 1, The mixed extract may be obtained by pulverizing dried olive leaves, lotus seeds, or lotus leaves and then adding 20 to 70% by weight of methanol; Repeating the step of mixing the crude pulverized product and methanol with the solution twice or more while refluxing at 60 to 80 ° C; Filtrating the extracted mixed solution, concentrating it under reduced pressure, and freeze-drying it; And optionally fractionating by adding water and an organic solvent, and concentrating the mixture at a reduced pressure in the range of 0 to 80 ° C, followed by lyophilization. delete 3. The method of claim 2, Wherein the organic solvent is at least one selected from the group consisting of hexane, chloroform, ethyl acetate and butanol. The antioxidant food composition according to claim 2, wherein the reflux cooling step is carried out at 60 to 80 ° C. The antioxidant food composition according to claim 2, wherein the reduced-pressure concentration step is carried out at 30 to 80 ° C. The method according to claim 1, Wherein the food composition is any one selected from the group consisting of powders, tablets, and capsules. The method according to claim 1, Wherein the food composition is any one selected from the group consisting of breads, cakes, cookies, crackers, noodles, dairy products, wire, carbonated or non-carbonated beverages, vegetable juices and fruit juices.

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