KR101429745B1 - functional fish oil composition using natural materials and food composition for improving liver function - Google Patents

Abstract

The present invention relates to a functional fish oil composition using a natural substance, and a food composition for improving liver function using the same. More specifically, the present invention relates to a fish oil composition for enhancing the quality of fish oil, To a fish oil composition and a food composition for improving liver function.
The functional fish oil composition using the natural substance of the present invention is characterized in that a fish oil extract is added to fish oil. The herbal extract is extracted by adding at least one extraction solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof.

Description

Technical Field [0001] The present invention relates to a functional fish oil composition using a natural substance and a food composition for improving liver function using the natural fish oil composition,

The present invention relates to a functional fish oil composition using a natural substance, and a food composition for improving liver function using the same. More specifically, the present invention relates to a fish oil composition for enhancing the quality of fish oil, To a fish oil composition and a food composition for improving liver function.

In recent years, the domestic food market has become increasingly interested in health by improving living standards and entering into an aging society. As interest in functional nature of foods that can maintain a healthy lifestyle away from simple food has increased, It is gradually expanding.

Fish oil is rich in n-3 highly unsaturated fatty acids, docosahexaenoic acid (DHA, C22: 6) and eicosapentaenoic acid (EPA, C20: 5) It has been reported that the function of lowering cholesterol concentration in the body and inhibiting coronary artery disease, thrombosis and cancer induction has been reported.

DHA, which is an important functional substance in fish oil, is a major component of retina and brain tissue. It contains about 11 ~ 13% of the lipids of sardine, saury, anchovy, mackerel and tuna, . In addition, fatty acids, which are the main constituent lipids of fish oil, are essential components of phospholipids, which are constituents of all cell membranes in the body. Fatty acids, which are the main constituents of lipids, are directly absorbed without further hydrolysis in the digestive tract, And thus has attracted attention as an effective functional lipid.

However, DHA and EPA, which are abundant in fish oil, have polyunsaturated fatty acids with double bonds of 5 and 6, which are easily corroded during distribution and storage. Therefore, there is an urgent need to improve the oxidation stability for high quality of fish oil and preservation of fish oil processed product.

Korean Patent Publication No. 2005-0029264 discloses a method for producing purified fish oil and a composition thereof as one of the measures for improving the oxidation stability of fish oil.

The above-described conventional technique provides citrus acid and magnesium silicate in the process of refining fish oil and improves the treatment method by the deodorization method by nitrogen, thereby providing purified fish oil having excellent storage stability and high catching effect.

However, the conventional technology has a problem that it is difficult to trust the stability by adding citric acid or magnesium silicate, which is a synthetic chemical substance, to the fish oil, and the existing fish oil production method can not be applied as it is, and a new facility must be provided. Further, there is a limitation in that the functionality of the fish oil itself can not be improved.

The present invention has been made to overcome the above-mentioned problems, and it is an object of the present invention to provide a functional fish oil composition and a liver oil composition capable of enhancing the preservability and enhancing the functionality of fish oil by effectively preventing rancidity of unsaturated fatty acids contained in fish oil by using natural materials safe for human body It is an object of the present invention to provide a food composition for improving function.

In order to accomplish the above object, the functional fish oil composition using the natural substance of the present invention is characterized in that a fish oil extract is added to fish oil.

The herbal extract is extracted by adding at least one extraction solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol or a mixture thereof.

The fish oil is characterized by being refined fish oil obtained by purifying sardine crude oil extracted from sardines.

The fish oil composition is characterized in that the herbal extract is added in an amount of 0.1 to 3.0 wt%.

The food composition for improving liver function of the present invention is characterized by containing the fish oil composition.

As described above, according to the present invention, it is possible to increase the preservability of fish oil by effectively preventing acidosis of unsaturated fatty acids contained in fish oil by adding uroguin extract to fish oil.

In addition, the functional fish oil composition of the present invention can provide a healthier food composition by significantly improving liver function.

1 and 2 are graphs showing the results of electron donating ability analysis for 48 kinds of natural extracts, respectively,
FIG. 3 is a graph showing the acid value measurement results for sardine refined oil,
4 to 9 are graphs showing the acid value measurement results for the test fish oil to which the six kinds of natural extracts were added,
10 is a graph showing a change in body weight of a male according to a mouse test result,
11 is a graph showing a change in body weight of a female according to a mouse experimental result,
12 is a graph showing the results of measurement of hemoglobin of a mouse,
13 is a graph showing the results of measurement of hematocrit of a mouse,
14 is a graph showing the results of measurement of glucose in mouse,
15 is a graph showing the results of measurement of Triglyceride in a mouse,
16 is a graph showing the results of measurement of total cholesterol of a mouse,
17 is a graph showing the results of measurement of HDL-cholesterol in a mouse,
18 is a graph showing the results of measurement of GOT of a mouse,
19 is a graph showing the results of GPT measurement of a mouse.
20 is a graph showing the results of measurement of lysozyme in a mouse,
21 is a graph showing the color change of the test fish oil to which the natural extract is added.

Hereinafter, a functional fish oil composition using a natural substance according to a preferred embodiment of the present invention and a method for producing the functional fish oil composition will be described in detail.

The functional fish oil composition according to one embodiment of the present invention is prepared by adding a natural extract to fish oil extracted from fish. The functional fish oil composition of the present invention may be composed of 95 to 99% by weight of fish oil and 1 to 5% by weight of natural extract.

Fish oil is extracted from fishes (including fish by-products) such as high herring, salmon, cod, mackerel, sardine, shark, tuna, squid and the like, which are higher in unsaturated fatty acids. (DHA, C22: 6) and eicosapentaenoic acid (EPA, C20: 5) which are highly unsaturated fatty acids. In particular, fish oil extracted from sardines is lower in price than fish oil extracted from other fishes, so it is preferable to use fish extracted from sardines in economic terms.

Fish oil is a main component of the functional fish oil composition of the present invention, and includes crude oil extracted primarily from fishes, and refined oil obtained by purifying crude oil.

Conventional methods of producing fish oil can be applied as an extraction method of crude oil. The extracted crude oil can be used as a main component of the functional fish oil composition of the present invention. However, since crude oil itself contains various impurities and there is odor such as fishy smell, purified oil purified from crude oil is preferably used. Especially, refined oil has a low acid value and is advantageous for storage.

The crude oil refining method is a well-known technique and a detailed description thereof will be omitted. As an example of the refining method, 20% of caustic soda (NaOH) solution is added to crude oil in a deoxidation tank, deoxidized by stirring, and then subjected to desalting through a stirrer. Then, it is transferred to a flushing tank, water is rinsed, and water is added. Then, active white clay is added in a decolorizing tank and discolored while stirring. Then, the decolorized crude oil is deodorized at a temperature of 170 ° C for 3 hours under reduced pressure by steam distillation, and filtered in a filter press to obtain purified oil.

Another method for refining crude oil is to employ supercritical fluid extraction. Supercritical Fluid Extraction is a supercritical fluid extraction method in which the supercritical fluid has a low viscosity and is highly permeable to the sample. Therefore, the extraction efficiency is high and the diffusion coefficient is high. Therefore, the extraction speed is fast, And the advantage of easy separation of the extraction residue and the solvent because the density difference between the sample and the supercritical fluid is large and the viscosity of the supercritical fluid is low.

Supercritical fluid means that a fluid under special conditions under high-temperature and high-pressure conditions is applied to supercritical substances, that is, substances such as carbon dioxide, water, alcohol, and helium, with a critical pressure and a critical temperature. This supercritical fluid has a value close to the density of the liquid, the viscosity is close to the gas, and its diffusion coefficient is about 100 times larger than that of the liquid. By penetrating the supercritical fluid into the extraction target and reacting with the desired content, extraction of high purity content becomes possible.

Carbon dioxide can be used as a supercritical fluid for supercritical extraction and can be extracted under extraction conditions at a temperature of 35 to 45 DEG C and a pressure of 100 to 300 bar for 30 to 240 minutes.

The natural extract to be added to fish oil is at least one of extracts selected from the group consisting of Ulgi, Odili, Sansui oil, White pear leaf, Seaweed and Green tea. Natural extracts extracted from Ulgum, Odili, Sansui, Leaf, Wakame, and Green tea are added to fish oil to effectively prevent rancidity of unsaturated fatty acids contained in fish oil, thereby enhancing preservation of fish oil and improving quality. Lt; / RTI >

Meanwhile, a method for producing the functional fish oil includes a fish oil extraction step for extracting fish oil from a fish and a natural extract extraction method for extracting natural extracts from at least one selected from the group consisting of fish, oil, marine oil, And an addition step of adding the natural extract to the fish oil.

First, crude oil is extracted from fish (including fish by-products) such as herring, salmon, cod, mackerel, sardine, shark, tuna and squid in the fish oil extraction step. The extracted crude oil can be used as fish oil, but it is preferable to use purified oil purified from crude oil as described above.

Next, in the step of extracting natural extracts, natural extracts are extracted from at least one selected from the group consisting of gilt, oak, corn oil, white pear leaf, seaweed and green tea. Natural extracts can be extracted by various methods.

For example, the extracting solvent is mixed in an amount of 2 to 20 times by weight of the extracting solvent to at least any one selected from the group consisting of ganoderma lucidum, olive oil, corn oil, Pyrrhizae leaf, seaweed, and green tea and then subjected to hot water extraction, A warm-up extraction method, or the like.

Further, a reflux cooling extraction method, an ultrasonic extraction method, or the like can be used. In addition to the above-described extraction method, an extract obtained through a conventional purification process is also included. For example, an active fraction obtained through various purification methods, such as separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies, etc., is also included in the extract.

As the extraction solvent, at least one selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof may be used. As the lower alcohol having 1 to 4 carbon atoms, methanol, ethanol and the like can be used. As the polyhydric alcohol, butylene glycol, propylene glycol, pentylene glycol and the like can be used. Mixtures of water and lower alcohols, mixtures of water and polyhydric alcohols, mixtures of lower alcohols and polyhydric alcohols, or mixtures of water and lower alcohols and polyhydric alcohols can be used as the mixture. It goes without saying that the extract obtained by using the above-mentioned extraction solvent can be obtained in the form of powder through filtration, concentration under reduced pressure or freeze-drying, spray drying or the like.

Next, the natural fish oil is added to the prepared fish oil to prepare a functional fish oil composition composed of 95 to 99% by weight of fish oil and 1 to 5% by weight of natural extract.

The functional fish oil composition of the present invention prepared as described above can be used as a raw material for foods, feed for livestock, cosmetics and the like.

The functional fish oil composition of the present invention can be added to food for the purpose of improving liver function. The food composition for improving liver function preferably contains the fish oil composition in an amount of 0.01 to 10% by weight based on the total weight of the food composition. Here, the food composition refers to a food that a person usually eats, and is a collective term for eating and drinking by a person. Therefore, the form of the food composition in the present invention is not particularly limited. It can be manufactured into various forms including beverages, granules, tablets, powders, rings, capsules, waxes, noodles, confectionery, beverages, meat, fish, herbs, stew or rice mixed with known ingredients, food additives and the like .

Examples of the food additives include sugars such as monosaccharides, disaccharides, polysaccharides and sugar alcohols and flavorings such as tau martin, stevia extract, saccharin and aspartame, and nutrients, vitamins, edible electrolytes, flavors, , Cheese, chocolate, etc.), pectic acid, alginic acid, organic acid, protective colloid thickening agent, pH adjusting agent, stabilizer, preservative, glycerin, alcohol and carbonating agent.

Preferably, the food composition of the present invention is formed into any one form selected from beverage, granule, tablet, powder, ring, and capsule. The food composition having beverages, granules, tablets, powders, capsules, and capsules is easy to carry and is easily ingested at anytime and anywhere.

As an example of the beverage which is in the form of a liquid, 0.01 to 60 wt% of fruit extract, 5 to 70 wt% of purified water, 0.1 to 5 wt% of taurine, 0.1 to 5 wt% of citric acid, 0.1 to 5 wt% of vitamin A, 10 to 20% by weight of carbohydrates, and 0.1 to 2.0% by weight of a fish oil composition. The carbohydrate may be a monosaccharide such as glucose, fructose, etc .; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and xylitol, sorbitol, erythritol, and the like.

In addition, various flavorings such as ordinary beverages may be contained as additional ingredients. Natural flavoring agents such as tau Martin and stevia extract, and synthetic flavoring agents such as saccharin and aspartame may be used as the flavoring agent.

Solid formulations such as granules, tablets, powders, rings, capsules and the like contain 0.1 to 10% by weight of the fish oil composition, and may further contain other adhesive agents, spices, vitamins, carbohydrates and the like.

Hereinafter, the present invention will be described with reference to the following experimental examples and the like, but the following experimental examples are only illustrative of the present invention, and the scope of the present invention is not limited to the following experimental examples.

<Experimental Example 1: Analysis of components of fish oil>

In order to select the fish oil to be used in the present invention, crude oil, refined oil and supercritical extract oil of sardine produced by Hanjin Industry Co., Ltd. were obtained, and general components, fatty acid, acid value and coliform group of each fish oil were investigated according to the food revolution .

As a result of the investigation, the general components of fish oil showed a tendency to decrease gradually in the order of crude oil, refined oil and supercritical extraction oil as shown in Table 1 below, and crude fat tended to increase gradually.

division Moisture (wt%) Crude fat (wt%) Ash (wt%) crude oil 0.18 99.91 0.1 Refined oil 0.15 99.93 0.1 Supercritical extraction oil
(200 bar, 40 &lt; 0 &gt; C, 3h) 0.13 99.96 0.1

As shown in Table 2, the contents of n-3 fatty acids were increased in the order of crude oil, refined oil and supercritical fluid.

division Crude oil (wt%) Refined oil (wt%) Supercritical extraction oil (wt%)
(200 bar, 40 &lt; 0 &gt; C, 3h) n-3 fatty acid 24.95 25.75 28.42

  As shown in Table 3 below, supercritical extraction oil showed the lowest value in the moisture of fish oil, but the acid value showed the lowest value of the purified oil. And there was no significant difference in crude fat when comparing purified oil with supercritical extraction oil. And crude oil, refined oil, and supercritical extraction oil were all negative for coliform bacteria.

Test Items
crude oil
Refined oil Supercritical extraction oil 200 bar, 40 &lt; 0 &gt; C (2h) 200 bar, 40 &lt; 0 &gt; C (3h) 200 bar, 40 &lt; 0 &gt; C (4h) Acid value 6.7 0.8 2.3 2.5 2.4 Moisture (wt%) 0.25 0.18 0.13 0.11 0.1 Crude fat (wt%) 99.95 99.93 99.95 99.94 99.96 Coliform group voice voice voice voice voice

Table 4 shows the results of the content (wt%) analysis according to the kinds of fatty acids.

constitutional formula General name crude oil Refined oil Supercritical extraction oil
200 bar, 40 &lt; 0 &gt; C (4h) C14: 0 Myristic acid 10.29 10.02 9.73 C16: 0 Palmitic acid 22.16 21.51 22.55 C16: 1 Palmitoleic acid 10.85 9.39 10.81 C18: 0 Stearic acid 1.88 2.42 0.62 C18: 1n9 Oleic acid 10.52 11.19 12.07 C18: 1n7 Vaccenic acid 3.25 2.96 0 C18: 2n6 Linoleic acid 1.22 1.02 1.33 C18: 4n3 Linolenic acid 2.1 2.16 2.35 C20: 1n9 11-Eicosenoic acid 0.87 0.96 2.52 C20: 5n3 Eicosapentaenoic (EPA) 15.52 17.24 17.52 C22: 1n9 Erueic acid 0 0 0 C22: 1n11 Docosanoic acid 0 0.82 0 C22: 5n3 Docosapentaenoic (DPA) 1.55 2.34 1.58 C22: 6n3 Docosahexaenoic (DHA) 7.88 6.17 9.32 Unsaturated fatty acid 53.76 54.25 57.5 Saturated fatty acid 34.33 33.95 32.9 n-3 fatty acid 24.95 25.75 28.42

Referring to Table 4, it was confirmed that omega-3 (or n-3) fatty acid, which is known to be highly functional, is contained in a large amount in crude oil, refined oil and supercritical extraction oil. Especially, docosahexaenoic acid (DHA, C22: 6) and eicosapentaenoic acid (EPA, C20: 5) of omega-3 fatty acids were found to be abundant. The content of unsaturated fatty acids and n-3 fatty acids in supercritical extracts was higher than that of unsaturated fatty acids and n-3 fatty acids in crude oils and refined oils.

When the crude oil, refined oil, and supercritical extraction oil of sardine were compared with each other, there was no significant difference in the characteristics of the components. Crude oil was considered to be difficult to use because of high impurity content and deep odor. And supercritical extraction oil has no odor but has a high production cost in extraction. Considering the quality and economical aspect, the use of refined oil is desirable. Especially, refined oil is preferable in terms of enhancing preservation of fish oil because the acid value is lower than that of supercritical extracted oil.

Experimental Example 2: Search for natural extracts &gt;

1. Experimental Method

There are a total of 48 kinds of natural materials (Ulgum, Green tea, Pepper, Tomato, Dodok, Hwangbyeon, Bokbunja, Black soybean, Kelp, Rhododendron, Green onion, carrot, wormwood, chlorella, strawberry, alpha-cone, corn syrup, gugija, saury, persimmon, mung bean, , Mussels, seaweed, shellfish, black rice) were selected.

The selected natural substances were prepared by drying and pulverized powder form. In order to evaluate the antioxidant function of fish oil, we firstly measured the electron donating ability of selected natural substances. 80 g of each natural substance was mixed with 650 mL of ethanol (99%) and stirred for 3 to 5 minutes. After storing for more than 1 day in a dark room, the mixture was extracted with a microfiltration filter and filter paper (Whatman, No. 2) . The extracted natural extract was concentrated to 50 mL (Tokyo rkiakikai, N-1000, Japan), diluted with distilled water to a concentration of 1000 ppm, and used as a sample for measurement of electron donating ability (EDA). L-ascorbic acid (Junsei, Japan) was used as a vitamin C control and α-tocoperol (Sigma, USA) was used as a vitamin E control.

To determine the electron donating ability, 0.5 mL of 0.2 mM 1.1-diphenyl-2-picrylhydrazyl (DPPH) was added to 1 mL of the sample using Blois's (1958) method. After stirring for 30 minutes, a spectrophotometer (Optizen 2120UV, Mecasys. Korea) was used to measure the absorbance at 517 nm. The absorbance was calculated by substituting the following formula, and the result was expressed as a percentage (%).

Electron donating ability (%) = (1 - absorbance of sample-added group / absorbance of sample-free group) x 100

SOD-like activity, Peroxynitrite, total phenol and flavonoid, and reducing power were measured by selecting the top 6 high-value candidates based on the results of electron donating ability. L-ascorbic acid (Junsei, Japan) and α-tocoperol (Sigma, USA) as a vitamin E control.

SOD-like activity was measured according to Marklund's method (1974). 2.6 ml of Tris-HCl buffer (50 mM Tris, 10 mM EDTA, pH 8.5) and 0.2 ml of 7.2 mM pyrogallol were added to 0.2 ml of each sample, followed by reaction at 25 ° C for 10 minutes. Then, 0.1 ml of 1 M HCl was added to stop the reaction The amount of pyrogallol oxidized in the reaction solution was measured at 420 nm using a spectrophotometer (Optizen 2120UV, Mecasys, Korea). The SOD-like activity was expressed as a percentage by substituting the absorbance values of the sample addition and no-added sections into the following formula.

SOD-like activity (%) = (1 - absorbance of sample-added group / absorbance of sample-free group) × 100

And Peroxynitirte was prepared by the following method according to Kooy's method (1994). 0.5 ml of peroxynitrite was added to 1 ml of each sample, and the mixture was allowed to stand for 30 minutes. Then, absorbance was measured at 302 nm using a spectrophotometer (Optizen 2120UV, Mecasys., Korea). The scavenging activity of peroxynitrite was expressed as a percentage by substituting the absorbance value of the sample without and with no sample into the formula below.

Peroxynitirte scavenging ability (%) = (1 - absorbance of sample-added group / absorbance of sample-free group) × 100

The total phenolic content was determined by adding 1 ml each of the samples, Folin-Cilcalteau reagent and 10% Na ₂ CO ₃ solution in turn, according to the Folin-Denis method of Swain and Hillis (1959) Absorbance was measured using a photometer (Optizen 2120UV, Mecasys., Korea). The concentration of caffeic acid (Sigma Co. Ltd., St Louis, USA) was adjusted to 10 ppm (2.46 mg) and 100 ppm (3.27 mg) / 100 g) was calculated.

Total flavonoids were prepared by adding 0.1 ml of 10% aluminum nitrate, 0.1 ml of 1 M potaasium acetate, and 4.3 ml of ethanol to a sample of 1 mg / ml in accordance with the method of Moreno (2000), and the mixture was allowed to stand at room temperature for 40 minutes, Absorbance was measured using a spectrophotometer (Optizen 2120UV, Mecasys., Korea). The total flavonoid content (mg / 100 g) of the sample was calculated from a standard calibration curve obtained at a concentration range of 10 ppm (0.18), 100 ppm (0.70), and 1000 ppm (3.71) using Quercetin (Sigma Co.) as a reference material.

The reducing power was measured by Oyaizu (1996), and 1 ml of 200 mM phosphate buffer (pH 6.6) and 1 ml of 1% potassium ferricyanide were added to 1 ml of each sample, followed by stirring in a thermostatic chamber at 50 ° C for 20 minutes. 1 ml of 10% trichloroacetic acid (TCA) solution was added to 1 ml of the supernatant obtained by centrifugation (13.500 rpm, 15 minutes), and 1 ml of each of the distilled water and ferric chloride was added to the mixture. The mixture was then measured with a spectrophotometer (Optizen 2120UV, Mecasys ., Korea). The reducing power of the sample is expressed by the absorbance value.

2. Experimental results

The results of electron donating ability analysis of 48 natural extracts are shown in FIG. 1 and FIG. 2, respectively.

1 and 2, the green tea extract was found to have a high value of more than 60% when compared to the control antioxidant vitamin C and E, and then the extracts of white rye leaves, oodi, % Or more. As a result, the natural extracts to be mixed with fish oil were selected as candidates of six kinds of extracts (Paeonia leaf, Odie, Sansui oil, Seaweed, Seaweed and Green tea extract) of 20% or more based on the electron donating ability. Perxynitrite (%), SOD - like activity, total phenol and flavonoid, and reducing power. The analytical results are shown in Tables 4, 5 and 6, respectively.

Item Vitamin C Vitamin E Kool Audi Corn oil White leaf Seaweed green tea Peroxynitrite 78.88 + - 8.23 45.59 + - 3.56 78.88 ± 3.16 51.11 + - 5.71 6.20 ± 6.84 72.12 + - 7.35 66.59 + - 9.14 15.49 + - 11.25 SOD-like activity 75.38 + - 1.32 15.77 ± 1.76 18.14 + - 3.44 17.28 ± 2.97 17.71 ± 1.34 20.95 ± 4.21 29.16 ± 2.22 19.22 + - 3.01

  As shown in Table 5, Perxynitrite showed a high value of 70% or more and a predominant level of vitamin C, which is a control group, in Ulgum and Paeoniae leaf extracts. In the SOD - like activity, the seaweed extract was the highest at 29%, while the other extracts showed less than 20%.

Item Kool Audi Corn oil White leaf Seaweed green tea Total phenol 2.20 ± 0.04 2.19 ± 0.00 2.37 ± 0.17 0.90 + 0.13 0.37 ± 0.09 3.10 0.04 Total flavonoid 0.04 0.01 0.01 ± 0.00 02. ± 0.00 0.14 + 0.02 0.03 0.03 0.11 + - 0.01

As shown in Table 6, the total amount of green tea extract was 3.10 ± 0.09 mg / 100 g in total phenol and 2.0 mg / 100 g or more in the extracts of Ulgum, Oddi and Sapporo. Total flavonoids showed the highest value with 0.14 ± 0.02 mg / 100g of Leaf Leaf Extract.

Item Vitamin C Vitamin E Kool Audi Corn oil White leaf Seaweed green tea
Reducing power 0.89 ± 0.15 0.89 0.22 0.18 + 0.03 0.25 0.08 0.46 ± 0.05 0.58 + - 0.12 0.27 + 0.04 1.55 0.26

 Referring to Table 7, the reducing power of green tea extract was 1.55 ± 0.26 higher than that of the control.

<Experimental Example 3: Effect on functional fish oil composition containing natural extract>

1. Acid value determination of functional fish oil composition containing natural extract

Six kinds of natural extracts selected from antioxidant functionalities were added to sardine oil to prepare a functional fish oil composition (hereinafter referred to as test fish oil). The acid value changes at room temperature for the tested fish oil containing 1, 3, and 5 wt% of natural extracts were analyzed at 2 - week intervals along with the control diet, sardine refined oil.

2. Test results of test fish oil

To investigate the safety and functionality of fish oil, the test fish oil supplemented with the extracts of Ulgum (1,5wt%), Green tea (5wt%), White Leaf (5wt%) and Odi (5wt%), . The stability of the tested fish oil was tested for the stability of cadmium (Cd), lead (Pb), mercury (Hg) and coliforms by selecting one test fish oil supplemented with 5wt% Total eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and total flavonoids were measured in the test fish oil to which the added fish oil was added.

3. Color change of test fish oil

To measure the color difference of fish oil, test fish oil containing 1, 3, and 5 wt% of natural extracts was added to each of L, a, b Respectively.

4. Results

 The results of the acid value measurement for sardine oil are shown in FIG. 3, and the acid value measurement results for the test fish oil to which the six kinds of natural extracts are added are shown in FIG. 4 to FIG.

Referring to FIGS. 3 to 9, the acid value change measured for 60 days of test fish oil showed an increase in acid value from 0.2 to 0.4. However, the test fish oil except for the fish oil added with uroguin extract showed the increase of the acid value immediately after the addition. The test fish oil supplemented with Ulgum and Pyrrhizae leaf extracts remained at a similar level compared with the purified fish oil until 5th day. However, the test fish oil added with Pyrrhizae extract increased significantly after 5 days. The test fish oil supplemented with uroglycine extract showed similar acid value up to 60 days except 5wt%. As a result, among the six kinds of natural extracts, only the test fish oil containing 1 to 3 wt% of the fish extracts did not increase the acid value.

The results of the test analysis for EPA (eicosapentenoic acid), DHA (docosahexaenoic acid) and total flavonoids, cadmium, total mercury, lead, and coliforms in test fish oil are shown in Table 8 below.

Item
FOTU FOMU (5 wt%)
FOWM (5 wt%)
FOGT (5 wt%)
1wt% 5 wt% EPA (mg / g) - 164.4 166.9 158.9 159.7 DHA (mg / g) - 34.6 31.9 29.3 31.2 Flavonoid (mg / g) 118.38 10.06 138.58 100.03 62.82 Cd (mg / g) - not detect - - - Pb (mg / g) - not detect - - - Hg (mg / g) - not detect - - - Coliform group - negative - - -

Fish oil + white mulberry (FOMU), fish oil + white magnolia (FOWM), fish oil + green tea (FOGT) , Fish oil + green tea extract), -: not measured]

 Referring to Table 8 above, the amount of EPA and DHA in FOTU, a test fish oil containing 5 wt% Flavonoids were detected in all tested fish oil, especially FOMU, which was the test fish oil to which Audi extract was added. In FOTU, the flavonoid content was significantly lower than that of 1wt% added with 5wt%. From these results, it is considered that the addition amount should be decided at 5wt% or less in order to mix fish oil and koi extract.

On the other hand, FIG. 21 shows the L, a, and b values of the test fish oil in terms of the color change by the concentration colorimeter. The L (brightness) value of the color difference meter shows a bright white color and a black color with a lower value. A value shows a red color with a higher value, a green color with a - value shows a green color, It shows yellow, and the more it is shifted to - value, the more blue.

21, the test fish oil having a darker color as the added concentration of the six candidate groups added to fish oil (acidol, green tea, ugum, Green tea, uroguan, and oat extract were added. In the test fish oil added with seaweed extract, there was no significant difference in 3wt% and 5wt%. The value of a in the colorimetric system was red when the added concentration was higher than that of the test fish oil with addition of corn oil, waxy green tea, The higher the value, the greener the value. The b value of the colorimetric system was blue when the concentration was higher in the test fish oil added with seaweed, green tea, ugum, and augy extract. The test fish oil with the addition of the marine oil extract was yellowish, The most abundant yellow was observed in 3 wt% of tested fish oil.

From the results of color measurement by colorimetric system, it was confirmed that the koi extract was beneficial for the addition of fish oil in terms of color change among 6 kinds of extracts. In other words, the Ulgum extract is expected to help improve the visual value of fish oil by making the color of transparent yellow fish oil brighter.

<Experimental Example 4: Effect of feeding fish oil on animal test>

1. Production of experimental animals and feed

In order to investigate the in vivo effects on test fish oil, 50 male and 50 female rats were purchased from a 3 to 4 week old experimental rat (C57BL / 6J mouse) in a central experimental animal (Korea), and 3 rats were housed in a breeding cage. Were classified as oil-based pens and stabilized for 8 weeks. After 2 weeks, high fat diet (Rodent diet with 60% Kcal Fat) was fed to the experimental animals to induce obesity.

(CJ, Korea) were powdered and then mixed with 5 wt% of each of six test fish oils. Then, the mixture was mixed with water and then dried with hot air (50 ° C) for 2 days so as to have a moisture content of 10% Were used as experimental feeds. Test fish oil containing 5% by weight of sardine oil, white pearl leaf, olive oil, corn oil, sea urchin, seaweed, and green tea extract were added to each. Soybean oil, a natural antioxidant, vitamin C, vitamin E, and BHT, a synthetic antioxidant, were blended to a concentration of 5 wt%, respectively. The mixture was mixed with water, (50 ° C) for 2 days. Then, the general diet (CJ, Korea) was powdered as a control diet and mixed with 5% by weight of sardine refined oil. After mixing with water, it was dried by hot air (50 ℃) for 2 days to have a moisture content of 10%. Eleven kinds of diets were fed and tested in one control group and 10 experimental groups. Control (control) is a group fed by adding sardine refined oil to feed.

And 10 experimental groups were added fish oil to soybean oil group, SO group, fish oil supplemented with SOHL leaf extract to feed, FOWM group to feed group, test fish oil to which oder extract was added to feed FOOD group and FISH group were added to the diets, and FOCU group and FISH group were added to the diets. The fish group was divided into FOTU group, Fish oil supplemented with sea mustard extract was added to the feed, FOBS group and green fish extract were added to the feed, and the feed group was supplemented with FOGT and vitamin C to feed FOVC group and vitamin E were added to the feed, FOVE group and BHT were added to the feed, and the feed group was classified into the FOBH group.

2. Breeding experiment method

  Feeds were fed at a rate of 25g per cage once every 3 days. Feed intake was measured before feeding, and the body weight change of each group was measured from time to time. During the breeding period, the floor sawdust for mouse was changed to a new cage and the water was free water. Breeding was carried out for 69 days. During the breeding period, the humidity was maintained at 25 ~ 35% and the temperature was maintained at 21 ~ 25 ℃. All studies were carried out according to the animal experiment regulations of Chonnam National University.

3. Blood collection and blood characteristics

Hematocrit and hemoglobin were measured immediately after each group of mice were anesthetized and dissected after anesthesia in order to determine the blood characteristics of the mouse. After the blood was centrifuged (10,000 rpm, 15 minutes) (Aasan Phamacy, Kyungki-do, Korea) were analyzed for glucose, total cholesterol, triglyceride, HDL-cholesterol, GOT and GPT. Lysozyme is an immune system metrics Parry et al ., (1966).

4. Statistical Analysis

Statistical analysis was performed using the Duncan's multiple range test (Duncan, 1955) using the ANOVA-test. Statistical analysis was performed using SPSS (SPSS Ver. Respectively.

5. Results

As a result of raising the mice for 69 days, the weight changes of the male and female were shown in FIGS. 10 and 11, respectively, and blood analysis results of the mice were shown in FIGS. 12 to 19 [Fish oil (control, (FOVC, fish oil + vitamin C), fish oil + BHT (FOBH, fish oil + synthetic antioxidant), soy oil (SO, soybean oil) Fish oil + white mulberry (FOMU), Fish oil + Korea cornnelian cherry (FOCC), Fish oil + white magnolia (FOCU) FOWM, fish oil + green tea (FOGT, fish oil + green tea extract)], fish oil + brown seaweed (FOBS, fish oil + seaweed extract).

10 and 11, both males and females showed a tendency to increase weight continuously in the SO group. And we observed the tendency of weight gain in control and SO groups in males. Male FOTU group fed fish oil supplemented with Eolgi extract tended to lose weight gradually while maintaining low body weight compared with the other groups. In females, the FOGT group fed the green tea extract supplemented with green tea extract showed a significant weight loss at the beginning, then gradually recovered, but was significantly lowered again at 10 weeks. The body weight of FOVC group fed fish oil supplemented with vitamin C was significantly decreased from 50 days in females. The FOWM group fed the test fish oil supplemented with Paeoniae leaf extract, FOBS group fed fish oil with test and FOBS group fed fish oil with sea mustard extract also showed a tendency to decrease after 50 days.

As a result of the mouse experiment, it was confirmed that the weight gain of the male fish was not greater than that of the fish oil to which the natural extract was added, when the fish oil (soybean oil) and the refined oil were fed. Especially, the lowest weight change of the test fish oil added with ugum was shown, and the weight gradually decreased after 60 days. However, in the case of females, weight loss was observed in all experimental groups except cooking oil after 56th day, and vitamin C, It is presumed that the cause of weight loss of females is composed of various causes, and that the weight change due to the addition of natural extracts is different from that of males.

12 and 20, there was no significant difference ( P <0.05 ) between Hemoglobin and Hemoglobin in the hematocrit ( P <0.05 ). In the hematocrit, the FOBS and FOGT groups were significantly higher than the control group . ( P <0.05 ), but FOD, FOCC, and FOGT were significantly lower in triglyceride than in the control ( P <0.05 ). There was no significant difference between HDL-Cholesterol and GOT , And GPT was significantly lower than that of control and antioxidants ( P <0.05 ).

And all treatment groups in the blood lysozyme analysis of mouse are did not show the control group and significantly different (P> 0.05), FOMU group, FOWM group FOVE group, FOBH group, exhibited a significantly higher value than the FOGT group (P &Lt; 0.05 ).

As a result of analysis of hematological characteristics, hemoglobin was significantly increased in FOGT group and hematocrit in FOBS group and FOGT group, and glucose was significantly decreased in experimental groups, and thus, carbohydrate metabolism activity It seems to be active. In the case of triglyceride, the tendency is similar to that of glucose, and the addition of antioxidants seems to affect the lipid metabolism of fish oil. Serum chlosterol levels are closely related to cardiovascular diseases such as heart disease, hyperlipidemia, and arteriosclerosis, which are influenced by genetic factors and the type and amount of dietary lipids. Polyunsaturated fatty acids are known to have a significant effect on cholesterol lowering (Remesha et al., 1980; Nancy Becker et al., 1983; Reiser et al., 1985). There was no significant difference in total cholesterol except for SO and FOVE groups. SO group showed significantly lower cholesterol - lowering effect than cholesterol - treated group. When hepatic injury is caused by long-term administration of a drug causing liver damage, enzymes such as GOT and GPT are generated in the blood. These enzymes are the normal enzymes necessary for the metabolism of the liver, but they are usually released from the liver due to abnormal deformation and damage of hepatocytes due to chronic hepatitis, acute hepatitis, fatty liver, alcoholic hepatitis and liver cancer. (Yoo and Shin, 2012). Although GPT is mainly elevated in blood levels only when hepatocytes are destroyed, GOT is present in red blood cells, heart and muscle cells in addition to hepatocytes, and may be increased by other symptoms. Most liver diseases have higher GPT than GOT, and GPT is known to be more useful for diagnosing liver disease than GOT (Friedman et al., 2003). In this experiment, GOT was not significantly different among the experimental groups, but GPT was significantly lower in the experimental group added with the natural extract. Therefore, it was confirmed that the test fish oil supplemented with the natural extract was effective for the improvement of liver function. These results were significantly lower than those of the FOVC and FOVE groups, indicating that the substances which are not antioxidant functionalities but can be applied to improve liver function in natural substances such as phenols, flavonoids, catechins of green tea, It is highly likely that the result is a result of a variety of substances. The lysozyme activity, which is an immune system active substance, does not affect the immune system since there is no significant difference between the control and experimental groups.

As a result of the above-mentioned experimental results, it was confirmed that the refined oil was most advantageous in terms of quality and economical efficiency in manufacturing a functional fish oil composition containing the natural extract in the refined oil. When six natural extracts selected from natural extracts were added to fish oil, functionalities such as flavonoids were added. In particular, animal experiments showed that inhibition of weight gain, increase of carbohydrate metabolism activity, Decrease in liver function, and improvement of liver function. However, among the six natural extracts, five kinds of natural extracts except for the extract of Eugyolin increased the acidity of the fish oil, and the Eugolin extract was most effective in preventing the acidosis of the fish oil. In particular, the addition of 5wt% of Ulgum resulted in a significant increase of the acid value. The addition of Ulgum Extract at 3wt% or less, for example, 0.1 ~ 3.0wt%, could increase the effect of Ulgus and fish oil.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation. Accordingly, the true scope of protection of the present invention should be determined only by the appended claims.

Claims (5)

A functional fish oil composition using a natural substance, wherein the fish oil extract is added in an amount of 0.1 to 3.0 wt%. The functional fish oil composition according to claim 1, wherein the extract is extracted by adding at least one extraction solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, a polyhydric alcohol, or a mixture thereof. The functional fish oil composition according to claim 1, wherein the fish oil is a purified fish oil obtained by purifying sardine crude oil extracted from sardines. delete A food composition for improving liver function, which comprises the fish oil composition according to any one of claims 1 to 3.

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