KR20200087900A - health food composition using pomegranate fermented by lactic acid bacteria and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a health food composition using a pomegranate lactic acid bacteria fermentation product, and a manufacturing method thereof. More specifically, the present invention relates to the health food composition with excellent anti-obesity and anti-inflammatory effects by containing the lactic acid bacteria fermentation product obtained by fermenting pomegranate with lactic acid bacteria. The health food composition using the pomegranate lactic acid bacteria fermentation product of the present invention contains: milk; and the lactic acid bacteria fermentation product fermented by the lactic acid bacteria by mixing with a pomegranate extract.

Description

Health food composition using pomegranate fermented by lactic acid bacteria and manufacturing method thereof

The present invention relates to a health food composition using a pomegranate lactic acid bacterium fermentation product and a method for manufacturing the same, and more particularly, it contains a lactic acid bacterium fermentation product fermented with pomegranate as a lactic acid bacterium, and a health food composition having excellent efficacy such as anti-obesity and anti-inflammatory. It relates to a manufacturing method.

As the living environment is speeding up, the eating habits accordingly have been instantiated mainly with high calories, and reports of obesity, high blood pressure, and various adult diseases are rapidly increasing. Nuclear family, individualism, population growth at night activities, mass consumption period, diversification of consumption patterns, convenience pursuit trends, and changes in socioeconomic environment such as women's social advancement are undergoing changes in dietary structure.

And this change in consciousness about the composition of the meal and the increase in consumer demand for convenience food began to appear as a tendency to pursue convenience. However, these modern people's eating habits are prone to malnutrition, malnutrition and malnutrition due to incorrect nutrition information. Malnutrition due to malnutrition causes various diseases such as anemia, and excessive intake of nutrients causes overweight and obesity, making it a health problem today, as well as the development of chronic degenerative diseases such as arteriosclerosis, diabetes and hypertension. Weighted.

In particular, an increase in the incidence of adult diseases such as arteriosclerosis, hypertension, heart disease, diabetes, and hyperlipidemia due to obesity, leading to serious conditions such as various cancers, malignant tumors, heart disease, kidney disease, etc., and shortened lifespan.

In addition, as scientific evidence that dietary habits are very closely related to diseases is rapidly emerging, consumers who want to improve their health through lifestyle changes including dietary life are rapidly increasing. Accordingly, research on health supplements and functional foods has been actively conducted to prevent various diseases and help health.

On the other hand, pomegranate is a deciduous broad-leaved tree belonging to the pomegranate family. It is a plant native to Afghanistan and the western North. It was used in religious rites, arts, and mythology in ancient Egypt and Greece.

Pomegranate has a sweet and sour taste and has effects such as sterilization and insecticide. In the past, it has been thought that it is used as a medicinal herb or discarded the skin or seeds and eat only the flesh surrounding the seeds. A lot of attention has been paid to development as an enhanced drinking water.

Looking at the pomegranate's efficacy, it has the effect of preventing hypertension or arteriosclerosis in the fruit and skin, and it also has effects on menopause, menstrual dysfunction, obesity, various gynecological diseases, and swelling due to estrogen in the Internet, etc. It is described as an error that is used to restore physical strength, support nutrition for healthy people, and improve physical strength.

However, since pomegranate has a natural plant-based herbal ingredient, it is important to make it easy to consume while maintaining its natural state because it may easily deteriorate in heat or acid, and domestic studies so far have not escaped the initial stage.

On the other hand, lactic acid bacteria fermentation has been used in the production of dairy products mainly in fermented milk and sausages in the West, and has been used in the manufacture of agricultural and fisheries foods including kimchi, soy sauce, and salted fish since the early days of the Three Kingdoms period in Korea.

Lactic acid bacteria not only produce substances such as aromas, spices, and enzymes that can affect the sensory properties of fermented foods, but also some lactic acid bacteria have physical properties, texture, viscous, saccharification, stability, and moisture binders. , Gelling, it has been reported to produce exopolysaccharides (EPS) that can affect enrichment properties.

Exopolysakiride has been reported to exhibit useful effects such as immune enhancement, anti-ulcer activity, and reduced blood cholesterol, which are useful for the human body as well as physical properties of industrial useful aspects, and have been generally recognized as stable substances.

Since the late 1990s, products with emphasis on the functionality of probiotics and lactic acid bacteria have been shipped like floods, but many products with exaggerated functionality have led to a decrease in consumer confidence. Recently, as research has been conducted on the functionality and efficacy of new biotics that combine probiotics with natural materials, consumers' interest in this trend is increasing.

In Korea, probiotics are used in three senses: formal medicines for human medicine, lactic acid bacteria preparations, probiotics as feed additives, and lactic acid bacteria foods, a type of health food defined in the food industry.

Most of the conventional lactic acid bacteria products are in the form of fermented milk products, and various materials need to be explored, and it is necessary to develop high-quality products through combination with other excellent food materials that can expect synergistic effects.

1. Republic of Korea Patent Publication No. 10-2006-0014731: Method for manufacturing healthy food using pomegranate 2. Korea Patent Publication No. 10-2017-0120264: Balm fermented puree using probiotic lactic acid bacteria and food composition containing the same

The present invention was created to improve the above-mentioned problems, and it is possible to provide new probiotics made by fermenting pomegranate useful for health as lactic acid bacteria as foods of various formulations, and thus has excellent anti-obesity and anti-inflammatory effects, and is useful for health. It is an object to provide a health food composition and a manufacturing method thereof.

The health food composition using the pomegranate lactic acid bacteria fermentation product of the present invention for achieving the above object contains a lactic acid bacteria fermentation product fermented by lactic acid bacteria by mixing milk and pomegranate extract.

The lactic acid bacteria are strains of the genus Lactobacillus.

The strain is any one selected from Lactobacillus acidophilus , Lactobacillus plantarum , Lactobaciilus fermentum .

In addition, the health food composition is formed of any one selected from beverages, granules, tablets, powders, pills, capsules.

In addition, the lactic acid bacteria fermentation product is fermented with the pomegranate extract, Garcinia cambogia extract.

And a method for producing a health food composition using the pomegranate lactic acid bacteria fermentation product of the present invention for achieving the above object is a mixing step of mixing the pomegranate extract with milk; A sterilization step of sterilizing the mixture obtained in the mixing step; And a culture step of inoculating the mixture with the lactic acid bacteria after the sterilization step to obtain a fermented product of lactic acid bacteria.

In addition, in the mixing step, Garcinia cambogia extract is further added to the milk.

As described above, the present invention provides a health food composition having excellent efficacy such as anti-obesity and anti-inflammatory, and a method for manufacturing the same, by providing new probiotics made by fermenting pomegranate useful for health as foods of various formulations Can.

In addition, the existing lactic acid fermented milk products have a relatively weak point in terms of preservation, such as shelf life, since most of them are in liquid form, but the present invention extends the shelf life while maintaining the taste of lactic acid bacteria fermented milk, and ferments natural materials such as pomegranate. By processing into various formulations, it is possible to provide probiotic products differentiated from conventional products.

1 is a table showing the results of measuring the yield after lyophilizing the sample of Example 1,
Figure 2 is a table showing the results of the analysis of the free sugar of the sample of Example 1,
3 is a table showing the sensory test results for the sample of Example 1,
Figure 4 is a table showing the results of sensory tests conducted to evaluate the palatability according to the concentration of pomegranate extract,
5 is a graph showing the results of measuring the cytotoxicity of the sample of Example 2,
6 is a graph showing the results of measuring the anti-inflammatory properties of the sample of Example 2,
7 is a graph showing the results of measuring only the anti-obesity of the sample of Example 2.

Hereinafter, a health food composition using a pomegranate lactic acid bacteria fermentation product according to a preferred embodiment of the present invention and a method for manufacturing the same will be described in detail.

The health food composition of the present invention contains a lactic acid bacteria fermentation product. For example, the health food composition of the present invention may contain lactic acid bacteria fermentation product in 1 to 99% by weight. In addition, of course, the health food composition of the present invention may contain 100% by weight of the fermented lactic acid bacteria.

The lactic acid bacteria fermentation product is a mixture of milk and pomegranate extract fermented by lactic acid bacteria. The lactic acid bacteria fermentation product may be liquid or powder.

You can use commercially available products as milk, or you can use crude oil.

Pomegranate extract is extracted from pomegranate. The pomegranate can use whole fruit, or the rest of the fruit except the skin.

Various extraction methods are applicable to extract pomegranate extract. For example, pomegranate can be extracted by adding an extraction solvent. At this time, at least one selected from water, lower alcohol having 1 to 4 carbon atoms, polyhydric alcohol, or a mixture thereof may be used as the extraction solvent.

As a lower alcohol having 1 to 4 carbons, methanol and ethanol can be used, and as the polyhydric alcohol, butylene glycol, propylene glycol, pentylene glycol, and the like can be used. And as the mixture, a mixture of water and lower alcohol, a mixture of water and polyhydric alcohol, a mixture of lower alcohol and polyhydric alcohol, or a mixture of water and lower alcohol and polyhydric alcohol may be used.

As an example of extraction, the extraction solvent may be added at a weight ratio of 2 to 20 times with respect to pomegranate, and then extracted at 20 to 120° C. for 1 to 48 hours and filtered to obtain a liquid pomegranate extract. In addition, it is of course possible to obtain pomegranate extract in the form of a concentrate by filtration and concentration. In addition, the concentrate can be obtained in powder form through lyophilization or spray drying.

Lactobacillus strains are used as lactic acid bacteria. These strains include Lactobacillus acidophilus , Lactobacillus plantarum , Lactobacillus fermentum fermentum ).

The fermented lactic acid bacteria fermented with pomegranate extract as described above can provide a new probiotic product using pomegranate. This provides a differentiated probiotic product because of its superior functionality compared to conventional probiotics.

The fermented lactic acid bacteria fermented pomegranate extract with lactic acid bacteria is very useful for health due to its excellent anti-obesity and anti-inflammatory effects.

In the health food composition of the present invention, a known food additive may be added in addition to the fermented product of lactic acid bacteria.

Examples of known food additives include saccharides such as monosaccharides, disaccharides, polysaccharides, and sugar alcohols, flavoring agents such as taumatin, stevia extract, saccharin and aspartame, nutrients, vitamins, edible electrolytes, flavors, colorants, and enhancers (eg , Cheese, chocolate, etc.), pectic acid, alginic acid, organic acid, protective colloidal thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonic acid, purified water and the like can be used.

The term'food composition' means a natural product or a processed product containing one or more nutrients, and preferably means a state that can be directly eaten after a certain process, and in a general sense, food It includes food additives, functional foods and beverages.

In addition,'health food composition' refers to the food group or food composition that has added value to act and express the function of the food for a specific purpose using physical, biochemical, and biotechnological methods, and prevent disease prevention and control. Refers to food products designed and processed to sufficiently express the body's regulating functions related to recovery.

The health food composition according to the present invention includes, for example, various foods, beverages, gum, tea, vitamin complexes, functional foods, and the like. Furthermore, the present invention provides various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, coloring agents and fillers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, It may contain a protective colloidal thickener, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonic acid used in carbonated beverages, etc., and the components may be used independently or in combination.

In addition, the health food composition may include a food-acceptable food supplement additive, and may further include suitable carriers, excipients, and diluents commonly used in the production of functional foods.

Since the health food composition of the present invention can be formulated in various forms, the form is not particularly limited. For example, the health food composition may be formed of any one selected from beverages, granules, tablets, powders, pills, and capsules. The health food composition having such drinks, granules, tablets, powders, pills, and capsules is easy to carry and easy to take at any time, anywhere.

On the other hand, the health food composition of the present invention may contain a fermented lactic acid bacteria fermented by a lactic acid bacteria garcinia cambogia extract with a pomegranate extract as another example.

Garcinia cambogia is a plant of the Clusiaceae family, which uses the skin part of the fruit, and the skin is rich in hydroxycitric acid (HCA), which has the ability to inhibit carbohydrates from being stored as fat. It is said to be.

Garcinia cambogia extract has a synergistic effect with pomegranate extract by inhibiting the synthesis of carbohydrates into fats, thereby further improving anti-inflammatory and anti-obesity functions.

Garcinia cambogia extract is extracted from the skin of Garcinia cambogia fruit. The extraction method is the same as the extraction method of the pomegranate extract described above, so a detailed description is omitted.

Hereinafter, a method for manufacturing the functional food of the present invention will be described.

The method of manufacturing the functional food of the present invention is largely a mixing step of mixing the pomegranate extract with milk, a sterilizing step of sterilizing the mixture obtained in the mixing step, and inoculating the mixture with lactic acid bacteria after the sterilization step to incubate to obtain a lactic acid bacteria fermentation product. Cultivation step. Each step is explained in detail.

1. Mixing step

First, prepare a pomegranate extract.

The pomegranate extract can be extracted by adding an extractant to the pomegranate as described above. As the extraction solvent, at least one selected from water, lower alcohol having 1 to 4 carbon atoms, polyhydric alcohol, or a mixture thereof can be used.

As an example of extraction, the extraction solvent may be added at a weight ratio of 2 to 20 times with respect to the pomegranate fruit, extracted at 20 to 120° C. for 1 to 48 hours, and filtered to obtain a liquid pomegranate extract. In addition, it is of course possible to obtain pomegranate extract in the form of a concentrate by filtration and concentration.

The prepared pomegranate extract is mixed with milk to obtain a mixture.

An example of the mixture may be obtained by mixing 10 to 100 parts by weight of pomegranate extract with respect to 100 parts by weight of milk.

In addition, another example of the mixture may be obtained by mixing pomegranate extract and garcinia cambogia extract in milk. For example, it can be obtained by mixing 10 to 100 parts by weight of pomegranate extract with respect to 100 parts by weight of milk and 10 to 100 parts by weight of Garcinia cambogia extract. Garcinia cambogia extract is extracted from the skin of garcinia cambogia fruit, and the extraction method is the same as the extraction method of the pomegranate extract described above.

In addition, sugar may be further added as another example of the mixture. For example, 10 to 100 parts by weight of pomegranate extract with respect to 100 parts by weight of milk and 0.1 to 5 parts by weight of sugar can be obtained by mixing. In addition, 10 to 100 parts by weight of pomegranate extract with respect to 100 parts by weight of milk, 10 to 100 parts by weight of Garcinia cambogia extract, and 0.1 to 5 parts by weight of sugar can be obtained.

As the sugar, monosaccharides such as glucose, disaccharides such as lactose or sucrose may be used.

2. Sterilization step

Next, the mixture obtained in the mixing step is sterilized.

To this end, the mixture is put into an autoclave and sterilized at 121°C and 1.2 atmospheres for 10 to 20 minutes. The sterilized mixture is allowed to cool in aseptic phase.

3. Culture stage

Next, lactic acid bacteria are inoculated into the sterilized 돤 mixture and cultured to obtain lactic acid bacteria fermentation products.

As a lactic acid bacteria, any one selected from Lactobacillus acidophilus , Lactobacillus plantarum , and Lactobaciilus fermentum is used.

Lactic acid bacteria are inoculated in MRS medium and liquid cultured for 24 hours in an incubator at 37° C. to obtain a culture medium for lactic acid bacteria. And after inoculating 0.5 to 5 parts by weight of the lactic acid bacteria culture solution with respect to 100 parts by weight of the sterilized mixture Lactic acid bacteria fermentation product can be finally obtained by incubating for 96 hours in a 37°C incubator using a low-temperature incubator.

The lactic acid bacteria fermentation product thus obtained may be a health food composition of the present invention as a liquid itself. In addition, the lactic acid bacteria fermentation product may be lyophilized to form a powder.

In addition, it may be formulated by adding a food-acceptable food supplement additive to the lactic acid bacteria fermentation product, or by adding an appropriate carrier, excipient and diluent. Examples of such formulations include drinks, granules, tablets, powders, pills, and capsules.

Hereinafter, the present invention will be described through the following examples and experiments. However, the following examples are intended to illustrate the present invention in detail, and the scope of the present invention is not limited to the following examples.

(Example 1)

10 times water was added to the pomegranate fruit in a weight ratio, and the extract was repeated twice at 80°C for 5 hours, and then concentrated to 10 brix to obtain a pomegranate extract.

And the mixture obtained by adding pomegranate extract and glucose to milk was sterilized at 121°C and 1.2 atmospheres for 15 minutes using an autoclave, and then allowed to cool in aseptic conditions.

Then, after inoculating 10 ml of lactic acid bacteria culture in 490 ml of the sterilized mixture, Lactic acid bacteria fermentation products were prepared by incubating for 96 hours in a 37°C incubator using a low-temperature incubator.

The culture of lactic acid bacteria is Lactobacillus even know Phil Ruth (Lactobacillus acidophilus) KCCM 41619, Lactobacillus Planta Room (Lactobacillus plantarum) KCCM 12116, Lactobacillus Pere momentum (Lactobaciilus fermentum ) Each of KCCM 35469 was inoculated in MRS medium and obtained by incubating for 24 hours in a 37°C incubator.

According to the composition of the mixture and the strain used during fermentation, as shown in Table 1 below, a total of nine lactic acid bacteria fermentation products were prepared.

division Mixture composition (wt%) Strain milk Pomegranate extract Glucose Sample 1 89 10 One L.acidophilus Sample 2 79 20 One L.acidophilus Sample 3 69 30 One L.acidophilus Sample 4 89 10 One L.plantarum Sample 5 79 20 One L.plantarum Sample 6 69 30 One L.plantarum Sample 7 89 10 One L.fermentum Sample 8 79 20 One L.fermentum Sample 9 69 30 One L.fermentum

<Yield of fermented lactic acid bacteria>

The results of measuring the yield after lyophilizing the sample of Example 1 are shown in FIG. 1.

Referring to FIG. 1, the lactic acid bacteria fermentation product cultured with L. fermentum by adding 30% of pomegranate extract was the highest with a yield of 13.9%, and the rest of the samples did not show a significant difference. Therefore, the amount of pomegranate extract does not appear to have a significant effect on the yield of lactic acid bacteria fermentation.

<Analysis of Free Sugars of Lactic Acid Bacteria Fermentation>

Free sugar analysis was performed according to the method of Wilson et al. The sample of Example 1 was centrifuged (3,000 rpm, 30 min), the supernatant was taken, filtered with filter paper (Whatman No. 2), purified with Sepak C18, and then filtered with a 0.45 μm membrane filter (Millipore Co., USA). The filtrate was analyzed using HPLC (High Performance Liquid Chromatography). The results of the free sugar analysis are shown in FIG. 2.

Referring to FIG. 2, the L. acidophilus strain fermented lactic acid bacteria fermented products showed the highest total sugar content. Therefore, the L. acidophilus strain is most likely to be fermented.

<Sensory test>

A sensory test was performed on the sample of Example 1. The sensory test consisted of 20 trained panels composed of panels, and the color, taste, aroma, and overall preference were conducted by a seven-step evaluation method.

The scoring criteria were very good 7, good 6, slightly good 5, normal 4, slightly rejected 3, rejected 2, strong rejected 1, and the sample number was changed every 2 hours to repeat the same panel 3 times. Average score was calculated except high and lowest score.

The SPSS statistical analysis program (SPSS, ver. 16.0, USA) was used to calculate the mean value and standard deviation between each experimental group, and the statistical significance was obtained with a significance level of p<0.05. It was verified by.

The sensory test results are shown in FIG. 3.

As a result of sensory test, L. fermenterim sample with 30% pomegranate extract showed the lowest preference in terms of flavor, color, taste, and general preference. Regardless of the type of lactic acid bacteria used, samples added with 30% of pomegranate extract showed low results due to the peculiar flavor and taste of pomegranate, and samples added with 10% and 20% pomegranate extract were not significantly lower than the control without added pomegranate extract. The result was not shown. Differences by individual preference are visible, but considering the content and preference of pomegranate, L. acidophilus lactic acid bacteria fermentation product with 20% pomegranate extract is considered to be suitable as a developed product.

A sensory test was additionally performed to evaluate the palatability according to the concentration of the pomegranate extract.

A sensory test was conducted using a fermented lactic acid bacteria fermented by adding L. acidophilus culture to a mixture of 20% of pomegranate extracts with different concentrations (1, 3, 5, 10 brix) in milk, and the results It is shown in FIG. 4.

Referring to FIG. 4, the degree of preference according to the degree of concentration was in the order of 5brix> 3brix >1brix> 10brix. Although there is a difference by individual preference, it is judged that the pomegranate extract, which is excellent in preference, is suitable as a development product.

(Example 2)

Water was added to the pomegranate fruit at a weight ratio of 10 times and extracted repeatedly at 80° C. for 5 hours twice, and then concentrated to 5 brix to obtain a pomegranate extract. Then, the garcinia cambogia fruit peel was added at a weight ratio of 10 times and extracted at 80°C for 6 hours, and then concentrated to 2brix to obtain a garcinia cambogia extract.

Then, a mixture of pomegranate extract, garcinia cambogia extract, and glucose added to milk was sterilized at 121°C and 1.2 atmospheres for 15 minutes using an autoclave, and then allowed to cool in aseptic conditions. Then, after inoculating 10 ml of lactic acid bacteria culture solution into 490 ml of the sterilized mixture, Lactic acid bacteria fermentation products were prepared by incubating for 96 hours in a 37°C incubator using a low-temperature incubator.

The lactic acid bacteria culture solution was obtained by inoculating Lactobacillus acidophilus KCCM 41619 in MRS medium and performing liquid culture for 24 hours in an incubator at 37°C.

According to the composition of the mixture, as shown in Table 2 below, lactic acid bacteria fermentation products were prepared in a total of five types.

division Mixture composition (wt%) milk Pomegranate extract Garcinia cambogia extract Glucose A sample 49 40 10 One B sample 49 30 20 One C sample 49 20 30 One D sample 49 10 40 One E sample 49 - 50 One

Cytotoxicity experiments, anti-inflammatory experiments, and anti-obesity experiments were performed on the samples of Example 2. And the 5brix pomegranate extract used in Example 2 was used as a comparative example.

<Cytotoxicity experiment>

Raw 264.7 was incubated in a 96 well plate at a concentration of 1 × 10 ⁵ cells/mL and used in the experiment. After the samples were treated at concentrations of 10, 50, 100, and 500 µg/mL for 24 hours, the medium was removed and 10 uL MTS was added to the new medium to react for 4 hours, and absorbance was measured at 540 nm. The cell viability of each sample was calculated relative to the sample-untreated group as 100%.

The results of measuring cytotoxicity are shown in FIG. 5.

Referring to FIG. 5, as a result of measuring cell viability of pomegranate extract and A, B, C, D, and E, cells of 96.2, 95.1, 97.3, 96.2, 98.1, and 97.0% at the highest concentrations of 500 μg/mL, respectively. The viability showed no toxicity in all samples.

<Anti-inflammatory experiment>

After dispensing Raw 264.7 into a 96-well plate at a concentration of 1 × 10 ⁵ cells/mL, each sample was treated for each concentration and cultured for 24 hours. 100 μL of cell culture supernatant and 50 μL of Griess reagent (A reagent: 1% sulfanilamide, B reagent: 0.1% naphthylethylendiamine in 25% phosphoric acid) were mixed and reacted in a 96-well plate for 10 minutes, then absorbed at 540 nm using a microplate reader. Was measured. NO ₂ -The standard curve was prepared using NaNO ₂ by concentration.

The results of the anti-inflammatory experiment are shown in FIG. 6.

Referring to FIG. 6, as a result of measuring the NO production rates of pomegranate extract and A, B, C, D, and E, when the treatment group inducing inflammation with LPS was 100%, each NO production rate was 500 µg/mL. In 98.1, 98.2, 96.2, 97.3, 91.2, and 95.3%, respectively, D showed the highest inhibition rate of NO production.

<Anti-obesity experiment>

ㅇ Confirm the inhibition of adipocyte differentiation of 3T3-L1 cells

3T3-L1 cells were seeded at 1 × 10 ⁴ cells/mL in 12 well plates, medium was exchanged every 2 days, and cultured for 4 days. Differentiation medium (MDI: 0.5 mM IBMX, 1 mg/mL Insulin, 0.25 μM dexamethasone) was treated to cause differentiation into adipocytes. In order to confirm the inhibitory effect of the differentiation of each sample, 10, 50, 100, and 500 μg/mL were simultaneously treated. After incubation for 48 hours, the medium was replaced with insulin medium (1 mg/mL Insulin) and further incubated for 48 hours. After the cultivation was completed, it was confirmed by staining with Oil red O to suppress differentiation into adipocytes.

ㅇ Oil red O staining

In order to confirm the effect of each sample on adipocytes, 3T3-L1 cell line was induced by differentiation and stained with Oil red O. After treating the extract with differentiated 3T3-L1 cells for 48 hours, the cells were washed 3 times with PBS and fixed with 4% paraform aldehyde at 4°C for 30 minutes. The fixed cells were washed 3 times with PBS, stained with 0.5% 　Oil red O staining reagent (solution filtered with 0.45 μm filter after dissolving in 60% isopropanol and 40% water) for 1 hour at 4° C., and then 3 with PBS. Washed once. The dyed fat was dissolved in DMSO and absorbance was measured at 500 nm. Fat content was calculated as a percentage of the average absorbance of the control group.

The results of the anti-obesity experiment are shown in FIG. 7.

Referring to FIG. 7, as a result of measuring the lipid accumulation of pomegranate extract and A, B, C, D, and E, a treatment for inducing differentiation into adipocytes (MDI: 0.5 mM IBMX, 1 mg/mL Insulin, 0.25 μM dexamethasone) When the absorbance was 100%, lipid accumulation was 100.0, 99.1, 98.2, 98.5, 87.6, and 91.8%, respectively, showing the highest anti-obesity effect in the sample of D.

As described above, the present invention has been described with reference to one embodiment, but this is only exemplary, and those skilled in the art will understand that various modifications and equivalent embodiments are possible therefrom. Therefore, the true protection scope of the present invention should be defined only by the appended claims.

Claims (7)

A health food composition using a pomegranate lactic acid bacteria fermentation product, comprising a lactic acid bacteria fermentation product fermented by lactic acid bacteria by mixing milk and pomegranate extract. The method of claim 1, wherein the lactic acid bacteria is a healthy food composition using a pomegranate lactic acid bacteria fermentation, characterized in that the strain of genus Lactobacillus. The method of claim 2, wherein the strain is Lactobacillus acidophilus ( Lactobacillus acidophilus ), Lactobacillus plantarum ( Lactobacillus plantarum ), Lactobacillus fermentum ( Lactobaciilus fermentum ) Pomegranate lactic acid bacteria fermentation characterized in that any one selected from Health food composition using water. The method of claim 1, wherein the health food composition is a health food composition using a pomegranate lactic acid bacteria fermentation product, characterized in that it is formed of any one selected from beverages, granules, tablets, powders, pills, capsules. The method of claim 1, wherein the lactic acid bacteria fermentation product together with the pomegranate extract Garcinia cambogia ( Garcinia cambogia ) Health food composition using pomegranate lactic acid bacteria fermentation product, characterized in that the extract is fermented by the lactic acid bacteria. A mixing step of mixing pomegranate extract with milk;
A sterilization step of sterilizing the mixture obtained in the mixing step;
Method of manufacturing a health food composition using a pomegranate lactic acid bacteria fermentation product comprising; a culture step of inoculating the mixture with the lactic acid bacteria after the sterilization step to obtain a fermented product of lactic acid bacteria. 7. The method of claim 6, wherein the mixing step further comprises adding Garcinia cambogia extract to the milk.

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