KR102512796B1 - Extract of Molokhia with high antioxidant effect, and use thereof - Google Patents

Abstract

The present invention relates to a Molochia extract whose antioxidant properties are enhanced by irradiating the fermented Molochia extract with radiation. The Molochia extract of the present invention is a natural product having high antioxidant activity such as inhibition of lipid oxidation, prevention of fatty acid oxidation of fatty foods, and inhibition of melanin production, and can be usefully used as a natural antioxidant in products such as oils, foods, and cosmetics.

Description

Molokhia extract with high antioxidant effect and use thereof {Extract of Molokhia with high antioxidant effect, and use thereof}

The present invention relates to a Molokhia extract improved in antioxidant properties by extracting Molokhia ( Corchorus olitorius L.) with a solvent, fermenting it with mushroom mycelium, and then irradiating it with radiation. More specifically, it relates to a Molokhia extract having excellent antioxidant activity by fermenting the Molokhia extract using Bovista plumbea and then irradiating radiation to sterilize the fermentation broth.

The present invention is to provide a Molochia extract having far superior antioxidant properties than conventional heat sterilization or filtration methods by irradiating the fermentation broth with radiation.

Oxidation is the state in which a substance combines with oxygen or loses hydrogen. In other words, it refers to a change due to a reaction that loses electrons or hydrogen, or a chemical change that follows it. Many of the products that use organic substances as main ingredients, such as food and cosmetics, are easily oxidized, and oxidation occurs for various reasons to deteriorate the product and in some cases to produce toxic substances. Oxidation is mainly caused by oxidases such as polyphenol oxidase and lipoxygenase, and sometimes by autoxidation. Organic components susceptible to oxidation include fats and oils, essential oils, vitamins, carotenoids, and polyphenols. Oxidation is accelerated by light, transition metals, heme pigments, etc., and inhibited by natural and synthetic antioxidants or synergists, but it is difficult to completely prevent oxidation even when stored at extremely low temperatures.

Agents used to prevent oxidation are called antioxidants, and ethoxyquin, butylated hydroxytoluen (BHT), and butylated hydroxyanisol (BHA) are chemically synthesized and used. Natural substances that prevent oxidation include tocopherols, flavone derivatives, phyllozurcins, gallic acid derivatives, catechin, nordihydroguaiaretic acid, gossypol, and lignan glycosides, which are also called natural antioxidants. Natural antioxidants include polyphenols, vitamin C, vitamin E, and β-carotene, which are present in living organisms. It has been found that they inhibit the activity of active oxygen involved in arteriosclerosis, brain/cardiovascular system disorders, aging, and carcinogenesis. In addition to existing antioxidants, the effects and efficacy of orally ingested antioxidants have recently attracted attention.

The action of antioxidants is not usually permanent, but indirect antioxidants that inactivate metals that act as catalysts for oxidation reactions, such as quinones, amines, and phenols, which prevent oxidation by being directly involved in oxidation reactions. there is. Phenyl-β-naphthylamine is generally used for synthetic rubbers, and aromatic amines and hydroquinone are also used. In natural rubber, amino acids in latex are natural antioxidants. Fats and oils contain vitamin E, sesamol, vitamin C, quercetin and the like as natural antioxidants. Food sanitation law permits the addition of fruit extract, cysteine hydrochloride, dibutylhydroxytoluene (BHT), nordihydroguazaretic acid, butylhydroxyanisole (BHA), propyl gallate, etc. , If phosphoric acid, citric acid, etc. are used in combination with these, the effect is even greater. In addition, phenols, aromatic amines, etc. are used for lubricating oil, and sulfur compounds and phosphorus compounds, which are metal deactivators, are used for those used at high temperatures, such as engine oil.

On the other hand, Molokhia ( Corchorus olitorius L.) is a greenish-yellow annual plant of the family Pinaceae known to originate from southern China. It is a leafy vegetable harvested from young leaves for food. It is becoming. Its scientific name is Corchorus Olitorius , which was widely used as a mysterious herb in the Arab Empire, and it is said that in the past, an Egyptian king ate this vegetable and was cured of a serious illness, so it is said that from then on, it began to be called Molokhia (the vegetable of the king) (Azuma K, Nakayama M, Koshioka M, Ippoushi K, Yamaguchi Y, Kohata K, Yamauchi Y, Ito H, Higashio H. 1999. Phenolic antioxidant from the leaves of Corchorus olitorius L. J Agric Food Chem 47, 3963-3966). Depending on the variety, Molokia is also used as an ingredient for holy oil. The leaves have no scent and when cut finely, a unique slime comes out. In Korea, a study has been published that Molokhia has a health-promoting effect of lowering cholesterol and triglyceride and improving constipation (Jung CH, Choi IW, Kim SR and Seog, HM. 2003. Effect of Molokhia ( Corchorus olitorius) . ) and its mucilage on choresterol fed rats. Kor. J. Food Sci. Technol. 35, 379-385). Molokia is rich in dietary fiber, various vitamins, minerals such as calcium, polyphenols, carotenoids, etc., and is rich in viscous polysaccharides, yet has resistance to diseases and pests (Oshodi AA. 1992. Comparison of protein, minerals and vitamin C contents of some dried leafy vegetables.Pak J Sci Ind Res 35(3):267-269). Compared to spinach, it contains 3.5 times more carotene, 8 times more calcium, 4 times more vitamin A, 20 times more vitamin B2, and 30% more dietary fiber than spinach.

Prior art related to the present invention is a cosmetic composition for skin improvement containing a mixed fermented extract of marjoram, molochia and lemon balm (Patent Registration No. 10-1926263), in which the mixed fermented extract of marjoram, molochia and lemon balm is used as an active ingredient. It relates to a cosmetic composition comprising a. When this mixed extract is fermented using Saccharomyces cerevisiae, it is reported that the fermented product has skin wrinkle improvement, skin whitening, skin moisturizing, skin irritation mitigation and antioxidant effects, and skin condition improvement and anti-aging by customized extraction method of natural materials The cosmetic composition (Patent Registration No. 10-2084158) relates to a cosmetic composition comprising a mature product of a mixture containing molochia, purple carrot, mate, and borage or an extract of the aged product. After maximizing the content of active ingredients by aging molochia and purple carrots, which are super vegetables containing essential amino acids, and mate and borage rich in minerals and antioxidants, the destruction of active ingredients is minimized by low-temperature ultrasonic waves. There are reports that can be extracted stably. However, it is different from the present invention in which the mycelium for fermentation presented in this specification is used for bioconversion and impurities are removed using ionizing radiation, and the activity thereof is also significantly different.

On the other hand, irradiation technology is widely used for the purpose of sterilization of food or medical supplies without significantly changing the product temperature. Accordingly, the inventors of the present invention completed the present invention by finding that when fermentation is stopped by irradiating the fermented Molochia extract with radiation, the antioxidant property of the fermented Molochhia extract is further increased, unlike other fermentation stopping methods.

Registered Patent Publication No. 10-1926263 Registered Patent Publication No. 10-2084158 Patent Publication No. 10-2019-0065127

The present inventors have intensively tried to develop antioxidants that do not cause side effects or drug resistance from natural extracts and have low toxicity and can be administered for a long time. As a result, it was confirmed that the fermented Molochia extract obtained after fermenting the Molochia extract using glutinous rice cake mycelium exhibited high antioxidant activity. The present invention was completed by confirming that the antioxidant properties of the fermented Molochia extract were significantly improved when heat treatment, microfiltration, and irradiation were performed to stop fermentation by selecting the final fermentation time.

Accordingly, an object of the present invention is to provide a Molokhia extract with improved antioxidant properties by irradiating mycelial fermentation and irradiation.

Another object of the present invention is to provide a method for producing a Molochia extract having an increased antioxidant effect by irradiating the Molochia extract fermented with mycelium with radiation.

Another object of the present invention is to provide a natural antioxidant containing the fermented Molochia extract as an active ingredient.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problem, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to achieve the object of the present invention, the present invention provides a Molokhia extract having high antioxidant activity.

More specifically, the present invention changes the structure of organic compounds such as polyphenols and glycosides contained in the Molochia fermented extract or useful By increasing the component content (Table 2), a Molochia extract with higher antioxidant activity (Table 3) than common Molochia extract fermentation products was obtained.

The present invention also

(a) fermenting an extract obtained by extracting Molochia root using a low-carbon alcohol solvent using glutinous rice cake mushroom; and

(b) providing a method for producing a molochhia extract with increased antioxidant effect, comprising the step of irradiating radiation to the fermented molochhia extract obtained in step (a).

In the following, the present invention is described in more detail.

In a previous study, the present inventors confirmed that the main component showing efficacy in Molokhia extract was polyphenolic compounds, and as a result of fermenting Molokhia extract with various mushroom mycelium and confirming the content of polyphenolic compounds, fermented with glutinous rice cake mushroom mycelium It was confirmed that the fermented Molochia extract fermented with Molochia extract or other types of mushroom mycelium was the most effective in increasing the total phenol content and total flavonoid content compared to the fermented Molochia extract fermented liquid.

When fermenting Molochia extract, physicochemical methods can be used to stop fermentation, but it is very important to select and apply a method to stably stop fermentation without changing the useful efficacy of the Molokhia fermented extract. . In addition, suppression of discoloration (browning phenomenon, etc.) caused by heat sterilization and the like and prevention of oxidation during storage should be considered.

Accordingly, when the present inventors performed heat treatment, microfiltration, and irradiation to stop fermentation, the irradiated group observed higher antioxidant activity than the untreated group. On the other hand, in the heat treatment group, the antioxidant property was rather slightly reduced, and in the case of microfiltration, there was no significant change in antioxidant property, but the clogging of the microfilter paper continued to occur, which had the disadvantage of requiring a lot of manufacturing cost and time.

In addition, according to the following examples of the present invention, the irradiated Molochia fermented extract of the present invention showed a very excellent fatty acid spoilage inhibitory effect in the case of oil or fat food, and in the case of cosmetics, the effect of inhibiting melanin pigment production to a very high level It has been shown to be useful as a natural antioxidant.

Accordingly, the present invention provides an antioxidant containing, as an active ingredient, a Molochia extract obtained by fermenting the Molochia extract using glutinous rice fungus and irradiating radiation to stop the fermentation.

"Moloki ( Corchorus olitorius L.)" of the present invention belongs to the Linaceae, and of course includes its subspecies and variants. In addition, Molochia of the present invention includes cognate herbal medicines that are obvious in the art and can be used for the same or similar purposes as those of the present invention.

In the present invention, "fermented molokia extract" refers to a fermented extract obtained after fermenting a molokia ( Corchorus olitorius L.) extract using Bovista plumbea .

In the present invention, "Irradiated Molochia fermented extract" is Molochia ( Corchorus olitorius L.) extract fermented with glutinous rice cake mushroom ( Bovista plumbea ) and then irradiated with ionizing radiation such as gamma rays, electron beams, or X-rays to stop fermentation Refers to the fermented extract.

The Molochia extract according to the present invention can be extracted with an ultrasonic extractor using a low-carbon alcohol solvent from Molochia leaves. Dried leaves or powders thereof commonly sold in the market for Molochia leaves may be used. It can also be prepared directly by cutting molokia leaves and then drying them.

Methanol, ethanol, propanediol, and mixed solvents thereof may be used as the low-carbon alcohol solvent, and ethanol is preferably used. When the Molochia extract is obtained using ethanol as an extraction solvent, there is an advantage in that various subsequent processes can be applied because the active ingredient does not dissolve or precipitate even when water is added to the extract.

In addition, the Molochia extract according to the present invention may be further fractionated by using an organic solvent having a different polarity from the primary extract obtained by extracting the Molochia leaves using a low-carbon alcohol solvent as described above. In addition, additionally purified fractions may be obtained using various chromatography and reverse osmosis-membrane separation.

Therefore, in the present invention, "Molochia extract" is a concept including all extracts, fractions, and purified products obtained in each step of extraction, fractionation, or purification as described above, as well as dilutions, concentrates, or dried products thereof. The Molochia extract of the present invention may be prepared in a powder state by additional processes such as distillation under reduced pressure and lyophilization or spray drying.

The glutinous rice cake mushroom used in the present invention ( Bovista plumbea ) has a size of 2-4 cm, is somewhat spherical, and grows mainly in Europe and California. It is used for food, but its efficacy has not been specifically known yet. In the present invention, the glutinous rice cake mushroom belongs to the family Wrinkle mushroom, and includes subspecies and variants thereof.

In the present invention, when using the mycelium of glutinous rice cake or glutinous rice cake to ferment the Molokia extract, but not limited to, 0.1 to 20% (v / v), preferably 1 to 10% (v / v) or 0.1 to 20% by weight, 1 to 10% by weight can be used for fermentation. If it is less than the above range, the Molochia extract may not be properly fermented, and if it exceeds the above range, the active ingredient may be destroyed due to over-fermentation. Volume unit is % by weight

In one embodiment of the present invention, it is preferable to use gamma rays, electron beams, or X-rays as the radiation irradiated to the fermented product extracted from Molochia, more preferably, gamma rays or electron beams can be used, and more preferably gamma rays. can be used.

In addition, the radiation is preferably irradiated with a dose of 5 kGy to 20 kGy, and more preferably irradiated with a dose of 10 kGy to 15 kGy. When irradiated with radiation at a dose of less than 5 kGy, there was no change in antioxidant properties, although not significantly, and when irradiated with a dose exceeding 20 kGy, there was a large difference compared to when irradiated with radiation at 20 kGy. It was found that there was no (Table 3, Table 4). Therefore, since irradiation of radiation exceeding 20 kGy is inefficient in terms of energy, it is preferable to irradiate radiation below 20 kGy.

In one embodiment of the present invention, when the visible color change of the samples before freeze-drying was observed, as the radiation dose increased, the color of the dark brown fermented extract was found to become thinner (FIG. 1), application as an additive It is expected that the level will increase.

According to a preferred embodiment of the present invention, the present invention provides an antioxidant containing, as an active ingredient, a fermented product extracted from Molochia that has been sterilized by irradiation. According to the present invention, the fermented product extracted from Molochia sterilized by irradiation of the present invention contains a higher content of total polyphenols and total flavonoids compared to other sterilization methods by ionization energy (Table 2), and The scavenging ability is very high (Table 3), and the effect of preventing oxidation during storage of oil or fat food is excellent (Table 4, Table 6). The extract of the present invention may be contained in the range of 0.5 to 5% by weight based on the total weight. If it is less than 0.5% by weight, it is difficult to expect a substantial anti-oxidation effect of Molochia extract, and if it exceeds 5% by weight, the color of the product may be changed or the physicochemical properties may be affected. The antioxidant according to the present invention may be prepared as a solution, powder, suspension, dispersion, emulsion, oil dispersion, paste, dust, dust, or granules, but is not limited thereto.

When the fermented product extracted from Molochia sterilized by irradiation of the present invention is used as a food additive, the fermented product may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).

There is no particular limitation on the type of food. Examples of foods to which the above substances can be added include meat, sausages, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes.

When using the composition as a food additive, the composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods.

The food composition may include a food chemically acceptable carrier.

The food composition may be a health functional food. Functional food is the same term as food for special health use (FoSHU), and refers to food with high medical and medical effects processed to efficiently display bioregulatory functions in addition to nutrient supply. , The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, and pills in order to obtain useful effects for preventing or improving skin inflammation.

At this time, the content of the strain or its culture medium contained in the food is not particularly limited thereto, but may be included in 0.01 to 100% by weight, more preferably 1 to 80% by weight based on the total weight of the food composition. If the food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g, based on 100 mL.

The health functional food can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and components commonly added in the art during the manufacture. In addition, unlike general drugs, there is an advantage in that there is no side effect that may occur when taking a drug for a long time by using food as a raw material, and it can be excellent in portability.

According to the present invention, the fermented product extracted from Molochia sterilized by irradiation of the present invention has a melanin production inhibitory effect (Table 7, FIGS. 2 and 3). The extract of the present invention may be contained in the range of 0.1 to 30% by weight based on the total weight. If it is less than 0.1% by weight, it is difficult to expect a practical whitening effect of the Molochia extract, and if it exceeds 50% by weight, the synergistic effect due to excess input is small and it is not economical.

In addition, the present invention provides a pharmaceutical composition for improving skin darkening comprising fermented Molochia extract sterilized by irradiation.

The pharmaceutical composition for improving skin darkening according to the present invention may further include a pharmaceutically acceptable carrier, and is formulated with the carrier to be used as medicines, foods (health functional foods), feed additives, drinking water additives, cosmetic compositions, etc. can be provided.

As used herein, the term 'pharmaceutically acceptable carrier' may refer to a carrier or diluent that does not inhibit the biological activity and properties of the administered compound while not stimulating living organisms.

The type of the carrier that can be used in the present invention is not particularly limited, and any carrier commonly used in the art and pharmaceutically acceptable can be used. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in combination of two or more.

In addition, if necessary, other conventional additives such as other antioxidants, buffers, and/or bacteriostatic agents may be added and used, and diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to form aqueous solutions, suspensions, emulsions, etc. It may be formulated into an injection formulation, pill, capsule, granule or tablet and the like.

The administration method of the pharmaceutical composition for improving skin darkening according to the present invention is not particularly limited, and may follow a method commonly used in the art. As a non-limiting example of the administration method, the composition may be administered by oral administration or parenteral administration. The pharmaceutical composition for improving skin darkening of the present invention may be prepared in various formulations according to a desired administration method.

The pharmaceutical composition of the present invention can be used as a single formulation, and can be prepared and used as a combination formulation by further including a drug known to have a recognized skin whitening effect, and is formulated using a pharmaceutically acceptable carrier or excipient to unite It can be prepared in dosage form or introduced into a multi-dose container.

As another aspect, the present invention provides a skin whitening treatment method comprising administering the pharmaceutical composition for skin whitening to a subject.

As used herein, the term "subject" may refer to all animals, including humans, that require skin whitening.

Specifically, the treatment method of the present invention may include administering a pharmaceutically effective amount of the fermented product extracted from Molochia that has been sterilized by irradiation to a subject in need of skin whitening.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and is generally administered in an amount of 100 mg to 10 g once a day to several times. It can be administered in divided doses. However, for the purpose of the present invention, a suitable total daily amount of the composition containing the extract can be determined by a treating physician within the scope of correct medical judgment, and can be administered once or divided into several times. However, for the purposes of the present invention, a specific therapeutically effective amount for a specific patient is determined by the type and extent of the response to be achieved, the specific composition, including whether other agents are used as the case may be, the patient's age, weight, general health condition, It is preferable to apply differently according to various factors including gender and diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used together with or concurrently used with a specific composition, and similar factors well known in the medical field.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be single or multiple administrations. It is important to administer the amount that can obtain the maximum effect with the minimum amount without causing side effects in consideration of all of the above factors, and can be easily determined by those skilled in the art.

As used herein, the term "administration" means introducing the pharmaceutical composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention is oral or parenteral as long as it can reach the target tissue. It can be administered through various routes.

The administration method of the pharmaceutical composition according to the present invention is not particularly limited, and may follow a method commonly used in the art. As a non-limiting example of the administration method, the composition may be administered by oral administration or parenteral administration. The pharmaceutical composition according to the present invention may be prepared in various formulations depending on the desired administration method.

The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be administered once a day or administered several times by dividing the dose.

In addition, the present invention provides a composition for food (health functional food) capable of preventing or improving skin darkening comprising fermented molochia extract sterilized by irradiation as an active ingredient. Since the intake of the strain of the present invention exhibits a blackening inhibitory effect in melanocytes, it can be provided as a food for preventing or improving skin blackening using it as an active ingredient. These health functional foods for preventing or improving skin melanin pigmentation can be consumed as edible cosmetics (Inner beauty).

According to another embodiment of the present invention, the present invention provides a quasi-drug composition for preventing, improving, or treating skin blackening comprising the fermented product extracted from Molochia that has been sterilized by irradiation. The fermented product extracted from Molochia sterilized by irradiation of the present invention may be added to a quasi-drug composition for the purpose of preventing, improving or treating skin inflammation.

In the present invention, the term "quasi-drugs" refers to textiles, rubber products or similar products used for the purpose of treating, mitigating, treating or preventing diseases of humans or animals, products that have weak effects on the human body or do not directly act on the human body, and devices or non-machines and similar items, products falling under one of the agents used for sterilization, insecticidal, and similar purposes to prevent infectious diseases, for the purpose of diagnosing, treating, mitigating, treating, or preventing human or animal diseases It may refer to items that are not instruments, machines or devices among items used and items other than instruments, machines or devices among items used for the purpose of pharmacologically affecting the structure and function of humans or animals.

In addition, the quasi-drugs may include external skin preparations and personal hygiene products. Preferably, it may be a disinfectant cleaner, shower foam, gargreen, wet tissue, detergent soap, hand wash, or ointment, but is not limited thereto.

When the composition according to the present invention is used as a quasi-drug additive, the fermented product extracted from Molochia sterilized by irradiation may be added as it is or used together with other quasi-drugs or quasi-drug ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined depending on the purpose of use.

According to another embodiment of the present invention, the present invention provides a drinking water additive for preventing or improving skin blackening comprising the fermented product extracted from Molochia that has been sterilized by irradiation. The drinking water additive of the present invention can be used by separately preparing the fermented molochia extract sterilized by irradiation in the form of a drinking water additive and mixing it into drinking water, or by directly adding it during preparation of drinking water. The drinking water additive of the present invention may be in a liquid or dry state, preferably in a dried powder form. A drying method for preparing the drinking water additive of the present invention in a dried powder form is not particularly limited, and a method commonly used in the art may be used. The drinking water additive of the present invention may further include other additives if necessary. Non-limiting examples of the usable additives include binders, emulsifiers, and preservatives added to prevent deterioration of drinking water quality; There are amino acids, vitamins, enzymes, probiotics, flavors, non-protein nitrogen compounds, silicates, buffers, coloring agents, extractants or oligosaccharides added to increase the effectiveness of feed or drinking water, and other feed additives are added. can be included with These may be used singly or two or more may be added together.

The fermented product extracted from Molochia that has been sterilized by irradiation of radiation according to the present invention exhibits an antioxidant effect even when used at a lower concentration, as the antioxidant effect is significantly higher than that of a general Molochia extract or fermented product extracted from Molochia. It can be used as a natural antioxidant by suppressing fatty acid spoilage during storage, and can be used as a pharmaceutical composition, a quasi-drug composition, a food (health functional food) composition, a drinking water additive, and a feed additive. It is effective in improving skin darkening when applied to cosmetics because of its excellent melanin production inhibitory effect in skin cells. In addition, the irradiated fermented product of Molochia extract of the present invention does not show hepatotoxicity and renal toxicity as a result of animal experiments, and is derived from natural products, so it can be safely and continuously used without applying serious stimulation to the body or causing harmful effects.

Figure 1 shows the change in color tone according to the irradiation dose of the irradiated fermented product extracted from Molochia.
Figure 2 visually shows the production of melanin produced in mouse melanocyte, Melan A cells.
Figure 3 shows the expression of whitening-related genes according to the irradiation dose of the fermented product extracted from Molochia irradiated in mouse melanocyte and Melan A cells.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited thereto.

[Comparative Example 1]

<Preparation of Molochia extract>

500 g of minced, shade-dried, and pulverized Molochia leaf powder was allowed to stand in a 50% (v/v) ethanol aqueous solution for 1 hour, and then 1,000 g using an ultrasonic extractor (SJC-II-50L, GREENAPPLE, China) fixed at 30 ° C. The first extraction was performed for 30 minutes at a frequency of 20 KHz at the output of W. After standing still for 30 minutes, the second extraction was performed again for 30 minutes under the above extraction conditions. After extraction, the extract was centrifuged at 5,000 rpm for 20 minutes, and the filtrate (1) was taken separately. . After mixing the filtrates (1) and (2), ethanol was removed using a vacuum concentrator, and lyophilization was performed to obtain Molochia leaf extract powder.

[Comparative Example 2]

<Preparation of fermented product extracted from Molochia>

In order to ferment the Molochia extract, the mycelium of glutinous rice cake was used as a spawn. First, the mycelium was cultured on a slope in a test tube containing potato dextrose agar medium, stored at 4 ° C, and subcultured every month for use. Aseptically homogenize the mycelia subcultured on the slope medium, and inoculate the Mushroom mycelium to 5% (v / v) of the medium in which the Molokia leaf extract and yeast-wort mixture obtained in Comparative Example 1 are mixed. The fermenter was cultured for 6 days under conditions of a temperature of 25° C., a rotation speed of 150 rpm, an aeration rate of 1.5 vm, and a pH of 7.0. After culturing, the culture broth was divided into 4 parts and used as untreated groups (Comparative Example 1), heat treatment (Comparative Example 2), microfiltration (Comparative Example 3), and radiation treated groups (Examples 1 to 8) for stopping fermentation.

[Comparative Example 3]

<Heat treatment and sterilization of fermented product extracted from Molochia>

The fermented Molochia extract prepared in Comparative Example 1 was put into an Erlenmeyer flask, the inlet was sealed with cotton, fixed in a stirring water bath fixed at 90 ° C, and heat treated for 30 minutes. After heat treatment, the Erlenmeyer flask was separated from the stirring bath, allowed to cool, and lyophilized and stored frozen.

[Comparative Example 4]

<Microfiltration treatment and sterilization of fermented product extracted from Molochia>

The fermentation broth of the Molochia extract prepared in Comparative Example 2 was filtered while being little by little poured into a vacuum filtration flask equipped with 0.45 μM filter paper. If the elution of the filtrate fell during filtration, the filter paper was exchanged and the filtration continued. The filtered filtrate was lyophilized and stored frozen.

[Examples 1 to 8]

<Irradiation treatment and sterilization of fermented product extracted from Molochia>

After evenly dispensing 100 mL of the fermented Molochia extract prepared in Comparative Example 1 into a test tube with a lid, the lid was closed and gamma rays and electron beams were irradiated, respectively. As shown in Table 1, the doses of gamma rays and electron beams were irradiated at room temperature so that the absorbed doses were 5, 10, 15, and 20 kGy. Montreal, Canada) was used at room temperature (20±1℃) at a dose rate of 70 Gy per minute, and the error of the total absorbed dose was calculated using a Ceric cerous dosimeter (Bruker Instruments, Rheinstetten, Germany) to confirm the absorbed dose. did For electron beam irradiation, an electron-beam accelerator (model ELV-8, 2.5 MeV, Eb-Tech, Daejeon, Korea) was used. It was irradiated with a dose rate of 5 mA acceleration current and 10 m/min velocity. The absorbed dose at this time was confirmed with a cellulose triacetate (CTA) dosimeter. Each of the irradiated Molochia extracts was lyophilized and stored frozen.

When the visible color change of the samples before freeze-drying was observed, as the irradiation dose increased, the color of the dark brown fermented extract became thinner (FIG. 1), and the applicability as an additive is expected to increase.

Radiation type and irradiation dose by sample sample name Gamma ray irradiation dose Electron beam irradiation dose Comparative Example 2 0 0 Example 1 5 0 Example 2 10 0 Example 3 20 0 Example 4 30 0 Example 5 0 5 Example 6 0 10 Example 7 0 20 Example 8 0 30

[Experimental Example 1] Measurement of antioxidant content contained in fermented product extracted from Molochia according to sterilization method

<Experiment method>

In Experimental Example 1, the fermented Molochia extract prepared in Comparative Example 1, the heat treatment group prepared in Comparative Example 2, the microfiltration treatment group of Comparative Example 3, the gamma ray treatment group of Examples 1 to 4, and the electron beams of Examples 5 to 8 The total polyphenol content and total flavonoid content of the treatment groups were compared.

<1-1> Measurement of total polyphenol content

After adding 10 mL of distilled water to each 1 g of each fermentation extract treatment sample, 2 mL of Folin-Ciocalteu phenol reagent was added and mixed, and then reacted at room temperature for 5 minutes. 2 mL of 20% sodium carbonate was added to the reaction mixture, mixed, and reacted at room temperature for 1 hour, and then absorbance was measured at 725 nm. At this time, tannic acid was used as the indicator material.

<1-2> Measurement of total flavonoid content

0.1 mL of 10% aluminum nitrate, 0.1 mL of 1 M potassium acetate, and 4.3 mL of ethanol were sequentially added to 0.5 g of each fermented extract treatment sample, mixed, reacted at room temperature for 40 minutes, and absorbance was measured at 415 nm. did At this time, quercetin was used as the indicator material.

<Experiment result>

The results of measuring the total polyphenol content and total flavonoid content according to the sterilization method of the fermented product extracted from Molochia are shown in Table 2 below.

Measurement results of total polyphenol content and total flavonoid content sample name Total polyphenol content
(μg/ml of extract) Total flavoid content
(μg/ml of extract) Comparative Example 1 79.5± ^1.9a 45.5± ^1.3a Comparative Example 2 96.5± ^5.3bc 74.4± ^2.8bc Comparative Example 3 88.2± ^1.4b 55.6± ^3.4ab Comparative Example 4 94.9± ^4.6bc 64.5± ^4.2b Example 1 102.2± ^4.7c 83.3± ^1.8c Example 2 113.8±4.3 ^cd 86.1±3.5 ^cd Example 3 124.1±4.5 ^d 94.5±2.6 ^d Example 4 123.5±2.2 ^d 93.3±1.5 ^d Example 5 102.6± ^3.7c 82.7± ^2.5c Example 6 114.6±4.2 ^cd 87.6±5.1 ^cd Example 7 123.8± ^1.4d 94.3±1.9 ^d Example 8 123.1± ^4.7d 94.1±2.1 ^d

The different English superscripts ( ^ad ) of the total polyphenol content and total flavonoid content values obtained according to the sterilization method of the fermented product extracted from Molochia indicate statistical significance (p<0.05).

Total polyphenol and total flavonoid contents were detected in a similar trend in all treatment groups. The heat treatment group (Comparative Example 3) showed the lowest content. It is considered that the total content is lowered because the heat treatment causes polymerization of organic substances present in the fermentation broth to lose water solubility and precipitate. All of the treatment groups irradiated with gamma rays or electron beams showed high content, but there was no statistically significant difference by the irradiation dose in the treatment groups of 10 kGy or more (Examples 2 to 4, Examples 6 to 8), but the increase in the irradiation dose It was found that the content increased as well. The microfiltration treatment group (Comparative Example 4) was detected slightly lower than the fermentation treatment group (Comparative Example 2). It is judged to be From the above results, it was confirmed that the total polyphenol content and the total flavonoid content could be increased when radiation of about 5 kGy was irradiated for sterilization of the fermented extract of Molochia leaf extract with glutinous rice fungus mycelium.

[Experimental Example 2] Antioxidant activity comparison test according to sterilization method

<Experiment method>

In this experimental example, the scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical was measured by slightly modifying the method used by Yoshida et al. to measure the antioxidant effect of the substances obtained in Examples 1 to 8, ABTS radical scavenging activity was measured by modifying the method of Roberta et al.

<2-1> Antioxidant effect test using DPPH radical

Since DPPH is a stable free radical and exhibits a purple color, the degree to which the purple color is removed is measured as the sample scavenges the radical. First, DPPH was dissolved in methanol at a concentration of 1×10 ^-4 M, and 0.15 mL of this solution was put into a test tube. 0.15 ml of each sample was put here, and the absorbance was measured at 565 nm after reacting at room temperature for 20 minutes. Vitamin C was used as a positive control. The free radical scavenging effect of each sample was expressed as SC ₅₀ by obtaining a concentration at which 50% of the radicals were scavenged. At this time, as a negative control, the reaction absorbance of the solution not treated with the sample was used.

<2-2> Antioxidant effect test using ABTS radical

ABTS radical solution was prepared by adding 7 mM ABTS and 2.45 mM potassium sulfate to distilled water to produce ABTS cations at room temperature for 16 hours. At this time, it was prepared by diluting the absorbance value at 734 nm to be 0.7 or less. After adding 50 μL of each concentration to 950 μL of ABTS solution and reacting for 10 minutes, absorbance was measured at 734 nm. As a positive control, butylic hydroxy aniline (BHA), a synthetic antioxidant, was used instead of the material in the examples. The free radical scavenging effect of each sample was expressed as SC ₅₀ by obtaining the concentration at which 50% of the radicals were scavenged. At this time, as a negative control, the reaction absorbance of the solution not treated with the sample was used.

<Experiment result>

<Antioxidant activity result>

The results of testing the DPPH radical scavenging activity and ABTS radical scavenging activity according to the sterilization process of the Molokia extract bioconverted into glutinous rice cake mushroom mycelium are shown in Table 3 below.

Antioxidant activity result according to sterilization process sample name DPPH radical scavenging activity
SC ₅₀ (μg/mL) ABTS radical scavenging activity
SC ₅₀ (μg/mL) Comparative Example 1 23.62± ^1.48e 27.14± ^2.51e Comparative Example 2 13.12± ^0.69d 12.77±0.23 ^d Comparative Example 3 19.64± ^0.84e 15.87± ^0.24e Comparative Example 4 14.18± ^1.02d 13.24±0.32 ^de Example 1 10.31± ^0.21c 10.17±0.14 ^cd Example 2 8.76± ^0.26c 8.85± ^0.13c Example 3 6.38± ^0.42b 6.13± ^0.51bc Example 4 5.42± ^0.37b 4.98± ^0.23b Example 5 10.51±0.45 ^c 10.24±0.26 ^cd Example 6 9.16± ^0.41c 8.81± ^0.24c Example 7 6.72± ^0.36b 6.22± ^0.44bc Example 8 5.91± ^0.25b 4.56± ^0.31b vitamin C 1.82±0.65 ^a BHA 1.52±0.95 ^a

The different English superscript ( ^ae ) of the DPPH radical scavenging activity value and the ABTS radical scavenging activity value obtained according to the sterilization method of the fermented Molochia extract indicate statistical significance (p<0.05).

In the simple extract of Molochia extract (Comparative Example 1), DPPH and ABTS radical scavenging activities were the lowest. The radical scavenging effect of each sterilization method for the fermented product extracted from Molochia was the lowest in the heat treatment group (Comparative Example 2) and the highest in the radiation irradiation group (Examples 1 to 8). In the case of the irradiated area, there was no significant difference between the doses at the irradiation dose of 15 kGy or more. The microfiltration treatment group (Comparative Example 4) showed similar results to the results of the sterilization non-treatment group (Comparative Example 2). On the other hand, in the case of DPPH radical scavenging activity, the SC ₅₀ value of vitamin C, which is a positive control, is 1.82 μg/mL, whereas the Molochia fermented extract of the present invention (Comparative Example 2) is 13.12 μg/mL, which is the same as that of a simple extract (Comparative Example 1). It showed higher activity than 23.62 μg/mL, and in the ABTS radical scavenging activity test, the SC ₅₀ value of BHA, a positive control, was 1.52 μg/mL, whereas the fermented product extracted from Molochia (Comparative Example 2) of the present invention was 12.77 μg/mL. It was detected as mL and showed higher activity than 27.14 μg/mL of the simple extract (Comparative Example 1). This is considered to have a very high radical scavenging activity considering the characteristics of the extract rather than a single substance.

[Experimental Example 3] Lipid rancidity inhibitory effect test of fats and oils

In this experimental example, in order to measure the lipid rancidity inhibitory effect of the samples obtained in Examples 1 to 8, pork fat was used as a model, and α-tocopherol, a natural antioxidant, and BHA, a synthetic antioxidant, were used as positive controls. Pork fat was supplied and used from Seoul Heinz Co., Ltd. Molochia extract samples (Comparative Examples 1 to 4, Examples 1 to 8) and positive control samples prepared at a concentration of 0.02% by weight were added to 100 g of pork fat, and then methanol was added and mixed for 2 minutes under a nitrogen stream. After removing methanol using a vacuum concentrator at 40 ° C, the lipid rancidity was measured while storing for 30 days in a thermostat fixed at 50 ° C.

Lipid rancidity of pork fat during storage was evaluated by measuring the peroxide value every 5 days. That is, after adding 35 ml of chloroform: acetic acid in a mixed solution prepared at a ratio of 2: 3 (V / V) to the pork fat sample and dissolving it, 0.5 mL of a saturated solution of potassium iodine (KI) was added and stirred sufficiently for 1 minute. It was stored in the dark for 5 minutes. Thereafter, 75 mL of distilled water and 1 mL of starch indicator were added, titrated with 0.01 N sodium sulfite solution (Na ₂ S ₂ O ₃ ), and the peroxide value was measured by Equation 1 below.

[Equation 1]

A: Titration value of sample (mL, Na ₂ S ₂ O ₃ )

B: Titration value of Control (mL, Na ₂ S ₂ O ₃ )

f : Factor

N: Specified concentration of Na ₂ S ₂ O ₃

S: amount of sample taken

Changes in rancidity during storage of pork fat sample name Change in peroxide value (meq/kg) during storage 0 days 5 days 10 days 15th 20 days 25 days 30 days Comparative Example 1 0 ^{12.4d 38.2d} 75.2 ^de 154.4 ^{f 213.3f 296.2g} Comparative Example 2 0 ^{8.1c 29.3c 69.8d 120.5e 154.6e 184.6e} Comparative Example 3 0 ^9.4c 34.5 ^{CDs 72.6d 124.7e 168.5e 204.6f} Comparative Example 4 0 ^{8.2c 31.1c 72.5d 121.6e 158.7e 188.4e} Example 1 0 7.6 ^{BC 18.1c 52.4c} 92.6 ^{d 110.8d 154.9d} Example 2 0 ^{6.7b 14.6b 31.2b} 79.8 ^c 96.5 ^{c 124.3c} Example 3 0 ^{6.3b 13.9b 32.4b} 80.5 ^c 97.7 ^c 125.4 ^c Example 4 0 ^{6.5b 13.2b 32.1b} 79.5 ^c 98.6 ^c 126.5 ^c Example 5 0 ^{7.5b 19.2c} 49.7 ^{c 87.8d 112.5d 160.1d} Example 6 0 ^{7.0b 14.5b 31.5b 80.4c} 96.5 ^{c 126.2c} Example 7 0 ^{6.8b 13.7b 32.6b 78.4c 98.1c} 125.8 ^c Example 8 0 ^{6.6b 14.0b 30.8b} 79.9 ^c 98.2 ^c 126.7 ^c tocopherol 0 4.3 ^{ab 12.8b 29.1b 67.3b 82.4b 106.5b} BHA 0 ^{2.5a 5.7a 10.6a} 22.8 ^a 29.7 ^a 35.9 ^a additive-free furniture 0 ^{15.2e 43.6e 82.8e 202.4g 318.1g 400.5h}

The other English superscript ( ^ah ) of the same storage days of the peroxide value production value during storage of the pig fat to which the sample obtained according to the sterilization method of the fermentation product extracted from Molochia was added indicates statistical significance (p<0.05).

As shown in Table 4, the effect of inhibiting fat rancidity during storage of pig fat added with samples treated by sterilization method for the fermented product extracted from Molochia was irradiated at an absorbed dose of 10 kGy or more (Examples 2 to 4, Comparative Examples 6 to 8) were found to be better than the non-irradiation group (Comparative Example 2), the heat treatment group (Comparative Example 4), and the microfiltration treatment group (Comparative Example 4), and it was observed that there was no difference between doses. Although the fat oxidation inhibitory effect was lower than that of the positive controls, tocopherol and BHA, considering that it is an extract, it is judged to have a very high fat oxidation inhibitory activity. On the other hand, the Molochia extract simple treatment group (Comparative Example 1) also appeared to have an effect of inhibiting rancidity during storage of pig fat. Therefore, it was confirmed that the radiation sterilization method for the fermented product extracted from Molochia of the present invention is effective in inhibiting oxidation of fatty foods.

[Experimental Example 4] Fatty acid spoilage inhibitory effect test during storage of hamburger patties

1 cm)를 이용하여 10 mm의 두께로 성형한 후 가열처리하였다. 가열은 85℃로 예열된 Cooker(NU-VU ES-3 cooker, Food Service System, Menominee, USA)에서 중심온도가 75℃가 될 때까지 실시한 후 상온에서 방냉하고 알루미늄과 PE가 라미네이팅된 멸균된 복합필름포장재(MULTIVAC, Wolfertxchwenden, Germany)에 포장기기(Leepack, Hanguk Electronic, Kyungi, Korea)를 이용하여 혼합가스(CO2 : N2 = 25 : 75,V/V)로 치환, 충전하여 포장하였다. 포장한 햄버커 패티 시료들은 8℃ 냉장고에 저장하면서 실험에 사용하였다.In this experimental example, in order to measure the effect of inhibiting fatty acid plaque of the samples obtained in Examples 1 to 8, hamburger patties containing a lot of fat were prepared as a model food, and the amount of malonaldehyde produced by fatty acid oxidation during storage was observed. As shown in Table 6, the main and sub-materials were prepared and mixed according to the mixing order. For sample mixing, each raw material was put into a mixer (M15 mixer, Falsf Co., Barcelona, Spain) in accordance with the mixing order, and at a speed of 60 rpm/min, the first mixing for 8 minutes, the second mixing for 2 minutes, and the third mixing was carried out for 1 minute and mixed, and the samples of Comparative Examples 1 to 3 and Examples 1 to 8 were added during the first mixing, respectively. 50 g of meat mixture was molded (5×5×

1 cm) was molded to a thickness of 10 mm and then heat treated. Heating is performed in a cooker preheated to 85℃ (NU-VU ES-3 cooker, Food Service System, Menominee, USA) until the center temperature reaches 75℃, then cooled at room temperature and sterilized composite laminated with aluminum and PE. The film packaging material (MULTIVAC, Wolfertxchwenden, Germany) was replaced with a mixed gas (CO2 : N2 = 25 : 75, V/V) using a packaging machine (Leepack, Hanguk Electronic, Kyungi, Korea), filled and packed. Packaged hambucker patty samples were used in the experiment while stored in a refrigerator at 8 ° C.

Fatty acid degradation during storage of hamburger patties was measured by the 2-thiobarbituric acid reactive substances (TBARS) method. That is, 50 μL of BHA (7.2% in ethanol) and 15 mL of distilled water were added to 5 g of the sample and then homogenized with a homogenizer (DIAX 900, Heidolph Co., Ltd., Germany). After adding 2 mL of TBA/TCA solution (20 mM TBA in 15% TCA) to 1 mL of the homogenate, it was heated in boiling water for 15 minutes. After cooling, centrifugation (2,000 rpm, 15 minutes) using a centrifuge (UNION 5KR, Hanil Science Industrial Co., Ltd, Incheon, Korea), and then taking 1 mL of the supernatant, measuring the absorbance at 532 nm, and measuring the calibration curve The concentration of malondialdehyde was calculated using . The result obtained at this time was expressed as μg malondialdehyde/g meat (wet weight basis).

Hamburger Patty Ingredients and Content Ratio material name content (%) order of addition ground beef 12.4 One ground pork 39.1 One ground pork back fat 16.35 One trash ice 6.1 One ginger 1.0 2 onion 8.5 2 egg white 4.3 2 tomato ketchup 1.6 2 soybean protein 4.1 2 bread crumbs 4.1 3 nutmeg powder 0.05 One salt 0.63 One cooking wine 0.41 One pepper powder 0.22 One phosphate 0.21 One sugar 0.83 One sample 0.05 One bout 100

Changes in fatty acid density during storage of hamburger patties sample name Changes in malonaldehyde (μg/g) during storage 0 days 10 days 20 days 30 days Comparative Example 1 ^1.91c 2.03 ^{CD 2.14d 2.31f} Comparative Example 2 ^{1.83b 1.84b 1.88b} 2.04 ^d Comparative Example 3 ^{1.91c 1.92c 1.96c 2.12e} Comparative Example 4 ^{1.84b 1.86b 1.89b} 2.06 ^d Example 1 ^{1.81b 1.84b 1.88b 1.95c} Example 2 1.78 ^ab 1.81 ^ab 1.83 ^{a 1.89b} Example 3 1.78 ^ab 1.79 ^a 1.82 ^{a 1.90b} Example 4 1.77 ^{ab 1.80ab 1.85ab 1.91b} Example 5 ^{1.80b 1.83b 1.90b 1.96c} Example 6 1.75 ^{ab 1.80ab 1.86ab 1.91b} Example 7 1.76 ^{ab 1.80ab} 1.87 ^{ab 1.94b} Example 8 1.77 ^ab 1.79 ^{a 1.86ab 1.94b} tocopherol 1.76 ^ab 1.79 ^a 1.83 ^{a 1.90b} BHA 1.73 ^a 1.76 ^a 1.81 ^a 1.83 ^a additive-free furniture ^{1.95c 2.12d 2.28e 2.51g}

During the storage of hamburger patties with samples obtained according to the sterilization method of the Molochia fermented extract, the other English superscript ( ^ag ) of the same storage days of the malonaldehyde production value during storage indicates statistical significance (p <0.05) .

As shown in Table 6, malonaldehyde production by fatty acid plaque during storage of hamburger patties to which samples treated by sterilization methods for Molochia-extracted fermented product were added were samples containing irradiated Molochia-extracted fermented product powder (Example 1 to 8) was confirmed to be very suppressed. Although it was less effective than BHA, a synthetic antioxidant used as a positive control, it showed almost similar antioxidant effect to the same amount of tocopherol. There was no difference between radiation types, and there was no significant difference at the dose of 10 kGy or more. The heat treatment group (Comparative Example 3) or the microfiltration treatment group (Comparative Example 4) showed a slightly lower fatty acid spoilage inhibitory effect than the radiation treatment group. On the other hand, in the case of the negative control, no additives, it was confirmed that rapid fatty acid spoilage occurred during storage. On the other hand, Molochia extract simple treatment group (Comparative Example 1) was also found to have a fatty acid spoilage inhibitory effect. From the above results, it was found that the fermented Molochia extract sterilized by irradiation of the present invention effectively suppresses fatty acid spoilage of meat products with high fat content such as hamburger patties.

[Experimental Example 5] Test of melanin production inhibitory effect in skin cells

In this experimental example, mouse melanocyte Melan A cells were used to observe the melanin production inhibitory effect of the samples obtained in Examples 1 to 8. That is, Melan A cells were dispensed in a 6-well plate at 2×10 ⁵ /well, and the culture solution was removed for 18 hours in a 37° C., 5% CO 2 incubator condition. At this time, arbutin as a positive control and samples were added for each concentration and cultured. After culturing for 48 hours, the medium was removed, and the bottom surface of 6 wells was evenly washed with PBS (phosphate buffered saline), treated with trypsin-EDTA, and the cells were collected in a micro tube. The recovered cells were added with 200 μL of 1 N NaOH, left in an 80 ° C water bath for 1 hour to dissolve intracellular melanin, and then left on ice for 1 hour. The absorbance at 405 nm was measured for the melanin content using ELISA. Each data is presented corrected for the amount of protein.

As shown in Table 7 below, the melanin synthesis inhibitory effect of the sample treated by the sterilization method for the fermented extract of Molochia of the present invention was irradiated at an absorbed dose of 10 kGy or more (Examples 2 to 4, Comparative Examples 6 to 8) was found to be better than Molokia simple extract (Comparative Example 1), fermentation treatment group (Comparative Example 2), heat treatment group (Comparative Example 2) and microfiltration treatment group (Comparative Example 3), and the difference between radiation types and radiation doses observed to be absent. In particular, it was found that the fermented Molochia extract of the present invention had an excellent melanin production inhibitory effect when irradiated with radiation for sterilization, as it exhibited a more excellent melanin production inhibitory effect than arbutin, which is a positive control group.

Inhibition of melanin production in skin cells Sample name (100 μg/ml) melanin concentration
(μg/10 ⁴ cells) Comparative Example 1 3.8± ^0.3c Comparative Example 2 2.2± ^0.8b Comparative Example 3 2.6± ^0.5b Comparative Example 4 2.3± ^0.6b Example 1 1.7± ^0.7ab Example 2 1.1±0.4 ^a Example 3 1.2±0.1 ^a Example 4 1.1±0.4 ^a Example 5 1.8± ^0.2ab Example 6 1.1±0.5 ^a Example 7 1.1±0.6 ^a Example 8 1.2±0.1 ^a Positive control group (Arbutin) 2.6± ^0.4b No additives (negative control group) 7.1±0.8 ^d

The other English superscript ( ^ad ) of the melanin production value generated by mouse melanocyte Melan A cells to which the sample obtained according to the sterilization method of the Molochia fermented extract was added indicates statistical significance (p<0.05).

[Statistical analysis]

Statistical analysis of the experimental results was performed using the 'Independent-Sample-T-Test method', an SPSS 13.0 statistical software, and expressed as the average value ± standard difference. In addition, multiple comparisons were indicated by the LSD method, P>0.05: the difference is not clear, P<0.05: means the difference is clear.

<Preparation Example 1> Preparation of powder formulation

Antioxidant preparations containing the fermented product obtained from radiation sterilized extract of Molochia obtained in Examples 1 to 8 were dried by a conventional freeze-drying method or a hot air drying method, and then mixed with an excipient.

<Preparation Example 2> Preparation of food and beverage (health functional food) composition

<2-1> Manufacture of beverages

The beverage is prepared by mixing the ingredients of the formulations presented below according to a method commonly practiced in the art. Honey 522 mg, thioctic acid amide 5 mg, nicotinic acid amide 10 mg, riboflavin sodium hydrochloride 3 mg, pyridoxine hydrochloride 2 mg, inositol 30 mg, ortic acid 50 mg, radiation-sterilized Molochia extract obtained in Examples 1 to 8 1 g fermented powder, 200 mL water.

<2-2> Preparation of powder

The powder is 2 g of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 according to a method commonly performed in the art, 2 g of a composite probiotic (1×10 ⁹ CFU/g), and 5 g of lactose. , 1 g of talc is mixed and filled in an airtight bag to prepare a powder agent.

<2-3> Manufacture of tablets

The tablet was prepared by mixing 250 mg of the fermented product powder obtained from radiation sterilized Molochia extract obtained in Examples 1 to 8, 150 mg of corn starch, 100 mg of lactose, and 20 mg of magnesium stearate according to a method commonly performed in the art. After that, tablets are prepared by tableting according to a conventional tablet manufacturing method.

<2-4> Preparation of capsules

Capsules are prepared by mixing 50 mg of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 according to a method commonly performed in the art, 3 mg of crystalline cellulose, 14.8 mg of lactose, and 0.2 mg of magnesium stearate. Capsules are prepared by mixing according to the method for preparing capsules and filling into gelatin capsules.

<2-5> Preparation of mixed powder formulation

The mixed powder is 5 g of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 according to a method commonly performed in the art, appropriate amounts of a vitamin mixture (vitamin A acetate 70 μg, vitamin E 1.0 mg, vitamin B1 0.13 mg, vitamin B2 0.15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 μg, vitamin C 10 mg, biotin 10 μg), nicotinamide 1.7 mg, folic acid 50 μg, calcium pantothenate 0.5 mg, appropriate amount of mineral mixture (sulfate 1 iron 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, monobasic potassium phosphate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg) to prepare a normal health functional food After mixing according to the method, granules are prepared, and can be used for preparing a health functional food composition according to a conventional method. The composition ratio of the above vitamin and mineral mixture was prepared by mixing ingredients suitable for a relatively health functional food in a preferred manufacturing example, but the mixing ratio may be arbitrarily modified.

<Manufacture Example 3> Manufacture of processed meat products

<3-1> Manufacture of hamburger patties

According to methods commonly practiced in the art, the hamburger patty was prepared by weight: 12.4% ground beef, 39.1% ground pork, 16.35% ground pork backfat, 6.1% ice water, 1.0% ginger, 8.5% onion. , Egg white 4.3% by weight, tomato ketchup 1.6% by weight, soybean protein 4.1% by weight, bread crumbs 4.1% by weight, nutmeg powder 0.05% by weight, salt 0.63% by weight, cooking wine 0.41% by weight, pepper powder 0.22% by weight, phosphate 0.21% by weight, It is prepared by mixing 0.83% by weight of sugar and 0.5% by weight of the fermented product powder extracted from Molochia obtained in Examples 1 to 8 and subjected to heat treatment.

<3-2> Manufacture of emulsified sausage

According to a method commonly practiced in the art, emulsified sausage contains 57.84% ground pork, 19.29% ground pork backfat, 19.29% ice water, 1.45% salt, 0.58% sugar, 0.58% phosphate. , 0.48% by weight of the composite spice, 0.5% by weight of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 are mixed, cut, emulsified, and then smoked and heated, or prepared by boiling in hot water at 80 ° C.

<3-3> Manufacture of block ham (including bacon)

According to a method commonly performed in the art, the meat block ham containing bacon contains 0.5% by weight of the fermented product powder extracted from radiation sterilized Molochia obtained in Examples 1 to 8 in 80.0% by weight of pork loin (or leg meat, etc.). After evenly injecting 20.0% by weight of the salting solution into the land mass using an injector, massage and dyeing for 6 hours or more in a vacuum tumbler fixed at 4 ° C., followed by heating by smoking. The salt solution is composed of 73.26% by weight of water, 1.45% by weight of salt, 0.58% by weight of sugar, 0.58% by weight of phosphate, 0.48% by weight of complex spice, and 0.5% by weight of fermented product powder extracted from Molochia.

<Preparation Example 4> Preparation of skin external preparation (ointment)

The ointment contains 5.00% by weight of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 according to a method commonly performed in the art, 10.00% by weight of caprine/capryltriglyceride, 10.00% by weight of liquid paraffin, and sorbate. Bitanesesquinoleate 6.00% by weight, octyldodeceth-25 9.00% by weight, cetylethylhexanoate 10.00% by weight, squalane 1.00% by weight, salicylic acid 1.00% by weight, glycerin 15.00% by weight, sorbitol 10.00% by weight, purified water remainder weight It is prepared by mixing %.

<Preparation Example 5> Preparation of feed composition

According to a method commonly performed in the art, the composition for feed is 45 g of corn, 30 g of soybean meal, 15 g of wheat, 4 g of beef tallow, 3 g of molasses, 2 g of calcium phosphate, 0.4 g of limestone, 0.2 g of salt, Examples 1 to 3 It is prepared by mixing 5 g of the radiation-sterilized Molochia extract fermented product powder obtained in 8.

<Production Example 6> Manufacturing of cosmetics

<6-1> Preparation of lotion

According to a method commonly performed in the field, the lotion was prepared by adding 0.05 g of polypyrrolidone, 0.1 g of polyol alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of perfume, 0.1 g of paraoxybenzoic acid methyl ester, and a small amount of 8 g of 95% ethanol. of antioxidants and a small amount of pigment are mixed and dissolved. Molochia extract fermented powder powder obtained in Examples 1 to 8 and 5 g of glycerin dissolved in 85.33 g of purified water were added to the mixed solution and stirred to obtain lotion having a skin whitening effect.

<6-2> Manufacture of Lotion

The lotion was prepared by using 0.10% by weight of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8 according to a method commonly performed in the art, 3.00% by weight of glycerin, 0.10% by weight of carbomer, 0.05% by weight of xanthan gum, 1,3-butylene glycol 3.00% by weight, polyglyceryl-3 methylglucose distearate 1.50% by weight, glyceryl stearate 0.50% by weight, cetylaryl alcohol 0.30% by weight, jojoba oil 3.00% by weight, liquid paraffin 2.00% by weight %, 3.00% by weight of squalane, 0.50% by weight of dimethicone, 0.20% by weight of tocopheryl acetate, 0.10% by weight of triethanolamine, preservatives, fragrances, trace amounts of pigments, and the remaining amount of purified water.

<6-3> Manufacture of emulsion

The emulsion was prepared according to a method commonly performed in the art, including 1.2 g of cetyl alcohol, 10 g of squalane, 2 g of petrolatum, 0.2 g of p-oxybenzoic acid ethyl ester, 1 g of glycerin monoester, and polyoxyethylene (20 moles added). 1 g of monooleate and 0.1 g of perfume were mixed and dissolved at 70 ° C., 0.05 g of the fermented product powder obtained from radiation sterilized molochia extract obtained in Examples 1 to 8, 5 g of dipropylene glycol, 2 g of polyethylene glycol 1500, 0.2 g of triethanolamine g, 76.2 g of purified water is heated to 75 ° C and dissolved by heating. After emulsifying by mixing the two, it is cooled to obtain a water/oil-based emulsion having a skin improving effect.

<6-4> Preparation of serum

According to a method commonly performed in the art, the essence is 95% ethyl alcohol 5 g, polyoxyethylene sorbitan monooleate 1.2 g, kitulose 0.3 g, sodium hyaluronate 0.2 g, vitamin E-acetate 0.2 g, licorice acetic acid By mixing 0.2 g of sodium, 0.1 g of p-hydroxybenzoic acid ester ester, 1 g of fermented molochia extract powder sterilized by radiation, and an appropriate amount of pigment, a skin-improving serum is obtained.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention may be embodied in other specific forms without changing its technical spirit or essential features. In this regard, the manufacturing examples described above should be understood as illustrative in all respects and not limiting. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later and equivalent concepts rather than the detailed description above are included in the scope of the present invention.

As described above, the fermented product extracted from radiation sterilized Molochia of the present invention has an excellent effect on antioxidant activity, and inhibits skin melanin pigmentation to effectively inhibit skin blackening, thereby providing natural antioxidants, health functional foods, and whitening functional cosmetics. It can be usefully used in development and production.

Claims (12)

Molochia extract ( Corchorus olitorius L.) Molokia extract obtained by fermenting the extract using glutinous rice cake mushroom ( Bovista plumbea ) or its mycelium and then irradiating the fermented product obtained from Molokhia extract with radiation. According to claim 1, Molokia extract characterized in that the glutinous rice cake mushroom or its mycelium is fermented using 0.1 to 20% (v / v). According to claim 1, wherein the extract is characterized in that the use of any one or more selected from the inhibition of lipid rancidity, prevention of oxidation of fatty foods, or inhibition of melanin production Molokhia extract. According to claim 1, wherein the radiation is characterized in that any one selected from the group consisting of gamma rays, electron rays and X-rays, Molokhia extract. According to claim 4, wherein the radiation is characterized in that irradiated with an absorbed dose of 5 to 20 kGy, Molokhia extract. A pharmaceutical composition for the prevention, improvement, or treatment of skin blackening comprising the Molochia extract of claim 1 as an active ingredient. A food composition for preventing or improving skin blackening comprising the Molochia extract of claim 1 as an active ingredient. A processed meat food composition comprising the Molochia extract of claim 1 as an active ingredient. A quasi-drug composition for the prevention, improvement, or treatment of skin blackening comprising the Molochia extract of claim 1 as an active ingredient. A drinking water additive comprising the Molochia extract of claim 1 as an active ingredient. A feed additive comprising the Molochia extract of claim 1 as an active ingredient. A cosmetic composition for preventing or improving skin darkening comprising the Molochia extract of claim 1 as an active ingredient.

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