KR20230080923A - Method for producing a ferment extract of the centella asiatica and composition for antiocidative comprising the ferment extract of the centella asiatica produced by the same - Google Patents

Abstract

The present application relates to a food composition for antioxidant comprising a fermented centella asiatica extract as an active ingredient, and has an excellent antioxidant effect.

Description

Manufacturing method of centella asiatica fermented extract and antioxidant food composition containing centella asiatica fermented extract prepared therefrom

The present invention relates to a method for preparing a fermented centella asiatica extract and a food composition using the same.

Basic triplet oxygen, a stable molecular state generally present in the air, is a free radical that is highly reactive due to environmental and biochemical factors such as enzymatic systems, reduction metabolism, chemicals, pollutants, and photochemical reactions in the body. It is converted into active oxygen and irreversibly destroys cellular components.

Active oxygen is an antioxidant enzyme such as superoxide dismutase (SOD), catalase, peroxidase, and glutathione, which are defense mechanisms in the body, and vitamin C (ascorbic acid), It can be minimized by the action of antioxidants such as vitamin E (tocopherol). However, when exposed to excessive active oxygen or when abnormality in biological defense occurs, the balance is broken, and active oxygen causes fatal oxygen toxicity to the living body.

Excessive active oxygen in the body reacts with lipids, proteins, sugars, DNA, etc., which are cellular constituents, and has a destructive effect, thereby preventing aging as well as cancer, brain diseases such as stroke and Parkinson's disease, heart disease, ischemia, arteriosclerosis, skin damage, and inflammation. , rheumatism, autoimmune diseases, etc. cause various diseases.

As such oxidative stress has been found to be an important cause of various diseases including aging, research to develop antioxidants that remove active oxygen in the body is being actively conducted.

BHA (butylated hydroxy anisol) and BHT (butylated hydroxy toluene), which are widely used as synthetic antioxidants, have been widely used because of their effectiveness, economy, and safety. It is known to cause serious toxic effects on the back, etc., and its usage is regulated by law.

In addition, research on natural antioxidants that are harmless to the human body and have excellent antioxidant power has been conducted for a long time, but natural antioxidants such as tocopherol are safe for the human body, but have the ability to block oxidative chain reactions alone and are expensive. there is

Therefore, there is an urgent need to develop new antioxidant foods that are safer, have excellent antioxidant effects, and are economically feasible.

Korean Registered Patent No. 10-1771702 (Announced on September 6, 2017)

The problem to be solved by the present application is to provide a method for producing a fermented centella asiatica extract with excellent antioxidant efficacy.

Another problem to be solved by the present application is to provide a food composition having an antioxidant effect and collagen absorption promoting effect by including the fermented Centella asiatica extract as an active ingredient.

One embodiment of the present invention comprises the steps of grinding centella asiatica powder at 90 to 120 ° C. for 10 minutes to 3 hours; Extracting by adding a mixed solvent of water and ethanol to the pulverized powder; And fermenting for 8 hours to 72 hours at 25 ~ 35 ° C. by adding a strain of the genus Leuconostoc to the Centella asiatica extract; It provides a method for producing a fermented centella asiatica extract, including.

The method according to an embodiment of the present invention includes sterilizing the fermented product at 115 ° C to 140 ° C after the fermentation; may further include.

In one embodiment of the present invention, the Leuconostoc stock ( Leuconostoc ) genus strain, Leuconostoc citrium ( Leuconostoc citreum ) Cares-1 (Accession No. KACC91931P) may be a strain.

One embodiment of the present invention provides a food composition for antioxidant comprising the fermented centella asiatica extract prepared by the above manufacturing method as an active ingredient.

In one embodiment of the present invention, the centella asiatica fermented extract may be included in 0.0001 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the composition may further include collagen having an average molecular weight of 1000 Da or less obtained from mammals or fish.

In one embodiment of the present invention, the collagen may be included in 0.0001 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the centella asiatica fermented extract may increase the absorption rate of collagen by 120% to 180% more than vitamin C.

The food composition of the present application has the advantage of excellent antioxidant effect because it contains a high content of asiaticoside.

In addition, the food composition of the present application has an effect of increasing the absorption rate of collagen by 120% to 180% compared to vitamin C.

1 shows the ABTS scavenging activity effect of a composition according to an embodiment of the present invention.
Figure 2 shows the results of measuring the polyphenol content of the composition according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail.

The following specific functional descriptions are only exemplified to explain embodiments according to the concept of the present invention, and embodiments according to the concept of the present invention can be implemented in various forms and are limited to the embodiments described herein. should not be interpreted as

Embodiments according to the concept of the present invention can apply various changes and can have various forms, so specific embodiments will be described in detail herein. However, this is not intended to limit the embodiments according to the concept of the present invention to a specific form disclosed, and should be understood to include all modifications, equivalents, or substitutes included in the spirit and scope of the present invention.

Terms used in this specification are only used to describe specific embodiments and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly dictates otherwise.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and are not interpreted in an ideal or excessively formal sense unless explicitly defined herein. .

Centella asiatica is a perennial creeping medicinal plant mainly found in South Asia, and in Korea, it grows in gregarious areas in Jeju Island and low-lying wetlands in the central and southern regions. The main stem extends to the side, and there are 2 degenerated scale-like leaves near the root node. Leaves grow densely at nodes, elongated, 2-5cm in diameter, glossy surface, with dull sawtooth on the edge, petiole 4-20cm long. Flowers are reddish-purple, 3-4 flowers hang in an umbel at the axilla, and are shaped like heads. The style is two-pronged, and the fruit is the oblong shape of the branch. In India, seeing a wounded tiger rolling around in a place where there are many centella asiatica plants for treatment, it is called tiger grass and has been used as a medicine for a long time. As it was discovered that the ingredient called madecassic acid in the leaves and stems of Centella asiatica heals inflammation and has the power to heal wounds such as tumors and ulcers, it is used as a raw material for ointments, toothpaste, or cosmetics.

An embodiment of the present invention comprises the steps of grinding centella asiatica powder at 90 to 120 ° C. for 10 minutes to 3 hours (S10); Extracting by adding a mixed solvent of water and ethanol to the pulverized powder (S20); and fermenting at 25 to 35° C. for 8 to 72 hours by adding a strain of the genus Leuconostoc to the Centella asiatica extract (S30); It provides a method for producing a fermented centella asiatica extract, including.

The method according to an embodiment of the present invention may further include the step of sterilizing the fermented product at 115 ° C to 140 ° C after the fermentation (S40).

In one embodiment of the present invention, the Leuconostoc stock ( Leuconostoc ) genus strain, Leuconostoc citrium ( Leuconostoc citreum ) Cares-1 (Accession No. KACC91931P) may be a strain.

The Leuconostoc citreum ( Leuconostoc citreum ) Cares-1 strain is characterized by having branched oligosaccharide production activity and antifungal activity, which is one of the recalcitrant prebiotics. In addition, the strain produces fructose, which is a by-product of the glycosyltransfer reaction, by producing glucansucase to enhance sweetness. The strain has a recalcitrant oligosaccharide-producing activity. The above "hardly degradable oligosaccharide" means a branched oligosaccharide that is not hydrolyzed and is a kind of prebiotics. The recalcitrant oligosaccharide may be selected from the group consisting of panose, isomaltosyl maltose, and isomaltotriosyl maltose, but is not necessarily limited thereto, and is widely known in the art. All of the hardly decomposable oligosaccharides may be included. The strain has antifungal activity. The fungus may be selected from the group consisting of Aspergillus ochraceus (KACC41810) and Cladosporium gossypiicola (KACC43795).

The grinding after roasting (S10) has an effect of increasing the antioxidant effect of the centella asiatica extract. If the grinding step after roasting is omitted, there is a significant difference in the efficacy of the final product.

In the step of extracting (S20), it is preferable to extract the centella asiatica using an aqueous ethanol solvent rather than a solvent of water alone, and then inoculate the extract with bacteria and ferment. Therefore, fermentation by inoculating bacteria into the alcohol extract using an aqueous ethanol solution solvent is much more effective in antioxidant than fermentation by inoculating bacteria into the hot water extract of Centella asiatica powder.

In addition, the fermentation step (S30) after the extraction step (S20); has the effect of improving the antioxidant efficacy of the centella asiatica fermented extract. If the order of the S20 and S30 steps is changed, the antioxidant efficacy is not good.

The fermentation may proceed at 25 to 35 ° C, specifically at 28 ° C to 32 ° C, more specifically at 30 ° C, for 8 to 72 hours, specifically for 8 to 48 hours, and more specifically for 8 to 24 hours. there is. After 24 hours, there is no significant difference in the efficacy of the fermented product. The fermentation may be selectively carried out under rotation at 100 to 300 rpm, and specifically under conditions of 120 rpm to 180 rpm.

If the fermentation temperature is less than the above range, it is not economical because the extraction yield of useful substances is low and equipment for lowering the temperature is required, and if the stirring temperature exceeds the above range, useful substances are destroyed or unuseful substances are high It is undesirable because it can be extracted together and a fermented product with reduced functionality can be produced. For example, when the fermentation temperature is in the temperature range of 25 ° C to 35 ° C, the plant cell tissue is softened to facilitate the elution of components such as phenols, which are effective substances having cosmetic activity, so that a fermented product with excellent efficacy can be obtained It works. In addition, when the culture is performed for less than 8 hours, it is difficult for the contained bioactive components to be completely eluted due to insufficient time, and when the fermentation time exceeds 72 hours, the contained physiologically active components do not increase to a significant level. Rather, due to excessive culture, a fermented product with reduced antioxidant and skin whitening activity may be produced.

After fermentation, it may be sterilized at 115 ° C to 140 ° C, specifically at 121 ° C so that fermentation does not proceed any further. Thereafter, impurities may be removed using a conventional filtration method or device, and for example, a fermented product from which impurities are removed may be obtained by filtering using a centrifugal separation method or a micro filter. For example, a usable microfilter may have a pore size of 0.3 μm to 5 μm. Specifically, it can be filtered using a filter having pores of 3㎛ size. As another example, centrifugation may be performed at 1000 rpm to 2000 rpm for 5 to 20 minutes. Specifically, by centrifuging at 1500 rpm for 10 minutes, the precipitate is discarded, and only the supernatant is collected and used as it is.

One embodiment of the present application provides a food composition for antioxidant comprising the fermented centella asiatica extract obtained by removing the impurities as an active ingredient.

The centella asiatica fermented extract may be included in an amount of 0.0001 to 10% by weight based on the total weight of the composition.

At this time, if the content of the fermented material is less than 0.0001% by weight, the antioxidant effect of the present application cannot be obtained, and if it exceeds 10% by weight, the effect may not be proportional to the increase in the content, which may be inefficient, and the stability of the formulation is secured. There is a problem that doesn't work.

In one embodiment of the present invention, the composition may further include a collagen peptide having a low average molecular weight obtained from mammals or fish, having a molecular weight of 1 to 1000 Da, 1 to 800 Da, or 1 to 500 Da. It may be a collagen peptide.

"Collagen" is a protein that occupies the largest part quantitatively and functionally in the human body, and is a major component of the extracellular matrix that forms the skeleton of all cells in the human body, more specifically, connective tissue. it means. Human tissue containing a large amount of collagen includes skin, bone tissue such as bone and cartilage, tendon, ligament, and the like. Aging of the tissues results in human aging diseases such as skin wrinkles and sagging and osteoporosis, which is known to be caused by a decrease in the amount and function of collagen in the body due to a decrease in collagen synthesis in tissues. In one aspect of the present invention, the collagen hydrolyzate includes those obtained from mammals or fish. In another aspect of the present invention, the collagen hydrolyzate includes one obtained from mammalian skin or fish young (scales) or fish skin.

In one embodiment of the present invention, the collagen may be included in 0.0001 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the centella asiatica fermented extract may increase the absorption rate of collagen by 120% to 180% more than vitamin C. Specifically, the absorption rate may be increased by 140% to 160% compared to vitamin C.

In the present application, the composition containing the above-described fermented product may also be provided as food for the purpose of efficacy as described above, for example, health supplement food, functional food, food additive, and the like.

The term "food" as used in this application means a natural product or processed product containing one or more nutrients, and specifically means a product that can be eaten directly through a certain degree of processing process. As the meaning, it is intended to include all foods, food additives, functional foods and beverages.

As used herein, the term "functional food" refers to a food group or processed food that has added added value so that the function of the food acts for a specific purpose and is expressed using physical, biochemical, and bioengineering techniques.

The functional food may further include a food-acceptable food supplement additive, and may further include appropriate carriers, excipients, and diluents commonly used in the manufacture of functional foods.

Foods include, for example, various foods, beverages, chewing gum, tea, vitamin complexes, and functional foods. In addition, food in this application includes special nutritional food (eg, formula milk, infant food, baby food, etc.), processed meat product, fish meat product, tofu, jelly, noodles (eg, ramen, noodles, etc.), health supplement food, seasoning food (eg, soy sauce, soybean paste, gochujang, mixed paste, etc.), sauces, confectionery (eg, snacks), dairy products (eg, fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various types of kimchi, pickles, etc.), Beverages (eg, fruit, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.), but are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.

Functional beverages are manufactured as functional beverages, and functional beverages are biological defenses of a beverage group or beverage composition that adds value to the beverage to act and express the function of the beverage for a specific purpose by using physical, biochemical, and bioengineering methods. It refers to a beverage designed and processed to sufficiently express body control functions related to rhythm control, disease prevention and recovery, etc. to the living body, and in particular, refers to a beverage for antioxidation, improvement of skin elasticity, and promotion of collagen synthesis.

The health beverage of the present invention may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrins and cyclodextrins; or sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the health beverage of the present invention.

Food compositions include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, It may contain a protective colloidal thickener, a pH adjuster, a stabilizer, a preservative, glycerin, alcohol, a carbonating agent used in carbonated beverages, and the like. These components may be used independently or in combination.

In addition, the health food of the present invention may contain fruit flesh for the production of natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health food of the present invention.

Hereinafter, the present application will be described in more detail through manufacturing examples, examples, and experimental examples, but the following examples are only intended to aid understanding of the present application and do not limit the scope of the present application.

<Preparation Example 1> Centella asiatica extract sample preparation

Centella asiatica grown in Korea was purchased, washed, dried at 50 ° C for 24 hours, roasted at 100 ° C for 30 minutes, and then pulverized. Ethanol was added to 10% in water 20 times the weight of the crushed Centella asiatica, and then extracted at 60° C. for 24 hours.

<Example 1> Preparation of fermented product of centella asiatica

As the strain, Leuconostoc citreum Cares-1 (Accession No. KACC91931P) strain was used. It was cultured at 30 ℃ with MRS Broth as a growth medium.

2% sugar, 4% maltose, and 1% strain were added to the extract of Preparation Example 1, shake-cultured at 30°C for 8 hours, and then autoclaved at 121°C for 10 minutes to prevent further fermentation. After the completion of the culture, it was filtered, and the precipitate was discarded, and only the supernatant was collected to prepare a fermented product.

<Example 2> Preparation of fermented product of centella asiatica

A fermented product was prepared in the same manner as in Example 1 except for culturing for 24 hours.

<Comparative Example 1> Centella asiatica Extract Preparation

An extract was prepared in the same manner as in Preparation Example 1 except that only water was extracted.

<Comparative Example 2> Preparation of centella asiatica extract

The extract of Preparation Example 1 was tested as Comparative Example 2.

<Experimental Example 1> Measurement of ABTS radical scavenging activity of fermented centella asiatica

ABTS assay was measured by modifying the method of Re et al. (1999) with the fermentation products of Comparative Example 2 and Examples 1 and 2. That is, after mixing 5 mL of 7 mM ABTS and 88 μL of 140 mM potassium persulfate, blocking light at room temperature for 16 hours to form ABTS cations. Then, this solution was diluted with PBS to obtain an absorbance value of 1.5 at 414 nm. 190 μl of the prepared diluted solution and 10 μl of the sample were mixed, reacted at room temperature for 6 minutes, and then absorbance was measured at 734 nm. As a solvent control group, the radical scavenging ability of the control group was expressed as a percentage.

Radical scavenging ability of Centella asiatica fermentation broth was shown in Table 1 and Figure 1 by confirming the concentration-dependent increase in activity. Centella asiatica fermented extracts, Example 1 and Example 2, compared to the ethanol and water extracts of Comparative Example 2, had a significant difference in ABTS scavenging activity, and showed similar effects to the control vitamin C.

ABTS scavenging ability control group 47.93 Comparative Example 2 22.75 Example 1 45.97 Example 2 51.77

<Experimental Example 2> Measurement of polyphenols of fermented centella asiatica

The total polyphenol content was analyzed by an authorized institution and proceeded with the total polyphenol test method in Article 4. 3-59 of 「Standards and Specifications for Health Functional Food」, and the fermented products of Comparative Example 2 and Examples 1 and 2 are shown in Table 2 below. and shown in Figure 2. Centella asiatica fermented extracts of Examples 1 and 2 were significantly different in polyphenol content compared to the ethanol and water extracts of Comparative Example 2.

Polyphenol content (ug/ml GAE) Comparative Example 2 100.91 Example 1 147.66 Example 2 157.46

<Experimental Example 3> Asticoside measurement of fermented centella asiatica

The contents of Asiaticoside, Madecassoside, Asiatic acid, and Madecassic acid of Comparative Examples 1 and 2 and Examples 1 and 2 were measured and shown in Table 3 below. showed up Compared to Comparative Examples, it was confirmed that the content of madecassoside and asiaticoside in Examples 1 and 2 was significantly higher, and madecassic acid was not detected, and the content of asiatic acid was also significantly higher.

Madecassoside
(μg/ml) Asiaticoside (μg/ml) Madecassic acid (μg/ml) Asiatic acid (μg/ml) Comparative Example 1 28±0.71 8.49±0.80 - 3.95±1.43 Comparative Example 2 163.82±4.16 112.31±2.86 - 4.29±0.95 Example 1 192.24±2.52 130.94±0.25 - 6.12±0.88 Example 2 193.50±12.37 129.40±14.62 - 5.49±0.86

Hereinafter, an example of preparing a food composition according to the present invention is described, but it can be applied to various formulations in addition to the following examples, which is only intended to be specifically described, not intended to limit the present invention.

<Production Example 2> Production of food

Foods containing the fermented centella asiatica extract of the present invention were prepared as follows.

1. Manufacture of flour food

1 part by weight of the Centella asiatica fermented extract was added to wheat flour, and bread, cakes, cookies, crackers, and noodles were prepared using the mixture to prepare health foods.

2. Manufacture of dairy products

1 part by weight of the Centella asiatica fermented extract was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

3. Manufacture of wire

Brown rice, barley, glutinous rice, and adlay were alphanized by a known method, dried, roasted, and then prepared into a powder having a particle size of 60 mesh using a grinder. Black beans, black sesame seeds, and perilla seeds were also steamed and dried by a known method, roasted, and then prepared into powder with a particle size of 60 mesh using a grinder. The fermented extract was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray or hot air dryer was pulverized with a grinder to a particle size of 60 mesh to obtain a dry powder.

The dry powders of the grains, seeds, and centella asiatica fermented extracts prepared above were blended in the following ratio with respect to 100 parts by weight of the mixed powder.

Cereals (brown rice 30 parts by weight, adlay 15 parts by weight, barley 20 parts by weight),

Seeds (7 parts by weight of perilla seeds, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds),

Ferment extract (1 part by weight),

Reishi (0.5 parts by weight),

Rehmannia glutinosa (0.5 parts by weight)

<Production Example 3> Production of beverage

1. Manufacture of health drinks

Centella asiatica fermented extract 5g

Citric Acid 1000mg

Oligosaccharide 100g

plum concentrate 2g

1 g taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and collected in a sterilized 2L-container, sealed and sterilized, and refrigerated. It was used for preparing the health drink composition of the present invention.

Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.

2. Manufacture of fruit juice

Centella asiatica fermented extract of the present invention 4.5g, passion fruit concentrate (Vietnam) 4g, mango concentrate (Israel) 1g, pineapple concentrate (Philippines) 1g, orange concentrate (US) 1g, jujube extract concentrate (domestic) 0.7g, 2 g of fructooligosaccharide, 1 g of low molecular weight fish collagen powder, and the remaining amount of purified water were added to prepare a total of 100 mL of vegetable juice for health promotion.

In the above, the present application has been described in detail with preferred embodiments and experimental examples, but the present application is not limited by the above embodiments and experimental examples, and those skilled in the art within the technical spirit and scope of the present application Various modifications and changes are possible by the person.

Those skilled in the art to which the present invention pertains will be able to make various applications and modifications within the scope of the present invention based on the above information.

Having described specific parts of the present invention in detail above, it is clear that these specific techniques are merely preferred embodiments for those skilled in the art, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (7)

Grinding centella asiatica powder at 90 ° C. to 120 ° C. for 10 minutes to 3 hours;
Extracting by adding a mixed solvent of water and ethanol to the pulverized powder; and
Adding a strain of the genus Leuconostoc to the centella asiatica extract and fermenting it at 25 ° C to 35 ° C for 8 to 72 hours; Method for producing a fermented centella asiatica extract, including.
The method of claim 1,
The Leuconostoc genus strain,
Leuconostoc citreum ( Leuconostoc citreum ) Cares-1 (accession number KACC91931P) method for producing a fermented centella asiatica extract.
The method of claim 1,
Sterilizing the fermented product at 115 ° C to 140 ° C after the fermentation step; Method for producing a fermented centella asiatica extract further comprising a.
A food composition for antioxidant comprising the fermented centella asiatica extract prepared by the method of any one of claims 1 to 3 as an active ingredient.
The method of claim 4,
The centella asiatica fermented extract is a food composition for antioxidant, characterized in that contained in 0.0001 to 10% by weight based on the total weight of the composition.
The method of claim 4,
The composition is
An antioxidant food composition further comprising collagen having an average molecular weight of 1000 Da or less obtained from mammals or fish.
The method of claim 4,
Antioxidant food composition, characterized in that the centella asiatica fermented extract increases the absorption rate of collagen by 120 to 180% more than vitamin C.

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