KR102239257B1 - Method for producing fermented extract of Artemisia annua with improved contents of physiological active ingredients and skin improvement effect and cosmetic composition comprising the same - Google Patents

Abstract

The present invention relates to a method for manufacturing a fermented product of Artemisia annua with improved content of physiologically active substances and improved skin improvement efficacy. According to the method for manufacturing the fermented product of Artemisia annua according to an aspect, it is possible to manufacture the fermented product of Artemisia annua with increased content of artemisinin and quercetin, which are physiologically active substances. Therefore, the fermented product of Artemisia annua manufactured by the method has an effect of being able to be usefully used for preventing or alleviating various skin-related conditions.

Description

TECHNICAL FIELD [Method for producing fermented extract of Artemisia annua with improved contents of physiological active ingredients and skin improvement effect and cosmetic composition] comprising the same}

It relates to a method for producing a fermented product of Mugwort with improved content of a physiologically active substance and skin improvement effect, and a cosmetic composition comprising the same.

Artemisia annua L. is one of 300 species of sagebrush and grows on roadsides, glades, and riversides. It is about 1m in height, and unlike ordinary mugwort, there is no hair on the whole grass and has a characteristic smell. The stem is green, and the branches are divided and the leaves are relatively thin. In herbal medicine, it is called Cheongho, and its actions such as antipyretic, anti-inflammatory, analgesic, antimalarial, immune regulation, antitumor, antibacterial, antiviral, and anticancer are being studied. Mugwort extract, as a cosmetic material, has been studied on antioxidant, antibacterial, whitening, and inhibitory effects on skin hypersensitivity reactions.

Artemisinin, Artemisinic acid, Arteannuin, Arteannuin B, Scopoletin, Coumarin, and Eupatin are known as major ingredients in Mugwort. In addition, components such as Chlorogenic acid, Rutin, Quercetin, Luteolin and Kaempferol have been isolated and identified. In particular, Dr. Toyouyou of China was awarded the Nobel Prize in Medicine in 2015 for discovering a component of artemisinin that exhibits high efficacy against the resistance of malaria pathogens, and artemisinin and its derivatives are used to treat malaria and parasitic infections. Was used.

On the other hand, pottery is a traditional food storage tool, made by kneading and baking soil, sand, and white clay. The characteristics of Onggi include breathability, storage, and fermentability. Onggi was used for storing food such as rice and water, fermenting liquor, kimchi, and soy sauce, and was also used for various purposes such as chimneys, sewage pipes, and fish tanks.

Fermentation generally refers to the action of generating useful metabolites using microbial metabolism, and is a complex chemical reaction and a natural process at the same time.

Solid fermentation refers to fermentation by culturing microorganisms on a solid substrate. A method of fermenting microorganisms directly on the surface of a solid substrate having a small amount of moisture. If the moisture content is appropriate, the growth of bacteria can be inhibited and useful fungi can be grown preferentially. Aerobic microorganisms are suitable and are mainly used to produce brewed foods or hydrolytic enzymes.

Liquid fermentation is fermentation using a liquid culture method that is cultured in a liquid medium. It is classified into batch culture, continuous culture, and fed-batch culture methods according to the method of supplying the substrate, and it is advantageous because the fermentation bacteria are not exposed to the air, so there is little possibility of contamination of various bacteria, and it is possible to uniformly disperse microorganisms.

Low-temperature and high-pressure extraction is a single component with high purity by using the principle that more components are eluted out of the cell by destroying the cell membrane under ultra-high pressure, and more components are eluted out of the cell. And an extract with less impurities is obtained. In particular, the high energy and pressure of the ultra-high pressure applied to natural products makes it possible to obtain useful physiologically active substances that cannot be obtained by ordinary hot water extraction, thereby re-evaluating the natural substances contained in plants.

Meanwhile, as described above, a method of fermenting plants in general has been disclosed, but a fermentation method capable of maintaining higher physiologically active ingredients contained in wormwood is not disclosed.

Korean Patent Publication 10-2019-0091161

One aspect is to provide a method of manufacturing a fermented wormwood (Artemisia annua).

Another aspect is to provide a cosmetic composition comprising the fermented wormwood prepared by the above method.

One aspect provides a method for producing a fermented wormwood wormwood comprising the step of solid fermenting artemisia annua.

The wormwood wormwood may be whole or part of the plant. The portion may be an above-ground portion or an underground portion. The above-ground part may be leaves, flowers, fruits, or a combination thereof. The basement part may be an underground stem, a root, or a combination thereof. The wormwood wormwood may be purchased commercially, or may be used by collecting or cultivating it in nature. In addition, the wormwood wormwood may be harvested in September to October. Mugwort harvested in September to October may have the highest content of Artemisinin and Quercetin, which are active ingredients contained in Mugwort.

The solid fermentation may be fermentation in an onggi. Onggi has fine pores on its surface, so it is breathable, and loess, the main component of the onggi material, inhibits the growth of other decaying and pathogenic microorganisms due to the antibacterial and far-infrared emission action, so when fermented in the onggi, fermentation can proceed effectively. . The onggi may be a chilyang onggi. The Chillyang Onggi refers to an onggi produced in Gangjin-gun, Jeollanam-do, or Chillyang-myeon, Gangjin-gun, Jeollanam-do, or an onggi produced by a nationally designated craftsman, onggijang (for example, a national intangible cultural property) or his successor as Chiliang Onggi I can.

The solid fermentation may include the step of inoculating the strain on the wormwood. The strain may be one or more selected from the group consisting of a strain belonging to the genus Lactococcus, a strain belonging to the genus Saccharomyces, and a strain belonging to the genus Lactobacillus.

Strains belonging to the genus Lactococcus include Lactococcus lactis, Lactococcusgarvieae, Lactococcus confusus, Lactococcus chungangensis, Lactococcus taiwanensis, and Lactococcus taiwan ensis. Lactococcus fujiensis and Lactococcus raffinolactis include, but are not limited to.

Strains belonging to the genus Saccharomyces include Saccharomyces cerevisiae, Saccharomyces rouxii, Saccharomyces obiformis, and Saccharomyces steineri (Saccharomyces steineri). ), Saccharomyces kleveri, Saccharomyces coreanus, and Saccharomyces sake.

Strains belonging to the Lactobacillus genus include Lactobacillus sakei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus acidophilus Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus lactis, Lactobacillus reutri, Lactobacillus brevis, Lactobacillus brevis, Lactobacillus brevis, Lactobacillus brevis Lactobacillus johnsonii, Lactobacillus buchneri, Lactobacillus fermentum, and Lactobacillus leuteri.

The strain may be inoculated at 0.1 to 10.0% (v/v). Specifically, the strain may be inoculated at 0.1 to 8.0% (v/v), 0.1 to 6.0% (v/v), 0.1 to 4.0% (v/v) or 0.1 to 2.0% (v/v) have.

The solid fermentation may include the step of inoculating the strain in the wormwood and fermenting it at a warmth of 20 to 45 °C. Specifically, it may be fermented at 18 to 40°C, 20 to 40°C, 20 to 35°C, 20 to 30°C, 20 to 25°C, 25 to 35°C, or 25 to 30°C.

In addition, the solid fermentation may further include fermenting in cold air of 0 to 19°C. Specifically, fermented at 0 to 22 ℃, 1 to 22 ℃, 6 to 22 ℃, 0 to 15 ℃, 1 to 15 ℃, 0 to 10 ℃, 5 to 19 ℃, 5 to 15 ℃ or 10 to 15 ℃ Can be. Accordingly, the solid fermentation may be performed by inoculating the strain in Mugwort, fermenting at 20 to 45°C, and then further fermenting at 0 to 19°C.

The warm fermentation and cold fermentation may be fermented for 30 to 70 days, 30 to 60 days, 40 to 70 days, or 40 to 60 days, respectively. The warm solid fermentation and the cold solid fermentation may be the same or different. Accordingly, it may be fermented at 20 to 45° C. for 40 to 60 days, and then fermented at 0 to 19° C. for 40 to 60 days. Specifically, the fermenting step is fermented for 40 to 55 days, 40 to 50 days, 45 to 60 days, 45 to 55 days, 45 to 50 days, 50 to 60 days, 50 to 55 days, or 55 to 60 days, respectively. Can be.

Another aspect provides a method for producing a fermented wormwood fermented wormwood further comprising the step of liquid fermentation.

The liquid fermentation may include the step of adding the solid fermented wormwood to the strain culture broth in a weight ratio of 1:10 and fermenting by adding a carbon source, a nitrogen source, an amino acid or a vitamin. Specifically, the solid fermented wormwood may be added to the strain culture medium in a weight ratio of 1:8, 1:6, 1:4 or 1:2. The carbon source may be glucose or xylose, in addition to sugars and carbohydrates such as sucrose, lactose, fructose, maltose, starch, and cellulose, oils and fats such as soybean oil, sunflower oil, castor oil, coconut oil, and the like, Fatty acids such as palmitic acid, stearic acid, linoleic acid, alcohols such as glycerol, ethanol, and organic acids such as acetic acid and ascorbic acid may be included. These carbon sources may be used alone or in combination. Examples of the nitrogen source include inorganic nitrogen sources such as ammonia, ammonium sulfate, ammonium chloride, ammonium acetate, ammonium phosphate, ammonium carbonate, and ammonium nitrate; Amino acids such as glutamic acid, methionine, glutamine, and organic nitrogen sources such as peptone, NZ-amine, meat extract, yeast extract, malt extract, corn steep liquor, casein hydrolyzate, fish or its degradation products, skim soybean cake or its degradation products, etc. I can. These nitrogen sources may be used alone or in combination. Essential growth substances such as amino acids and vitamins may be added in addition to the carbon source or nitrogen source. In one aspect, glucose or ascorbic acid may be added as a carbon source, and both glucose and ascorbic acid may be added. The capacity of the carbon source, nitrogen source, amino acid or vitamin may be added in a general amount known in the art, and the step of fermenting by adding a carbon source, nitrogen source, amino acid or vitamin may be fermentation by a general method known in the art. have. In one embodiment, the glucose may be added in an amount of 5 to 15 g per 1 L of the strain culture solution to which the wormwood is added, and the ascorbic acid is added in an amount of 0.1 to 2.0 g per 1 L of the strain culture solution to which the wormwood is added. It can be. In addition, the liquid culture may be fermented at 30 to 40° C. for 1 to 3 days by maintaining at 50 to 150 rpm in a shaking incubator.

In addition, the liquid fermentation may further include a step of fermenting by repeating the strain inoculation 1 to 5 times. The strain may be inoculated at 0.1 to 10.0% (v/v). Specifically, the strain may be inoculated at 0.1 to 8.0% (v/v), 0.1 to 6.0% (v/v), 0.1 to 4.0% (v/v) or 0.1 to 2.0% (v/v) have. In addition, the step of inoculating and fermenting the strain may be fermenting for 1 to 5 hours. Specifically, the step of inoculating and fermenting the strain may be fermenting for 1 to 4 hours, 1 to 3 hours, 2 to 5 hours, 2 to 4 hours, and 3 to 5 hours. In one embodiment, the strain inoculation may be repeated 3 times to ferment for 3 hours. When fermentation is repeated several times as described above, the particle size of the fermented product is reduced to a fine size, so that the skin absorption power of the effective substance may be increased.

Another aspect provides a method for producing a fermented wormwood fermented wormwood further comprising the step of extracting the fermented wormwood wormwood.

The extraction may be extracted by a hydrophilic solvent, for example, alcohol, water, or a combination thereof. The alcohol may be a compound having one or more -OH groups of C1 to C10. The alcohol may be a C1 to C6 alcohol or a C3 to C6 polyhydric alcohol. The alcohol may be methanol, ethanol, n-propanol, isopropanol, n-butanol, sec-butanol, isobutanol, tert-butanol, n-pentanol, n-hexanol, or a mixture thereof. The solvent may be, for example, a mixture of water and alcohol, that is, an aqueous alcohol solution. The alcohol concentration of the aqueous alcohol solution is 1 to 100% (w/w), for example, 1 to 99.5% (w/w), 10 to 100% (w/w), 20 to 100% (w/w), 30 to 100% (w/w), 40 to 100% (w/w), 50 to 100% (w/w), 60 to 100% (w/w), 70 to 100% (w/w), 75 to 100% (w/w), 60 to 90% (w/w), 60 to 80% (w/w), 65 to 75% (w/w), or 70% (w/w) have. The aqueous alcohol solution may be an aqueous methanol, ethanol, or butanol solution. In the composition according to an embodiment, the extract may be an ethanol extract.

The extract may be extracted by a conventional method in the art, such as warm extraction, pressurized extraction, ultrasonic extraction, hot water extraction, reflux cooling extraction, subcritical extraction, or supercritical extraction.

In one embodiment, the extraction may be a low temperature and high pressure extract.

In detail, the step of extracting may be to extract the solid fermented or liquid fermented wormwood fermented product at 15 to 30°C. Specifically, the step of extracting may be extraction at 15 to 25 °C, 15 to 20 °C, 20 to 30 °C, or 20 to 25 °C. In addition, the step of extracting may be to extract the solid fermented or liquid fermented wormwood fermented product at a pressure of 1 to 5 Mpa. Specifically, the extracting step is 1 to 4 Mpa, 1 to 3 Mpa, 1 to 2 Mpa, 2 to 5 Mpa, 2 to 4 Mpa, 2 to 3 Mpa, 3 to 5 Mpa, 3 to 4 Mpa or 4 to It may be extracted with a pressure of 5 Mpa. In addition, the extracting may be extracted for 1 to 5 hours. Specifically, the extracting may be performed for 1 to 4 hours, 1 to 3 hours, 2 to 5 hours, 2 to 4 hours, or 3 to 5 hours. In one embodiment, the step of extracting may be extracted for 3 hours under a pressure of 20 to 25 ℃ and 3 Mpa. The low-temperature and high-pressure extraction method as described above has a high extraction yield and takes less time to extract, compared to a low-temperature extraction method that takes a long time for extraction and has a low extraction yield, or a high-pressure extraction method in which the loss of active ingredients occurs due to heat due to high pressure. There is an effect that can reduce the loss of active ingredients.

In one embodiment, the fermented wormwood prepared by the above manufacturing method may be usefully used in conditions related to skin.

Examples of conditions related to the skin include skin aging, wounds, dermatitis, atopic dermatitis, pruritus, eczematous skin disease, dry eczema, erythema, urticaria, psoriasis, weak rash, papule swelling disease, insect and parasite mediated diseases, superficial skin. Mycosis, bacterial infections, viral diseases, adult diseases, autoimmune blistering diseases, connective tissue diseases, dyslexia, scleroderma pigmentosa, acne, and the like. Accordingly, the composition can be usefully used for skin improvement, such as skin moisturizing or whitening effect, skin aging inhibition, skin barrier strengthening, skin texture improvement, skin wound improvement, and skin inflammation suppression by having the above effects.

Another aspect provides a cosmetic composition comprising a fermented wormwood prepared by the manufacturing method.

The fermented wormwood is one containing quercetin in a concentration of 25 mg to 50 mg, 28 mg to 50 mg, 28 mg to 48 mg, 28 mg to 42 mg, 34 mg to 46 mg per 1 g of fermented wormwood I can. In addition, the fermented wormwood is 6 mg to 15 mg, 8 mg to 15 mg, 6 mg to 12 mg, 8 mg to 12 mg, 9 mg to 12 mg, or 10 mg of tannic acid per 1 g of fermented wormwood It may be included in a concentration of 14 mg.

The fermented wormwood is 0.0001% to 99.0% by weight based on the total weight of the composition, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% by weight To 20% by weight, 0.01% to 10% by weight, 0.01% to 5% by weight, 0.05% to 60% by weight, 0.05% to 40% by weight, 0.05% to 30% by weight, 0.05% to 20% by weight Wt%, 0.05 wt% to 10 wt%, 0.05 wt% to 5 wt%, 0.1 wt% to 60 wt%, 0.1 wt% to 40 wt%, 0.1 wt% to 30 wt%, 0.1 wt% to 20 wt% , 0.1% to 10% by weight, or 0.1% to 5% by weight may be included.

The cosmetic composition is composed of softening lotion, nourishing lotion, nourishing cream, moisture cream, massage cream, essence, ampoule, gel, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, powder, gel, lotion, and ointment. It may have a formulation selected from the group. The cosmetic composition may additionally include one or more cosmetically acceptable carriers formulated in general skin cosmetics, and as common ingredients, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelates Agents, pigments, preservatives, fragrances, and the like may be appropriately blended, but are not limited thereto.

The cosmetic composition may include other additives such as excipients, carriers, etc. in addition to the fermented wormwood, and it may be possible to apply and mix ordinary ingredients to be blended in general skin cosmetics as needed.

In addition, the cosmetic composition includes medicinal ingredients such as higher fatty acids and vitamins, and sunscreens, antioxidants (butylhydroxyanisole, propyl gallic acid, lysorbic acid, tocopheryl acetate, butylated hydroxytoluene, etc.), if necessary. ), preservatives (methylparaben, butylparaben, propylparaben, phenoxyethanol, imidazolidinylurea, chlorphenesin, etc.), colorants, pH adjusters (triethanolamine, citric acid, citric acid, sodium citrate, malic acid, sodium malate) , Pmalic acid, sodium fmarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.), moisturizing agents (glycerin, sorbitol, propylene glycol, butylene glycol, hexylene glycol, diglycerin, beta Ingredients such as phosphorus, glyceres-26, methylgluces-20, etc.), and lubricants may be further added.

In addition, the cosmetic composition further includes a substance capable of auxiliaryly providing essential nutrients to the skin, and preferably, a natural fragrance, a cosmetic fragrance, or an herbal medicine may be included, but may contain an adjuvant, which is not limited thereto.

The terms and methods described for the above invention are applied equally to each of the inventions.

According to the method for producing a fermented wormwood according to an aspect, it is possible to prepare a fermented wormwood with an increased content of artemisinin and quercetin, which are physiologically active substances, so that the fermented wormwood prepared by the above method has various skin-related conditions. There is an effect that can be usefully used in the prevention or improvement of.

1 is a graph showing NO (Nitric oxide) radical inhibitory activity of fermented wormwood fermentation in order to confirm the anti-inflammatory effect of fermented wormwood wormwood.
2 is a graph measuring the moisture content over time using a moisture meter in order to check the moisturizing effect of fermented wormwood fermented artemisiae.

It will be described in more detail through the following examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

Example 1. Preparation of fermented wormwood wormwood 1

For the preparation of fermented wormwood fermented wormwood, fermented wormwood fermented wormwood was prepared by fermentation in an Onggi for 100 days, followed by triple liquid fermentation, followed by low-temperature and high-pressure extraction. The specific method is as follows.

First, solid fermentation was performed using Onggi. Specifically, artemisinin and quercetin, which are the active ingredients contained in wormwood, were washed and dried naturally until the water disappears from the surface after washing the artemisinin and quercetin, which have the highest content of the artemisinin and quercetin. After adjusting the pulverization. The crushed wormwood was put in a domestically produced chiliyang pot that had undergone a sterilization process, and the wormwood and purified water were immersed in a weight ratio of 1: 0.5, and then Lactococcus lactis strain was inoculated with 1%. After sealing the junction by closing the lid (cover) of the pottery, the warm solid fermentation was carried out for 50 days while maintaining 25 to 30°C, and then the pottery was moved as it is, and the cold solid fermentation was carried out for 50 days in a cool place at 10 to 15°C.

Then, triple liquid fermentation was performed. Specifically, wormwood obtained by solid fermentation in warm and cold air for 100 days was added to the Lactococcus lactis strain culture solution at a ratio of 1:10 by weight. Here, as a carbon source, 7 to 13 g/L of glucose and 0.5 to 1.5 g/L of ascorbic acid were additionally added and cultured. The culture was fermented for 1 to 2 days while maintaining 100 rpm at 37° C. in a shaking incubator, and 1% Lactococcus lactis strain inoculation was performed 3 times in accordance with the death period, and fermentation was repeated for 3 hours each time. Finally, after removing the wormwood solid matter, a liquid fermented product was obtained.

Finally, low temperature high pressure extraction (LTPE) was performed. Specifically, the fermented wormwood obtained by repeating liquid fermentation three times was extracted by low-temperature high-pressure treatment for 3 hours at 20 to 25° C. and 3 MPa pressure using a high-pressure extractor, and then 0.2 μm with Whatman filter paper. A final composition was obtained by filtering the fermented wormwood having a size of to 0.8 μm.

As a control, wormwood wormwood was fermented by a general fermented wormwood fermentation method. Specifically, after washing with purified water, dried wormwood was added to the Lactococcus lactis strain culture solution at a ratio of 1:10 by weight. Here, as a carbon source, 7 to 13 g/L of glucose and 0.5 to 1.5 g/L of ascorbic acid were additionally added and cultured. The culture was fermented for 1 to 2 days while maintaining 100 rpm at 37° C. in a shaking incubator, and finally, a liquid wormwood fermentation was obtained by removing the solids of the wormwood (Control 1).

Example 2. Preparation 2 of fermented wormwood wormwood

For the preparation of fermented wormwood fermented wormwood, fermented wormwood fermented wormwood was fermented in an Onggi for 100 days, followed by liquid fermentation in a conventional method, followed by low-temperature and high-pressure extraction to prepare fermented wormwood wormwood. The liquid fermentation step is different from that of the manufacturing method of Example 1. The specific method is as follows.

First, solid fermentation was performed using Onggi. Specifically, artemisinin and quercetin, which are the active ingredients contained in wormwood, were washed and dried naturally until the water disappears from the surface after washing the artemisinin and quercetin, which have the highest content of the artemisinin and quercetin. After adjusting the pulverization. The crushed wormwood was put in a sterilized domestically produced pottery, and the wormwood and purified water were immersed in a weight ratio of 1:0.5, and then Lactococcus lactis strain was inoculated with 1%. After sealing the junction by closing the lid (cover) of the pottery, the warm solid fermentation was carried out for 50 days while maintaining 25 to 30°C, and then the pottery was moved as it is, and the cold solid fermentation was carried out for 50 days in a cool place at 10 to 15°C.

Then, liquid fermentation was carried out. Specifically, wormwood obtained by solid fermentation in warm and cold air for 100 days was added to the Lactococcus lactis strain culture solution at a ratio of 1:10 by weight. Here, as a carbon source, 7 to 13 g/L of glucose and 0.5 to 1.5 g/L of ascorbic acid were additionally added and cultured. The culture was fermented for 1 to 2 days while maintaining 100 rpm at 37° C. in a shaking incubator, and then the solids of wormwood were removed to obtain a fermented product in a liquid state.

Finally, low temperature high pressure extraction (LTPE) was performed. Specifically, the fermented artemisiae fermented product obtained by liquid fermentation was extracted by low temperature and high pressure treatment for 3 hours at 20 to 25° C. and 3 MPa pressure using a high pressure extractor, and then 0.2 μm to 0.8 μm with Whatman filter paper. The final composition was obtained by filtering the fermented wormwood of the size.

Example 3. Preparation 3 of fermented wormwood wormwood

For the preparation of fermented wormwood fermented wormwood, fermented wormwood fermented wormwood was fermented in Onggi for 100 days, followed by liquid fermentation in triplicates, and extracted to prepare fermented wormwood wormwood. The extraction step is different from that of the manufacturing method of Example 1 above. The specific method is as follows.

First, solid fermentation was performed using Onggi. Specifically, artemisinin and quercetin, which are the active ingredients contained in wormwood, were washed and dried naturally until the water disappears from the surface after washing the artemisinin and quercetin, which have the highest content of the artemisinin and quercetin. After adjusting the pulverization. The crushed wormwood was put in a sterilized domestically produced pottery, and the wormwood and purified water were immersed in a weight ratio of 1:0.5, and then Lactococcus lactis strain was inoculated with 1%. After sealing the junction by closing the lid (cover) of the pottery, the warm solid fermentation was carried out for 50 days while maintaining 25 to 30°C, and then the pottery was moved as it is, and the cold solid fermentation was carried out for 50 days in a cool place at 10 to 15°C.

Then, triple liquid fermentation was performed. Specifically, wormwood obtained by solid fermentation in warm and cold air for 100 days was added to the Lactococcus lactis strain culture solution at a ratio of 1:10 by weight. Here, as a carbon source, 7 to 13 g/L of glucose and 0.5 to 1.5 g/L of ascorbic acid were additionally added and cultured. The culture was fermented for 1 to 2 days while maintaining 100 rpm at 37° C. in a shaking incubator, and 1% Lactococcus lactis strain inoculation was repeated 3 times for each 3 hours according to the death period, and fermentation was repeated . Finally, after removing the wormwood solid matter, a liquid fermented product was obtained.

Finally, after adding an aqueous medium of butylene glycol 50% at 1:20 to the fermented wormwood obtained by repeating liquid fermentation three times, heated extraction at 80 to 90° C. for 2 hours, and then Whatman A final composition was obtained by filtering the fermented wormwood fermented wormwood having a size of 0.2 μm to 0.8 μm with filter paper.

Example 4. Preparation 4 of fermented wormwood wormwood

For the preparation of fermented wormwood fermented wormwood, fermented wormwood fermented wormwood was fermented in an Onggi for 100 days, followed by liquid fermentation in a conventional method, and hot water extraction to prepare fermented wormwood wormwood. Compared to the manufacturing method of Example 1, the step of fermenting pottery was the same, the step of fermenting liquid was the same as that of Example 2, and the step of extracting was the same as that of Example 3. The specific method is as follows.

First, solid fermentation was performed using Onggi. Specifically, artemisinin and quercetin, which are the active ingredients contained in wormwood, were washed and dried naturally until the water disappears from the surface after washing the artemisinin and quercetin, which have the highest content of the artemisinin and quercetin. After adjusting the pulverization. The crushed wormwood was put in a sterilized domestically produced pottery, and the wormwood and purified water were immersed in a weight ratio of 1:0.5, and then Lactococcus lactis strain was inoculated with 1%. After sealing the junction by closing the lid (cover) of the pottery, the warm solid fermentation was carried out for 50 days while maintaining 25 to 30°C, and then the pottery was moved as it is, and the cold solid fermentation was carried out for 50 days in a cool place at 10 to 15°C.

Then, liquid fermentation was carried out. Specifically, wormwood obtained by solid fermentation in warm and cold air for 100 days was added to the Lactococcus lactis strain culture solution at a ratio of 1:10 by weight. Here, as a carbon source, 7 to 13 g/L of glucose and 0.5 to 1.5 g/L of ascorbic acid were additionally added and cultured. The culture was fermented for 1 to 2 days while maintaining 100 rpm at 37° C. in a shaking incubator, and then the solids of wormwood were removed to obtain a fermented product in a liquid state.

Lastly, after adding an aqueous medium of butylene glycol 50% at a ratio of 1:20 to the fermented artemisiae fermented product obtained by liquid fermentation, the extract was heated for 2 hours at 80 to 90°C, and then 0.2 with Whatman filter paper. The final composition was obtained by filtering the fermented artemisiae fermentation product having a size of µm to 0.8 µm.

The manufacturing methods of Examples 1 to 4 are shown in Table 1 below.

100-day Onggi fermentation
(50 days warm fermentation,
50 days cold fermentation) Triple liquid fermentation Low temperature high pressure extraction Control 1 Fermentation of common wormwood Example 1 O O O Example 2 O - O Example 3 O O - Example 4 O - -

Experimental Example 1. Confirmation of antioxidant effect of fermented wormwood wormwood

In order to confirm the antioxidant effect of the prepared wormwood fermentation, the DPPH radical (1,1-diphenyl-2-picrylhydrazyl radical) scavenging ability of the fermented wormwood was analyzed.

Specifically, 50 µl of a 50 mM DPPH solution was added to 100 µl of the prepared fermented artemisiae fermented product sample, and then reacted at room temperature for about 30 minutes to measure absorbance at 517 nm. As a positive control, 500 μg/ml of L-ascorbic acid was used. The DPPH radical scavenging ability of each sample was calculated by the following formula:

DPPH radical scavenging ability (%) = (1-absorbance of sample added group/absorbance of untreated group) * 100

The measured DPPH radical scavenging ability (%) is shown in Table 2 below.

As a result, in the case of L-ascorbic acid as a positive control, about 89.48% of DPPH radical scavenging ability was shown, and in the case of the fermented wormwood according to Example 1, about 82.26% of DPPH radical scavenging ability was shown at a concentration of 1%, and the control group showed a DPPH radical scavenging ability of about 89.48%. It was confirmed that the antioxidant activity effect was excellent compared to the fermented wormwood according to 1 and Comparative Examples 1 to 3.

Sample concentration 0.1% 0.5% One% Control 1 4.39 13.26 23.30 Example 1 25.27 58.51 82.26 Example 2 22.67 42.83 66.93 Example 3 32.62 56.10 61.28 Example 4 8.24 46.95 57.53 Positive control - - 89.48

Experimental Example 2. Confirmation of flavonoid content of fermented wormwood wormwood

The content of flavonoids, which are effective substances contained in the prepared fermented wormwood, was measured using quercetin as a standard substance.

Specifically, it was diluted by dissolving in distilled water so that the final concentration of the prepared fermented mugwort fermented product sample was 100 µl/ml. To 200 µl of the diluted sample, 2,000 µl of diethylene glycol and 2,000 µl of 1N NaOH were added and reacted at 37° C. for 1 hour, and then 200 µl of the sample was added to 96 wells and the absorbance was measured at 420 nm. After preparing a standard curve for each concentration using quercetin as a standard material, the total flavonoid content was calculated and shown in Table 3 below.

As a result, in the case of the fermented wormwood according to Example 1, it was confirmed that the content of flavonoids was high compared to the fermented wormwood according to the control 1 and Examples 2 to 4.

Total Flavonoid Content in Fermented Mugwort (Quercetin, mg/g) Control 1 Example 1 Example 2 Example 3 Example 4 21.62 40.50 32.37 30.61 29.99

Experimental Example 3. Confirmation of polyphenol content of fermented wormwood wormwood

The content of polyphenol, an effective substance, contained in the prepared fermented wormwood was measured using tannin acid as a standard material.

Specifically, it was diluted by dissolving in distilled water so that the final concentration of the fermented mugwort fermented product sample was 100 µl/ml. 50 µl of the diluted sample was added to 96 wells, 50 µl of Folin & ciocaltean reagent was added, and the mixture was reacted for 5 minutes. Then, 50 µl of 10% Na ₂ CO ₃ was added, mixed, and reacted for 1 hour, and the absorbance was measured at 640 nm. After preparing a standard curve for each concentration using tannic acid as a standard material, the total phenol content was calculated and shown in Table 4 below.

As a result, it was confirmed that the fermented wormwood wormwood according to one embodiment had a higher content of polyphenol compared to the control group.

Total polyphenol content in fermented wormwood (tannic acid, mg/g) Control 1 Example 1 Example 2 Example 3 Example 4 4.52 11.97 10.39 11.41 8.81

Experimental Example 4. Confirmation of anti-inflammatory effect of fermented wormwood wormwood

In order to confirm the anti-inflammatory effect of the fermented wormwood prepared according to Example 1, the ability to inhibit NO (Nitric oxide) radicals of the fermented wormwood was analyzed.

Specifically, 100 µl of a sample of fermented wormwood prepared according to Example 1 was diluted with 1, 3 and 5%, and 50 µl of a 10 mM sodium nitroprusside solution was added, and then at 25°C. It was reacted for 2 hours and 30 minutes in a state of being shielded from light at temperature. After the reaction, 50 µl of a Griess reagent solution was added, followed by reacting for 10 minutes in a light-shielding state at room temperature, and absorbance was measured at 540 nm. L-ascorbic acid (500 μg/ml) was used as a positive control, and the NO radical inhibitory ability of each sample was calculated by the following equation:

NO radical inhibitory ability (%) = {1-(NO production amount of sample added group/NO production amount of positive control group)}*100

As a result, it was confirmed that the NO radical inhibitory ability of the fermented wormwood fermented wormwood according to Example 1 was significantly increased compared to that before fermentation (FIG. 1), indicating that the fermented wormwood fermented wormwood according to Example 1 had excellent anti-inflammatory activity.

Experimental Example 5. Confirmation of moisturizing effect of fermented wormwood

In order to confirm the moisturizing effect of the fermented wormwood prepared according to Example 1, the moisture content was measured using a moisture meter (corneometer).

Specifically, the fermented wormwood prepared according to Example 1 was applied to the selected test site of the forearm for 20 adult men and women, and the moisture content over time was measured by a Corneometer CM 825 (Courage & Khazaka, Germany) device. It was measured using.

As a result, it was confirmed that the water retention effect of the fermented wormwood wormwood according to Example 1 was superior compared to the control and before fermentation (FIG. 2).

Claims (10)

Inoculating a strain of Lactococcus lactis in Artemisia annua;
Solid fermentation of the wormwood inoculated with the strain at a temperature of 20 to 45° C.;
Solid-fermenting the warm fermented wormwood at a low temperature of 1 to 19°C;
Liquid fermentation by repeating the inoculation of Lactococcus lactis strains 1 to 5 times the solid fermented wormwood at the warmth and low temperature; And
Including the step of extracting the liquid fermented Mugwort under 15 to 30°C and 1 to 5 Mpa, and the step of solid fermenting at warm and low temperature is solid fermentation in Onggi, or Chilliyang Onggi. Way. The method of claim 1, wherein the step of fermenting solids at warm and low temperature is fermented for 30 to 70 days, respectively. delete delete delete delete delete The method of claim 1, wherein the wormwood wormwood is the above-ground part of wormwood wormwood. A cosmetic composition comprising a fermented wormwood prepared by the method of claim 1. The cosmetic composition of claim 9, wherein the fermented artemisiae fermented product contains quercetin in a concentration of 25 mg to 50 mg, or tannic acid at a concentration of 6 mg to 15 mg per 1 g of the fermented artemisiae.

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