KR102167786B1 - manufacturing method of fermented extract of Centella asiatica, Althaea rosea, Blue lotus, Swertia japonica Griseb using Lactobacillus and cosmetic composition using that fermented extract - Google Patents

Abstract

A manufacturing method of a fermented product of Centella asiatica, Alcea rosea, blue lotus, and Swertia japonica using Lactobacillus according to the present invention comprises the steps of: mixing and extracting Centella asiatica, Alcea rosea, blue lotus, and Swertia japonica to obtain a mixed extract; inoculating the mixed extract with Lactobacillus curvatus J2K-01 (accession number: KCTC 13494BP), followed by fermenting; and removing a precipitate and lactic acid bacteria to manufacture the fermented product. The present invention provides a cosmetic composition effective in skin soothing, inflammation amelioration, wound healing and regeneration.

Description

A cosmetic composition having a wound healing, skin rejuvenation, soothing, and anti-inflammatory effect, including the fermented product for centella, hollyhock, blue lotus, and bitter herb cosmetics using lactobacillus and its manufacturing method and the fermented product for cosmetics BACKGROUND OF THE INVENTION (manufacturing method of fermented extract of Centella asiatica, Althaea rosea, Blue lotus, Swertia japonica Griseb using Lactobacillus and cosmetic composition using that fermented extract}

The present invention relates to a cosmetic composition, a fermented product for cosmetics, and a method for manufacturing the same, and in particular, Lactobacillus kurbatus J2K-01 is used as an inoculation strain, and the fermentation target is soothing the skin using centella, hollyhock, blue lotus, and bitter grass. Regarding the manufacturing method of fermented centella, hollyhock, blue lotus and bitter grass using Lactobacillus, which enhances wound healing and regeneration, and skin inflammation improvement effect, and wound healing and skin regeneration, soothing, anti-inflammatory cosmetic composition containing fermented products will be.

When in contact with the external environment, the skin plays a very important role in protecting the body from physical and chemical irritation and pathogens.

The cross section of the skin is divided into three layers: epidermis, dermis, and subcutaneous tissue. Among them, the stratum corneum of the epidermis contains about 10-20% of moisture and exists on the outermost part of the human body to inhibit water evaporation from the body. Blocks excessive penetration of material from

However, due to environmental changes, lifestyle habits, environmental pollution due to various stresses and pollutants from the social environment, frequent cleansing according to makeup habits, and natural skin deterioration due to aging, the skin becomes dry, the surface becomes rough, and the skin becomes shiny. As phenomena such as loss of skin and dull appearance occur, problems related to skin health are becoming increasingly important.

As a method of maintaining skin health from skin stress, much effort has been devoted to the development of moisturizing materials to prevent drying of skin moisture and improve moisture retention in the stratum corneum.

This is because when dry skin is formed, the barrier function of the skin surface is lost, and irritants or allergens from the outside can easily invade the skin, and the skin reacts to such intrusions.

Rejection of the skin is a cytokine produced and released by keratinocytes, which constitute the majority of the surface layer, by activating a small number of cells such as Langerhans and melanocytes. It appears as an inflammatory phenomenon caused by (cytokine).

The cytokine functions to attract inflammatory cells, and in particular, by stimulating the constituent cells of the skin, it causes an inflammatory reaction centered on the upper layer of the skin and forms an eczema reaction, which is called a non-allergic skin disease.

Therefore, substances with strong skin soothing and skin moisturizing effects have the potential to extend to anti-inflammatory effects, and on the contrary, substances with anti-inflammatory effects can be predicted to be effective in skin soothing and skin moisturizing. There has been a problem of having to go through a very difficult process to find.

Centella asiatica is a plant that has been discovered by necessity.

Centella asiatica is a family of parsley, also called tiger grass, and has long been widely used as a folk remedy, known to have anti-inflammatory, antipyretic, and antibacterial effects.

Specifically, the leaves and stems of Centella asiatica have an ingredient called madecassic acid, which soothes and gives vitality to tired skin, helps protect skin and manages traces, and heals damaged skin (inflammation). It is known to be effective in treating wounds.

Accordingly, the "cosmetic composition for preventing and improving skin troubles" (Korean Patent Application Publication No. 10-2019-0142007, Patent Document 1) contains extracts such as Eoseongcho, willow branch, chamomile, green tea, etc. in addition to centella asiatica extract. Is presented.

Chok-gyu-hwa refers to the flower of a hollyhock of the mallow family. The flower shape resembles a plate, so it is called a hollyhock, white flowers are called white flowers, and red flowers are called red flowers.

Chok-gyu-hwa has been used as a medicinal material for symptoms such as baekdaeha, lower abdominal coldness, dyspnea, and hematopoiesis, but "cosmetic composition containing herbal medicinal mixture extract" (Korean Registered Patent Publication No. 10-1135264, Patent Document 2) , An example used in the manufacture of a cosmetic composition having effects on collagen biosynthesis, collagenase expression suppression, elastase expression suppression, melanin production suppression, etc. in combination with native grass (branch root), chinensis (dianthus) and the like is disclosed.

On the other hand, in the case of the simple extract of the natural material described above, there is a problem in that the active ingredient is physically and chemically denatured during the extraction process, making it difficult to absorb in the body or reducing the effect.

In order to solve this problem, in recent years, a number of technologies for fermenting extracts using various strains and using them as raw materials for cosmetics have been disclosed.

As an example, in "A method for manufacturing fermented fruit and vegetables and a cosmetic composition containing fermented fruit and fermented fruit as an active ingredient" (Korean Patent Publication No. 10-1905517, Patent Document 3), after inoculation of Lactobacillus sp. It has been cultivated and fermented under conditions to obtain a fermented product, which has been used as a raw material for cosmetics.

However, studies on skin soothing and inflammation improvement through inoculation and fermentation of specific plant extracts and specific strains applied thereto are still insufficient. In particular, studies on bitter grass or blue lotus are still insufficient.

KR 10-2019-0142007 (2019.12.26) KR 10-1135264 (2012.04.03) KR 10-1905517 (2018.10.01)

The present invention has been researched and developed to solve the above problems, and an object of the present invention is to provide a method for producing a fermented product effective in soothing skin, improving inflammation, and healing wounds.

In addition, the present invention is to provide a cosmetic composition excellent in effect by providing a method for producing a fermented product fermented with Lactobacillus curbatus J2K-01 (Accession No.: KCTC 13494BP).

In order to solve the above problems, the manufacturing method of a fermented product for cosmetics using Lactobacillus kurbatus J2K-01 strain and bitter grass of the present invention is to obtain a mixed extract by mixing and extracting centella, hollyhock, blue lotus, and bitter grass. , After fermentation by inoculating Lactobacillus curbatus J2K-01 (accession number: KCTC 13494BP) to the mixed extract, the fermentation product is prepared by removing the precipitate and lactic acid bacteria.

In the above configuration, the raw material for the preparation of the mixed extract is characterized in that, based on 100 parts by weight of centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass are mixed. .

In addition, the fermented product of the present invention is characterized in that it is produced by the above manufacturing method.

The cosmetic composition using the Lactobacillus curbatus J2K-01 strain and bitter grass of the present invention is characterized in that the fermentation product contains 0.001 to 10% by weight based on the total content of the cosmetic composition.

At this time, the cosmetic composition is characterized in that for improving skin inflammation.

Or, the cosmetic composition is characterized in that for skin soothing.

Alternatively, the cosmetic composition is characterized in that for wound healing and regeneration.

According to the present invention, there is provided a cosmetic composition effective in skin soothing, inflammation improvement, wound healing and regeneration.

Figure 1 is a process diagram showing a method of manufacturing a fermented product in the present invention.

Centella asiatica is a family of parsley, also called tiger grass, and has long been widely used as a folk remedy, known to have anti-inflammatory, antipyretic, and antibacterial effects.

Specifically, the leaves and stems of Centella asiatica have an ingredient called madecassic acid, which soothes and gives vitality to tired skin, helps protect skin and manages traces, and heals damaged skin (inflammation). It is known to be effective in treating wounds.

The hollyhock is also called chokgyuhwa, and it refers to the flower of hollyhock.

A hollyhock is called a hollyhock because its flower shape resembles a plate, and a white flower is called a white flower, and a red flower is called a red flower.

[0003] Chok-gyuhwa has conventionally been used as a medicinal agent for symptoms such as baekdaeha, lower abdominal coldness, dyspepsia, and hematopoiesis.

Blue lotus is a perennial plant native to South Africa, and includes Thai water lily (Nymphaea coerulea) and Indian water lily (Nymphaea, stellata).

The water lily family is a wild aquatic plant that grows very large with leaves of about 30cm in diameter, and grows on islands near southern and eastern Asia. This plant, usually called water lily, is called blue lotus because it has blue flowers.

Bitter grass is a herbaceous herb called Swertia japonica MAKINO or gocho. It contains a strong bitter taste in its roots.

The stalk with hair is standing upright and pruned at the upper part. The stem is purple and the height is about 15~30cm.

The leaves are lanceolate, do not have petioles, and two are facing each other. Flowers gather in the shape of a root at the end of the stem and bloom in order from top to bottom.

It has 5 petals of an elongated oval shape, and the petals are fully spread out as they bloom. The diameter of the flower is 1.5~2cm, and it looks light purple because there are purple lines on the white background. Five stamens are located in the middle of the flower, and the pollen pouch is dark purple.

Distribution is mainly distributed in Jeju Island and southern regions, and it grows in sunny meadows in mountains and fields.

In the field of private medicine, it has been used as a medicine for indigestion, anorexia, and anthelmintic.

Recently, there have been reports of being used in the field of hair cosmetics, but there is no known one used for soothing skin and alleviating inflammation.

Lactobacillus Courbatus J2K-01 (Accession No.: KCTC 13494BP) is a new Lactobacillus Courbatus strain recently published in “a cosmetic composition for improving skin elasticity and wrinkles” (Korean Patent Publication No. 10-1930342), The nucleotide sequence and the deposit certificate are disclosed in the above patent publication.

Centella asiatica, hollyhock, blue lotus, and bitter grass can be used by extracting all or part of leaves, stems, flowers, and roots as needed.

More preferably, centella and bitter plants use outposts, and hollyhock and blue lotus flowers are preferably used.

The manufacturing method of a cosmetic fermentation product using the Lactobacillus kurbatus J2K-01 strain and bitter grass of the present invention, as shown in Fig. 1, mixed and extracted raw materials, sterilized, and then cultured and fermented using microorganisms, It is produced by vacuum drying through centrifugation and vacuum concentration steps.

More specifically, centella asiatica, hollyhock, blue lotus, and bitter grass are used as raw materials, and extracts may be mixed after individual extraction, or mixed and then extracted to obtain a mixed extract.

At this time, it is preferable to extract the raw materials after mixing.

The content ratio of the raw material may consist of 0.1 to 30 mass ratio of centella, 0.1 to 30 mass ratio of hollyhock, 0.1 to 30 mass ratio of blue lotus, and 1 to 50 mass ratio of bitter grass.

More specifically, based on 100 parts by weight of centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass may be mixed.

As for the extraction method, various methods such as hot water extraction, distillation extraction, ethanol extraction, ultrasonic extraction, etc. can be applied, but it is appropriate to extract for 1 to 6 hours using purified water at 80 ~ 100℃.

More preferably, it is good to extract for 3 to 5 hours at a temperature of 85 to 95 °C.

In addition, when the mixed extract is prepared by extraction, it is cooled and filtered, and sterilized for 10 to 20 minutes at a temperature of about 120°C.

In addition to the preparation of the mixed extract, Lactobacillus curbatus J2K-01 of accession number KCTC 13494BP was inoculated, cultured and fermented.

The culture and fermentation conditions are appropriate at a temperature of 25 to 32°C, and a culture period is appropriate for about 3 to 5 days.

When the fermentation broth is obtained by cultivation and fermentation, it is added to a centrifuge and centrifuged for 15 to 25 minutes under the conditions of 8,000 to 1,2000 rpm to obtain a fermented product from which precipitates and lactic acid bacteria are removed.

Then, the fermented product is concentrated under reduced pressure and vacuum-dried.

Meanwhile, the fermented product prepared as described above is included as a raw material for the cosmetic composition, and an appropriate content is preferably contained in 0.001 to 10% by weight based on the total content of the cosmetic composition.

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

Example 1: Preparation of fermented product

20g Centella asiatica, 20g flowers, 20g blue lotus flowers, and 50g bitter grass were placed in an extraction container, mixed with 1000g of purified water, and extracted for 3 hours at 90°C to prepare a mixed extract, cooled, and filtered.

Then, the mixed extract was sterilized at 121°C for 15 minutes for culture.

The sterilized mixed extract was inoculated with Lactobacillus kurbatus J2K-01 and cultured and fermented for 3 days at a temperature of 32° C. and stationary culture conditions to prepare a fermentation broth.

Subsequently, the fermentation broth was centrifuged at 10,000 rpm for 20 minutes to obtain a fermented product from which precipitates and lactic acid bacteria were removed.

This was concentrated under reduced pressure and dried under vacuum to obtain a powder.

Example 2: Preparation of fermented product

Proceed in the same manner as in Example 1, but 9g of hollyhock was used.

Example 3: Preparation of fermented product

Proceed in the same manner as in Example 1, but 31g of hollyhock was used.

Example 4: Preparation of fermented product

Proceed in the same manner as in Example 1, but 9g of blue lotus was used.

Example 5: Preparation of fermented product

Proceed in the same manner as in Example 1, but 31g of blue lotus was used.

Example 6: Preparation of fermented product

Proceed in the same manner as in Example 1, but 39g of bitter grass was used.

Example 7: Preparation of fermented product

Proceed in the same manner as in Example 1, but 61g of bitter grass was used.

Comparative Example 1: Preparation of mixed extract

Centella asiatica 20g, Phalaenopsis 20g, blue lotus 20g, and bitter grass 50g were placed in an extraction container, and the extract mixed with 1000g of purified water was extracted for 3 hours at 90°C, cooled, and filtered. The extract was concentrated under reduced pressure and dried under vacuum to obtain a powder.

Comparative Example 2: Preparation of fermented product

Proceed in the same manner as in Example 1, but inoculated with Lactobacillus pentosus J2K-185 as a strain.

Comparative Example 3: Preparation of fermented product

Proceed in the same manner as in Example 1, but inoculated with Lactobacillus brevis J2K-55 as a strain.

Comparative Example 4: Preparation of fermented product

Proceed in the same manner as in Example 1, but inoculated with Lactobacillus curbatus AMI-1214 as a strain.

Comparative Example 5: Preparation of fermented product

Proceeding in the same manner as in Example 1, except for excluding centella from the raw materials.

Comparative Example 6: Preparation of fermented product

Proceed in the same manner as in Example 1, except for the hollyhock among the raw materials.

Comparative Example 7: Preparation of fermented product

Proceed in the same manner as in Example 1, except for the blue lotus flower among the raw materials.

Comparative Example 8: Preparation of fermented product

Proceed in the same manner as in Example 1, except for bitter grass among the raw materials.

<Experimental Example 1> Evaluation of skin improvement effect

Using the extracts of Examples and Comparative Examples, a cosmetic composition in the form of a cream having the composition shown in Table 1 below was prepared.

In the table, the extracts prepared according to Examples 1 to 7, Comparative Examples 1 to 8 were adjusted to be 1% by weight of the total composition of the cosmetic.

Raw material Composition (% by weight) Purified water Balance glycerin 8 Beta glucan 5 Carbomer 0.1 Stearic acid cholesterol 2 Squalane 4 Cetearyl alcohol 6 Polyoxyethylene alcohol ester 3 Caprylic/Capric Triglyceride 8 Extracts of Examples or Comparative Examples One Propylene glycol 5 Sum 100

Then, when the extracts of Examples and Comparative Examples were used, the degree of skin regeneration and wound healing improvement, skin moisturizing, whitening effect, and skin soothing effect were evaluated.

Specifically, for subjects who have undergone laser treatment or dermabrasion treatment, use the cosmetics of Examples and Comparative Examples for 5 people each for a week (twice a day (in the morning and evening)), and then the 10-point scale method (1: very bad) --> 10: Very good), and then the significance was tested with Duncan's multiple range test.

As a result, the results shown in Table 2 were obtained.

division Skin regeneration Wound healing
(Anti-inflammatory effect) Skin moisturizing Whitening effect Skin soothing effect Example 1 9.5±0.21 9.4±0.32 9.15±0.14 9.25±0.33 9.4±0.57 Example 2 8.2±0.35 8.12±0.42 8.4±0.72 8.3±0.4 8.3±0.24 Example 3 9.4±0.72 9.36±0.35 7.9±0.45 8.1±0.3 8.1±0.25 Example 4 8.1±0.11 8.2±0.41 8.0±0.12 8.2±0.34 7.9±0.15 Example 5 8.2±0.32 8.7±0.25 8.2±0.64 8.3±0.28 9.3±0.66 Example 6 8.2±0.7 8.1±0.7 7.9±0.63 8.3±0.88 8.1±0.74 Example 7 7.9±0.54 8.2±0.33 8.1±0.76 7.9±0.91 8.2±0.81 Comparative Example 1 7.1±0.32 6.7±0.85 6.7±0.52 6.1±0.24 5.2±0.34 Comparative Example 2 7.4±0.17 7.1±0.41 7.0±0.87 6.5±0.55 5.8±0.21 Comparative Example 3 7.2±0.35 7.0±0.52 6.7±0.48 6.6±0.81 5.9±0.44 Comparative Example 4 7.2±0.8 6.7±0.68 6.8±0.33 6.8±0.67 5.9±0.56 Comparative Example 5 7.2±0.42 5.5±0.74 7.0±0.67 6.3±0.31 5.8±0.72 Comparative Example 6 5.1±0.56 6.6±0.54 6.6±0.87 6.5±0.54 5.7±0.34 Comparative Example 7 6.9±0.17 6.5±0.34 6.7±0.45 6.2±0.41 5.1±0.67 Comparative Example 8 7.1±0.52 5.1±0.57 5.4±0.72 6.6±0.21 5.6±0.33

As shown in Table 2 above, the Examples showed superior overall skin regeneration and wound healing effects compared to the Comparative Examples, and overall preference was found to be higher, including skin moisturizing, whitening, and skin soothing effects.

<Experimental Example 2> Skin irritation test

After preparing a cream-shaped cosmetic as in Experimental Example 1, a skin patch test was performed to investigate the skin irritation test for each cosmetic.

Subjects were selected as those without skin disease at the test site of 15-30 years old, and a total of 15 subjects were conducted. After wiping and drying the upper arm, which is the patch area, with 70% ethanol, 01 g of the product was added to the human upper arm for 24 hours, and the skin reaction was examined within 1 hour after removal of the patch, and the next day (after 48 hours) was examined again. The skin reaction was determined according to the following test criteria. Table 4 shows the results of the skin patch test.

* Skin reaction test criteria

- no response

± weak positive reaction (erythema)

+ Positive reaction (erythema)

++ strong positive reaction (erythema, edema)

+++ Severe positive reaction (erythema, swelling, blisters)

division - ± + ++ +++ Example 1 15 0 0 0 0 Example 2 15 0 0 0 0 Example 3 15 0 0 0 0 Example 4 15 0 0 0 0 Example 5 15 0 0 0 0 Example 6 15 0 0 0 0 Example 7 15 0 0 0 0 Comparative Example 1 11 2 One One Comparative Example 2 13 2 0 0 0 Comparative Example 3 13 One One 0 0 Comparative Example 4 13 0 2 0 0 Comparative Example 5 13 One One 0 0 Comparative Example 6 12 2 One 0 0 Comparative Example 7 13 One One 0 0 Comparative Example 8 13 0 2 0 0

As shown in Table 3, in the case of the Example, it was found that there was little skin irritation.

<Experimental Example 3> Measurement of the inhibitory effect of hyaluronidase activity

In order to measure the anti-inflammatory effect of the extracts of Examples and Comparative Examples, the inhibitory effect of hyaluronidase activity, an inflammatory enzyme, was measured. Hyaluronidase is an enzyme that hydrolyzes hyaluronic acid and causes inflammation.

In this experimental example, the anti-inflammatory effect was determined by applying a method of measuring the anti-inflammatory effect by inhibiting the activity of hyaluronidase (Kakegawa Y: Japanese J. of Inflammation, 4, 437-438, 1984). .

100 µl of sample and 50 µl of hyaluronidiase solution (type IV-S, Sigma, 400U/ml) were added and reacted at 37°C for 20 minutes, followed by enzyme activation solution (compound 48/80 CaCl ₂ ㆍ2H ₂ O, Sigma , 0.1 mg/ml) was added and reacted again at 37°C for 20 minutes.

250 µl of hyaluronic acid solution (0.4 mg/ml) was added and reacted at 37° C. for 40 minutes, and 100 µl of 0.4N NaOH was added to terminate the reaction. 100 µl of potassium borate solution was added, reacted at 95° C. for 3 minutes, cooled, 3 ml of ρ-dimethylaminobenzaldehyde solution was added, and reacted at 37° C. for 20 minutes, followed by color development. The absorbance was measured at 585 nm to measure the hyaluronidase activity inhibition rate.

The hyaluronidase activity inhibition rate (%) was calculated by Equation 1, and the IC ₅₀ value is the concentration of a substance that inhibits the hyaluronidiase enzyme activity by 50%.

<Equation 1>

Hyaluronidase activity inhibition rate (%) = [(A-B)/A] × 100

A: Enzymatic activity of wells without addition of samples (extracts of Examples and Comparative Examples)

B: Enzymatic activity of wells to which samples (extracts of Examples and Comparative Examples) were added

Sample name Hyaluronidase activity inhibitory effect (IC ₅₀ ) Example 1 0.36% Comparative Example 1 0.24% Comparative Example 4 0.27% Comparative Example 5 0.24% Comparative Example 6 0.28% Comparative Example 7 0.27% Comparative Example 8 0.25%

As a result of measuring the activity inhibitory effect of hyaluronidase (Table 4), the IC ₅₀ value of the mixed extract of Example 1 was 0.36%, indicating superior anti-inflammatory effect compared to the comparative examples.

Claims (7)

In the manufacturing method of fermented product for cosmetic,
Centella asiatica, hollyhock, blue lotus, and bitter grass were mixed and extracted to obtain a mixed extract,
After fermentation by inoculating Lactobacillus curbatus J2K-01 (accession number: KCTC 13494BP) to the mixed extract, the fermentation product is prepared by removing the precipitate and lactic acid bacteria,
A method for producing fermented products for cosmetic products of centella, hollyhock, blue lotus and bitter grass using lactobacillus.
The method of claim 1,
The raw material for preparing the mixed extract,
Based on 100 parts by weight of Centella asiatica, 50 to 150 parts by weight of hollyhock, 50 to 150 parts by weight of blue lotus, and 200 to 300 parts by weight of bitter grass are mixed.
A method for producing fermented products for cosmetic products of centella, hollyhock, blue lotus and bitter grass using lactobacillus.
In the cosmetic fermented product,
It characterized in that it is produced by the manufacturing method of any one of claims 1 to 2,
Fermented product for cosmetic products of centella, hollyhock, blue lotus, and bitter grass using lactobacillus.
In the cosmetic composition,
The cosmetic fermented product of claim 3 is characterized in that it contains 0.001 to 10% by weight based on the total content of the cosmetic composition,
A cosmetic composition having a wound healing, skin regeneration, soothing, and anti-inflammatory effect containing fermented products for centella, hollyhock, blue lotus, bitter herb cosmetics using Lactobacillus.
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