KR102512471B1 - Method for extracting high-molecular polysaccharides from yam, okra, lotus root and aloe vera leaves, and cosmetic composition containing them for skin moisturizing and skin inflammation relief - Google Patents

Abstract

The present invention relates to a method for extracting high-molecular polysaccharides from Dioscorea japonica, Abelmoschus esculentus, Nelumbo nucifera and Aloe Barbadensis leaves using a porous polymer, wherein the method comprises the steps of: adding purified water to the Dioscorea japonica, Abelmoschus esculentus, Nelumbo nucifera, Aloe Barbadensis leaves and porous polymer, and then obtaining an extraction liquid; filtering the extraction liquid; obtaining eluate by eluting the polymeric polysaccharide adsorbed on the porous polymer taken in the filtering step; and freeze-drying the eluate to obtain the high-molecular polysaccharide in dry powder form.

Description

Method for extracting high-molecular polysaccharides from yam, okra, lotus root and aloe vera leaves, and cosmetic composition for skin moisturizing and skin inflammation relief containing the same composition containing them for skin moisturizing and skin inflammation relief}

The present invention relates to a method for extracting polymeric polysaccharide from yam, okra, lotus root and aloe vera leaves using a porous polymer and a method for preparing a cosmetic composition containing polymeric polysaccharide as an active ingredient.

Skin is a tissue that protects internal organs, and acts as an essential barrier to protect the body by protecting moisture in the body and blocking invading factors from the outside. The skin serves functions such as protecting the body from pathogens against the external environment, regulating body temperature, and feeling sensations.

Inflammation is a defense mechanism in vivo against external stimuli such as tissue damage or external infectious agents (bacteria, fungi, viruses, etc.) through various pathways or internal stimuli such as metabolites in vivo. Various inflammatory mediators and immune cells in cells It is related to not only symptoms such as erythema, edema, heat, and pain, but also causes various diseases such as allergy, atopy, brain disease, circulatory disorder, and cancer. Various biochemical phenomena are involved in the causes of inflammation, and in particular iNOS (Induced NOS) induced by stimuli such as various cytokines, TNF-α (Tumor necrosis factor), LPS (Lipopolysaccharide) in macrophages Inflammation occurs as nitric oxide (NO) and prostaglandin E2 (PGE2) are produced by cyclooxygenase, an enzyme related to the synthesis of prostaglandins, from arachidonic acid and arachidonic acid. In addition, NO produced from L-arginine by NO synthase (NOS) is a biogenic molecule with high reactivity and plays a role in neurotransmission, blood coagulation, and immune function. However, when produced more than necessary, it exhibits harmful effects to the living body such as excessive vasodilation, cytotoxicity and tissue damage, and inhibition of wound healing. Since almost all diseases appearing on the skin are accompanied by inflammation, efforts are being made to develop materials with anti-inflammatory effects that can improve or treat them.

On the other hand, the skin contains about 65-70% of the water that the human body has, which carries various physiologically active substances necessary for the human body and helps the skin to maintain a moist state. In addition, the skin plays an important role in controlling the evaporation of this moisture outside the body.

On the other hand, research on cosmetic compositions for the purpose of moisturizing skin and relieving skin inflammation is being continuously conducted. In particular, studies using a number of natural substances as raw materials to prepare extracts using various extraction methods and use them as active ingredients in cosmetics have been actively conducted recently. The method of obtaining the extract according to the prior art has a problem of poor stability.

Registration No. 10-1308669

In order to solve the above problems, the present invention extracts and purifies yam, okra, lotus root, and aloe vera leaves using a porous polymer in order to secure the stability of the polymeric polysaccharide component and expand the applicability, thereby improving the stability of the polymeric polysaccharide. It is an object of the present invention to provide a method for extracting high-molecular polysaccharide.

In addition, an object of the present invention is to provide a method for producing a cosmetic composition containing the polymeric polysaccharide extracted through the extraction method as an active ingredient.

The tasks of the present invention are not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the method for extracting high molecular polysaccharides from yams, okra, lotus root and aloe vera leaves using a porous polymer according to an embodiment of the present invention is to add purified water to yams, okra, lotus root, aloe vera leaves and porous polymers. After adding, obtaining an extract; filtering the extract; obtaining an eluate by eluting the polymeric polysaccharide adsorbed on the porous polymer taken in the filtering step; and freeze-drying the eluate to obtain a polymeric polysaccharide in a dry powder form.

The filtering step is characterized in that, after removing impurities from the extract and collecting the filtrate in which the porous polymer is mixed, the porous polymer is taken using filter paper.

According to an embodiment of the present invention, the method for producing a cosmetic composition containing a polymeric polysaccharide extracted from yam, okra, lotus root, and aloe vera leaf as an active ingredient using a porous polymer is After adding purified water, obtaining an extract; filtering the extract; obtaining an eluate by eluting the polymeric polysaccharide adsorbed on the porous polymer taken in the filtering step; Freeze-drying the eluate to obtain a dry powder; and preparing a cosmetic composition containing the polymeric polysaccharide in the form of a dry powder as an active ingredient.

The preparing step is characterized by preparing a cosmetic composition of an oil-in-water emulsion type cream formulation.

The preparing step is characterized by preparing a cosmetic composition for moisturizing the skin and relieving skin inflammation.

Details of other embodiments are included in the detailed description and drawings.

According to the present invention, there is one or more of the following effects.

First, the production of high-molecular polysaccharides using porous polymers containing active ingredients of yam, okra, lotus root, and aloe vera leaves is less costly and stable against heat than existing hot water extraction methods, organic solvent extraction methods, supercritical extraction methods, and ultrasonic extraction methods. It has the effect of obtaining the active ingredient efficiently by securing.

Second, cosmetics containing a polymer polysaccharide obtained through extraction and purification using a porous polymer as an active ingredient have superior moisturizing and anti-inflammatory effects than methods using conventional extraction methods.

The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description of the claims.

1 is a view showing a method for extracting a polymeric polysaccharide according to an embodiment of the present invention.

Hereinafter, the embodiments disclosed in this specification will be described in detail with reference to the accompanying drawings, but the same or similar elements are given the same reference numerals regardless of reference numerals, and redundant description thereof will be omitted. The suffixes "module" and "unit" for components used in the following description are given or used together in consideration of ease of writing the specification, and do not have meanings or roles that are distinct from each other by themselves. In addition, in describing the embodiments disclosed in this specification, if it is determined that a detailed description of a related known technology may obscure the gist of the embodiment disclosed in this specification, the detailed description thereof will be omitted. In addition, the accompanying drawings are only for easy understanding of the embodiments disclosed in this specification, the technical idea disclosed in this specification is not limited by the accompanying drawings, and all changes included in the spirit and technical scope of the present invention , it should be understood to include equivalents or substitutes.

Terms including ordinal numbers, such as first and second, may be used to describe various components, but the components are not limited by the terms. These terms are only used for the purpose of distinguishing one component from another.

It is understood that when an element is referred to as being "connected" or "connected" to another element, it may be directly connected or connected to the other element, but other elements may exist in the middle. It should be. On the other hand, when an element is referred to as “directly connected” or “directly connected” to another element, it should be understood that no other element exists in the middle.

Singular expressions include plural expressions unless the context clearly dictates otherwise.

In this application, terms such as "comprise" or "have" are intended to designate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, but one or more other features It should be understood that the presence or addition of numbers, steps, operations, components, parts, or combinations thereof is not precluded.

1 is a view showing a method for extracting a polymeric polysaccharide according to an embodiment of the present invention.

Referring to the drawings, a method for extracting polymeric polysaccharide from yam, okra, lotus root, and aloe vera leaf using a porous polymer (S100) (hereinafter referred to as an extraction method) using a porous polymer includes an extraction step (S110), It may include a filtration step (S120), an elution step (S130) and a lyophilization step (S140).

In the extraction step, after adding purified water to yam, okra, lotus root, aloe vera leaf, and porous polymer, an extract solution is obtained at 20° C. for 24 hours (S110). Here, the porous polymer is PMMA (Polymethyl Methacrylate), and a pore size of 10 nm to 30 nm may be used.

In the filtration step, after removing impurities from the extract and collecting the filtrate in which the porous polymer is mixed, the porous polymer is taken using filter paper (S120).

In the elution step, an eluate is obtained by eluting the polysaccharide polymer adsorbed on the porous polymer taken in the filtration step (S130).

In the freeze-drying step, the eluate is freeze-dried to obtain a polymeric polysaccharide in the form of a dry powder (S140).

The manufacturing method of a cosmetic composition containing a polymeric polysaccharide extracted from yam, okra, lotus root, and aloe vera leaf as an active ingredient using a porous polymer according to an embodiment of the present invention (hereinafter, referred to as a manufacturing method) includes an extraction step (S110) , Filtering step (S120), elution step (S130), lyophilization step (S140), and the porous polymer extract prepared through the polymeric polysaccharide extraction method (S100) as an active ingredient to prepare a cosmetic composition. .

Steps S110 to S140 in the method for producing a cosmetic composition containing polymeric polysaccharide extracted from yam, okra, lotus root and aloe vera leaves using a porous polymer as an active ingredient, from yams, okra, lotus root and aloe vera leaves using a porous polymer The contents described in the high-molecular polysaccharide extraction method (S100) may be applied.

In the step of preparing a cosmetic composition using the porous polymer extract prepared through the polymeric polysaccharide extraction method (S100) as an active ingredient, a cosmetic composition containing the polymeric polysaccharide in dry powder form as an active ingredient is prepared.

The preparation step is characterized by preparing a cosmetic composition of a cream formulation of an oil-in-water emulsion (O / W emulsion) type.

On the other hand, the cosmetic composition according to an embodiment of the present invention can be prepared according to a conventional method, and is a lotion, lotion, nutrient cream, massage cream, essence, gel along with pharmaceutically, cosmetically, or dermatologically acceptable excipients. , It can be applied to the formulation of serum, pack, cleanser or emulsion.

The manufacturing step is characterized by preparing a cosmetic composition for moisturizing the skin and relieving skin inflammation.

The extraction method according to an embodiment of the present invention solves the instability of the polymeric polysaccharide due to heat during extraction with the extraction solvent and provides a cosmetic composition for moisturizing or anti-inflammatory containing these components.

The extraction method according to the embodiment of the present invention secures the stability of polymer polysaccharide components having various functions, and in particular, extracts and purifies yam, okra, lotus root, and aloe vera leaves using a porous polymer to improve the stability of polymer polysaccharide components. By securing, the applicability can be further expanded. In addition, the extraction method according to the embodiment of the present invention can ensure excellent moisturizing and anti-inflammatory efficacy.

[Example 1] - Polymeric polysaccharide extraction method from yam, okra, lotus root and aloe vera leaves using porous polymer

50 g of each of yam, okra, lotus root, and aloe vera leaf and 200 g of porous polymer were put into the prepared extractor, and then 1,000 g of purified water was added thereto, followed by extraction at 20° C. for 24 hours.

After the extraction was completed, impurities were removed and the filtrate mixed with the porous polymer was collected.

The porous polymer was taken using filter paper, and the polysaccharide adsorbed on the porous polymer was eluted using a 10% ethanol solution as an eluent, and then the eluate was freeze-dried to obtain 15.2 g of dry powder.

Porous polymer materials are used for material separation, storage, and adsorption by using many pores and a large surface area. A hole in a porous polymeric material is called a pore. Pores are divided into micropores smaller than 2 nm, mesopores 2 nm to 50 nm, and macropores larger than 50 nm according to their diameter.

The porous polymer used in this test was PMMA (Polymethyl Methacrylate) with a pore size of 10 nm to 30 nm.

[Comparative Example 1]

For comparison, 50 g of each of yam, okra, lotus root, and aloe vera leaves were put into a prepared extractor, 1,000 g of purified water was added, extracted at 50 ° C for 12 hours, and then concentrated under reduced pressure to obtain 22.1 g.

[Comparative Example 2]

For comparison, an ultrasonic extract was prepared in the same manner as in [Comparative Example 1] and then concentrated under reduced pressure to obtain 25.5 g.

[Experimental Example 1] Skin cell proliferation effect test

Toxicity to skin cells was measured using the extracts prepared in [Example 1], [Comparative Example 1], and [Comparative Example 2] as samples.

The cell lines used for this experiment were three types of cells including human dermal fibroblast (Lonza, Basel, Switzerland). The above cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco-BRL, Invitrogen; Life Technologies, Gaithersburg, MD, USA) in 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA), 1% Using a medium containing penicillin/streptomycin (penicillin 100 IU/mL, streptomycin 100 μg/mL; Life Technologies), the cells were cultured at 37° C. in a 5% CO2 cell incubator.

WST-1 (Water-soluble tetrazolium salt) assay is a method to measure and find out the activity of mitochondrial NADH-dehydrogenase in cells. When living cells are treated with tetrazolium salt, Colored formazan is formed by dehydrogenase in the mitochondria. The concentration of living cells can be confirmed by confirming the absorbance of the formed formazan at a wavelength of 450 nm. Cells were divided into 2×10 3 cells/well in a 96-well plate and cultured for 24 hours, and then the samples were treated according to the appropriate concentration and cultured for another 24 hours. After that, the WST-1 assay solution was treated with 10% of the medium amount and reacted at 37°C for additional 0.5 to 1 hour. Then, using an iMark microplate reader (Bio-Rad, Hercules, CA, USA), the absorbance was measured at 450 nm. was measured, and the reference absorbance was measured at 650 nm to correct the resulting value. The resultant values were expressed as mean values ± standard deviations from three independent experiments, and are shown in <Table 1> below. P<0.05 was verified through Student's t-test and indicates the significance of the result.

<Table 1>

Through the results of <Table 1>, it can be seen that each extract is a safe component that does not cause toxicity to skin cells.

[Experimental Example 2] Antioxidant effect test

The extracts prepared in [Example 1], [Comparative Example 1], and [Comparative Example 2] were used as samples and their antioxidant capacity was measured by DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity.

DPPH radical scavenging activity was referred to the method of Mensor et al. After adding 20 μl of 80 μl of 0.2 mM DPPH ethanol solution to [Example 1], [Comparative Example 1], and [Comparative Example 2], the samples were stirred for 1 minute, reacted in an incubator at 25 ° C. for 10 minutes, and then using an ELISA reader. Absorbance was measured at 517 nm.

The DPPH radical scavenging activity was expressed as a percentage according to <Calculation 1> below, and the results are shown in <Table 2> below.

<Calculation 1>

<Table 2>

As shown in <Table 2>, in the case of [Example 1], a very high DPPH radical scavenging activity value was measured, and it can be confirmed that the extract using the porous polymer of the present invention exhibits excellent antioxidant effect.

[Experimental Example 3] Anti-inflammatory effect test

NO (nitric acid) of macrophages (Raw 264.7 cell) induced by LPS using the porous polymer extract, hot water extract and ultrasonic extract prepared in [Example 1], [Comparative Example 1] and [Comparative Example 2] as samples. oxide) production was measured to confirm the anti-inflammatory effect.

Raw 264.7 cells were divided into 2×105 cells/well in a 96-well plate, and the cells were cultured in DMEM (Dulbecco's modified Eagle's medium) supplemented with penicillin (10 U/ml) and 10% FBS (Fetal bovine serum) at 37°C. , cultured in a 5% CO2 incubator. After culturing, 20mg/ml of LPS (1mg/ml) and [Example 1], [Comparative Example 1] and [Comparative Example 2] were added to the samples and cultured for 24 hours. After culturing, the supernatant was taken and centrifuged at 15,000 rpm for 2 minutes to collect the supernatant, and then reacted with Griess reagent 1: 1, and the absorbance was measured at 570 nm, and the results are shown in <Table 3> below.

<Table 3>

As shown in <Table 3>, it can be confirmed that [Example 1] has a higher anti-inflammatory effect than [Comparative Example 1] and [Comparative Example 2].

[Example 2]

Using the porous polymer extract prepared in [Example 1] as an active ingredient, an oil-in-water emulsion type cosmetic product of [Example 2] was prepared, and the composition of [Example 2] And components are shown in <Table 4> below.

<Table 4>

[Comparative Example 3] and [Comparative Example 4]

[Comparative Example 1] and [Comparative Example 2] using the hot water extract and the ultrasonic extract as active ingredients to prepare an oil-in-water emulsion (O / W emulsion) type cream formulation cosmetic, [Comparative Example 3] In Comparative Example 4, the components and composition ratios added except for the content of the active ingredient are the same as in [Example 2], and the composition ratios are shown in <Table 5> below.

<Table 5>

[Experimental Example 4] Skin irritation test

[Example 2], [Comparative Example 3] to [Comparative Example 4] in order to compare the degree of skin irritation to the oil-in-water type emulsion (O / W emulsion) type of cream formulation cosmetics, a closed patch test on the human body (closed patch test) was performed.

For 20 healthy adult men and women, 0.2g of each sample was applied to the inside of the lower arm, and after 24 hours, the pin chamber was removed, and after 4 hours, the skin condition was visually checked, and the results are shown in <Table 6> below. shown in The degree of skin irritation was calculated by the following <Calculation 2>.

<Calculation 2>

<Table 6>

As shown in Table 6, it was confirmed that the cosmetic composition containing the active ingredients of the porous polymer extract, hot water extract and ultrasonic extract had very low skin irritation.

[Experimental Example 5] Moisturizing effect test

The moisturizing effect was tested using samples prepared through [Example 2], [Comparative Example 3] and [Comparative Example 4].

Ten healthy adult males and females were constituted as test subjects under conditions of maintaining constant temperature and humidity. The forearms of the subjects were thoroughly washed and naturally dried for 10 minutes, and then the skin moisture content of the subjects was measured after 1 hour, 2 hours, 4 hours, and 8 hours. The measurement was measured using a corneometer CM 820 (Corage Khazaka Electronic GmbH, Germany), a moisture meter. The results are shown in <Table 7> below.

<Table 7>

As shown in <Table 7>, it was found that the moisturizing effect was excellent in the case of [Example 2].

The above detailed description should not be construed as limiting in all respects and should be considered illustrative. The scope of the present invention should be determined by reasonable interpretation of the appended claims, and all changes within the equivalent scope of the present invention are included in the scope of the present invention.

S100: Polymer polysaccharide extraction method

Claims (5)

Obtaining an extract after adding purified water to yam, okra, lotus root, aloe vera leaf and porous polymer;
filtering the extract;
obtaining an eluate by eluting the polymeric polysaccharide adsorbed on the porous polymer taken in the filtering step; and
Freeze-drying the eluate to obtain a polymeric polysaccharide in the form of a dry powder;
The porous polymer,
Method for extracting polymeric polysaccharide from yam, okra, lotus root and aloe vera leaves using a porous polymer characterized in that it is PMMA (Polymethyl Methacrylate) having a pore size of 10 nm to 30 nm. According to claim 1,
The filtering step is
After removing impurities from the extract and collecting the filtrate in which the porous polymer is mixed, the porous polymer is taken using a filter paper. From yam, okra, lotus root and aloe vera leaves using a porous polymer Polymeric polysaccharide extraction method. After adding purified water to yam, okra, lotus root, aloe vera leaf and porous polymer, obtaining an extract;
filtering the extract;
obtaining an eluate by eluting the polymeric polysaccharide adsorbed on the porous polymer taken in the filtering step;
Freeze-drying the eluate to obtain a dry powder; and
Preparing a cosmetic composition containing the polymeric polysaccharide in the form of a dry powder as an active ingredient;
The porous polymer,
Method for producing a cosmetic composition containing polymer polysaccharide extracted from yam, okra, lotus root and aloe vera leaf as an active ingredient using a porous polymer characterized in that it is PMMA (Polymethyl Methacrylate) having a pore size of 10 nm to 30 nm. According to claim 3,
The manufacturing step is
A method for producing a cosmetic composition comprising a polymeric polysaccharide extracted from yam, okra, lotus root and aloe vera leaf as an active ingredient using a porous polymer, characterized in that for preparing a cosmetic composition of oil-in-water emulsion type cream formulation. According to claim 3,
The manufacturing step is
A method for producing a cosmetic composition containing a polymeric polysaccharide extracted from yam, okra, lotus root and aloe vera leaf as an active ingredient using a porous polymer, characterized in that for preparing a cosmetic composition for moisturizing skin and relieving skin inflammation.

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