KR101814268B1 - Composition with Antibacterial Effect on Oral Bacteria Containing Purified Extract of Red Ginseng Ethanol Extract - Google Patents
Composition with Antibacterial Effect on Oral Bacteria Containing Purified Extract of Red Ginseng Ethanol Extract Download PDFInfo
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Abstract
The present invention relates to a composition for inhibiting oral bacteria including a column-purified fraction of a red ginseng ethanol extract, wherein the column-purified fraction of an ethanol extract of red ginseng has an antibacterial effect against S. mutans and F. nucleatum which cause tooth decay and periodontal disease, respectively The composition of the present invention can be usefully used as an effective ingredient in a composition or food for preventing or treating periodontal disease or tooth decay.
Description
The present invention relates to a composition for inhibiting oral bacteria, which comprises a column-purified fraction of red ginseng ethanol extract.
Periodontal disease and tooth decay are caused by bacteria that collectively grow in the gum area. If periodontal disease or tooth decay occurs, it can cause not only damage of the teeth but also severe pain and bad breath, and if the food intake is difficult due to the pain, the immunity can be lowered because it can not receive proper nutrition supply.
Periodontal disease is caused by inflammation of the gingiva called "gingivitis", which is a growing area of the teeth where gingival tissue meets. Gingivitis is characterized by easily reddened, bleeding gums, and is known to be the leading cause of tooth loss. As a bacterium that causes periodontal disease, Fusobacterium nucleatum is known.
Dental caries is also referred to as dental caries and is classified into oral diseases caused by the interaction of germs, foods, and saliva in the teeth, and carbohydrates such as sugar or starch of food residues attached to the teeth by fermentation of bacteria resident in the oral cavity The lactic acid that is generated by the decomposition causes the hard tissue lime of the teeth to be dislodged. Among the microorganisms in the oral cavity, Streptococcus mutans is known as a cause of tooth decay. Glucan is synthesized in the metabolic process, and the glucan increases the binding between the bacteria that grow on the tooth surface and induces the tooth decay.
In order to prevent such periodontal disease or tooth decay, sodium fluoride, benzethonium chloride, polyoxyethylene, polyoxypropylene polymer, cetylpyridinium chloride, sodium saccharin, sodium benzoate, and triclosan having antibacterial activity are used. However, swallowing of these ingredients may cause side effects such as vomiting, nervous breakdown, coma, and diarrhea. Excessive swallowing may cause fatal injury and damage to the oral cavity during long-term use. In particular, the use of triclosan has been reported to increase the resistance of bacteria to antibiotics, and there is a great concern about the abuse of triclosan acid.
On the other hand, the physiological activity of ginseng is due to saponin, polysaccharides and peptides, and many studies have been conducted centering on ginsenoside. Thus, the central nervous system control, memory, improvement of learning efficacy and antistress action, As the various pharmacological actions such as proliferation and transfer inhibition action, antioxidative action, antidiabetic action, liver function improvement, radiation protection action and inhibition of thrombogenesis have been scientifically proved, the demand for medicines and functional supplements Is increasing. Related to the inhibition of oral bacteria by using ginseng, Patent Document 1 discloses compositions for oral anti-inflammatory, antibacterial and deodorant containing red ginseng extract and natural plant-derived extract.
The present inventors have studied a raw material which shows an excellent antibacterial effect similar to the antimicrobial agent conventionally used for oral bacteria but does not cause resistance to bacteria, and the purified fraction of red ginseng extract has an excellent antibacterial effect against oral bacteria The present invention has been completed.
It is an object of the present invention to provide a composition for inhibiting oral bacteria, which comprises a column-purified fraction of red ginseng ethanol extract.
In order to solve the above-mentioned problems, the present invention provides a composition for inhibiting oral bacteria comprising a purified fraction of a column of ethanol extract of red ginseng.
The purified fraction of the column of the red ginseng ethanol extract of the present invention contains F. nucleatum , It can be used for the prevention or treatment of periodontal diseases or dental caries because it has an excellent antimicrobial effect against the cavity-inducing bacteria S. mutans .
Hereinafter, the present invention will be described in detail.
One aspect of the present invention provides a composition for inhibiting oral bacteria, which comprises a column-purified fraction of red ginseng ethanol extract.
The extract may be obtained by extracting from a raw material or a dried product thereof, and the raw material of the extract may be cultivated or commercially available.
When the extract is obtained by extracting from the raw material, any conventional extraction methods known in the art such as a solvent extraction method, an ultrasonic extraction method, a filtration method and a reflux extraction method may be used as the extraction method. Preferably, the extraction method or the reflux extraction method can do. The extraction process can be repeated several times, and then the step such as concentration or freeze-drying can be further roughened. Specifically, the obtained extract is concentrated under reduced pressure to obtain a concentrated liquid, the concentrated liquid is lyophilized, and a concentrated powder can be produced using a pulverizer.
The aqueous ethanol solution may be 50 to 100% (v / v), preferably 55 to 95% (v / v), more preferably 60 to 95% (v / v) But not limited to this. When the concentration of ethanol aqueous solution is less than 50% (v / v), the killing effect is relatively low for the dental caries-inducing bacteria or periodontal bacteria.
A filler such as silica gel, activated alumina, synthetic polymer, magnesium silicate, activated carbon, cellulose, ion exchange resin and the like may be used as the column used in the production of the purified fraction, and it is preferable that aromatic synthetic resin is used as a filler, The column may be an aromatic type synthetic adsorbent column, and more preferably, but not exclusively, a Diaion HP-20 synthetic adsorbent is used as a filler. The separation using the column can be carried out once to several times until the fraction of the desired purity is purified, and concentration and recrystallization can be carried out if necessary.
In the preparation of the purified fraction, the desorption solvent may be any one selected from the group consisting of water, organic solvents, and mixtures thereof. The organic solvent may be selected from the group consisting of alcohol, hexane (n-hexane), ether, glycerol, propylene glycol, butylene glycol, ethyl acetate, methyl acetate, dichloromethane, chloroform, ethyl acetate, benzene, acetone, acetonitrile, And the organic solvent may be any one selected from the group consisting of acetone, acetonitrile, C 1 to C 4 alcohols, and a mixed solvent thereof. More preferably, the organic solvent is selected from the group consisting of The organic solvent may be at least one of ethanol or methanol, but is not limited thereto.
When the desalting solvent is ethanol in the production of the purified fraction, the desorption solvent may be 70 to 100% (v / v) ethanol aqueous solution, preferably 75 to 99% (v / v) ethanol aqueous solution, (V / v) ethanol aqueous solution, and more preferably 85 to 95% (v / v) ethanol aqueous solution, but is not limited thereto. When the concentration of the aqueous ethanol solution used as the desorption solvent is less than the lower limit value, there is a problem that the obtained antibacterial effect of the oral bacteria is lowered in the obtained fraction.
When adding the aqueous ethanol solution, which is a desorption solvent in the production of the purified fraction, an aqueous solution of ethanol is flowed into the column, for example, it is pressurized and flowed.
In one embodiment of the present invention, a red ginseng ethanol extract is prepared by adding an aqueous 70% (v / v) ethanol solution and the obtained extract is adsorbed on an HP-20 column. The adsorbed components on the column are washed with various concentrations of ethanol When fractions were obtained by desorption using a solvent (30, 50, 70, 85, 95% (v / v)), the fraction obtained using a 95% (v / v) aqueous ethanol solution as a desorbing solvent, Concentrate, 50% (v / v) ethanol aqueous solution as a desorption solvent, and an antimicrobial effect against S. mutans and F. nucleatum more than the propolis (see Tables 1 to 3).
In another embodiment of the present invention, a red ginseng ethanol extract is prepared by adding an aqueous 70% (v / v) ethanol solution and the obtained extract is adsorbed on an HP-20 column. The adsorbed components on the column are washed with methanol When fractions were obtained by desorption using a solvent (30, 50, 70, 85, 95% (v / v)), the fraction obtained using a 95% (v / v) aqueous methanol solution as a desorbing solvent, Concentrate, 50% (v / v) methanol aqueous solution as a desorption solvent, and an antimicrobial effect against S. mutans and F. nucleatum than propolis (see Tables 4 to 6).
The oral bacteria may be one which causes periodontal disease or cavities. For example, the oral bacteria may be selected from the group consisting of Streptococcus mutans , S. sanguinis , S. sobrinus , Streptococcus latifi , Streptococcus mutans , Wherein the microorganism is selected from the group consisting of S. criceti , S. anginosus , S. gordonii , and F. nucleatum . Preferably, the oral germ can be at least one of S. mutans and F. nucleatum , and more preferably S. mutans , but is not limited thereto.
The purified fraction of the red ginseng ethanol extract may be contained in an amount of 0.01 to 30% by weight, preferably 0.01 to 20% by weight, more preferably 0.01 to 10% by weight in the composition for inhibiting oral bacteria , And even more preferably from 0.01 to 5 wt%. If the content is less than 0.01% by weight, the oral bacteria-killing effect may be insignificant. If the content is more than 30% by weight, the oral bacteria-killing effect may be relatively low compared to the input concentration.
The composition for oral anticancer composition of the present invention may be any one selected from the group consisting of toothpaste, oral cleanser, aerosol spray, chewing gum, lozenge, and topical composition, but is not limited thereto.
The compositions of the present invention may be formulated in a variety of ways, for example, as liquids, dry masses, toothpastes, mouthwashes, liquid suspensions, topical preparations, powdered food supplements, pastes, gels, emulsions, aerosol sprays, capsules, Food, wafers, lozenges, chewing gum, and the like. Other formulations may be used without limitation as are known to those of ordinary skill in the art. The compositions of the present invention may include nutritional supplement components and may include any of a wide variety of well known nutritional supplements including vitamins, minerals, essential and non-essential amino acids, carbohydrates, lipids, foods, dietary supplements and the like.
In addition, the oral antimicrobial composition of the present invention can be formulated into a temperature-sensitive sol-gel phase transition oral formulation. The temperature-sensitive sol-gel phase transition form is a state in which the polymer dissolves in water to dissolve in water to form a sol state, and when it exceeds a certain critical temperature, the solubility of the polymer rapidly decreases to become a gel state and solidify. Lt; RTI ID = 0.0 > reversible < / RTI > Therefore, the temperature-sensitive sol-gel phase transition oral formulation is in the form of a sol-like liquid before application to the oral cavity (room temperature), and is applied to the oral cavity (body temperature) and then transferred to the gel state, The increased adhesion to teeth, tongue, and gum surface has the advantage of allowing the active ingredient to exert long-lasting effects in the oral cavity. In addition, when it is rinsed with cold water, it is transferred to a sol state again in a liquid phase and can be easily washed out in the oral cavity.
The composition for inhibiting oral bacteria of the present invention may be combined with a component that is usually added to oral gavages such as toothpastes. These components may include, but are not limited to, any one selected from the group consisting of abrasives, wetting agents, binders, surfactants, flavoring agents and thickening agents.
Examples of the abrasive agent include silica gel, silica precipitate, dicalcium phosphate, dicalcium phosphate dihydrate, alumina trihydrate, calcium pyrophosphate, calcium carbonate and insoluble sodium metaphosphate. Examples of the wetting agent include sorbitol, glycerin, and polyethylene glycol. Examples of the thickener include silica airgel, pyrogenic silica, silica precipitate, carboxymethyl cellulose, carboxyvinyl polymer, xanthan gum and carrageenan. Examples of the surfactant include sodium lauryl sulfate and dodecylbenzenesulfonate. The flavoring agent can be used without limitation as long as it provides a palatable flavor or another refreshing flavor such as cinnamon which is usually used.
When the composition for oral antimicrobial control of the present invention is in a liquid form, i.e., an oral cleanser, these oral cleansers may be in the form of an aqueous, injectable emulsion. Oral cleansing agents may include flavorants, surfactants that maintain flavor in suitable dispersants, wetting agents, thickeners, and actives. Surfactants may also be included to provide foaming.
Hereinafter, the present invention will be described in detail with reference to examples.
However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention by the following production examples, examples and experimental examples.
Production of red ginseng ethanol extract and purified fraction thereof
[1-1] Preparation of Ethanol Extract of Red Ginseng
To 1 kg of red ginseng, 5.5% of 70% (v / v) ethanol was added and extracted once at 50 ° C for 10 hours. The extract was concentrated to a concentration of 50 brix or more using a reduced pressure concentrator, and residual ethanol was kept at 1% or less. Using a vacuum concentrator, more than 50 g of concentrated red ginseng ethanol extract was obtained.
[1-2] Preparation of purified fraction of red ginseng ethanol extract (using ethanol as desorption solvent)
In order to purify the hydrophobic fraction in the red ginseng ethanol extract concentrate prepared in the above Example [1-1], purified water was diluted by adding 3 times of purified water to the red ginseng ethanol extract concentrate, and 150 g of the diluted solution was added to HP-20 ion exchange resin (Mitsubishi Chemical Industry) Was passed through a column packed with 100 g. The column was washed with purified water corresponding to 5 times of the diluted solution, and then the column was subjected to various concentrations of aqueous ethanol solution (30, 50, 70, 85, 95% (v / v) The efficacious component fractions were recovered. After the solvent of the obtained fractions was completely evaporated, purified water was added to the fractions, and the mixture was frozen at -70 ° C, and then placed in a freeze dryer and dried to obtain a solid content according to each desorption solvent used.
[1-3] Preparation of purified fraction of red ginseng ethanol extract (using methanol as desorption solvent)
A solid fraction of methanol fraction was obtained in the same manner as in Example 1-2 except that an aqueous methanol solution (30, 50, 70, 85, 95% (v / v)) was used as a desorption solvent.
[1-4] Preparation of butanol fraction of red ginseng ethanol extract
50 g of the red ginseng ethanol extract concentrate prepared in the above Example [1-1] was diluted in 100 g of purified water and stored at room temperature for 24 hours. The diluted concentrate was mixed with butanol at a ratio of 3: 2. The mixture of the diluted concentrate and the solvent was put in a fraction funnel and separated at room temperature for 24 hours. The supernatant of the separated layer was recovered and the solvent was completely evaporated. Then, purified water was added thereto, and the resultant was frozen at -70 ° C., then placed in a freeze dryer for 72 hours and dried to obtain a butanol fraction.
Comparison of antimicrobial effects against oral bacteria when ethanol was used as a desorption solvent in the preparation of purified fractions
[2-1] Streptococcus mutans ( Streptococcus mutans ) Compared with triclosan as a positive control.
The red ginseng ethanol extract obtained in Example 1, the HP-20 column fraction of red ginseng ethanol extract (fraction obtained by using ethanol as the desorption solvent of Example [1-2]) and the oral bacteria of the butanol fraction of red ginseng ethanol extract The killing effect was confirmed. The solid content of each extract or fraction obtained in Example 1 was dissolved in purified water at a concentration of 50 to 6.25 mg / ml, and the antibacterial activity against S. mutans as a cavity-inducing microorganism was confirmed by paper disk method. S. mutans (KCTC 3065, Microbiology Resource Center, Daejeon) were used in the enrichment culture of Brain Heart Infusion Agar. The prepared S. mutans broth on a plate medium with the same medium as the culture medium was inoculated evenly with a sterilized cotton swab. Clear zones, a range in which oral bacteria did not grow, were measured and the effects of each sample were compared. As a positive control, triclosan (3 mg / ml) used as an antimicrobial agent was used. The experimental results are shown in Table 1 below.
concentrate
30% ethanol fraction
50% ethanol fraction
70% ethanol fraction
85% ethanol fraction
95% ethanol fraction
As a result, it was fractionated with 30% (v / v) and 50% (v / v) ethanol in the red ginseng ethanol extract concentrate, the butanol fraction of the red ginseng ethanol extract concentrate and the purified fraction of the red ginseng ethanol extract concentrate among the extracts obtained in Example 1 The obtained fractions had no antibacterial effect on S. mutans . On the other hand, the fraction obtained by fractionation of ethanol with 70% (v / v) or more of the purified fraction of red ginseng ethanol extract concentrate showed antimicrobial effect against S. mutans , and the fraction of ethanol extract of red ginseng was 95% v) ethanol showed 50% higher antimicrobial effect than the positive control triclosan.
[2-2] S. mutans Compared with propolis as a positive control group
The antimicrobial activity of the 95% (v / v) ethanol fraction of the red ginseng ethanol extract, which showed antimicrobial effect similar to that of the positive control triclosan, against S. mutans in Example 2-1, Respectively. Propolis was used as a water soluble component and a lipid soluble component, respectively, and they were all purchased from Seoul Propolis (Daejeon). Table 2 shows the clear zones for each sample.
Water-soluble component
Lipophilic component
95% ethanol fraction
As a result, the ethanolic fraction of 95% (v / v) of the red ginseng ethanol extract of the present invention showed an excellent antibacterial effect compared to propolis at a sample concentration of 12.5 mg / ml to 50 mg / ml. Therefore, a 95% (v / v) ethanol fraction of the red ginseng ethanol extract can be usefully used as an antibacterial agent for oral bacteria.
[2-3] Peptide Bacterium Nuclearum ( Fusobacterium nucleatum ) Compared with triclosan as a positive control.
The ethanolic fraction of 85% (v / v) and 95% (v / v) of the red ginseng ethanol extract of the present invention exhibited an antibacterial effect against F. nucleatum (periodontal bacteria) at a sample concentration of 6.25 mg / . The antimicrobial effect was evaluated in the same manner as in the above Example [2-1] except that the strains used were different. Reinforced Clostridial Medium was used for enrichment culture of F. nucleatum (KCTC 3314, Microbiology Resource Center, Daejeon). The results for F. nucleatum are shown in Table 3.
concentrate
30% ethanol fraction
50% ethanol fraction
70% ethanol fraction
85% ethanol fraction
95% ethanol fraction
As a result of the antimicrobial effect evaluation, F. nucleatum was fractionated with 70% (v / v), 85% (v / v) and 95% (v / v) ethanol after the red ginseng ethanol extraction. The antimicrobial effect was shown in 70% (v / v) ethanol fraction after red ginseng ethanol extraction, the antimicrobial effect was improved in the 85% (v / v) ethanol fraction and the best antimicrobial effect was obtained in the 95% (v / v) ethanol fraction Respectively. F. nucleatum showed more than 40% of the antimicrobial activity of the positive control triclosan when treated with 85% (v / v) or 95% (v / v) ethanol fraction of red ginseng ethanol extract.
On the other hand, the embodiment as compared with those of [2-1], when processing the ginseng ethanol extract concentrate to the same concentration to the bacteria causing dental caries in S. mutans, F. nucleatum bacteria causing periodontitis, antimicrobial effect on S. mutans , But the antimicrobial effect was observed when treated with F. nucleatum . From these results, it can be seen that the antibacterial effect is different depending on the type of the strain even when the same sample is treated with the carotid bacterium, S. mutans , and F. nucleatum , a periodontitis inducing bacterium.
Comparison of antimicrobial effects against oral bacteria when methanol was used as a desorption solvent in the preparation of purified fractions
[3-1] S. mutans Against Triclosan as a Positive Control
The red ginseng ethanol extract obtained in Example 1, the HP-20 column fraction of red ginseng ethanol extract (fraction obtained by using methanol of Example [1-3] as the desorption solvent), and the oral bacteria of the butanol fraction of red ginseng ethanol extract The killing effect was confirmed. The same procedure as in Example [2-1] was carried out except that the samples used for confirmation of the effect were different. The experimental results are shown in Table 4 below.
concentrate
30% methanol fraction
50% methanol fraction
70% methanol fraction
85% methanol fraction
95% methanol fraction
As a result, the extracts obtained in Example 1 were fractionated with 30% (v / v) and 50% (v / v) methanol in the red ginseng ethanol extract concentrate, the butanol fraction of the red ginseng ethanol extract concentrate and the purified fraction of the red ginseng ethanol extract concentrate The obtained fractions had no antibacterial effect on S. mutans . On the other hand, the fraction obtained by methanol fractionation of 70% (v / v) or more of the purified fraction of the red ginseng ethanol extract concentrate showed antimicrobial effect against S. mutans and the extract fraction of red ginseng ethanol was 95% v) The fraction recovered with methanol showed 50% higher antimicrobial effect than the positive control triclosan.
[3-2] S. mutans Compared with propolis as a positive control group
In Example 3-1, the antimicrobial effect of the 95% (v / v) methanol fraction of the ethanol extract of red ginseng showed an antibacterial effect similar to that of triclosan, which is a positive control against S. mutans , Respectively. Propolis was used as a water soluble component and a lipid soluble component, respectively, and they were all purchased from Seoul Propolis (Daejeon). Table 5 shows the clear zones for each sample.
Water-soluble component
Lipophilic component
95% methanol fraction
As a result, the 95% (v / v) methanol fraction of the red ginseng ethanol extract of the present invention exhibited an excellent antimicrobial effect over propolis in the sample concentration range of 12.5 mg / ml to 50 mg / ml. Therefore, a 95% (v / v) methanol fraction of the red ginseng ethanol extract can be usefully used as an antibacterial agent for oral bacteria.
[3-3] F. nucleatum Against Triclosan as a Positive Control
The 85% (v / v) and 95% (v / v) methanol fractions of the red ginseng ethanol extract of the present invention exhibited an antibacterial effect against F. nucleatum (periodontal bacteria) at a sample concentration of 6.25 mg / . The antibacterial effect was evaluated in the same manner as in the above Example [2-1], except that the strains and samples used were different. Reinforced Clostridial Medium was used for enrichment culture of F. nucleatum (KCTC 3314, Microbiology Resource Center, Daejeon). The results for F. nucleatum are shown in Table 6.
concentrate
30% methanol fraction
50% methanol fraction
70% methanol fraction
85% methanol fraction
95% methanol fraction
As a result of the antimicrobial effect evaluation, F. nucleatum was fractionated with 70% (v / v), 85% (v / v) and 95% (v / v) methanol fractions after extracting red ginseng ethanol. The antimicrobial effect was obtained from the 70% (v / v) methanol fraction after red ginseng ethanol extraction, the antimicrobial effect was improved from the 85% (v / v) methanol fraction and the most effective from the 95% (v / v) methanol fraction Respectively. F. nucleatum showed more than 40% of the antimicrobial effect of the positive control triclosan in the fraction of methanol extract of 85% (v / v) or 95% (v / v) of ethanol extract of red ginseng.
Claims (11)
Wherein the column is a column comprising a synthetic adsorbent of aromatic type,
Wherein the desorbing solvent is 70 to 100% (v / v) methanol or ethanol aqueous solution for preparing the purified fraction.
Wherein the ethanol for preparing the red ginseng ethanol extract is an aqueous 50 to 100% (v / v) ethanol solution.
Wherein the desalting solvent is 80 to 97% (v / v) methanol or ethanol aqueous solution for preparing the purified fraction.
The oral bacteria may be selected from the group consisting of Streptococcus mutans , Streptococcus sanguinis , Streptococcus sobrinus , Streptococcus ratti , Streptococcus criceti , Wherein the composition is at least one selected from the group consisting of Streptococcus anginosus , Streptococcus gordonii , and Fusobacterium nucleatum .
Wherein the oral bacterium is Streptococcus mutans .
Wherein the oral bacterium is Fusobacterium nucleatum .
Wherein the composition for oral-bacterial-inhibiting composition is any one selected from the group consisting of a toothpaste, a mouthwash, an aerosol spray, a chewing gum, a lozenge, and a composition for topical use.
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| KR101965983B1 (en) * | 2017-12-04 | 2019-04-04 | 주식회사 한국인삼공사 | Antibiotic composition containing panaxynol or panaxydol |
| CN113749969A (en) * | 2020-06-01 | 2021-12-07 | 美慕(北京)科技有限公司 | Oral care composition and preparation with oral micro-ecological balance maintaining and dental plaque reducing effects and preparation method thereof |
| KR102547208B1 (en) * | 2021-06-10 | 2023-06-26 | 주식회사 한국인삼공사 | A Composition For Enhancing Skin Immunity Comprising Column-purified Fraction Of Ginseng Extract |
| KR102452255B1 (en) | 2022-02-11 | 2022-10-07 | (주)엔에스티바이오 | Composition comprising deacetyl asperulosidic acid and/or asperulosidic acid as active ingredient for prevention or treatment of oral disease |
| KR102438800B1 (en) | 2022-02-11 | 2022-08-31 | (주)엔에스티바이오 | Composition comtaining lactic acid fermented noni extract as active ingredient for prevention or treatment of oral disease |
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