Manufacturing method of bamboo multi-complex with antibacterial properties

Abstract

The present invention relates to an antibacterial and cosmetic composition containing a bamboo multi-complex, and in particular to an antibacterial, anti-inflammatory and cosmetic composition containing a bamboo shoot skin multi-complex, and relates to an antibacterial, anti-inflammatory and cosmetic composition containing bamboo shoot skin, bamboo sludge, elm root, Houttuynia cordata, licorice, young trifoliate orange, celandine, Japanese guar root, Japanese guar leaf, Centella asiatica, Sanbaigou, fermented green tea and bamboo extract.

Description

[Technical field]

[0001] The present invention relates to antibacterial, anti-inflammatory and cosmetic compositions containing a bamboo multi-complex, and in particular to a method for producing a bamboo multi-complex containing bamboo shoot bark extract, bamboo extract, elm root, Gyoseiso, licorice, Citrus aurantium (young trifoliate orange), Celandine, Hariguwa root, Hariguwa leaf, Centella asiatica, Sanbaigou, and fermented green tea extract, and a cosmetic composition using the same. [Background technology]

[0002] Currently, many types of antibacterial products are commercially available and are widely used for the prevention and treatment of diseases.

[0003] Antibacterial agents can be divided into antifungal agents and antibacterial agents based on their antibacterial power, and antifungal agents have the function of inhibiting the growth of or killing higher organisms that have fungal cells, such as mold. Also, antibacterial agents refer to antibiotics that have the function of inhibiting the growth of or killing lower organisms that have prokaryotic cells, such as bacteria. To date, antibacterial agents (antifungal and antibacterial agents) have mainly been obtained from mold or chemically synthesized.

[0004] These antifungal agents include, for example, Amphotericin B, Clotrimazole, Crinipan AD, Fluconazole, Griseofulvin, Haloprogin, Hexachlorophene, Ichthyol, Itraconazole, Ketoconazole, Myconazole, Nystatin (Mycostatin), Pimaricin (Natamycin), Terbinafine, and the like. Examples of antibacterial agents include adapalene gel, benzoyl peroxide, fusidic acid, resorcinol, retinoic acid, salicylic acid, sulfonamides, tetracycline, triclosan, and penicillin.

[0005] However, these synthetic antifungal and antibacterial agents have high manufacturing costs, are difficult to purify, and have side effects such as increased microbial resistance when used in commercialized products. In addition, because their cosmetic properties have not been considered, they can cause undesirable skin problems (contact dermatitis) and side effects when applied to the skin as external preparations (ointments, etc.). For this reason, it is known that many researchers are currently developing antibiotics that have the desired antifungal and antibacterial properties without side effects. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] KR10-0465831 B [Patent Document 2] KR10-2009-0004295 A Summary of the Invention [Problem to be solved by the invention]

[0007] In order to solve the above-mentioned problems, the present invention aims to provide an antibacterial and cosmetic composition containing a bamboo multi-complex, and in particular to provide an antibacterial, anti-inflammatory and cosmetic composition containing a bamboo shoot skin multi-complex.

[0008] Another object of the present invention is to provide a method for producing a bamboo multi-complex containing bamboo shoot bark extract, bamboo extract, elm root, Gyoseiso, licorice, Citrus aurantium (young trifoliate orange), Celandine, Hariguwa root, Hariguwa leaf, Centella asiatica, Sanbaigou, and fermented green tea extract. [Means for solving the problem]

[0009] In order to achieve the above object, the present invention The method for producing a bamboo multi-complex with antibacterial properties includes the steps of: heating bamboo, cooling the gas generated when the temperature of the heating furnace is 90°C to 280°C, storing the vinegar liquid obtained, and leaving it to stand for more than three years to produce a bamboo extract; steaming bamboo shoot skins at 80°C to 120°C, drying at low temperature at a temperature of 15°C to 80°C for 20 to 80 hours, and extracting it to produce a bamboo shoot skin extract; mixing sugar and citronella in a ratio of 0.5:1, maturing for more than three months, and extracting it to produce a saccharified citronella extract; extracting under reduced pressure, and concentrating it under reduced pressure to produce a plant extract; and preparing a bamboo multi-complex containing the bamboo extract, the bamboo shoot skin extract, the saccharified citronella extract, and the plant extract.

[0010] The plant extract is at least one of bamboo extract, elm root extract, ginseng extract, licorice extract, celandine extract, scutellaria root extract, scutellaria leaf extract, Centella asiatica extract, Sanbai grass extract, and fermented green tea extract. It is characterized by:

[0011] The bamboo shoot skin extract is dried at low temperature. 、3 The alcohol extract is characterized by being obtained by adding alcohol at a concentration of 80% to 50% (v / v) as a solvent and extracting at a temperature of 80°C to 100°C for 3 to 5 hours. Effect of the Invention

[0019] As described above, according to the present invention, a bamboo multi-complex containing bamboo shoot bark extract, bamboo extract, elm root, Gyokuseisou, licorice, Citrus aurantium (young trifoliate orange), Celandine, Hariguwa root, Hariguwa leaf, Centella asiatica, Sanbaigou, and fermented green tea extract can be produced to provide an antibacterial composition and a cosmetic composition.

[0020] The present invention also provides a cosmetic composition containing a bamboo extract, particularly a bamboo shoot bark extract, which can be used in antibacterial and antifungal cosmetic products. [Brief description of the drawings]

[0021] [Figure 1] The results of evaluating the cytotoxicity of a 5% alcohol extract of bamboo shoot skin are shown. BEST MODE FOR CARRYING OUT THEINVENTION

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention belongs. Generally, the nomenclature used herein is that which is well known and commonly used in the art.

[0023] The term "about" refers to an amount, level, value, number, frequency, percent, dimension, size, amount, weight or length that varies by about 30, 25, 20, 25, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% relative to a reference amount, level, value, number, frequency, percent, dimension, size, amount, weight or length. Throughout this specification, unless otherwise required by context, the terms "comprising" and "comprises" should be understood to encompass the inclusion of the stated step or component, or group of steps or components, but not the exclusion of any other step or component, or group of steps or components.

[0024] In the present invention, the term "bamboo shoots" refers to all bamboos taxonomically classified as bamboos, i.e., bamboos belonging to the Poaceae family of the Poaceae order, regardless of the type of bamboo from which they are obtained. Regardless of the type of bamboo, bamboo shoots obtained from bamboos belonging to the genus Phyllostachys, bamboo shoots obtained from bamboos belonging to the genus Arundinaria, bamboo shoots obtained from bamboos belonging to the genus Sasa, or bamboos belonging to the genus Pseudosasa can all be used in the present invention.

[0025] The present invention will be specifically described below. The bamboo multi-complex of the present invention is characterized by containing bamboo extract, bamboo skin extract, saccharified citron fruit extract, and reduced pressure concentrated plant extract, which are obtained by distilling the vinegar obtained by cooling the gas generated during the bamboo pyrolysis process at a furnace temperature of 90°C to 280°C and leaving it to stand for more than three years. The bamboo multi-complex of the present invention contains bamboo skin extract, bamboo extract, elm root extract, Gyokuseisou extract, licorice extract, Citron fruit (young trifoliate orange) extract, Celandine extract, Hariguwa root extract, Hariguwa leaf extract, and Centella asiatica fermented green tea extract.

[0026] The method for producing the bamboo extract includes a bamboo drying step (S110), a heat extraction step (S120), a bamboo sap maturing step (S130), and a bamboo sap filtering step (S140).

[0027] The bamboo drying step (S110) is a step of cutting the bamboo to a length suitable for drying and heating. The heating and extraction step (S120) is a step of heating the bamboo and obtaining the extract emitted from the heated bamboo. Here, the bamboo is heated by indirect heating, and the vinegar obtained by cooling the gas generated when the temperature of the heating furnace is 90 to 280°C is stored. The bamboo vinegar ripening step (S130) is a step of naturally ripening and oxidizing the produced vinegar for at least three years. The bamboo vinegar filtration step (S140) is a step of performing precision filtration using a carbon filter or a medical filter.

[0028] The method for producing the bamboo shoot skin extract is characterized by using a raw material fermented at low temperature and extracting it with hot water. The method for producing the bamboo shoot skin extract uses raw bamboo shoots from which the skin has been removed. The method for producing the bamboo shoot skin extract includes a material preparation step (S210), a drying step (S220), and an extract separation step (S230).

[0029] In the material preparation step (S210), the bamboo shoot skin is washed, cut, dried or crushed according to a conventional method. At this time, washing, cutting and drying can be performed in that order, and after cutting, sterilization is performed before the drying step (S220), and steaming treatment is performed at 80 to 120°C to adjust the functionality of the raw material. The steaming treatment can be performed in a steamer to neutralize the effect of oxalic acid, a pungent substance that gives the bamboo shoots their bitter taste. The steaming treatment includes treatment in a steamer for 5 to 10 minutes.

[0030] The drying step (S220) can be carried out by any one of air drying, hot air drying, low temperature drying, and freeze drying. In the case of the steamed material, it includes low temperature drying at a temperature of 15 to 80° C. for 20 to 80 hours.

[0031] In the extract separation step (S230), the method for separating the bamboo shoot bark extract is not particularly limited, and may be a method commonly used in the art for producing an extract. For example, a method using an extraction device such as hot water extraction, alcohol extraction, immersion extraction, reflux cooling extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction, ultrasonic extraction, or a method using an adsorption resin including XAD and HP-20 may be used, but is not limited thereto. Preferably, the extract is obtained by treating the raw material with an extraction solvent, extracting by heating, and concentrating under reduced pressure.

[0032] A polar solvent can be used as the extraction solvent. Examples of the polar solvent include, but are not limited to, at least one selected from the group consisting of water, alcohol, acetic acid, DMFO (dimethyl-formamide) and DMSO (dimethyl sulfoxide). Examples of the alcohol include, but are not limited to, at least one selected from the group consisting of methanol, ethanol, propanol, butanol, normal propanol, isopropanol, normal butanol, 1-pentanol, 2-butoxyethanol and ethylene glycol. Preferably, water or alcohol is used as the polar solvent in consideration of the stability to the human body and the effect when applied to the skin. The alcohol can be diluted alcohol mixed with water. Preferably, alcohol at a concentration of 3 to 50% (v / v) is used as the polar solvent in order to maximize the antioxidant and moisturizing effects of the composition on the skin, and more preferably, alcohol at a concentration of 3 to 10% (v / v) is used in order to maximize the effectiveness of the bamboo shoot skin extract to be separated. When bamboo shoot skin is used as the extraction raw material, in order to improve the extraction yield and increase productivity, it is preferable to use alcohol with a concentration of 3 to 30% (v / v) as the polar solvent, and in consideration of both the extraction yield and the application effect on the skin, it is preferable to use alcohol with a concentration of 3 to 10% (v / v).Preferably, the alcohol is proethanol.

[0033] The saccharified betel nut extract is a saccharified extract obtained by mixing sugar and the original material in a ratio of 0.5:1, aging the mixture for more than three months, and then extracting the mixture.

[0034] The plant extract concentrated under reduced pressure is characterized in that it is concentrated after extraction under reduced pressure.

[0035] In order to aid in the understanding of the present invention, the present invention will be described in more detail below based on examples. However, these examples are merely illustrative of the present invention and do not limit the scope of the appended claims. It is clear to those skilled in the art that various changes and modifications can be made to these examples within the scope of the scope and technical ideas of the present invention, and it is natural that these changes and modifications fall within the scope of the appended claims.

[0036] Test 1 (cytotoxicity test) - 5% alcohol extract of bamboo shoot skin The extraction solvent used was 5% (v / v) fermented alcohol. The bamboo shoots were chopped and dried, and the extractant was treated with 20 times the weight of the raw material. The extractant was then extracted at 100°C for 4 hours, and concentrated in a concentrator to a Brix of 30 or more. The extract was then freeze-dried to obtain a powdered extract with a yield of 22%.

[0037] FIG. 1 shows the results of evaluating the cytotoxicity of a 5% alcohol extract of bamboo shoot skin.

[0038] Furthermore, Table 1 below shows that a cytotoxicity test (MTT assay) was performed using CCD-986SK (Fibroblast) cells to evaluate the cytotoxicity of the 5% alcohol extract of bamboo shoot skin.

[0039] [Table 1]

[0040] For cell dispensing, add 3 x 10 cells to a 96-well plate. 4 (100 μL / well) and incubated at 37°C in a 5% CO2 incubator for 24 hours. For treatment with test substances, the medium was replaced with serum-free medium in which the test substances were diluted according to concentration, and incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0041] For the MTT assay, the plate was washed twice with PBS, and 200 μL of MTT solution (0.5 mg / mL, serum-free medium) was dispensed into each well, followed by incubation at 37°C in a 5% CO2 incubator for 3 hours.

[0042] For absorbance measurements, the medium was removed after incubation, and 200 μL of DMSO solution was added to each well and extracted for 20 to 30 minutes on a plate shaker. The absorbance was then measured using ELISA at a wavelength of 570 nm.

[0043] In the results, if the cell viability was reduced to less than 70% of that of the sample-free group, it was determined that there was potential cytotoxicity. The cell viability was calculated using the following formula 1.

[0044]

number

[0045] The significance of the experimental results was confirmed by performing a t-test (paired t-test, two-sided verification) using the SPSS (Ver. 19) statistical program.

[0046] [Table 2]

[0047] As a result of the cytotoxicity test, in the case of 5% bamboo shoot skin alcohol extract, cell viabilities of 116.98±7.27%, 127.81±7.25%, 161.08±15.56%, 83.47±8.85%, and 45.47±6.46% were observed at concentrations of 0.1%, 0.2%, 0.5%, 1.0%, and .0%.

[0048] That is, the 5% bamboo shoot skin extract in alcoholic spirits showed a cell viability of 70% or more at a concentration of 1.0% or less. If the cell viability falls below 70% of the control group, it is judged to have potential cytotoxicity, so the 5% bamboo shoot skin extract in alcoholic spirits does not appear to have cytotoxicity at a concentration of 1.0% or less.

[0049] Unlike existing antibacterial agents (antifungal, antibacterial), the inventors extracted and purified the above complex extract, and applied it to skin trouble-causing bacteria (acne, athlete's foot, skin resident bacteria) and obtained significant results. This test was conducted by the Korea Chemical Fusion Testing and Research Institute (KTR), an official testing organization.

[0050] The vinegar liquid obtained by cooling the gas generated during the bamboo pyrolysis process at furnace temperatures between 90℃ and 280℃ was distilled, and the bamboo extract obtained by leaving it to stand for more than three years was used as a sample for testing.

[0051] The pathogenic microorganisms (skin trouble-inducing bacteria) used in the test are as follows. (1) Acne-causing bacteria (one type): Propionibacterium acnes (gram-positive) (2) Athlete's foot (ringworm)-causing fungus (one species): Trichophyton rubrum (3) Skin resident bacteria (2 types): Staphylococous aureus, Staphylococous epidermidis

[0052] Test 2 (Antibacterial and antifungal effects) For preculture of the test bacteria, Trichophyton rubrum strain was inoculated into Sabouraud dextrose agar and cultured at (25±1)℃ for 10-14 days. Propionibacterium acnes strain was inoculated into reinforced clostridial medium (RCM) broth and cultured at (35±1)℃ for 3 days.

[0053] The cultured bacterial solution was inoculated into fresh RCM broth and cultured under the same conditions as above, and this process was repeated once more. Anaerobic conditions were maintained throughout the entire process of the strain using sterile liquid paraffin.

[0054] To prepare the test fungus solution, 0.05% polysorbate 80 solution was dispensed onto the solid medium on which Trichophyton rubrum was grown, and the spores were removed using a spreader, after which the mycelium was removed by passing it through gauze. The spore solution thus prepared was diluted with (1.0-9.9) x 10 7 The cultured Propionibacterium acnes solution was adjusted to a concentration of 1.0 to 9.9 x 10 viable bacteria / mL with sterile saline and used as a test bacteria solution. 7 The test bacteria solution was diluted with sterile saline to a concentration of CFU / mL.

[0055] 0.2 mL of test bacteria solution was added to 20 mL of sample (concentration: undiluted solution) and mixed, then left at room temperature for 24 hours for Trichophyton rubrum and 2 hours for Propionibacterium acnes. The viable bacteria count was then measured to determine the reduction rate for each control group. The first dilution was performed using D / E neutralizing broth. The neutralized test solution was diluted stepwise and 1 mL was dispensed into two Petri dishes for each concentration. For Trichophyton rubrum, 15 to 25 mL of Sabouraud dextrose agar prepared in advance was dispensed into Petri dishes, and for Propionibacterium acnes, reinforced clostridial medium agar was used to solidify the mixture at room temperature. The solidified Trichophyton rubrum dish was cultured at (25±1)℃ for 10 to 14 days, and the Propionibacterium acnes Petri dish was cultured anaerobically at (35±1)℃ for 3 days. Initial and control viable counts were performed using sterile saline.

[0056] Table 3 shows the Trichophyton rubrum antifungal test results, and Table 4 shows the Propionibacterium acnes antibacterial test results.

[0057] [Table 3]

[0058] The initial inoculum size was 6.0×10 5 CFU / mL, control group bacterial count after 24 hours was 5.5 × 10 5 CFU / mL, and test group bacterial counts after 24 hours were observed to be <10 CFU / mL.

[0059] [Table 4]

[0060] The initial inoculum size was 5.7 x 10 5 CFU / mL, the number of bacteria in the control group after 2 hours was 5.0 × 10 5 CFU / mL, and the test group bacterial counts after 2 hours were observed to be <10 CFU / mL.

[0061] After incubation, observations were made on Petri dishes that showed (8-80) fungi and (30-300) bacteria. When viable bacteria were observed only at the lowest dilution stage, they were counted regardless of the observation range. When bacteria grew, the number of bacteria on the medium was multiplied by the dilution factor to calculate the number. When bacteria did not grow on the medium, the number was multiplied by the dilution factor performed at the neutralization stage and shown as "less than 10" (<10).

[0062] The viable cell count was calculated as N (viable cell count) = C (number of colonies, average number of colonies on two plates) x D (dilution factor).

[0063] The reduction rate (%) was measured as Log reduction (LR) = log(A)-log(B). (A: logarithm of viable bacteria count after a certain time in the control group, B: logarithm of viable bacteria count after a certain time in the test group)

[0064] [Table 5]

[0065] The test results showed that the antifungal test showed a score of >4.74 for Trichophyton rubrum and the antibacterial test showed a score of >4.70 for Propionibacterium acnes. A log reduction of 3 or more is 99.9%, and a log reduction of 4 or more is 99.99% or more.

[0066] Test 3 (Antibacterial effect) For pre-culturing the test bacteria, the test strain was collected from the slant medium, inoculated into tryptic soy broth, and cultured at 35±1°C for 18 to 24 hours. In preparing the test bacteria solution, the viable cell count is (1-9.9) x 10 8 The solution was diluted with sterile saline to give a CFU / mL and used as the test bacteria solution.

[0067] 0.2 mL of the test bacteria solution was added to 20 mL of the sample (undiluted solution) and mixed, and then the mixture was left to stand at (22±2)°C for 60 and 90 minutes.

[0068] For the antibacterial effect test against the test strains Staphylococcus aureus and Staphylococcus epidermidis, the initial dilution was performed using D / E neutralizing broth. The neutralized test solution was serially diluted and 1 mL was dispensed into two Petri dishes for each concentration. (15-25) mL of pre-prepared tryptic soy agar at (45-50)°C was dispensed into Petri dishes and allowed to solidify at room temperature. The solidified Petri dishes were inverted and incubated at (35±1)°C for (24-48) hours. The test was performed twice for each strain, and the initial and control viable counts were measured using sterile saline.

[0069] After incubation, the viable cell count was observed by selecting Petri dishes showing (30-300) cells. When viable cell counts were observed only at the lowest dilution stage, they were counted regardless of the observation range. When bacteria grew, the number of bacteria on the medium was multiplied by the dilution factor to calculate the number. When bacteria did not grow on the medium, it was multiplied by the dilution factor performed at the neutralization stage and expressed as "less than 10" (<10).

[0070] Table 6 below shows the antibacterial test results for Staphylococcus aureus.

[0071] [Table 6]

[0072] The initial inoculum size was 3.1 x 10 6 CFU / mL, 60-min and 90-min control bacterial counts were 3.2 × 10 6 CFU / mL and 4.5×10 6 After 60 and 90 minutes, the test counts were <10 CFU / mL in both replicates.

[0073] Table 7 below shows the antibacterial test results for Staphylococcus epidermidis.

[0074] [Table 7]

[0075] The initial inoculum size was 3.2 x 10 6 CFU / mL, 60-min and 90-min control bacterial counts were 3.5 × 10 6 CFU / mL and 3.7 × 10 6 After 60 and 90 minutes, the test counts were <10 CFU / mL in both replicates.

[0076] Table 8 below shows a summary of the antibacterial test results in Tables 6 and 7. (Unit: meam log)

[0077] [Table 8]

[0078] L-R *= Log reduction test results showed that the antibacterial test values ​​for Staphylococcus aureus were >5.49 and >5.49 at 60 and 90 minutes, respectively, and the antibacterial test values ​​for Staphylococcus epidermidis were >5.51 and >5.51 at 60 and 90 minutes, respectively. A log reduction of 4 or more means 99.99% or more, and 5 or more means 99.999%.

[0079] A body wash product was manufactured using bamboo mulch complex as an active ingredient and the additives shown in Table 9.

[0080] [Table 9]

[0081] In addition, a shampoo containing 2.0% bamboo extract and 2.5% composite extract was prepared by adding the additives shown in Table 10. In Table 10 below, the addition ratio refers to the wt% content of each ingredient based on a total weight of 100wt%.

[0082] [Table 10]

[0083] A facial cream was also produced with the ingredients shown in Table 11. In Table 11 below, RM or ingredient % in RM means the ingredient content, and ingredients % in RM means the ingredient content in the ingredients.

[0084] [Table 11] JPEG0007601526000013.jpg191158

[0085] Although the specific parts of the present invention have been described in detail above, the present invention is not limited to those illustrated in the drawings, and it is clear to those skilled in the art that such specific techniques are merely preferred embodiments and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention should be defined by the appended claims and their equivalents.

Claims

1. A process of heating the bamboo, cooling the gas generated when the temperature of the heating furnace is 90°C to 280°C, storing the vinegar liquid obtained, and leaving it to stand for more than three years to produce a bamboo extract; A step of subjecting the bamboo shoot skin to steam treatment at 80°C to 120°C, followed by low-temperature drying at a temperature of 15°C to 80°C for 20 hours to 80 hours, and then extracting the bamboo shoot skin extract; mixing sugar and citrus fruit in a ratio of 0.5:1, aging the mixture for at least 3 months, and then extracting the mixture to produce a saccharified citrus fruit extract; After the extraction under reduced pressure, the plant extract is concentrated under reduced pressure; and preparing a bamboo multi-complex containing the bamboo extract, the bamboo shoot skin extract, the saccharified bamboo fruit extract, and the plant extract; A method for producing a bamboo multi-complex having antibacterial properties, comprising:

2. The method for producing a bamboo multi-complex with antibacterial function described in claim 1, wherein the plant extract is at least one of bamboo extract, elm root extract, ginseng extract, licorice extract, celandine extract, Japanese laurel root extract, Japanese laurel leaf extract, Centella asiatica extract, Sanbai extract, and fermented green tea extract.

3. The bamboo shoot skin extract is an alcohol extract obtained by adding alcohol at a concentration of 3% to 50% (v / v) as a solvent after low-temperature drying and extracting at a temperature of 80°C to 100°C for 3 to 5 hours. This is the method for producing a bamboo multi-complex with antibacterial properties described in claim 1.

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