KR20200018075A - microcapsule including Citrus unshiu, Torreya nucifera, and Chamaecyparis obtusa - Google Patents

Abstract

The present invention relates to an anti-oxidant, antibacterial and anti-atopic composition comprising Citrus unshiu, Torreya nucifera, and Chamaecyparis obtusa oils. Highly volatile oil containing the composition can be manufactured into a microcapsule to prevent the composition from volatilizing, thereby maintaining effects, and the microcapsule can be used to manufacture functional fibers, fabrics, and paper having antibacterial and anti-atopic effects.

Description

Microcapsule including Citrus unshiu, Torreya nucifera, and Chamaecyparis obtusa}

The present invention relates to microcapsules for antimicrobial, antioxidant and anti-atopy including Wenzhou, Persimmon, Cypress oil and the fiber coated with the microcapsules.

Wenzhou Citrus ( Citrus unshiu ) is an evergreen sub-tree in Korea, Japan, Latin America and the Black Sea. Wenzhou mandarin fruits are spherical, 5-8cm in diameter, mature to yellowish red, and are called tangerine, citrus, or mandarin. In addition, the fruit peels well, and the fruit reproduces. It is cultivated in the southern coastal climatic zone, and there are about 10 kinds of crude species, mesozoic species and late species.

Torreya nucifera is a conifer that belongs to the coniferous family Kabinzawa and is native to southern Japan. The fruit of the visa contains seeds called visa, which has been widely used as an insect repellent since ancient times.

Chamaecyparis obtusa is an evergreen tree native to Japan. Its height is 30 ~ 40m, its width is about 1 ~ 2m, the bark is reddish brown, and small needle-shaped leaves grow in the branches. Small flowers bloom on the branches in spring, and green conifers ripen in red in October. Leaves and wood contain 1% essential oil and are used for medicinal purposes. Cypress is known to have excellent bactericidal action because it contains a large amount of natural antibacterial substance called phytoncides.

The natural plant resources are limited and the yield of natural oils is not high. Therefore, there is a need for a technique capable of improving the natural function of natural oil and using natural oil efficiently.

Republic of Korea Patent No. 10-0905386

One object of the present invention is antimicrobial, antioxidant or anti-atopic dermatitis comprising a mixed oil containing hot-wheat citrus, non-visible, cypress oil in a weight ratio (w / w) of 7-8: 0-1: 2-3 It is to provide a microcapsules for.

Another object of the present invention is antimicrobial, antioxidant or anti-atopic dermatitis comprising a mixed oil containing hot-glut, persimmon, cypress oil in a weight ratio (w / w) of 7-8: 0-1: 2-3 It is to provide a biochemical composition comprising microcapsules.

Still another object of the present invention is antimicrobial, antioxidant or anti-atopic dermatitis comprising a mixed oil containing hot-wheat citrus, non-visa, and cypress oil in a weight ratio (w / w) of 7 to 8: 0 to 1: 2 to 3 as an active ingredient. It is to provide a fiber product coated with microcapsules.

In order to achieve the above object, the present invention includes a mixed oil containing, in one embodiment, hot-alcoholic, non-visible, cypress oil in a weight ratio (w / w) of 7-8: 0-1: 2-3 It provides antimicrobial, antioxidant or anti-atopic microcapsules.

Terms of this invention "Satsuma Mandarin (Citrus unshiu)" is sangrokseong arborescent distributed in Korea, Japan and Latin America, and elsewhere Black Sea. Wenzhou citrus fruit is oblong, 5-8cm in diameter, matured yellowish red, and is called tangerine, citrus, or citrus. In addition, the fruit peels well, and the fruit reproduces.

The term “visa ( Veraya nucifera )” of the present invention is an evergreen conifer tree belonging to the family of the family of attention, and is distributed only in Korea and Japan worldwide. Visa is used for edible, ornamental, industrial and medicinal purposes. Seeds are eaten or squeezed oil. In oriental medicine and private sector, fruit is used as medicine for insect repellent, hair growth, health, landscaping, intestinal bleeding, and wood is used for building materials, equipment materials, and marine materials.

The term " Chamaecyparis obtusa " of the present invention is an evergreen tree native to Japan. It is 30 ~ 40m high, about 1 ~ 2m wide, and the bark is reddish brown, and small needle-shaped leaves grow in the branches. Small flowers bloom on the branches in spring, and green conifers ripen in red in October.

In the present invention, the term "oil" refers to an essential oil extracted from hot sake, visa or protein. The extraction method of the essential oil may be a method known in the art such as steam distillation, compression, water distillation, solvent extraction.

Steam distillant (Steam Distillantion) is the most commonly used method of extracting essential oil, almost 80% or more is using steam distillation. Steam is generated when the raw plant is heated in a steam reactor. Hot heat above 100 ° C destroys the cell walls of the plant cells, the mixtures of the destroyed plants turn into steam and collect in the pipes, and the vapor-like mixtures turn into liquids by the chiller. The oil is extracted by distilled water in the upper part and the lower part.

Expression is used when the distillation extraction method cannot be used because it is sensitive to heat in the raw materials used to extract essential oils. It is recommended that these raw materials not be heated as much as possible, but the expression is one of the most widely used low temperature extraction methods for extracting citrus essence oil. Mainly lemon, orange. It is a method of extracting essential oils by compressing the secretory sac cells on the outer surface of citrus fruits such as bergamot and mandarin, and treating unstable citrus fruits at low temperature. Commonly called cold press method, this press method is used to extract a lot of citrus-based essential oils. Since most citrus peels contain oil, it is a way to remove and extract inorganic salts from the peels.

Water distillation is an extraction method in which the plant is completely immersed in water and boiled at a temperature below 100 ° C. and low pressure. Used for plants that are heat sensitive but not affected by the time of extraction.

Solvent extraction is an extraction method in which a large container is filled with a plant, hot solvent is poured, and the essential oil component is eluted. Semi-solid material from which solvent is removed after oil extraction by solvent extraction is called concrete. Absolute is a double distillation of concrete, a mixture of plant's natural waxes, resins and essential oils in hexane, followed by alcohol purification. In the process of extracting resinous essential oils such as jasmine, a lot of solvent extraction methods are used, and solvents used in the extraction include hexane, hexane, alcohol and alcohol.

In the present invention, the Wenzhou citrus oil can be prepared using the flesh or peel of the fruit of Wenzhou persimmon.

The cypress oil in the present invention can be prepared using the leaf or stem of the cypress.

In the present invention, the non-oil can be prepared using the leaves or stems of the non-tree.

Antioxidant, antimicrobial and subjective fragrance evaluation was confirmed using 12 kinds of mixed oils (Table 1) with different solenoid oils and mixed ratios of Wenzhou sensation, cypress or non-oil in the examples of the present invention. The mixing ratio was designed using response surface analysis.

Antioxidant activity was confirmed by performing a DPPH assay. As a result, it was confirmed that in the case of hot jujube persimmon, visa or protein oil alone oil, each of the non-visa and proteinaceous oil exhibited less than 10% free radical scavenging activity, and the wine juju has a free radical scavenging activity of about 15%. .

Next, as a result of analyzing the antioxidant activity of the mixed oil, it was confirmed that the antioxidant activity increases when the mixed oil of the same concentration is treated than when treating the single oil. Specifically, when the sample 2 (50% hot spring, 50% visa), sample 10 (75% hot spring, 10%, 15% visa), and sample 12 (75% hot spring, 25% visa), antioxidant activity Especially, in case of sample 12, free radical scavenging activity was confirmed to be 32.9%, and sample 2 also exhibited free radical scavenging activity of 23.8% or more. It was confirmed.

Next, to check the antimicrobial effect of mixed oils according to Wenzhou tightness, cypress, visa oil alone and mixing ratio, Propionibacterium acne ( P. acne ) known as acne-causing bacteria and Staphylococcus aureus ( S ) known as causative bacteria aureus ) was evaluated for antimicrobial activity.

Concentrations of P. acne and S. aureus at 0.05% non-oil, 0.01% cypress oil, and 0.01% Wenzhou persimmon (citrus) oil were analyzed. As a result, it was confirmed that the growth rate of bacteria was reduced compared to the positive control group containing only three kinds of oils, among which 75 to 70% of P. acne and S. aureus treated with visa 0.05% Measured as it was confirmed that the most excellent antibacterial activity of the three oils.

Next, as a result of analyzing the antimicrobial activity of the above 12 kinds of mixed oils, it was confirmed that the antimicrobial activity increased than the single oil according to the mixing ratio. Specifically, when the mixed oil was treated in comparison with the positive control, the growth rate of P. acne was measured to be 78.1 to 48.7%, and the growth rate of S. aureus was 78.6 to 38.5%. When double sample 2 was treated, the growth rate of bacteria in P. acne 58.1%, 63.3% in S. aureus , and bacterial growth rate was 48.7% in P. acne and 38.5% in S. aureus when sample 12 was treated. That is, it was confirmed that the antimicrobial activity is different according to the mixing ratio, the double antimicrobial activity of Sample 2 and Sample 12 was confirmed to be the best (Fig. 3b).

Subjective fragrance evaluation was conducted for women in their 20s, and the satisfaction with the fragrance was scored between 0 and 10.

As a result of the evaluation of isolated oil of Wenzhou citrus, visa or cypress oil, the fragrance preference for visa oil was the highest and the preference for cypress oil was lowest.

Statistical analysis of the results analyzed in the above example to derive the best point of the mixing ratio with the best antioxidant, antibacterial, and subjective satisfaction.

As a result of deriving the optimal point of antioxidant, antimicrobial and subjective satisfaction, 70% of Wenju's secret, 10% of Cypress and 20% of Visa were common. In order to verify the optimum mixed oil ratio, sample 13 (70% hot spring and 10% visa) and sample 14 (70% hot spring and 30% visa), sample 15 (80% hot spring and 20% visa) ) And sample 16 (80% hot spring, 10% white and 10% non-visible) were prepared and analyzed for antioxidant, antibacterial and subjective satisfaction.

As a result of the analysis of antioxidant activity, free radical scavenging activity of Sample 15 was 40% or more, and Sample 13 and Sample 14 were also measured to be more than 30%, and it was confirmed to have excellent antioxidant activity. Sample 16 showed less than 20% scavenging activity, confirming that the antioxidant activity was slightly weak (FIG. 5B).

Antimicrobial effect was also similar to the above, in all samples 13 to 15 P. acne and S. aureus and bacteria growth rate was reduced by more than 30% on the basis of the positive control group was confirmed to have excellent antimicrobial activity, but in the case of sample 16, the survival rate of bacteria about 75% It was determined that the antimicrobial effect is slightly weak.

The evaluation result of the subjective fragrance sensitivity is as shown in FIG. Sample 13, which consists of 70% Wenju Sensation, 10% Cyclones, and 20% Visa, suggested by the optimal point derivation, is “comfortable (F = 1.08)”, “fresh (F = 0.54)”, “like” (F). = 1.15) ”was higher than the other three samples. However, there was no statistically significant difference. For “strong” and “new”, the proposed sample 13 score was not as high as for the other samples. Overall, Sample 13, which was selected as the optimal mixing ratio, was subjectively positively evaluated for “comfortable”, “fresh”, and “like”, and confirmed that subjective satisfaction was high.

In addition, to determine the anti-atopic effect of the oil according to the mixing ratio, HaCaT cells were treated with LPS and 12 mixed oils (0.01%) in Table 1 and analyzed for expression of inflammation or atopy biomarkers after 24 hours. Specifically, skin inflammation biomarkers were analyzed for interleukin-1β (IL-1β) and interleukin-8 (IL-8), and atopy biomarkers were analyzed for chemokine (CCL17, CCL22) genes.

As a result, iNOS, IL-1β, IL-8, CCL17, and CCL22 were not expressed in the cells that did not process the sample, but iNOS, IL-1β, IL-8, CCL17, and CCL22 genes were all expressed in LPS-treated cells. It was confirmed that expression was induced.

It was confirmed that expression of all iNOS, IL-1β, IL-8, CCL17, and CCL22 genes was suppressed except Sample 6 in 12 mixed oils having different mixing ratios.

Next, using the mixed oil (samples 13 to 16) derived from the optimum ratio derived from the statistical analysis above, the effects of the mixed oil according to the mixing ratio were further confirmed. In a further experiment, 1 μg / ml LPS was treated with 0.05% of each mixed oil.

As a result, it was confirmed that the expression of IL-1β, iNOS, CCL17 and CCL22 induced by LPS was not inhibited in the cells treated with 0.05% of sample 16. It was confirmed that iNOS, IL-1β, IL-8, CCL17 expression inhibitory effect is excellent. That is, it was confirmed that Sample 13 to Sample 15 except Sample 16 had excellent inflammation or atopy suppression effects.

Therefore, the microcapsules of the present invention include a mixed oil containing hot-alcohol, non-visible, and white oil in a weight ratio of 7-8: 0-1: 1-2-3. Specifically, the mixed oil may contain Wenju, Persimmon, and Cypress Oil in a weight ratio of 7: 1: 2 or 8: 0: 2.

It was confirmed that the mixed oil is excellent in antibacterial, antioxidant or anti-atopic dermatitis, the microcapsules of the present invention has an antimicrobial, antioxidant, or anti-atopic effect.

In addition, the subjective fragrance sensitivity evaluation results confirmed that the satisfaction is superior to the single oil, the microcapsules of the present invention can be used as a fragrance.

Wet citrus, non-visible or proteinaceous oils are volatile and most of them are volatilized when suspended on the surface. Therefore, the anti-oxidation, antimicrobial and / or anti-atopic effect of using the warmth, non-visible or cypress oil is not long, and when the oil is applied, it may cause a problem of using excess oil to maintain the effect. have. Therefore, the present invention solves the problem of volatilization by preparing the mixed oil of Wenzhou, Persimmon and Cypress in microcapsules.

The microcapsules of the present invention are characterized in that the microcapsules burst when a pressure is applied.

In the embodiment of the present invention, the microcapsules were prepared using melamine and formaldehyde as wall materials of the microcapsules, and styrene was used as an emulsifier. In addition, by adjusting the stirring speed to prepare a microcapsules having an average diameter of 3 or 5㎛. Specifically, the average diameter of the microcapsules prepared by stirring at 6,000 rpm was measured at 3.450 μm, and the average diameter of the microcapsules prepared by stirring at 4,000 rpm was measured at 5.000 μm.

Therefore, the microcapsules of the present invention may have an average diameter of 3㎛ 5㎛. The microcapsules may be prepared by stirring at 6,000 rpm to 4,000 rpm.

In another embodiment of the present invention, antimicrobial, antioxidant or anti-atopic microcapsules comprising mixed oil containing hot-glut, persimmon and cypress oil in a weight ratio of 7-8: 0-1: 1-2-3 as an active ingredient It provides a biochemical composition.

In the present invention, description of the hot spring, visa, protein, oil is as described above.

As used herein, the term biochemical may include home care products, hair products, skin cleansers or oral products.

Specifically, the home care product may include a laundry detergent, a fabric softener, a dish cleaner, a dishwasher cleaner, a household space cleaner, a wet tissue, a deodorant, an air cleaner, an indoor air freshener, a fragrance agent, a multipurpose cleaner, a disinfectant or a disinfectant. have.

Specifically, the hair product may include a shampoo, rinse, treatment, hair essence or scalp pack.

Specifically, the skin cleanser may include a body wash, body gel, soap, and cleansing foam.

Specifically, the oral product may include a toothpaste or a mouthwash.

In another embodiment of the present invention is coated microcapsules for antimicrobial, antioxidant or anti-atopic skin containing mixed oil containing a weight ratio of hot jujube persimmon, non-visa, cypress oil in a weight ratio of 7 to 8: 0 to 1: 2 to 3 as an active ingredient Textile products.

In the present invention, description of the hot spring, visa, protein, oil is as described above.

As used herein, the term “fiber” refers to a natural or artificial linear object used as a raw material of textile products such as woven fabrics, knitted fabrics, ropes, nets, felts, and the like.

The natural fiber includes plant fiber, animal fiber and mineral fiber, and the plant fiber includes seed fiber including cotton, kapok cotton and coir, and leaf vein fiber including manila hemp and sisal hex, and flax, ramie, jute, hemp There is a bast fiber comprising a. Animal fibers include wool fibers including wool, goat wool, camel hair and cashmere, and silk fibers including gaze and night-dogs. And mineral fiber has asbestos.

Artificial fibers include inorganic fibers and organic fibers, and inorganic fibers include gold fibers, metal fibers including silver yarns, silicate fibers including glass fibers, rock fibers and slag fibers. Organic fibers include regenerated fibers, semisynthetic fibers, and synthetic fibers. Regenerated fibers include fibrous and protein fibers including viscose rayon and cupra rayon. Semisynthetic fibers include acetate and triacetate. , Polyester, polyurethane, polyurea, polyethylene, polyvinyl chloride, polyvinylidene, polytetrafluoroethylene, polyvinyl alcohol, polyacrylonitrile, polypropylene fiber, and the like. .

 In the present invention, the fiber product is a product made of natural fibers or artificial fibers, including paper and fabric.

The fiber product may be a paper fabric manufactured using fibers coated with microcapsules, or the microcapsules may be coated on at least one surface of paper or fabric.

The textile products may include fashion products including wallpaper, scarves, ties and bags, wallpaper, packaging materials, and interior fabrics.

The microcapsule-coated fabric may break the microcapsules under pressure applied from friction with the user's skin, and mixed oil in the microcapsule may come into contact with the skin to produce an antioxidant, antibacterial or anti-atopic effect.

The present invention relates to a composition for antioxidant, antibacterial, and anti-atopic skin containing Wenzhou, Persimmon, and Cypress oils, wherein the highly volatile oil containing the composition is prepared as a microcapsule to prevent volatilization of the composition to maintain the effect. The microcapsules may be used to produce functional fibers, fabrics, and papers having antibacterial and anti-atopic effects.

Figure 1a is a result of analyzing the cytotoxicity of the concentration of non-leaf, cypress, Wenzhou citrus (citrus) oil.
Figure 1b is the result of analyzing the cytotoxicity according to the treatment concentration of 12 mixed oils according to the mixing ratio of non-leaf, cypress, citrus (citrus) oil.
Figure 2a is a result of measuring the free radical scavenging activity according to the concentration of ascorbic acid.
Figure 2b is a result of measuring the free radical scavenging activity according to the treatment concentration of each single oil of non-leaf, cypress, citrus, citrus.
Figure 2c is the result of measuring the free radical scavenging activity of the mixed oil of non-leaved leaves, cypress, citrus fruits (citrus fruits).
Figure 3a is an analysis of the antimicrobial effect at the concentration of non-leaf oil of 0.05%, cypress oil 0.01%, Wenzhou persimmon (citrus) oil 0.01%, (A) P.acne and (B) S. aureus each single oil After treatment, the growth rate of the bacteria was measured.
Figure 3b is the result of measuring the growth rate of the bacteria after 0.01% treatment of 12 kinds of mixed oil (A) P.acne and (B) S. aureus .
Figure 4a is a result of analyzing the subjective fragrance satisfaction of the sole oil of non-leaf, cypress, Wenzhou citrus (citrus fruits).
Figure 4b is the result of analyzing the subjective fragrance satisfaction of 12 kinds of mixed oil of non-leaf, cypress, Wenzhou citrus (citrus fruits).
Figure 5a is a result of analyzing the cytotoxicity according to the concentration of four mixed oils (samples 13 to 16) of the optimum mixing ratio derived through the statistics.
Figure 5b is the result of analyzing the antioxidant activity of the four mixed oils (samples 13 to 16) of the optimum mixing ratio derived through the statistics.
Figure 5c is the result of analyzing the antimicrobial effect on P.acne of the four mixed oils (samples 13 to 16) of the optimum mixing ratio derived through the statistics.
Figure 5d is a result of analyzing the antimicrobial effect of S. aureus of the four mixed oils (samples 13 to 16) of the optimum mixing ratio derived through the statistics.
5E is a result of analyzing the subjective fragrance satisfaction of four mixed oils (samples 13 to 16) having an optimum mixing ratio derived through statistics.
Figure 6a is a result of analyzing the skin inflammation and atopic inhibition of 12 mixed oils of non-leaved leaves, cypress, citrus fruits (citrus fruits).
Figure 6b is a result of analyzing the skin inflammation and atopic inhibition of the four mixed oils (samples 13 to 16) of the optimum mixing ratio derived through the statistics.
7A is a scanning electron microscope image of a microcapsule (3 μm) of the present invention.
7B is a scanning electron microscope image of a microcapsule (5 μm) of the present invention.
Figure 7c is the result of the characterization of the 3㎛ microcapsules.
Figure 7c is a result of the characterization of the 5㎛ microcapsules.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention.

1. Preparation of materials

Natural oils of the flesh of Citurs unshiu , the leaves and stems of Chamaecyparis obtusa , and the leaves of Torreya nucifera were used.

The three oils were designed in the composition of Table 1 using reaction surface analysis to prepare 12 kinds of mixed oils having different mixing ratios of different weights.

Sample Number Wenzhou Citrus
(%) White
(%) Visa
(%) One 50 50 0 2 50 0 50 3 0 50 50 4 50 25 25 5 50 25 25 6 50 30 20 7 50 20 30 8 25 37.5 37.5 9 75 15 10 10 75 10 15 11 75 25 0 12 75 0% 25

2. Cytotoxicity Assessment

To confirm cytotoxicity, each sample was treated with HaCaT cells incubated with DMEM medium, and MTT assay was performed to measure cell viability.

Single oil

First, we analyzed the cytotoxicity according to the concentrations of individual oils of Wenzhou persimmon, cypress, and visara. Cytotoxicity assays were performed at concentrations of 0.01, 0.05, 0.1, 0.2, 0.5, 1, 2% (w / w). The cells used human dermal fibroblasts (HaCaT).

It was confirmed that non-oil oil had a cytotoxicity at a concentration of 0.1% or more from a concentration of 0.1% or more decreased to less than 28.3%, and a decrease in cell survival rate of 20.8% at 0.05% of cypress oil. In addition, it was confirmed that Wenzhou citrus oil also reduced the cell survival rate to 28.6% at 0.05%, it was confirmed that cypress and Wenzhou citrus oil have cytotoxicity at a concentration of 0.05% or more (Fig. 1a)

Mixed oil

Next, the cytotoxicity of 12 kinds of mixed oils prepared by varying the mixing ratio of oils of Wenzhou persimmon, cypress and visara in Table 1 was evaluated. Compared with the single oil, when all 12 mixed oils were treated with a concentration of 0.1% or more, it was confirmed that the cell viability was significantly reduced, and the mixed oil was found to be somewhat stronger in cytotoxicity than the single oil at the same concentration. However, all 12 kinds of mixed oils did not show cytotoxicity when treated with a concentration of 0.01% or less (FIG. 1B).

3. Antioxidant Activity

Antioxidant activity was analyzed using DPPH assay. Ascorbic acid was used as a positive control for verifying antioxidant activity.

First, it was confirmed that free radical scavenging activity was increased as the concentration of ascorbic acid increased, and it was confirmed that the free radical scavenging activity was 40% when 100 µg / ml was treated (FIG. 2A).

Single oil

Free radical scavenging activity was measured according to the concentrations of isolated oils of Wenzhou persimmon, cypress and non-visa. The treatment concentration of oil was 0.005% and 0.01%, the concentration without the cytotoxicity based on the results in the cytotoxicity.

As a result, it was confirmed that non-radical scavenging activity was less than 10% at 0.01% concentration of non-visa and cypress oil. It showed activity, but showed higher free radical scavenging activity than non-leaf and cypress oil, but it was confirmed that the antioxidant effect is weak compared to ascorbic acid (about 40%) (Fig. 2b).

Mixed oil

Next, the antioxidant activity of 12 mixed oils was confirmed. A sample at a concentration of 0.01% was found to be free of cytotoxicity. As a result of the measurement, it was confirmed that the antioxidant activity was increased during the mixed oil treatment than when the same concentration of the monotreated oil was treated.

Specifically, in the case of Samples 2, 10, and 12, it was confirmed that the free radical scavenging activity was higher at the same concentration than that of the Wenzhou dense persimmon (0.01%) alone oil, which was excellent in antioxidant activity. In particular, in the case of sample 12, the inhibitory activity was 32.9%, and the free radical scavenging activity was increased by about 2 times than that of the hot jujube persimmon treatment. It was confirmed that mixed oil having a high content of non-oil oil had excellent antioxidant activity (FIG. 2C).

4. Antimicrobial Evaluation

Antimicrobial evaluation was performed on propionibacterium acne ( P. acne ), which is known to be the cause of acne , and S. aureus , which is known to be the cause of dermatitis.

Based on the cytotoxicity evaluation, the antimicrobial activity was evaluated at a concentration without cytotoxicity. Concentrations of P. acne and S. aureus at 0.05% non-leaf oil, 0.01% Cypress oil, and 0.01% Wenzhou persimmon (citrus) oil were analyzed.

As a result of the analysis, the growth rate of P. acne bacteria treated with the leaves of cypress leaves, cypress and Wenzhou citrus was 78.1% when treated with Visa 0.05% and 88.1% when treated with 0.01% Cypress, compared to the control group (100%). , Citrus fruits were measured as 86.2% at 0.01% treatment. The growth rate of the bacteria was reduced from 11.9 to 21.9%, confirming that all three oils had antimicrobial activity against P. acne .

In addition, when the oil is treated with S. aureus bacteria, the viability of the bacteria is measured as 73.1% when treated with Visa 0.05%, 78.1% when treated with 0.01% Cypress, and 80.2% when treated with 0.01% Citrus. , The bacterial growth rate was reduced to 19.8 to 26.9%, it was confirmed that all three oils have an antimicrobial activity against S. aureus (Fig. 3a).

Mixed oil

Next, the antimicrobial activity of 12 mixed oils was evaluated. Based on the cytotoxicity evaluation results, the antimicrobial activity was evaluated using a mixed oil having a concentration of 0.01%.

As a result of antimicrobial evaluation, the growth rate of P. acne was 78.1 to 48.7% and the growth rate of S. aureus was 78.6 to 38.5%. When the sample 2 was treated, the bacterial growth rate was measured as P. acne 58.1%, S. aureus 63.3%, and when sample 12 was treated, the bacterial growth rate was measured as 48.7% P. acne and S. aureus 38.5%.

That is, it was confirmed that the antimicrobial activity in the mixed oil is generally higher than the single oil, the antimicrobial activity was different according to the mixing ratio, it was confirmed that the best antimicrobial activity of Sample 2 and Sample 12 (Fig. 3b).

5. Subjective fragrance sensitivity evaluation

The subjective fragrance sensitivity evaluation of 18 women in their 20s with normal sense of smell was performed. The subject participated in the experiment without drinking or smoking for 24 hours before the experiment. A mixed oil was applied to the smell paper to close the subject's nose and smelled for 3 seconds, the smell paper was removed, and the satisfaction with the smell was answered by a score between 0 and 10. The average value was calculated by performing two replicates for each subject.

Single oil

As a result of fragrance sensitivity evaluation using Wenzhou citrus, visa or cypress single oil, we felt strong about cypress oil, and the satisfaction with comfort, liking and freshness was the lowest. Responsive, refreshed, new and liked.

In other words, the subjective fragrance evaluation was confirmed that the preference for the non-oil oil fragrance is high, and relatively satisfactory for the cypress oil was confirmed that the results are significantly lower than the other two oils (Fig. 4a).

Mixed oil

The fragrance sensitivity of 12 mixed oils was evaluated in 18 women in their 20s with normal sense of smell. As a result, in the case of mixed oil samples 2, 4, 5, 6, 10, and 12, the subjective satisfaction with the fragrance was calculated to be 5 or more, and it was confirmed that the fragrance oil had the higher satisfaction than the highest non-fragrant oil in the single oil ( 4b).

6. Screening of the optimum mixed oil ratio

Design of optimum mixed oil ratio

The regression model was obtained by statistical analysis of antimicrobial activity, antioxidant activity and subjective satisfaction. Each individual estimate was derived and the optimal point was derived, and the optimal mixing ratio was derived by comprehensively judging the results of the three variables. In this case, Wenju's citrus was set to be 50% or more in consideration of production value and by-product utilization value.

Table 2 to Table 5 are the results of calculating the optimal point by statistical analysis of antimicrobial, antioxidant, subjective satisfaction analysis results.

Optimal point of antibacterial-1 ( P. acne ) Wenzhou Citrus
(%) White
(%) Visa
(%) Cell survival rate (%) Opinion 60 0 40 65.9132 Best 70 0 30 62.6711 Best of best 70 10 20 67.5955 best

Optimal point of antibacterial-2 ( S. aureus ) Wenzhou Citrus
(%) White
(%) Visa
(%) Cell survival rate (%) Opinion 60 0 40 64.5716 Best 70 0 30 58.1471 Best of best 70 10 20 66.7937 best

Optimum point of antioxidant Wenzhou Citrus
(%) White
(%) Visa
(%) Radical scavenging ability (%) Opinion 70 0 30 19.0762 Best 70 10 20 18.2551 Best 80 0 20 22.0125 Best of best 80 10 10 18.2953 Best

Optimum point of subjective satisfaction Wenzhou Citrus
(%) White
(%) Visa
(%) Subjective Satisfaction Opinion 60 10 30 5.4661 Best 60 20 20 5.6664 Best of best 70 10 20 5.5247 best

As a result of derivation of optimum points, considering antibacterial and antioxidant properties, (1) 70% Wenzhou citrus, 10% Pycnometer, 20% Visa, and (2) 70% Wenzhou Citrus, 0% Pyramid and 30% Visa It was selected as the optimal ratio with excellent performance. Finally, as a result of judging the antimicrobial, antioxidant, and subjective satisfaction, (1) 70% Wenju Sensation, 10% Cyclones, and 20% Visa were selected as the optimal mixing ratio.

Verification of optimal mixed oil ratio

In order to analyze the physiological activity of the mixed oil according to the ratio of the optimum mixed oil derived above, four mixed oils of the mixing ratio as shown in Table 6 were prepared, and the cytotoxicity, antioxidant and antibacterial activity were confirmed.

Sample name Wenzhou Citrus
(%) White
(%) Visa
(%) Sample 13 70 10 20 Sample 14 70 0 30 Sample 15 80 0 20 Sample 16 60 20 20

Cytotoxicity Assessment

Cytotoxicity was performed MTT assay using HaCaT cells in the same manner as described above. Each sample was treated at a concentration of 0.01, 0.02, 0.05, 0.1, 0.2, 0.5 or 1%. As a result, it was confirmed that the cell survival rate of sample 13 was halved at 0.1% concentration, and in case of sample 15, the survival rate of cells treated at 0.5% concentration was rapidly decreased. Therefore, in the subsequent antioxidant and antimicrobial evaluation, the whole sample used a concentration of 0.05% that does not exhibit cytotoxicity (Fig. 5a).

Antioxidant evaluation

Antioxidant activity was evaluated by analyzing DPPH, and vitamin C was used as a positive control. As a result of analysis, Sample 15 showed excellent free radical scavenging activity of 40% or more, Sample 13 and Sample 14 showed about 30% of antioxidant activity, and Sample 16 exhibited less than 20% of scavenging activity, compared with Samples 13 to 15. It was confirmed that the antioxidant effect is slightly weak (Fig. 5b).

Antimicrobial activity

The antimicrobial effect of P. acne and S. aureus was treated by 0.05% of Samples 13 to 16, respectively. As a result, the sample 16 was confirmed that the cell viability was measured to be 75% or more, but the antimicrobial effect was weak, but the samples 13 to 15 had an excellent antimicrobial effect against P. acne and Saure (Fig. 5c-5d) .

Subjective satisfaction

As a result of evaluating subjective fragrance sensibility, the sample 13 composed of 70% of hot jujube sensation, 10% of white powder, and 20% of non-visible mixture suggested by derivation of optimum point was “comfortable (F = 1.08)”, “fresh (F = 0.54). And scores for “like” (F = 1.15) were higher than the other three samples. However, there was no statistically significant difference. For “strong” and “new”, the proposed sample 13 score was not as high as for the other samples. Overall, Sample 13, which was selected as the optimal mixing ratio, was highly evaluated for “comfortable”, “fresh”, “like”, and confirmed that subjective satisfaction was high (FIG. 5E).

7. Anti-Atopy Activity

Next, the anti-atopic activity of the mixed oil according to the mixing ratio was analyzed. HaCaT cells were treated with 1 μg / ml LPS and 12 mixed oils at a concentration of 0.01%, and 24 hours later, interleukin-1β (IL-1β) and interleukin-8 (IL-8), a dermatological biomarker, were treated. The effect of inhibiting the expression of the chemokine (CCL17, CCL22) genes, atope and biomarkers, was analyzed.

6 is a result of confirming the inflammation and atopy inhibitory effect of the mixed oil according to the mixed ratio of LPS and 12 mixed oils with different mixing ratios inducing inflammation at the same time. Gene expression of iNOS, interleukin-8, interleukin-1β, CCL17 and CCL22 was confirmed in 1% agarose gel, and GAPDH was used as intra cellular control.

Each gene was checked by electrophoresis after RT-PCR was loaded on 1% Agarose gel.

As a result, iNOS, IL-1β, IL-8, CCL17, and CCL22 were not expressed in the cells that did not process the sample, but iNOS, IL-1β, IL-8, CCL17, and CCL22 genes were all expressed in LPS-treated cells. It was confirmed that expression was induced.

It was confirmed that expression of all iNOS, IL-1β, IL-8, CCL17, and CCL22 genes was suppressed except for Sample 6 in 12 mixed oils having different mixing ratios (FIG. 6A).

The mixed oils (Samples 13 to 16) derived from the optimal ratios derived from statistical analysis were further confirmed to inhibit the inflammation and atopy of the mixed oils according to the mixing ratios. In a further experiment, 1 μg / ml LPS was treated with 0.05% of each mixed oil.

As a result, it was confirmed that the expression of IL-1β, iNOS, CCL17 and CCL22 induced by LPS was not suppressed in the cells treated with 0.05% of sample 16. It was confirmed that the iNOS, IL-1β, IL-8, CCL17 expression inhibitory effect is excellent (Fig. 6b).

That is, it was confirmed that Sample 13 to Sample 15 except Sample 16 had excellent inflammation or atopy suppression effects.

8. Preparation and Evaluation of Submicrocapsules Containing Mixed Oils

Wall materials of the microcapsules were prepared using melamine (Junsei Chemical, Japan) and formaldehyde (Junsei Chemical, Japan), and styrene (Styren, Junsei Chemical, Japan) was used as an emulsifier.

After dissolving 5% of styrene (Styren) as an emulsifier in distilled water, mixed oil was added to prepare a mixed solution. The mixed solution was strongly stirred using a homogenizer (homogenizer, T.K Mark II 2.5, Japan) to prepare an oil in water (O / W) emulsion. The O / W emulsion was added to the melamine-formaldehyde prepolymer, acetic acid was added to adjust the pH to 4.8, and submicrocapsules were prepared by varying the stirring speed using a homogenizer at 70 ° C. for 90 minutes. .

To prepare the average diameter of the microcapsules in two species of 3㎛ and 5㎛, it was prepared by varying the stirring speed. At this time, the stirring speed was performed at 6,000 rpm to produce 3 μm and at 4,000 rpm to produce 5 μm.

The average particle size and distribution change of the two submicrocapsules were measured by particle size analysis by laser diffraction using a particle size analyzer (Mastersizer 2000, Malvern Instrument, U.K.).

The shape and surface of the dried submicrocapsules were observed at various magnifications using a field emission scanning electron microscope (JSM-7500F, JEOL, Japan) (FIG. 7A), and the thermal properties of the submicrocapsules were measured by a thermogravimetric analyzer (TGA7). , Perkin Elmer, USA). The temperature of the thermogravimetric system was held at 30 ° C. for 1 minute, and then heated up to 50 ° C. at 10 ° C. per minute.

7c to 7d are the results of analyzing the particle size of the microcapsules. Red line means the percentage (%) of the total microcapsules according to the size of the microcapsules on the x-axis, based on the right side of the Y-axis (diff%). The blue line represents the cumulative percentage (%) and was based on the left Y axis (cumu%).

In the cumulative distribution of the mixed oil-containing microcapsules, the size values corresponding to 10%, 50%, and 90% of the maximum diameters were D10, D50, and D90, respectively. And D50 was analyzed as the diameter average value of all the particles.

Figure 7c is a result of measuring the average diameter of 3㎛ microcapsules prepared by stirring at 6,000rpm, particles with a diameter of 2㎛ 5㎛ or less occupied 55.28% of the particles, of which the particle diameter of 4.222-4.699㎛ It was confirmed that it occupies the highest ratio with 8.85% of the whole capsule. In addition, D50 was analyzed to be 3.450 μm.

Figure 7d is a result of measuring the microcapsules of 5㎛ the average diameter prepared by stirring at 4,000rpm, it was confirmed that the particles of 5㎛ to 10㎛ diameter occupy 47.41%, the capsule of 5㎛ size It occupies about 9% of the total capsule. In addition, D50 was analyzed to be 5.000 μm.

Claims (8)

Antimicrobial, antioxidant or anti-atopic microcapsules comprising mixed oil containing Wenju, persimmon and cypress oil in a weight ratio (w / w) of 7-8: 0-1: 2-3. Biochemical composition comprising the microcapsules of claim 1. The method of claim 2,
The biochemical product is a biochemical composition comprising a home care product, a hair product, a skin cleanser or an oral product. The method of claim 3, wherein
The home care product includes a laundry detergent, a fabric softener, a dishwasher, a dishwasher cleaner, a household space cleaner, a wet tissue, a deodorant, an air cleaner, an indoor fragrance, a fragrance agent, a multipurpose cleaner, a disinfectant or a disinfectant. Life chemical composition. The method of claim 3, wherein
The hair product is a biochemical composition comprising a shampoo, conditioner, treatment, hair essence or scalp pack. The method of claim 3, wherein
The skin cleanser comprises a body wash, body gel, soap, biochemical composition comprising a cleansing foam. The method of claim 3, wherein
The oral product is biochemical composition comprising a toothpaste or mouthwash. Claim 1 microcapsules coated textile product.

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