Mosquito repellent composition and method for producing the same

Abstract

The mosquito repellent composition can be produced by mixing 100 to 500 parts by weight each of cinnamon, Chinese laurel, chrysanthemum, Japanese jasmine, angelica root, nutmeg, peppermint, and fennel to form a mixture, mixing 100 parts by weight of the mixture with 100 to 700 parts by weight of 70 to 95% ethanol as a solvent to form a mixed liquid, and then subjecting the mixed liquid to low-temperature ultrasonic extraction to extract the active ingredient extract.

Description

[Technical Field]

[0001] The present invention relates to a mosquito repellent composition and a method for producing the same. More specifically, the present invention relates to a mosquito repellent composition that is composed of natural ingredients that are harmless and safe to the human body, has a skin soothing effect, has excellent extractability of active ingredients, and is suitable for mass production, and a method for producing the same. [Background technology]

[0002] Mosquitoes not only cause direct and indirect harm to humans and livestock by sucking their blood, disturbing their sleep, and restricting their outdoor activities, but are also pests that transmit various diseases such as dengue fever, yellow fever, encephalitis, and malaria.

[0003] Various solutions have been researched to protect the human body from such mosquito attacks, and a representative method is to spray repellents on the skin or clothes, which contain substances that mosquitoes dislike to prevent them from biting, although they have little effect in killing mosquitoes.

[0004] However, considering that repellents are used by being sprayed directly on the human body, the chemical ingredients of the repellents must have a repellent function that stimulates the mosquito's sensory organs, must not be toxic to the human body, and must minimize the impact and side effects on humans and the ecosystem, making the requirements for their chemical ingredients very difficult.

[0005] There are many different mosquito repellents available, but N,N-diethyl-m-toluamide (DEET) is widely used due to its excellent mosquito repellent effect. However, DEET has an unpleasant odor and is highly penetrating into the skin, so its use is restricted for children, pregnant women, people with low blood pressure, and those with sensitive skin. In particular, the United States restricts the production of mosquito repellents containing DEET at concentrations of 20% or more.

[0006] Recently, in response to the need for the development of natural mosquito repellents that are safe for the human body, have a mild scent, and are non-irritating, natural ingredient extracts from cinnamon and jasmine are being considered. However, coumarin, the main ingredient in cinnamon, has a boiling point of over 300°C, making it difficult to extract by thermal extraction. Patchouli alcohol, the active ingredient in jasmine, has a boiling point lower than that of water, making it difficult to extract by thermal extraction. Instead, it must be extracted using a distillation method, in which the distilled steam is cooled to obtain the active ingredient. In other words, the extraction method for the active ingredient varies depending on the ingredient, making it difficult to standardize the manufacturing process. Since the extraction efficiency of the active ingredient varies depending on the ingredient, it is difficult to guarantee the same effectiveness even for the same product.

[0007] Therefore, the present applicant proposes a mosquito repellent composition and a manufacturing method thereof that can extract various natural mosquito repellent materials using a single method, thereby improving manufacturing efficiency and ensuring uniform extraction of active ingredients contained in natural materials. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Korean Patent Publication No. 10-2011-0126888 (Natural mosquito repellent composition with excellent long-lasting effect) [Patent Document 2] Korean Patent Publication No. 10-2020-0137703 (Natural mosquito repellent composition) [Non-patent literature]

[0009] [Non-Patent Document 1] Korean Journal of Food and Nutrition Science, Vol. 41, No. 7, July 2012, pp. 914-920 Summary of the Invention [Problem to be solved by the invention]

[0010] An object of the present invention is to provide a mosquito repellent composition and a method for producing the same, which can be produced by a single process using natural materials with different extraction and production conditions to produce an effective and uniform active ingredient. Another object of the present invention is to provide a mosquito repellent composition that uses natural materials that are harmless to humans to repel mosquitoes, and a method for producing the same. Yet another object of the present invention is to provide a mosquito repellent composition that, when applied to the skin, heals skin damage caused by physical irritation or mosquito bites without causing any skin trouble, and a method for producing the same. [Means for solving the problem]

[0011] The above object can be achieved by the present invention by mixing 100 to 500 parts by weight of each of cinnamon, Chinese laurel, chrysanthemum, green alder, angelica root, nutmeg, peppermint, and fennel to form a mixture, This is achieved by a mosquito repellent composition comprising an active ingredient extract obtained by mixing 100 parts by weight of the mixture with 100 to 700 parts by weight of 70 to 95% ethanol as a solvent to produce a mixed liquid, and then subjecting the mixed liquid to low-temperature ultrasonic extraction.

[0012] Here, the mixture may contain the same parts by weight of cinnamon, shanin, chrysanthemum, kawamidori, angelica root, nutmeg, mint and fennel.

[0013] Here, it is preferable that the concentration of the active ingredient extract is diluted by mixing with any one of 25% to 85% ethanol, purified water, and a mixture thereof.

[0014] Here, the low-temperature ultrasonic extraction is preferably carried out by applying ultrasonic waves of 700 kHz to 1400 kHz to the mixture in an atmosphere of 45 to 55 degrees Celsius for 24 to 96 hours to extract the active ingredient extract.

[0015] The above object can be achieved by the present invention by the steps of: preparing a mixture of 100 to 500 parts by weight of any one of cinnamon, Chinese laurel, chrysanthemum, Japanese laurel, angelica root, nutmeg, peppermint, and fennel; mixing 100 parts by weight of the mixture with 100 to 700 parts by weight of 70 to 95% ethanol as a solvent to prepare a mixed liquid; and applying ultrasound in the range of 700Khz to 1400Khz to the mixed liquid in an atmosphere at a temperature of 45 to 55 degrees Celsius to extract the active ingredient from the mixed liquid.

[0016] Here, after extracting the active ingredient, The method may further include the step of diluting the concentration by mixing with any one of 25% to 85% ethanol, purified water, and mixtures thereof.

[0017] Here, the extracting step is carried out for 24 to 96 hours, and the method may further include a step of filtering out by-products after the ultrasonic extraction. [Effects of the Invention]

[0018] According to the present invention, natural materials with different extraction and manufacturing conditions for the active ingredients can be used in a single process to ensure uniformity of the active ingredients while increasing the extraction efficiency, making it suitable for effective mass production. This makes it possible to obtain a uniform mosquito repellent effect similar to products made with chemicals, while eliminating the harmful effects on the human body compared to existing mosquito repellents made with chemicals.

[0019] Furthermore, according to the present invention, the mosquito repellent composition produced from naturally occurring substances maintains a mosquito repellency rate of 95% or more even at an extract concentration of 33%, demonstrating excellent mosquito repellency and being harmless to humans, unlike chemical preparations. [Brief explanation of the drawings]

[0020] [Figure 1] 1 shows a flow chart for a method of manufacturing a mosquito repellent composition according to one embodiment of the present invention. [Figure 2]Reference images are shown for test methods carried out by the applicant. [Figure 3] Reference images are shown for test methods carried out by the applicant. [Figure 4] Reference images are shown for test methods carried out by the applicant. [Figure 5] The repellent rates of Culex pipiens and Aedes albopictus when the mosquito repellents according to the examples were applied to test subjects are shown. [Figure 6] 1 shows the mosquito repellency rate of Culex pipiens mosquitoes at different concentrations for the mosquito repellents of the examples. [Figure 7] The results of the probit analysis for Figure 6 are shown. [Figure 8] 1 shows the mosquito repellency rate of Aedes albopictus at each concentration for the mosquito repellent according to the embodiment. [Figure 9] The results of the probit analysis for Figure 8 are shown. [Figure 10] 1 shows a reference diagram for the duration of mosquito repellent effect of the mosquito repellent according to the embodiment. [Figure 11] 1 shows experimental images of skin stability and skin soothing effect when the mosquito repellent prepared in this example is applied to the skin. DETAILED DESCRIPTION OF THE INVENTION

[0021] Before describing the present invention, the main materials mentioned in the present invention will be briefly described as follows. Cinnamon contains components such as phenylpropanoids, tannins, phenolic acids, diterpenoids, and coumarin, and we would like to clarify that the term "cinnamon" used in this specification does not refer to cinnamon, which is used as a food ingredient in the West.

[0022] Cinnamon can refer to one of the following: Chinese cassia, Vietnamese Saigon cinnamon, and Indonesian cinnamon. While the term cinnamon in Korea usually refers to cassia varieties, Saigon cinnamon and Indonesian cinnamon also qualify as "cinnamon" in this specification. Cinnamon also refers to the bark of the aforementioned cassia, Saigon cinnamon, and Indonesian cinnamon trees.

[0023] The cinnamon used in Western cuisine, also known as cinnamon bark, contains only 1 / 100 to 1 / 250 of the cinnamaldehyde and coumarin components, which are active ingredients that repel insects, compared to cassia, Saigon cinnamon, and Indonesian cinnamon. To extract the same active ingredients, 100 to 250 times more material is required than the cinnamon mentioned in this specification, which requires excessive material costs and therefore cannot be used commercially.

[0024] The cinnamon used in Western cooking contains only 0.004% cinnamaldehyde per kg, while the cinnamon contains 1% per kg.

[0025] The term "Angelica dahurica" ​​(Angelica dahurica Benth. et Hooker) referred to herein is a medicinal herb made from the dried root of the angelica tree, a plant in the Umbelliferae family. It is known for its effectiveness in expelling colds, eliminating dampness, reducing swelling, and relieving pain. Angelica dahurica root contains byakangelicin, a coumarin derivative, and oxypeucedanin essential oil. Because the efficacy of angelica dahurica is greatest in the root, the term "Angelica dahurica" ​​referred to herein refers to the root of the plant. However, the terms "Angelica dahurica" ​​and "Angelica dahurica root" may be used interchangeably herein.

[0026] The pharmacological effects of Angelica sinensis mentioned in this specification include antibacterial effects against Escherichia coli, Shigella, and Haemophilus influenzae, and stimulating effects on the vasomotor center, respiratory center, and vagus nerve. In particular, coumarin contained in the roots of Angelica sinensis is a substance with a simple structure containing a phenol group that gives off a sweet scent, but also has hostile properties against insects, mold, and bacteria.

[0027] Fennel (Foeniculum vulgare Miller) is the dried mature fruit of the perennial herb Foeniculum vulgare, a plant belonging to the Apiaceae family. It contains volatile compounds (essential oils), phenylpropanoids, monoterpenes, stilbenes, and other compounds. Fennel has antibacterial and antiviral properties, as well as bactericidal, anti-mite, and insecticidal properties. The fennel referred to in this invention refers to fennel fruit, and will be referred to as fennel, rather than separately referring to fennel fruit.

[0028] Nutmeg is the dried ripe fruit of the nutmeg tree, an evergreen broadleaf tree in the Myristicaceae family of the Ranunculaceae order. Nutmeg contains large amounts of fat (28-34%) and essential oil (3-9%). The fat is primarily composed of myristic acid and trimyristin, while the essential oil is composed of d-camphene (60-80%), d-pinene, limonene, d-borneol, l-terpineol, geraniol, safrole, and myristicin. Trimyristin, which is found in large amounts in nutmeg, is classified as a compound hostile to insects.

[0029] Shanin (Amomum villosum Lour) is a medicinal herb made from the fruit of Amomum villosum, Amomum villosum var. xanthioides, or Amomum longiligulare TLWu, a perennial herbaceous plant belonging to the Zingiberaceae family. Its medicinal properties include 1.7% to 3% aromatic essential oil components, the main components of which include d-camphor, dibornaneol, bornyl acetate, linalool, neroldal, and monoterpenes.

[0030] Mint (Mentha arvensis var piperascens) is a dicotyledonous perennial plant in the Lamiaceae family. The plant itself is aromatic, growing upright and branching from the top. Mint emits a clean, refreshing scent and has bactericidal and antibacterial properties against pathogenic fungi that cause eczema and scabies on the skin and scalp, as well as microorganisms such as E. coli and staphylococcus. When concentrated, limonene, one of the substances contained in mint, can be used as a natural insecticide, paralyzing the nervous systems of small organisms such as insects, killing them.

[0031] Chrysanthemum is a perennial herb in the family Asteraceae, which contains mainly essential oils, flavonoids, sesquiterpenes, and triterpenoids. Chrysanthemum is known to have antipyretic, anti-inflammatory, antibacterial, antiviral, and antioxidant properties.

[0032] Patchouli japonica, also known as patchouli japonica, is a perennial herb belonging to the mint family that is widely cultivated in the Philippines, India, southern China, and other regions. It is known to contain approximately 15% essential oils, including patchouli alcohol, eugenol, cinnamic aldehyde, pagostol, and patchouli pyridine. Patchouli alcohol, one of the components of patchouli japonica, is also effective in repelling pests, and patchouli leaves were once used as an insect repellent by placing them between clothes to protect against pests.

[0033] Patchouli extracts have been extracted by distillation and by adding water to dried leaves for direct extraction. When using distillation, the cell membrane of the patchouli leaves can be destroyed before distillation. For example, patchouli essential oil with a high content of patchouli alcohol can be obtained by distilling dried patchouli leaves and stems to extract low-molecular-weight, highly volatile components.

[0034] A typical Kawamidori extract can be obtained in liquid form by removing impurities through filtration using conventional methods, or the extract can be concentrated through a vacuum concentration process or obtained in powder form through a drying process such as vacuum drying, freeze drying, or spray drying. The present invention will be described in detail below with reference to the drawings.

[0035] The mosquito repellent of the present invention is made from natural materials, including cinnamon, Chinese laurel, chrysanthemum, Japanese jasmine, angelica root, nutmeg, peppermint, and fennel, and the composition ratio of each material can be determined by the following items.

[0036] 1) Mixing cinnamon, Chinese laurel, chrysanthemum, green alder, angelica root, nutmeg, peppermint, and fennel in amounts ranging from 100 to 500 parts by weight to form a mixture; 2) A mixture can be prepared by using equal parts by weight of cinnamon, Chinese laurel, chrysanthemum, green alder, angelica root, nutmeg, peppermint and fennel.

[0037] The composition ratio of the mixture mentioned in item 1) is 400 parts by weight of angelica and cinnamon, and 100 parts by weight of the remaining materials, and the mixture can be produced in a way that increases the content of coumarin and coumarin derivatives, which have a high mosquito repellent effect, thereby enhancing the mosquito repellent effect.

[0038] On the other hand, the mixture may have a composition ratio of 300 to 500 parts by weight of angelica, 200 to 400 parts by weight of cinnamon, and 200 to 400 parts by weight of fennel, respectively, 100 to 300 parts by weight of kandol, 200 to 300 parts by weight of shantung, 100 to 300 parts by weight of nutmeg, 100 to 300 parts by weight of chrysanthemum, and 100 to 300 parts by weight of mint.

[0039] A mosquito repellent with such a composition ratio has a mosquito repellent effect of 95% or more, and when the mosquito repellent according to the embodiment is applied to the skin of a user, it exhibits an even more excellent skin soothing effect on skin affected by external irritation or mosquito bites. The mosquito repellent obtained by the mixture according to item 2) corresponds to the composition ratio that provides the optimum mosquito repellent effect in the present invention.

[0040] When the concentration of the active ingredient extract is diluted with a 1:1 mixture of 83% alcohol and purified water, the mosquito repellency rate for each concentration of the diluted active ingredient extract is as follows:

[0041] Here, the diluted active ingredient extract can be described as a mosquito repellent. For ease of explanation and understanding, it may be referred to as a diluted active ingredient extract or a mosquito repellent, or when the dilution ratio is different, it may be referred to as a diluted mosquito repellent or a diluted active ingredient extract.

[0042] When the concentration of the active ingredient extract was 38.9%, the repellent effect against Culex pipiens mosquitoes reached 95%, and when the concentration of the active ingredient extract was 28.4%, the repellent effect against Aedes albopictus mosquitoes reached 99.9%. The extraction characteristics of the main components of the composition according to the present invention are as follows:

[0043] Cinnamaldehyde, the main active ingredient in cinnamon, has a boiling point above 100°C and is highly heat-resistant, so it can be extracted using the thermal extraction method, which uses water as a solvent.

[0044] The active ingredient in jasmine is patchouli alcohol, which can weaken or even kill insects if it comes into direct contact with them. However, because patchouli alcohol has a lower boiling point than water, it cannot be extracted by thermal extraction. To extract patchouli alcohol from jasmine, the plant is usually heated to produce steam, which is then cooled and distilled to extract the active ingredient.

[0045] Menthol, the active ingredient in mint, is highly volatile and has a very low boiling point, so it can be extracted by distillation.

[0046] The roots of Angelica dahurica contain active ingredients, including angelicin and oxypoisedanin, which are active coumarin compounds. The active ingredients in Angelica dahurica are coumarin compounds, which have a high boiling point of 301.7℃ and cannot be extracted by distillation.

[0047] Flavonoids, the active ingredient found in chrysanthemums, are a group of substances with a structure in which two benzene rings (C6) are linked by three carbon atoms (C3). Flavonoids include a variety of pigments, such as anthocyanins, flavones, flavonols, and isoflavones. The boiling point of flavonoids ranges from 80°C to 87°C, and the closer they are to the boiling point of water (100°C), the more easily their active ingredients are destroyed. Flavonoids are not suitable for extraction using thermal extraction or distillation, and attempts to do so may result in the loss of most of the active ingredients.

[0048] As described above, cinnamon and angelica can be extracted by thermal extraction, while peppermint and jasmine can be extracted by distillation, while chrysanthemum is unsuitable for thermal extraction or distillation. This means that three or more extraction methods are required for natural ingredients that make up mosquito repellents. Due to this issue, natural-based mosquito repellents have been developed using thermal extraction methods using only cinnamon and angelica, while mosquito repellents that combine cinnamon, angelica, or chrysanthemum with herbal plants have been researched. Therefore, the present applicant aims to extract active ingredients from six natural ingredients in a single process using the following method, thereby ensuring uniformity of the active ingredients and the ability to mass-produce them.

[0049] FIG. 1 shows a flow chart of a method for producing a mosquito repellent composition according to one embodiment of the present invention. Referring to FIG. 1, a method for producing a mosquito repellent composition according to one embodiment includes the steps of: A mixture of equal parts by weight of cinnamon, Chinese laurel, chrysanthemum, Japanese laurel, angelica root, nutmeg, peppermint, and fennel is prepared (S10). At this time, the mixture can be in the form of chips or powder.

[0050] The reason for forming the mixture into chips or powder is to efficiently extract the active ingredients of the ingredients (cinnamon, chinese ginseng, chrysanthemum, jasmine, angelica root, nutmeg, peppermint, and fennel), which corresponds to the process of finely crushing the mixture into chips or powder. When crushing into chips, the size of the chips may be in the range of 1 mm to 10 mm, and the chips may be crushed into a regular or irregular shape. The shape of the chips does not necessarily need to be uniform.

[0051] Next, 100 parts by weight of the mixture is mixed with 100 to 700 parts by weight of 70 to 95% ethanol as a solvent to produce a mixed solution (S20). At this time, the ethanol solvent may be heated at a low temperature of 45 to 55°C, which corresponds to a temperature range in which the volatile and non-volatile active ingredients as well as the alcohol solvent are not damaged or volatilized.

[0052] The temperature at which the mixture is heated is at least 40°C and at most 80°C, but temperatures lower or higher than this are not suitable for extracting the active ingredients.

[0053] When ethanol and the mixture are mixed, the mixture exists in the form of particles in the ethanol. When the mixture of ethanol and the mixture is made into a mixed liquid, the mixed liquid can have a temperature range of 45°C to 55°C. The temperature of the mixed liquid can be determined by the following two conditions.

[0054] 1) If the active ingredients are volatile or easily destroyed by high temperatures, such as patchouli alcohol, menthol, and flavonoids, the temperature of the mixture must be lower than the temperature at which these active ingredients volatilize or are destroyed.

[0055] 2) When extracting active ingredients from a mixed solution, the higher the temperature of the mixed solution, the greater the extraction efficiency of the active ingredients. The higher the temperature of the mixed solution, the greater the amount of active ingredients extracted per unit time (e.g., 1 hour). This has also been mentioned in a paper explaining the relationship between the extraction of herbal medicine ingredients and temperature.

[0056] The Journal of the Korean Society of Plant Nutrition, Vol. 41, No. 7 (July 2012), pp. 914-920, points out that the extraction efficiency of active ingredients from most herbal medicines increases in proportion to the temperature of the solution (or solvent).

[0057] The Journal of the Korean Society of Plant Nutrition, Vol. 41, No. 7 (July 2012), pp. 914-920, points out that the active ingredients of herbal medicines can be extracted to the maximum extent by using a thermal extraction method in which the solution (or solvent, hereafter abbreviated) is heated. It also points out that extraction using ultrasound in the 22-40 kHz range at room temperature (24°C) shows two to four times the extraction efficiency of active ingredients compared to thermal extraction.

[0058] The mosquito repellent according to the present invention contains a large number of non-volatile compounds (e.g., cinnamaldehyde, angelicin, oxypoisedanin, etc.) and volatile compounds (menthol, flavonoids, patchouli alcohol, etc.) that affect mosquitoes, such as cinnamaldehyde, patchouli alcohol, and menthol compounds, so there is a high risk of loss of the volatile active ingredients when using a thermal extraction method. Therefore, the applicant has devised a method of applying mild heat to the mixture to increase extraction efficiency while preventing the active ingredients from volatilizing, thereby significantly reducing the time required to produce an environmentally friendly mosquito repellent with a high content of active ingredients.

[0059] For this purpose, the applicant set the maximum temperature at which volatile compounds are not damaged to 80°C, and considered a method of heating the mixture below the maximum temperature of 80°C and then performing ultrasonic extraction.

[0060] At this time, the temperature of the mixture to which the low-temperature ultrasonic extraction method is applied may have the temperature range described below, and the lower the temperature range, the longer the extraction time, and vice versa. -41℃ or higher and 79℃ or lower, -42℃ or higher and 78℃ or lower, -43℃ or higher and 77℃ or lower, -44℃ or higher and 76℃ or lower, -45℃ or higher and 75℃ or lower, -46℃ or higher and 74℃ or lower, -47℃ or higher and 73℃ or lower, -48℃ or higher and 72℃ or lower, -49℃ or higher and 71℃ or lower, -50℃ or more and 70℃ or less, -51℃ or higher and 69℃ or lower, -52℃ or higher and 68℃ or lower, -53℃ or higher and 67℃ or lower, -54℃ or higher and 66℃ or lower, It may be between -55°C and 65°C.

[0061] Preferably, the mixture is heated to a temperature in the range of 45°C to 55°C to minimize destruction or damage to the volatile active ingredients and to minimize volatilization of the volatile active ingredients, and the mixture is heated at a temperature higher than room temperature (24°C).

[0062] Next, the mixture is subjected to low-temperature ultrasonic extraction (S30). Instead of the 40Khz range of ultrasonic waves typically applied to herbal medicines, the ultrasonic range in this embodiment has a frequency range of 700Khz to 1400Khz. The ultrasonic frequency band corresponds to 700kHz to 1400kHz, and the mixture is strongly vibrated to extract the active ingredients.

[0063] Ultrasonic extraction is carried out in an atmosphere with a temperature of 45°C to 55°C, which is a relatively low temperature compared to the usual heating of natural materials at 100°C or higher. Therefore, the applicant has decided to refer to this as "low-temperature ultrasonic extraction (method)." The mixed liquid may be subjected to low-temperature ultrasonic extraction for about 24 to 96 hours.

[0064] When the temperature range of the mixed liquid is 45°C to 55°C, the extraction time is approximately 96 hours. When the temperature range of the mixed liquid is 55°C to 80°C, the time for low-temperature ultrasonic extraction is reduced, and the extraction time can be reduced to 24 hours or closer.

[0065] Using low-temperature ultrasonic extraction, the active ingredients from chips or powdered natural materials (cinnamon, chinese ginseng, chrysanthemum, jasmine, angelica root, nutmeg, peppermint, and fennel) contained in the mixture are extracted into the solvent alcohol. The active ingredients extracted into the alcohol dissolve in the alcohol to form an aqueous solution, which can then be put into containers and used as a product.

[0066] At this time, the temperature of the mixture to which the low-temperature ultrasonic extraction method is applied may have the temperature range described below, and the lower the temperature range, the longer the extraction time, and vice versa.

[0067] The mixture is then vibrated by ultrasonic waves at a temperature (45°C to 55°C) higher than room temperature (24°C), which is too low for the active ingredient to volatilize, and the active ingredient is dissolved.

[0068] Finally, the mixture is filtered to remove the active ingredients and by-products such as dregs generated during the active ingredient extraction process (S40), and the active ingredient extract is produced, which can then be placed in a container and commercialized.

[0069] At this time, the extracted active ingredient extract can be diluted by mixing with 25% to 85% ethanol to be shipped at the appropriate concentration before commercialization. The active ingredient extract can be diluted by mixing with 25% to 85% ethanol or distilled water to adjust the concentration of the active ingredient extract.

[0070] Such dilution may be performed after step S40 as an additional step of diluting the concentration of the active ingredient extract, which is a step of diluting the active ingredient extract to a concentration required to obtain a mosquito repellent effect, and the concentration is lower than that of the original solution of the active ingredient extract, thereby reducing the burden on the human body.

[0071] Furthermore, in the concentration dilution step, the active ingredient extract can also be diluted with a 1:1 mixture of 25% to 85% ethanol and distilled water, but this is not limited thereto.

[0072] Hereinafter, the mosquito repellent effect of the mosquito repellent prepared according to the examples will be explained through specific experiments. ■ Experiment 1 1. Experimental Conditions 1) Mosquito selection The mosquitoes used in the experiment, 200 adult female Culex pipiens or 50 adult female Aedes albopictus, were placed in a screened cage (80cm x 40cm x 40cm) with a visible interior and allowed to stabilize one hour before the experiment. All test conditions for the two species of mosquito were the same, but because mosquito species have different preferences for feeding on human blood, 200 adult female Culex pipiens and 50 adult female Aedes albopictus were used to ensure smooth experiments. The temperature inside the mosquito cage was 27.2±2°C and the relative humidity was 65±10%.

[0073] 2) Subject conditions The subjects wore masks to prevent their breath from affecting the mosquitoes' blood-sucking. The test was conducted after 6:00 PM, when the nocturnal Culex taeniorhynchus is most active in blood-sucking, and around 10:00 AM or between 3:00 PM and 6:00 PM, when the diurnal Aedes albopictus is most active in blood-sucking.

[0074] Approximately 100cm from the subject's forearm between the wrist and elbow 2 The subjects were then wrapped in cotton gloves and cotton cloth, with the exception of the exposed skin of the area. The left arm was used as the control group, and the right arm as the treatment group, and the same mosquitoes were used on the same person. The concentration of ethanol solution used in the control group was 25%, in accordance with WHO (World Health Organization) standards.

[0075] The control arm was then used as a control group, with 1 mL of 25% ethanol evenly applied to the exposed skin and allowed to dry for approximately one minute. The ethanol-coated arm was placed in a mosquito cage containing 200 female mosquitoes when conducting experiments with Culex pipiens pallens, and 50 female mosquitoes when conducting experiments with Aedes albopictus, and the number of mosquitoes that landed on the skin for three minutes at one-minute intervals was recorded, depending on the intensity of their attempts to suck blood.

[0076] If the rate at which mosquitoes landed on the subject's arm was two or more within 30 seconds, or one within 30 seconds, then one within 30 seconds, the group was considered to be ready to feed, and the test began. 3) Mosquito repellent dilution conditions

[0077] A powdered mixture containing equal parts by weight of cinnamon, Chinese laurel, chrysanthemum, jasmine, angelica root, nutmeg, peppermint, and fennel was prepared, and 100 parts by weight of the mixture was mixed with 100 to 700 parts by weight of 83% ethanol as a solvent to prepare a mixed solution. The mixed solution was then subjected to low-temperature ultrasonic extraction to extract the active ingredient extract, and 25% ethanol or purified water was added to adjust the concentration of the extracted active ingredient extract, thereby producing five concentrations of active ingredient extract (hereinafter referred to as mosquito repellent).

[0078] The concentrations of the mosquito repellent were 1%, 3%, 10%, 20%, and 33.3%, and we observed how the mosquito repellent effect and duration changed as the concentration went from low (1%) to high (33%).

[0079] 2. Experimental Method As shown in Figure 2, the subject's forearm area between the wrist and elbow was approximately 100 cm 2 The area was wrapped in gloves and cotton cloth except for the exposed skin. The experimental method shown in Figure 2 differs in part in terms of experimental time and method depending on whether the mosquitoes used are diurnal or nocturnal.

[0080] When conducting experiments on the diurnal Aedes albopictus mosquito, the experiment is conducted around 10:00 a.m. or between 3:00 p.m. and 6:00 p.m., as shown in Figure 3. However, when conducting experiments on the nocturnal Culex pipiens mosquito, as shown in Figure 4, only the lights are left on to observe the Culex pipiens mosquito inside the mosquito cage, and the area around the mosquito cage is kept dark according to the experimental time.

[0081] The subject's left arm was defined as the control group, and the right arm as the treatment group. The subjects' arms were used as the control and treatment groups, respectively, ensuring that the same subjects were exposed to the same mosquito population. The ethanol solution used for the control group was 25%, in accordance with WHO (World Health Organization) standards. The control arm was prepared by applying 1 mL of 25% ethanol evenly to the exposed skin and letting it dry for approximately one minute.

[0082] When conducting experiments with Culex pipiens mosquitoes, 200 female mosquitoes were used, and when conducting experiments with Aedes albopictus mosquitoes, 50 female mosquitoes were used. The subjects' arms, which had been coated with ethanol, were placed in mosquito cages containing 200 female mosquitoes, and the number of mosquitoes landing on the skin was recorded for three minutes, one minute at a time, depending on the degree of their attempt to suck blood. When the ratio of mosquitoes landing on the subject's arm was two or more within 30 seconds, or one within 30 seconds, and then one within the next 30 seconds, the group was considered to be ready to suck blood, and the test began.

[0083] The treatment group was treated with the sample solution on the exposed forearm in the same manner as the control group, and then tested in the same manner. Each test was completed within one day using the same mosquitoes on the same subject. In repeat tests, this process was repeated using mosquitoes in different locations over several days. For statistical analysis, a minimum of three repeat tests were performed. 3. Calculation of mosquito repellent effect

[0084] For statistical analysis of the experimental data, the mosquito repellency rate was calculated by taking into account the blood-feeding attempt rate of the control group and correcting the repellency rate of the treated group using the modified Abbott formula (Abbott 1925). The modified Abbott formula is as follows: Repellency rate (%) = 100 x (number of landings in the control group - number of landings in the treated group) / number of landings in the control group

[0085] To confirm the effective concentration range of the mosquito repellent, a diluted solution of the mosquito repellent was prepared using 25% ethanol as a solvent. The concentration that produced a repellent response from mosquitoes was used, and two to three concentrations that produced a repellent response of 50% or less and two to three concentrations that produced a repellent response of 50% or more were selected for use.

[0086] The selected concentrations of repellent were applied to the test forearm of each subject, and experiments were conducted to confirm repellent efficacy. At least five concentrations were used. Each test was completed within one day using the same mosquitoes for the same subject. For replicate tests, this process was repeated using mosquitoes in different locations over several days. At least three replicate tests were conducted per subject. Repellent efficacy was calculated by substituting the data into the modified Abbott formula described above. The effective concentrations (EC50, EC95, and EC99.9) of the mosquito repellents were determined with 95% confidence limits using probit analysis.

[0087] 4. Calculation of mosquito repellent effect duration (Complete Protection Time, CPT) This test was conducted to examine the duration of repellency (CPT) of the mosquito repellent according to the embodiment. The experimental method was the same as the mosquito repellent effect test described above. After applying the mosquito repellent to the subjects, the mosquito repellent effect was measured in one-hour increments up to five hours. However, the test was terminated when the mosquito repellent effect fell below 20%. To measure mosquito repellency, the subjects were exposed to the mosquito for three minutes each time, and the number of mosquitoes landing on the subjects was counted. This test was repeated at 60-minute intervals.

[0088] To evaluate the mosquito repellent durability of the mosquito repellent, Duncan's Multiple Range Test statistical method was used to test the difference in the time elapsed after mosquito repellent application, and the analysis was performed at the 95% probability level using the SPSS PC statistical program.

[0089] 5. Experimental Results 1) Mosquito repellent effect of mosquito repellents The repellency rate of a 33.3% concentration of the active ingredient extract (original herbal medicine solution: 83% ethanol: purified water = 1:1:1) against Culex pipiens mosquito and Aedes albopictus was measured. After checking the repellency rate for the two mosquito species for 3 minutes, the average was 97.2% for Culex pipiens mosquito and 97.6% for Aedes albopictus, as shown in Figure 5.

[0090] 1-1) Repellency rate of Culex pipiens mosquitoes To determine the effective concentrations (EC50, EC90, and EC99.9) of mosquito repellents against Culex pipiens mosquitoes, the repellency was measured at five or more concentrations, namely 1%, 3%, 10%, 20%, and 33.3%. The measurement results are shown in Figure 6 below.

[0091] A probit analysis was performed with 95% confidence limits using the mosquito repellency rates derived for each concentration of mosquito repellent (1%, 3%, 10%, 20%, and 33.3%), resulting in R2 = 0.751 and y = 2.232x + 7.982, as shown in Figure 7.

[0092] In Figure 6, when the mosquito repellent concentration is between 1% and 20%, the repellency rate against Culex pipiens mosquitoes is 22% to 35%, but when the mosquito repellent concentration reaches 33%, the mosquito repellency effect reaches 97%.

[0093] Although not shown in Figure 6, the applicant's experiments confirmed that when the mosquito repellent concentration was 18.8%, the effective concentration (EC50) for a 50% mosquito repellent rate was reached, when the mosquito repellent concentration was 38.9%, the effective concentration (EC90) for a 95% repellent rate was reached, and when the mosquito repellent concentration was 40.8%, the effective concentration (EC99.9) for a 99% repellent rate was reached.

[0094] 1-2) Repellency rate against Aedes albopictus To determine the effective concentrations (EC50, EC90, and EC99.9) of the mosquito repellent against Aedes albopictus, the mosquito repellency was measured at five concentrations (2%, 5%, 10%, 20%, and 33.3%), and the results are shown in Figure 8.

[0095] A probit analysis was performed with a 95% confidence limit using the mosquito repellency rates derived for each concentration (2%, 5%, 10%, 20%, and 33.3%). The results were R2 = 0.738, y = 2.201x + 35.421, as shown in Figure 9.

[0096] On the other hand, it was confirmed that the effective concentration (EC50) showing a 50% repellency rate for Aedes albopictus corresponds to a mosquito repellent concentration of 6.94%, the effective concentration (EC90) showing a 95% repellency rate corresponds to a mosquito repellent concentration of 28.4%, and the effective concentration (EC99.9) showing a 99% repellency rate for Aedes albopictus corresponds to a mosquito repellent concentration of 30.3%.

[0097] 2) Duration of mosquito repellent effect To evaluate the durability of mosquito repellent, the repellency rate was measured at hourly intervals after treatment. The repellent concentration was selected based on a previous experiment that was calculated to provide a repellency rate of 95% or higher. Therefore, experiments were conducted on 200 female Culex pipiens mosquitoes using a 33.3% repellent, and on 50 female Aedes albopictus mosquitoes using a 30% repellent.

[0098] The experimental results are shown in Figure 10. Immediately after treatment with the mosquito repellent, the repellency rates for Culex pipiens mosquito and Aedes albopictus were 97.2% and 90.3%, respectively. After three hours, the repellency rates for Culex pipiens mosquito and Aedes albopictus were 13.3% and 0.5%, respectively.

[0099] As shown in Figure 10, the repellency rate for both mosquito species exceeded 90% immediately after treatment, but dropped to below 20% one hour after treatment. The difference in mosquito repellency over time was analyzed using Duncan's Multiple Range Test with a 95% confidence limit. For Culex pipiens, the mosquito repellency rate immediately after treatment was higher than that after 1, 2, and 3 hours, but there was no difference in the mosquito repellency rates between 1, 2, and 3 hours. For Aedes albopictus, the mosquito repellency rate immediately after treatment was higher than that after 1 hour, and the mosquito repellency rate after 1 hour was higher than that after 2 hours, but there was no difference in the mosquito repellency rates between 2 and 3 hours. FIG. 11 shows experimental images of the skin stability and skin soothing effect when the mosquito repellent prepared according to this example was applied to the skin of a subject.

[0100] The study was conducted on subjects aged 19 to 60 who had not experienced any skin diseases or undergone any cosmetic skin treatments within the past six months. The images shown in Figure 11 were taken using the Antera 3D device.

[0101] The experimental method was to apply physical stimulation to the subject's forearm to induce erythema, and then to apply a 1cm 2 2 ml of mosquito repellent was applied per area and the results were photographed. To ensure the fairness of the experiment, one of the subjects' forearms was applied with mosquito repellent to serve as the test group, and the other forearm was used as the control group without mosquito repellent.

[0102] In other words, the results were obtained from the same subjects, whose reactions were the same when the mosquito repellent was applied and when it was not applied. This experiment was conducted on a total of 22 people, and the applicant presented a representative image.

[0103] Referring to Figure 11, the test group and the control group of subjects were given physical stimulation to cause redness (erythema) on their forearms. Looking at the images of the test group in Figure 11, the image in the "Before physical stimulation" category shows the Antera 3D image before the physical stimulation was given to the subject, the "After physical stimulation" category shows the Antera 3D image after the physical stimulation was given to the subject, and the "After one use of test product" category shows the image after the mosquito repellent according to this example was applied to 1 cm 2 2ml was applied per skin, and the image shown is taken 6 hours later.

[0104] As shown in Figure 11, the control group after physical stimulation did not receive the mosquito repellent of this example, and therefore had almost no soothing effect on the skin, and there was no significant change in the average redness of the skin.

[0105] In contrast, the test group showed a decrease in average skin redness after applying the mosquito repellent of this example. All 22 subjects showed an improvement in average skin redness, with a minimum of 9.29% and a maximum of 51.3%. Furthermore, none of the subjects experienced any adverse skin reactions 24 hours after applying the mosquito repellent. This is because the mosquito repellent of this example is made of naturally derived substances and a concentration of 33% was used as the appropriate concentration. Through Experiment 2, it can be seen that the mosquito repellent according to the embodiment is harmless to the human body and has a skin soothing effect without inducing side effects.

[0106] The present embodiment and the drawings attached to this specification only clearly show a part of the technical ideas included in the present invention, and all various modifications and specific embodiments that can be easily inferred by a person skilled in the art within the scope of the technical ideas included in the specification and drawings of the present invention are included in the scope of the present invention.

Claims

1. forming a mixture by mixing 100 to 500 parts by weight each of cinnamon, Chinese laurel, chrysanthemum, green alder, angelica root, nutmeg, peppermint, and fennel; The active ingredient extract is obtained by mixing 100 parts by weight of the mixture with 100 to 700 parts by weight of ethanol having a concentration of 70 to 95% as a solvent to produce a mixed solution, and then subjecting the mixed solution to low-temperature ultrasonic extraction. A mosquito repellent composition comprising:

2. The mixture comprises: The cinnamon, the shanin, the chrysanthemum, the kawamidori, the angelica root, the nutmeg, the mint and the fennel are mixed in equal parts by weight. The mosquito repellent composition of claim 1.

3. The active ingredient extract is Dilute the concentration by mixing with 25% to 85% ethanol, purified water, and any one of their mixtures. The mosquito repellent composition of claim 1.

4. The low-temperature ultrasonic extraction The mixture is subjected to ultrasonic waves of 700 kHz to 1400 kHz in an atmosphere of 45 to 55 degrees Celsius for 24 to 96 hours to extract the active ingredient extract. The mosquito repellent composition of claim 1.

5. producing a mixture of 100 to 500 parts by weight of any one of cinnamon, Chinese laurel, chrysanthemum, Japanese laurel, angelica root, nutmeg, peppermint, and fennel; Mixing 100 parts by weight of the mixture with 100 to 700 parts by weight of ethanol having a concentration of 70 to 95% as a solvent to produce a mixed liquid; and applying ultrasonic waves in the range of 700 Khz to 1400 Khz to the mixture in an atmosphere of 45 to 55 degrees Celsius to extract the active ingredient from the mixture. A method for producing a mosquito repellent composition, comprising:

6. After extracting the active ingredient, and diluting the concentration by mixing with any one of 25% to 85% ethanol, purified water, and mixtures thereof. A method for producing the mosquito repellent composition according to claim 5.

7. The extracting step includes: It takes place over 24 to 96 hours. and filtering the by-products after the ultrasonic extraction. A method for producing the mosquito repellent composition according to claim 5.

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