Finger-root liposomes containing finger-root extract and manufacturing method therefor

Abstract

The present invention relates to finger-root liposomes containing a finger-root extract and a manufacturing method therefor and, more specifically, to finger-root liposomes containing a finger-root extract and a manufacturing method therefor, wherein the finger-root liposomes are manufactured by highly concentrating the finger-root extract into liposomes, thereby reducing ginger flavor and exhibiting no toxicity.

Description

Fingerroot liposome containing fingerroot extract and method for preparing the same

[0001] The present invention relates to a fingerroot liposome containing a fingerroot extract and a method for producing the same, and more particularly, to a fingerroot liposome containing a fingerroot extract that does not exhibit toxicity while reducing ginger flavor by producing the fingerroot liposome by highly concentrating the fingerroot extract into liposomes, and a method for producing the same.

[0002] Fingerroot is a plant of the ginger family native to Southeast Asia. It has been used for various purposes, including in traditional Asian dishes such as pickles, curries, and drinks, as a tonic for childbirth, to treat stomachaches, diarrhea, flatulence, indigestion, ulcers, leukocyte prevention, and as a cost-effective agent.

[0003] These fingerroots contain flavonoids that inhibit infection by Helicobacter pylori bacteria, including those that cause indigestion and gastritis. They also contain pinostrobin, which has cytoprotective effects, which increases gastric mucus and reduces the area where gastric ulcers form, making them effective in treating peptic ulcers.

[0004] In addition, fingerroot contains flavonoid derivatives, pinocembrin, pinostrobin, ipinetin, tadamonin, pandurata, and panduratin A, which can inhibit the growth of cancer cells. In particular, the pandurata component is known to inhibit the growth of cancer cells, and the panduratin A component is known to have a preventive effect against prostate cancer and colon cancer.

[0005] Accordingly, attempts have recently been made to add fingerroot extracts extracted from fingerroot as raw materials for cosmetic compositions and foods.

[0006] However, because the ginger flavor of fingerroot extract is too strong, there was a problem of lowering consumers' flavor preference when added in large quantities as a raw material in cosmetic compositions and foods.

[0007] Additionally, fingerroot extract is highly toxic, so there are concerns about side effects when used in high concentrations.

[0008] The purpose of the present invention is to provide a fingerroot liposome containing a fingerroot extract that does not exhibit toxicity while reducing ginger flavor, and a method for producing the same by producing a fingerroot liposome by highly concentrating the fingerroot extract into liposomes.

[0009] A method for producing a fingerroot liposome containing a fingerroot extract according to an embodiment of the present invention is characterized by comprising the steps of: washing and hot-air drying a fingerroot; pulverizing the hot-air dried fingerroot to obtain a fingerroot powder, then adding the fingerroot powder to a solvent and performing a first extraction to form a fingerroot extract; filtering the fingerroot extract and then performing a second extraction using a rotary concentrator to form a fingerroot extract; and mixing a liposome base with a fingerroot extract dilution solution obtained by diluting the fingerroot extract in purified water and stirring to form a fingerroot liposome.

[0010] The above hot air drying is performed at 30 to 50°C for 12 to 60 hours.

[0011] The above solvent is selected from water, methanol, ethanol, phenoxyethanol, propylene glycol, and dipropylene glycol, or a mixture of two or more thereof.

[0012] The above fingerroot extract dilution solution uses the fingerroot extract diluted in purified water at a weight ratio of 1:8 to 1:12.

[0013] The above liposome base is mixed in an amount of 50 to 150 parts by weight per 100 parts by weight of the above fingerroot extract dilution.

[0014] The above liposome base comprises 70 to 90 wt% of a moisturizer, 10 to 30 wt% of an emulsifier, and 0.1 to 0.5 wt% of an oil.

[0015] The above stirring is performed at a room temperature of 1 to 40°C and a speed of 500 to 1,500 rpm for 1 to 3 hours.

[0016]

[0017] The fingerroot liposome containing the fingerroot extract according to an embodiment of the present invention is manufactured by concentrating the fingerroot extract into liposomes, and is characterized in that it exhibits no toxicity while reducing the ginger flavor.

[0018] The fingerroot liposome containing a fingerroot extract according to the present invention and the method for producing the same produce the fingerroot liposome by highly concentrating the fingerroot extract into liposomes, thereby reducing the ginger flavor and preventing toxicity.

[0019] As a result, the fingerroot liposome containing the fingerroot extract according to the present invention and the method for producing the same can make it possible to contain a large amount of the effective ingredient panduratin A when concentrating the fingerroot extract into liposomes.

[0020] In addition, the fingerroot liposome containing the fingerroot extract according to the present invention and the method for producing the same do not exhibit cytotoxicity, and not only does the fingerroot liposome increase both collagen and elastin production in human skin fibroblasts, but it also exhibits excellent wound healing efficacy.

[0021] Figure 1 is a process flow diagram showing a method for manufacturing a fingerroot liposome containing a fingerroot extract according to an embodiment of the present invention.

[0022] Figure 2 is a photograph showing a fingerroot extract and fingerroot liposome prepared according to Example 1.

[0023] Figure 3 is an SEM photograph showing a fingerroot liposome manufactured according to Example 1.

[0024] Figure 4 is a graph showing the results of the flavor preference and sensory evaluation of the fingerroot extract and fingerroot liposome manufactured according to Example 1.

[0025] Figure 5 is a graph showing the results of a cytotoxicity evaluation of fingerroot extract and fingerroot liposome prepared according to Example 1 on human skin fibroblasts.

[0026] Figure 6 is a graph showing the results of evaluating collagen production of fibroblasts for fingerroot liposomes according to Example 1 and general liposomes according to Comparative Example 1.

[0027] Figure 7 is a graph showing the results of evaluating elastin production by fibroblasts for fingerroot liposomes according to Example 1 and general liposomes according to Comparative Example 1.

[0028] Figure 8 is an SEM photograph showing the results of performing cell proliferation efficacy and wound healing efficacy of fingerroot liposomes according to Example 1 and general liposomes according to Comparative Example 1 on human skin fibroblasts.

[0029] Figure 9 is a graph showing the results of performing cell proliferation and wound healing efficacy of fingerroot liposomes according to Example 1 and general liposomes according to Comparative Example 1 on human skin fibroblasts.

[0030] Hereinafter, preferred embodiments of the present invention and the properties of each component will be described in detail. However, this is intended to be a detailed description to enable a person having ordinary knowledge in the technical field to which the present invention pertains to easily carry out the invention, and does not mean that the technical idea and scope of the present invention are limited thereby.

[0031]

[0032] Fingerroot liposome containing fingerroot extract according to an embodiment of the present invention is manufactured by highly concentrating fingerroot extract into liposomes, thereby reducing ginger flavor and exhibiting no toxicity.

[0033] To this end, a fingerroot liposome containing a fingerroot extract according to an embodiment of the present invention was prepared by diluting the fingerroot extract in purified water, adding it to a liposome base, and stirring at room temperature.

[0034] In this way, the fingerroot liposome containing the fingerroot extract according to the embodiment of the present invention can reduce toxicity while reducing the ginger flavor by manufacturing the fingerroot liposome by highly concentrating the fingerroot extract into liposomes.

[0035] As a result, the fingerroot liposome containing the fingerroot extract according to the embodiment of the present invention can contain a large amount of the effective ingredient panduratin A when the fingerroot extract is concentrated into liposomes.

[0036] In addition, the fingerroot liposome containing the fingerroot extract according to the embodiment of the present invention does not exhibit cytotoxicity, and not only increases both collagen and elastin production in human skin fibroblasts by the fingerroot liposome, but also exhibits excellent wound healing efficacy.

[0037]

[0038] This will be described in more detail through a method for producing fingerroot liposomes containing fingerroot extract according to an embodiment of the present invention.

[0039] Figure 1 is a process flow diagram showing a method for manufacturing a fingerroot liposome containing a fingerroot extract according to an embodiment of the present invention.

[0040] As shown in Fig. 1, a method for preparing fingerroot liposomes containing fingerroot extract according to an embodiment of the present invention includes a washing and hot air drying step (S110), a first extraction step (S120), a second extraction step (S130), and a dilution and stirring step (S140).

[0041]

[0042] Washing and hot air drying steps

[0043] In the washing and hot air drying step (S110), the finger root is washed and then dried with hot air.

[0044] Here, fingerroot contains flavonoid derivatives, pinocembrin, pinostrobin, ipinetin, tadamonin, pandurata, and panduratin A, which can inhibit the growth of cancer cells, and in particular, panduratin A has a preventive effect against prostate cancer and colon cancer.

[0045] Cleaning of these finger roots can be accomplished by using, but is not limited to, water washing, ultrasonic washing, and vibration washing.

[0046] It is recommended that hot air drying be performed at 30 to 50°C for 12 to 60 hours.

[0047] If the hot air drying temperature is lower than 30℃ or the hot air drying time is lower than 12 hours, there is a risk that the finger root may not be completely dried. If the hot air drying temperature exceeds 50℃ or the hot air drying time exceeds 60 hours, it may only increase manufacturing cost and time without any further increase in effectiveness, making it uneconomical.

[0048]

[0049] 1st extraction stage

[0050] In the first extraction step (S120), hot-air-dried fingerroot is pulverized to obtain fingerroot powder, and then the fingerroot powder is added to a solvent and first extracted to form a fingerroot extract.

[0051] Here, the grinding may be performed using one or more methods selected from a jet mill, a ball mill, a roller mill, etc., but is not limited thereto.

[0052] The fingerroot powder preferably has an average diameter of 100 to 1,000 nm, more preferably 300 to 500 nm. If the fingerroot powder has an average diameter of less than 100 nm, the process cost for grinding into fine particles may be excessively incurred, which is not preferred. In addition, if the fingerroot powder has an average diameter exceeding 1,000 nm, it may aggregate when mixed with a solvent, which is not preferred.

[0053] In addition, the solvent may be one selected from among water, methanol, ethanol, phenoxyethanol, propylene glycol, and dipropylene glycol, or a mixture of two or more thereof, and among these, it is more preferable to use ethanol.

[0054] It is desirable to carry out this primary extraction at room temperature of 1 to 40°C for 12 to 48 hours.

[0055]

[0056] Second extraction stage

[0057] In the second extraction step (S130), the fingerroot extract is filtered and then extracted a second time using a rotary concentrator to form the fingerroot extract.

[0058] Here, it is preferable to perform filtration of the fingerroot extract using a 200 to 600 mesh filter.

[0059] In this way, through the first and second extractions, a highly concentrated fingerroot extract can be obtained.

[0060]

[0061] Dilution and stirring steps

[0062] In the dilution and stirring step (S140), the liposome base is mixed with the fingerroot extract dilution solution, which is prepared by diluting the fingerroot extract in purified water, and stirred to form fingerroot liposomes.

[0063] Purified water is used to dilute the fingerroot extract, and may be water obtained through one or more distillations or purified through a filter.

[0064] Here, it is preferable to use a fingerroot extract dilution solution obtained by diluting the fingerroot extract with purified water in a weight ratio of 1:8 to 1:12.

[0065] It is preferable to mix the liposome base in an amount of 50 to 150 parts by weight per 100 parts by weight of the fingerroot extract dilution.

[0066] If the liposome base is mixed in an amount less than 50 parts by weight per 100 parts by weight of the fingerroot extract dilution, it may be difficult to control the fingerroot extract to be stably included inside the sphere, and if it is mixed in an amount exceeding 150 parts by weight, there is a concern that the content of the fingerroot extract may be relatively reduced and thus may not be highly concentrated, which is not desirable.

[0067] In this way, in the present invention, by strictly controlling the mixing ratio of the liposome base and the fingerroot extract dilution solution to highly concentrate the fingerroot extract into liposomes, fingerroot liposomes can be manufactured, thereby reducing toxicity while reducing the ginger flavor.

[0068] Accordingly, when the fingerroot extract is concentrated into liposomes, it becomes possible to contain a large amount of the active ingredient panduratin A.

[0069] In addition, it was experimentally proven that the fingerroot liposome manufactured by the method according to the embodiment of the present invention does not exhibit cytotoxicity, and not only increases both collagen and elastin production in human skin fibroblasts by the fingerroot liposome, but also exhibits excellent wound healing efficacy.

[0070] Here, the liposome base comprises 70 to 90 wt% of a moisturizer, 10 to 30 wt% of an emulsifier, and 0.1 to 0.5 wt% of an oil.

[0071] Humectants provide moisturizing benefits. These humectants include at least one selected from the following: butylene glycol, polyethylene glycol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, diglycerin, propanediol, 1,2-hexanediol, and sodium hyaluronate. Among these, propanediol is more preferred as a moisturizer.

[0072] If the amount of such moisturizer is less than 70% by weight, the amount may be insufficient to provide adequate moisturizing effects. Conversely, if the amount exceeds 90% by weight, the viscosity may increase, resulting in stickiness, which is not desirable.

[0073] In addition, emulsifiers play a role in maintaining high usability and emulsion stability.

[0074] These emulsifiers include Glyceryl Stearate, Ceteareth-20, Ceteareth-12, Cetearyl Alcohol, Cetyl Palmitate, Glyceryl Stearate, PEG-100 Stearate, Polyhydroxystearic Acid, Lecithin, Ethylhexyl Palmitate, Isopropyl Myristate, Isostearic Acid, Polyglyceryl-3 Polyricinoleate, Lauryl PEG-9 Polydimethylsiloxyethyldimethicone. It may include at least one selected from polydimethylsiloxyethyl dimethicone and PEG-10 dimethicone. Among these, it is more preferable to use lecithin as an emulsifier.

[0075] Oil plays a role in dissolving oil-based components and forming a strong interfacial film to improve spreadability, flowability, and absorbency, and can be used without restriction as long as it is commonly used in the relevant technical field. For example, various oils such as hydrocarbon oil, ester oil, silicone oil, and animal or vegetable oil can be used, and various other oils can also be applied. For example, as the oil, one or more selected from CEH (Cetyl 2-ethyl hexanoate), MCT (Medium Chain Triglyceride), BASF (Cetiol Ultimate), SF0015Z, SF9902E, Jojoba Oil, sunflower oil, Arlacel-165, Lecimul-LCR, stearic acid, Erocol-6850, etc. can be used. Among these, sunflower oil is preferably used as the oil.

[0076] If the amount of oil added is less than 0.1 wt%, it is difficult to properly exhibit the effects of improving spreadability, flowability, and absorbency, and if it exceeds 0.5 wt%, there is a concern that the content of other effective ingredients may be relatively low, so it is not desirable.

[0077] Additionally, in the dilution and stirring step (S140), it is preferable that stirring be performed at a room temperature of 1 to 40°C and a speed of 500 to 1,500 rpm for 1 to 3 hours.

[0078] If the stirring speed is less than 500 rpm, there is a risk that the liposome base may not be uniformly mixed into the fingerroot extract dilution solution, and if the stirring speed exceeds 1,500 rpm, it may only increase the manufacturing cost without any further effect, making it uneconomical.

[0079] In addition, it is more preferable to perform ultrasonic treatment at the same time with a frequency of 10 to 25 KHz and an output voltage of 3 to 10 W during stirring, in order to further improve the dispersibility between the fingerroot extract dilution and the liposome base.

[0080] When applying such ultrasound, if the frequency is less than 10 KHz or the output voltage is less than 3 W, the cavitation effect due to the application of ultrasound is minimal, making it difficult to properly exhibit the effect of improving dispersibility. In addition, if the frequency of the ultrasound exceeds 25 KHz or the output voltage exceeds 10 W, there is a risk that the liposome base may be damaged due to excessive application of ultrasound, which is not desirable.

[0081]

[0082] According to the above, the fingerroot liposome containing the fingerroot extract according to the embodiment of the present invention and the method for producing the same produce the fingerroot liposome by highly concentrating the fingerroot extract into liposomes, thereby reducing the ginger flavor and not exhibiting toxicity.

[0083] As a result, the fingerroot liposome containing the fingerroot extract according to the present invention and the method for producing the same can make it possible to contain a large amount of the effective ingredient panduratin A when concentrating the fingerroot extract into liposomes.

[0084] In addition, the fingerroot liposome containing the fingerroot extract according to the present invention and the method for producing the same do not exhibit cytotoxicity, and not only does the fingerroot liposome increase both collagen and elastin production in human skin fibroblasts, but it also exhibits excellent wound healing efficacy.

[0085]

[0086] Example

[0087] Hereinafter, the structure and operation of the present invention will be described in more detail through preferred embodiments of the present invention. However, these are presented as preferred examples of the present invention and should not be construed as limiting the present invention in any way.

[0088] Anything not described here is technically inferable enough to be understood by those skilled in this field, so its description will be omitted.

[0089]

[0090] 1. Liposome preparation

[0091]

[0092] Example 1

[0093] Preparation of fingerroot extract

[0094] The finger root was washed and dried with hot air at 40°C for 48 hours.

[0095] Next, 100 g of fingerroot powder obtained by crushing hot-air-dried fingerroot was added to 1 L of 70% ethanol and primary extraction was performed at room temperature (20°C) for 24 hours to prepare a fingerroot extract.

[0096] Next, the prepared fingerroot extract was filtered through a 400-mesh polyethylene net and then subjected to secondary extraction using a rotary concentrator (N-1300, Sunil Aira Co., Ltd.) to prepare a fingerroot extract.

[0097]

[0098] Preparation of fingerroot liposomes containing fingerroot extract

[0099] A fingerroot extract dilution solution was prepared by diluting the fingerroot extract with purified water at a weight ratio of 1:10, and then the fingerroot extract dilution solution was mixed with a liposome base at a weight ratio of 4:6. At this time, the liposome base was used as a mixture of 79.8 wt% propanediol, 20 wt% lecithin, and 0.2 wt% sunflower oil at a weight ratio.

[0100] Next, a mixture of the fingerroot extract dilution and liposome base was mixed and stirred at 1,000 rpm at room temperature (20°C) for 2 hours using a homodisper (Poonglim Tech Co., Ltd.) to prepare fingerroot liposomes.

[0101]

[0102] Comparative Example 1

[0103] 70% ethanol was mixed with liposome base at a weight ratio of 4:6. At this time, the liposome base was used in a weight ratio of 79.8% by weight of propanediol, 20% by weight of lecithin, and 0.2% by weight of sunflower oil.

[0104] Next, a mixture of 70% ethanol and liposome base was mixed and stirred at 1,000 rpm at room temperature (20°C) for 2 hours using a homodisper (Poonglim Tech Co., Ltd.) to prepare general liposomes.

[0105]

[0106] 2. Observation of fingerroot liposomes

[0107] Table 1 shows the results of measuring the average diameter size of fingerroot liposomes manufactured according to Example 1. In addition, Fig. 2 is a photograph showing a fingerroot extract and fingerroot liposomes manufactured according to Example 1, and Fig. 3 is a SEM photograph showing a fingerroot liposome manufactured according to Example 1.

[0108]

[0109] Distinctive diameter sizeAverage diameter sizeFinger root liposome3 ~ 9㎛6.2㎛

[0110] As shown in Table 1 and FIGS. 2 and 3, the fingerroot extract and fingerroot liposome manufactured according to Example 1 were observed, and it was confirmed that the fingerroot extract exhibited a red color, while the fingerroot liposome exhibited a yellow color. In addition, it was confirmed that the fingerroot liposome manufactured according to Example 1 had spherical particles uniformly dispersed and arranged. At this time, it was confirmed that the particle size of the fingerroot liposome had a diameter of 3 to 9 μm, and the average particle size had a diameter of 6.2 μm.

[0111]

[0112] Meanwhile, Table 2 shows the results of measuring the content of panduratin A, an active ingredient in the fingerroot extract and fingerroot liposome prepared according to Example 1. At this time, panduratin A represented by the following chemical formula 1 was used.

[0113]

[0114] [Chemical Formula 1]

[0115]

[0116]

[0117] Fingerroot extract Fingerroot liposome concentration (㎍ / mL) 221.2470

[0118] As shown in Table 2, the analysis results of the content of panduratin A, an effective ingredient in the fingerroot extract and fingerroot liposome prepared according to Example 1, confirmed that a large amount of panduratin A, an effective ingredient, was contained in the fingerroot liposome compared to the fingerroot extract.

[0119] Table 3 shows the results of content analysis of panduratin A, an active ingredient in the fingerroot extract and fingerroot liposome manufactured according to Example 1. At this time, HPLC (Agilent Technology infinity 1200) analysis was performed to analyze the content of the active ingredient in the fingerroot extract and fingerroot liposome. After dissolving the standard substance panduratin A in methanol at a concentration of 1.2 mg / mL, test solutions were prepared at concentrations of 3.675, 36.750, 73.5, 147, 294, and 588 μg / mL, which were used as standard solutions for the calibration curve. Fingerroot extract and fingerroot liposome were centrifuged at 13,000 rpm for 1 minute, and the supernatant was used as a test solution. Eclipse XDB-C18 (4.6 x 250 mm, 5 μm) was used as the column, the column temperature was maintained at 55°C, and UV-294 nm was used as the detector. The solvent concentration gradient was used as shown in the following table, and the flow rate was 1.0 mL / min.

[0120]

[0121] Time(min)%A%B-406054060130100180100204060254060

[0122] 3. Fragrance preference and sensory evaluation

[0123] Table 4 shows the results of the flavor preference and sensory characteristic intensity evaluation conducted based on the standard operating procedure (SOP) of the Korea Food Research Institute for the flavor preference evaluation of the fingerroot extract and fingerroot liposome manufactured according to Example 1. In addition, Fig. 4 is a graph showing the results of the flavor preference and sensory evaluation of the fingerroot extract and fingerroot liposome manufactured according to Example 1.

[0124] Specifically, the fragrance preference and sensory evaluation were conducted by 54 male and female test subjects in their 20s to 40s. The fragrance preference for each sample was evaluated using a 9-point preference scale (1 point: very dislike, 9 points: very like). The overall fragrance intensity, ginger fragrance, herbal medicine fragrance, and bitter fragrance were evaluated using a 9-point item scale (1: none, 9: very strong). The samples were prepared the day before the evaluation by aliquoting 2 mL into amber vials (16 ml, SciLab Korea, Korea) labeled with a 3-digit random number. The samples were presented to each panelist one sample at a time in a randomized order, and the temperature of the evaluation room during the evaluation was 23℃.

[0125]

[0126] Sample sensory characteristics Overall odor intensity Ginger aroma Herbal aroma Bitter aroma Fingerroot extract 6.02 6.11 4.7 3.19 Fingerroot liposome 3.67 3.67 3.42 76 P-value < 0.0001 < 0.0001 0.0001 0.01

[0127] ※ T-test results show significant differences between samples ( * p<0.005; ** p<0.01; *** p<0.001)

[0128] As shown in Table 4 and Figure 4, the results of the flavor preference and sensory evaluation of the fingerroot extract and fingerroot liposome according to Example 1 showed that the overall flavor intensity, ginger flavor, herbal medicine flavor, and bitter flavor of the fingerroot liposome were all evaluated as weaker than those of the fingerroot extract.

[0129]

[0130] 4. Cytotoxicity evaluation

[0131] Figure 5 is a graph showing the results of cytotoxicity evaluation of the fingerroot extract and fingerroot liposome prepared according to Example 1 in human skin fibroblasts. At this time, in order to evaluate the intracellular toxicity of the fingerroot extract and fingerroot liposome prepared according to Example 1, human skin fibroblasts, Detroit 551 cells, were treated with the fingerroot extract and fingerroot liposome at various concentrations. Specifically, 3,000 Detroit 551 fibroblast cells were placed in a 96-well dish and cultured for 24 hours. Thereafter, the fingerroot extract and fingerroot liposome were diluted to 1%, 0.5%, 0.25%, 0.125%, 0.0625%, and 0.03125%, treated to the cells, and cultured at 37°C. To analyze the number of viable cells after 48 hours, MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide, ThermoFisher Scientific) was dissolved in PBS (phosphate buffered saline, ThermoFisher Scientific) at 5 mg / mL and added to Detroit 551 cell culture at a weight ratio of 1:10 and reacted for 1 hour. Afterwards, the culture supernatant was removed, and 200 μl of DMSO (Dimethyl sulfoxide, Sigma Aldrich) was added to each well to dissolve the MTT-formazan crystals produced in viable cells. The absorbance was measured at 549 nm using an absorption spectrophotometer (spectrometer, Biochrom).

[0132] As shown in Fig. 5, Fig. 5 shows the results of cytotoxicity evaluation of the fingerroot extract and fingerroot liposome prepared according to Example 1, and the fingerroot extract showed cytotoxicity to cells at a concentration of 0.125% or higher, but the fingerroot liposome did not show cytotoxicity.

[0133]

[0134] 5. Evaluation of collagen and elastin production

[0135] Fig. 6 is a graph showing the results of evaluating collagen production in fibroblasts for the fingerroot liposomes according to Example 1 and the general liposomes according to Comparative Example 1, and Fig. 7 is a graph showing the results of evaluating elastin production in fibroblasts for the fingerroot liposomes according to Example 1 and the general liposomes according to Comparative Example 1. At this time, in order to confirm whether the fingerroot liposomes according to Example 1 and the general liposomes according to Comparative Example 1 affect the production of collagen and elastin in fibroblasts, Detroit 551 cells, which are human skin fibroblasts, were treated with each sample of the general liposomes and the fingerroot liposomes, and then an ELISA assay was performed. The measurement conditions and procedures were as follows.

[0136] Procollagen type I, elastin ELISA assay

[0137] Detroit 551 (1x10 5 Cells / well) were seeded into 6-well plates and cultured for 24 hours. The culture medium was replaced with fresh medium supplemented with each sample and cultured for 24 hours. Afterwards, the medium was collected from each well, and the expression levels of elastin and type I procollagen were measured using an ELISA assay kit (R&D Systems).

[0138] As shown in Figures 6 and 7, the general liposome according to Comparative Example 1 showed no change in the amount of collagen and elastin produced.

[0139] In contrast, it was confirmed that the fingerroot liposome according to Example 1 showed the highest collagen production efficacy, with collagen production increasing by 155.8% and elastin production increasing by 145.6%. Thus, it was confirmed that the fingerroot liposome according to Example 1 increased the amount of collagen and elastin production in human skin cells.

[0140]

[0141] 6. Evaluation of cell proliferation efficacy

[0142] Figure 8 is an SEM photograph showing the results of performing the cell proliferation efficacy and wound healing efficacy of the fingerroot liposome according to Example 1 and the general liposome according to Comparative Example 1 in human skin fibroblasts, and Figure 9 is a graph showing the results of performing the cell proliferation efficacy and wound healing efficacy of the fingerroot liposome according to Example 1 and the general liposome according to Comparative Example 1 in human skin fibroblasts. At this time, in order to confirm whether the fingerroot liposome according to Example 1 and the general liposome according to Comparative Example 1 affect the activity and proliferation of human skin fibroblasts, Detroit 551 cells were treated with the fingerroot liposome at a concentration of 1%, and then a wound-healing assay was measured. At this time, the measurement conditions and process were as follows. The fibroblasts were seeded in a 6-well plate at a density of 2x10 4 Cells were seeded at a density of 1 / cm2 and cultured in growth medium for 24 hours. The center of each plate was scratched to remove cells, and growth medium containing 1% each of regular liposomes and fingerroot liposomes was added and cultured. After 24 hours of culture, the cells were observed under an optical microscope to confirm the efficacy in wound healing through verification of increased cell activity and migration, and the number of cells that migrated to the wound site was measured.

[0143] As shown in FIGS. 8 and 9, the general liposome according to Comparative Example 1 did not affect the activity and proliferation of human skin fibroblasts.

[0144] In contrast, the fingerroot liposome according to Example 1 significantly affected the activity and proliferation of human skin fibroblasts and was found to exhibit excellent cell proliferation efficacy.

[0145] In addition, it was found that the fingerroot liposome according to Example 1 exhibited a wound healing effect more than twice that of the general liposome according to Comparative Example 1.

[0146]

[0147] 7. Skin irritation evaluation

[0148] Table 5 shows the results of the skin irritation evaluation on test subjects using the fingerroot liposome according to Example 1. The test subjects were 21 adult women aged 30 to 55 years. The fingerroot liposome according to Example 1 was applied to the face (around the eyes) and the effect was evaluated using a PRIMOS Lite (GFMesstenchnik GmbH, Germany) device. For the evaluation, the test subjects washed their faces and rested for 30 minutes in a waiting room under constant temperature and humidity (22±2℃, RH40~60%) to allow the skin surface temperature and humidity to adapt to the environment of the measurement space. The presence or absence of adverse reactions such as erythema, edema, scaling, itching, stinging, burning sensation, stiffness, tingling, or other adverse reactions was closely observed, and when an adverse skin reaction occurred, it was graded according to severity and the test findings were recorded.

[0149]

[0150] Erythema, Edema, Scaling, Itching, Stinging, Burning, Tightness, Prickling After 2 weeks of using the product-------- After 4 weeks of using the product--------

[0151] As shown in Table 5, the results of a questionnaire survey of test subjects using fingerroot liposomes according to Example 1 showed that no adverse skin reactions to the test product were observed in any of the test subjects. In addition, no adverse skin reactions were observed in the visual evaluation after use of the test product.

[0152]

[0153] 8. Eye wrinkle assessment

[0154] Table 6 shows the results of measuring the Ra analysis value and improvement rate of wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1, Table 7 shows the results of measuring the Rq analysis value and improvement rate of wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1, and Table 8 shows the results of measuring the Rmax analysis value and improvement rate of wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1. In addition, Fig. 10 is a graph showing the results of analyzing wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1, Fig. 11 is a graph showing the results of analyzing wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1, and Fig. 12 is a graph showing the results of analyzing wrinkles around the eyes in test subjects using the fingerroot liposome according to Example 1.

[0155] At this time, for the evaluation of wrinkles around the eyes, the fingerroot liposome according to Example 1 was applied to the facial (eye) area and used for 4 weeks, and measured with a PRIMOS Lite (GFMesstenchnik GmbH, Germany) device, and then analyzed with PRIMOS Lite version 5.8E. It means that wrinkles around the eyes have improved when the selected analysis variables Ra, Rq, and Rmax values ​​become smaller, and the unit is ㎛.

[0156] Additionally, the improvement rate (%) of wrinkles around the eyes was calculated according to the formula below.

[0157]

[0158] The results of the analysis of wrinkles around the eyes were analyzed using the parametric method, the Paired t-test, and the non-parametric method, the Wilcoxon signed rank test, after a normality test for changes before and after use of the fingerroot liposome according to Example 1. All statistical results were considered statistically significant when the significance level was 5% (p<0.05), which is the most frequently used level in biological statistical analysis.

[0159]

[0160] Time-PointMean(SD)Ratiop-value * Wrinkle(Ra)0W20.028(3.321)2W19.191(3.359)+4.180.006 1)** 4W18.921(3.341)+5.530.022 2)*

[0161] * p-value is a statistical result of changes before and after using the product; 1) Wilcoxon signed rank test, 2) Pairedt-test; ** p-value<0.01, * p-value<0.05※ Unit: Analysis value (㎛), improvement rate (%)

[0162]

[0163] Time-PointMean(SD)Ratiop-value * Wrinkle(Rq)0W24.803(4.026)2W23.763(4.072)+4.190.005 1)** 4W23.507(4.019)+5.230.024 1)*

[0164] * p-value is a statistical result of changes before and after using the product; 1) Wilcoxon signed rank test, 2) Pairedt-test; **p-value<0.01, * p-value<0.05※ Unit: Analysis value (㎛), improvement rate (%)

[0165]

[0166] Time-PointMean(SD)Ratiop-value * Wrinkle(Rmax)0W136.931(21.744)2W133.389(22.254)+2.590.114 1) 4W130.810(20.139)+4.470.039 1)*

[0167] * p-value is a statistical result of changes before and after using the product; 1) Pairedt-test; * p-value<0.05※ Unit: Analysis value (㎛), improvement rate (%)

[0168]

[0169] As shown in Tables 6 to 8 and Figures 10 to 12, the results of evaluating the effect of improving wrinkles around the eyes in test subjects using the finger root liposome according to Example 1 showed that, compared to before use of the product (week 0), after 2 weeks of use, the variable Ra showed an improvement rate of 4.18%, the variable Rq showed an improvement rate of 4.19%, and the variable Rmax showed an improvement rate of 2.59%, and after 4 weeks of use, the variable Ra showed an improvement rate of 5.53%, the variable Rq showed an improvement rate of 5.23%, and the variable Rmax showed an improvement rate of 4.47%.

[0170] Additionally, there was a statistically significant difference in variables Ra, Rq, and Rmax after 4 weeks of product use compared to before product use (0 weeks) (p<0.05).

[0171]

[0172] 9. Survey Results

[0173] 1) Results of the validity evaluation survey

[0174] Table 9 shows the results of a questionnaire evaluating the effectiveness of the test subjects who used the fingerroot liposome according to Example 1.

[0175]

[0176] Number of test subjects (percentage, %)Average standard deviationPositive response rate (%)*5*4*3*2*1Effect of improving wrinkles around the eyes1(5)15(71)5(24)0(0)0(0)3.80.5100*5: Very satisfied, 4: Satisfied, 3: Average, 2: Dissatisfied, 1: Very dissatisfied

[0177] ※ Positive response rate (%): The number of test subjects who answered 3 points (average) or higher is expressed as a percentage. ※ The percentage is calculated with a margin of error of ±1%.

[0178]

[0179] As shown in Table 9, the results of the descriptive survey conducted on measurement items after 4 weeks of product use are presented.

[0180] According to the survey evaluation results, 100% of the test subjects evaluated the product's effect on improving wrinkles around the eyes as average or higher.

[0181]

[0182] 2) Results of the preference survey

[0183] Table 10 shows the results of a preference questionnaire conducted on test subjects using fingerroot liposomes according to Example 1.

[0184]

[0185] Number of test subjects (percentage, %)Average standard deviationPositive response rate (%)*5*4*3*2*1Product scent7(33)10(48)4(19)0(0)0(0)4.10.7100Product color7(33)13(62)1(5)0(0)0(0)4.30.6100Product viscosity9(43)10(48)2(10)0(0)0(0)4.30.7100Spreading / appliance10(48)10(48)1(4)0(0)0(0)4.40.6100Skin absorption10(48)8(38)3(14)0(0)0(0)4.30.7100Freshness Degree 7(33)9(43)5(24)0(0)0(0)4.10.8100Softness 8(38)10(48)3(14)0(0)0(0)4.20.7100Moisture 9(43)10(48)2(10)0(0)0(0)4.30.7100Elasticity 5(24)11(52)5(24)0(0)0(0)4.00.7100*5:Very satisfied, 4:Satisfied, 3:Average, 2:Dissatisfied, 1:Very dissatisfied

[0186] ※ Positive response rate (%): The number of test subjects who answered 3 points (average) or higher is expressed as a percentage. ※ The percentage is calculated with a margin of error of ±1%.

[0187]

[0188] As shown in Table 10, the results of a survey on the test subjects' level of preference for product satisfaction, usability, and changes after use after 4 weeks of product use are shown.

[0189] According to the survey results, 100% of the test subjects rated the product's fragrance, color, and viscosity as average or higher in the product satisfaction section. Regarding the feel of the product, 100% of the test subjects rated the product's spreadability / application, skin absorption, and freshness as average or higher. Regarding the post-use changes section, 100% of the test subjects rated the product's softness, moisture, and elasticity as average or higher.

[0190]

[0191] Although the embodiments and comparative examples of the present invention have been described in detail as above, the scope of the present invention is not limited thereto, and the scope of the present invention extends to a range substantially equivalent to the embodiments of the present invention.

Claims

1. Step of washing and drying the finger root with hot air; A step of crushing the hot-air-dried fingerroot to obtain fingerroot powder, then adding the fingerroot powder to a solvent and performing a first extraction to form a fingerroot extract; A step of filtering the above fingerroot extract and then performing a second extraction using a rotary concentrate to form a fingerroot extract; and A step of forming a fingerroot liposome by mixing a liposome base into a fingerroot extract dilution solution obtained by diluting the fingerroot extract in purified water and stirring the mixture; A method for producing fingerroot liposome containing a fingerroot extract, characterized in that it comprises:

2. In paragraph 1, The above hot air drying A method for producing a fingerroot lysoform containing a fingerroot extract, characterized in that the method is performed at 30 to 50°C for 12 to 60 hours.

3. In paragraph 1, The above solvent is A method for producing fingerroot liposome containing a fingerroot extract, characterized in that it uses one selected from water, methanol, ethanol, phenoxyethanol, propylene glycol, and dipropylene glycol, alone or in a mixture of two or more thereof.

4. In paragraph 1, The above diluted fingerroot extract is A method for producing fingerroot liposome containing a fingerroot extract, characterized in that the fingerroot extract is diluted in purified water in a weight ratio of 1:8 to 1:

12.

5. In paragraph 1, The above liposome base is A method for producing fingerroot liposome containing fingerroot extract, characterized in that 50 to 150 parts by weight are mixed with respect to 100 parts by weight of the diluted fingerroot extract.

6. In paragraph 1, The above liposome base is A method for producing fingerroot liposome containing a fingerroot extract, characterized in that it comprises 70 to 90 wt% of a moisturizer, 10 to 30 wt% of an emulsifier, and 0.1 to 0.5 wt% of an oil.

7. In paragraph 1, The above stirring A method for producing fingerroot liposomes containing fingerroot extract, characterized in that the method is performed at a room temperature of 1 to 40°C and a speed of 500 to 1,500 rpm for 1 to 3 hours.

8. A fingerroot lysoform containing a fingerroot extract manufactured by a method according to any one of claims 1 to 7, The above fingerroot lysoform is a fingerroot extract-containing fingerroot lysoform characterized by having a reduced ginger flavor and no toxicity, by being manufactured by concentrating the fingerroot extract into liposomes.

Images