A wild apple fruit extract with skin barrier repair efficacy, and a preparation method and application thereof

Abstract

The present application relates to a Malus sieboldii extract with skin barrier repair efficacy and a preparation method and application thereof, the preparation method comprising the following steps: mixing Malus sieboldii raw materials with water, then performing microwave treatment, and then performing vacuum reduced pressure reflux extraction treatment on the treated mixture to obtain an extract; filtering the extract, concentrating the filtrate to obtain a concentrated solution; mixing the concentrated solution with an alcohol solution to perform alcohol precipitation treatment; filtering the alcohol precipitation treatment solution, drying the precipitate to obtain the Malus sieboldii extract. The Malus sieboldii extract has significant skin repair efficacy and is safe and mild.

Description

Technical Field

[0001] This invention belongs to the field of plant extraction technology, and relates to a skin barrier repairing extract of *Cyprinus pungens* fruit, its preparation method, and its application. Background Technology

[0002] As research into the physiological structure and mechanisms of the skin continues to expand, our understanding of the skin barrier is constantly growing. A healthy skin barrier is a crucial factor in maintaining healthy skin. The most common skin manifestation caused by a damaged skin barrier is sensitive skin. Furthermore, most skin diseases also involve skin barrier damage, such as acne, seborrheic dermatitis, melasma, rosacea, and steroid-induced dermatitis. Additionally, mesotherapy and laser treatments, which are most widely used in the medical aesthetics field, are also accompanied by skin barrier damage.

[0003] Human skin is generally considered to be a metabolic system, and the skin barrier is also a multi-dimensional and multi-faceted system that interacts and influences each other. From the outermost sebum film to the most important brick-and-mortar structure, to the water regulation mechanisms such as aquaporins and natural moisturizing factors, to the inflammation and immune regulation mechanisms, and finally to the activation and remodeling of the dermis, each layer has its own appropriate regulatory mechanism. If a problem occurs in any link, it will induce problems in other links, thus forming a vicious cycle.

[0004] There are many skin barrier repair products on the market, most of which focus on repairing the skin's "brick wall" structure and addressing aging in the dermis, but their targets are not comprehensive. Some products simply provide a non-irritating, moisturizing environment for the skin to repair itself, and these products often feel heavy and oily, failing to provide a comfortable user experience.

[0005] Some products illegally contain added hormones, causing new skin problems such as steroid-induced dermatitis. A significant number of products also contain harmful chemicals (such as irritating chemical preservatives), which not only irritate the skin and easily trigger allergic reactions like redness and swelling, but also damage the skin's barrier system and reduce its ability to retain moisture. As people's living standards improve, consumers have increasingly higher demands for cosmetics, requiring products to not only have skin-repairing effects but also to be natural, safe, healthy, and gentle.

[0006] Therefore, it is particularly important to develop a raw material that can repair the skin barrier, soothe skin discomfort and repair damage in the short term, maintain a healthy skin barrier in the long term, prevent the recurrence of allergic reactions, and be safe and gentle. Summary of the Invention

[0007] In view of the shortcomings of the existing technology, the purpose of this invention is to provide a skin barrier repairing extract of sea buckthorn fruit, its preparation method and application.

[0008] To achieve this objective, the present invention employs the following technical solution:

[0009] In a first aspect, the present invention provides a method for preparing a *Prickly ash* fruit extract with skin barrier repair effects, the preparation method comprising the following steps:

[0010] (1) Mix the raw material of the prickly pear fruit with water and then microwave it. Then, vacuum decompression reflux extraction is performed on the treated mixture to obtain the extract.

[0011] (2) Filter the extract, concentrate the filtrate, and obtain the concentrate;

[0012] (3) Mix the concentrated liquid with the alcohol solution for alcohol precipitation;

[0013] (4) Filter the alcohol precipitation solution, dry the precipitate, and obtain the prickly pear fruit extract.

[0014] This invention creatively employs microwave treatment combined with vacuum reflux extraction and alcohol precipitation to prepare *Prickly pear* fruit extract. This extract exhibits significant skin repair effects, specifically: it promotes increased epidermal thickness, reduces water loss from damaged skin, accelerates wound healing, and significantly promotes the expression of repair-related genes. Furthermore, its safety has been well-verified, making it safe for use in related products.

[0015] Preferably, the raw material of *Pteris vittata* is selected from any one or a combination of at least two of the following: *Pteris vittata* seed meal powder, *Pteris vittata* kernel, *Pteris vittata* fruit, *Pteris vittata* leaves, or *Pteris vittata* roots.

[0016] The raw material of *Cyprinus sarmentosus* used in this invention is not limited, but *Cyprinus sarmentosus* seed meal and *Cyprinus sarmentosus* kernels are preferred. The *Cyprinus sarmentosus* seed meal is the residue powder left after pressing the *Cyprinus sarmentosus* kernels for oil. This invention provides a new strategy for the secondary development of *Cyprinus sarmentosus* seed meal, which can realize waste utilization and is more economical and environmentally friendly.

[0017] Preferably, the ratio of the raw material of the prickly ash fruit to water is 1:(5-20)g / mL, for example, 1:5g / mL, 1:7g / mL, 1:8g / mL, 1:10g / mL, 1:12g / mL, 1:14g / mL, 1:16g / mL, 1:20g / mL, etc.

[0018] Preferably, the microwave processing conditions include: microwave power of 150-400W, such as 150W, 180W, 200W, 220W, 250W, 300W, 350W, 400W, etc.; and microwave time of 30-90min, such as 30min, 40min, 50min, 60min, 70min, 80min, 90min, etc.

[0019] The preparation method involved in this invention performs microwave-assisted treatment before vacuum decompression reflux extraction, which can further promote the extraction efficiency of the target component. Within the specific microwave treatment parameter range mentioned above, both extraction efficiency and the stability of the target component can be taken into account, thereby maximizing the technical effect of the final product.

[0020] Preferably, the vacuum degree of the vacuum decompression reflux extraction is 0.03-0.08 MPa, such as 0.03 MPa, 0.04 MPa, 0.05 MPa, 0.06 MPa, 0.07 MPa, 0.08 MPa, etc.

[0021] Preferably, the temperature of the vacuum decompression reflux extraction is 35-45℃, such as 35℃, 36℃, 37℃, 38℃, 39℃, 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, etc.

[0022] Preferably, the vacuum reflux extraction process is performed 1-3 times, for example, 1 time, 2 times, or 3 times; each extraction time is independently 60-120 min, for example, 60 min, 70 min, 80 min, 90 min, 100 min, 110 min, or 120 min.

[0023] Preferably, the filtrate is concentrated to 1 / 6 to 1 / 2 of its volume, for example, 1 / 6, 1 / 5, 1 / 4, 1 / 3, 1 / 2, etc.

[0024] Concentrating the filtrate to the specific range defined above allows for better coordination with subsequent alcohol precipitation. If the volume is further concentrated, the remedial effect of the alcohol precipitation product will be negatively affected.

[0025] Preferably, the alcohol solution is an anhydrous alcohol solution or an aqueous alcohol solution, and the alcohol concentration of the aqueous alcohol solution is 90-99%, such as 90%, 92%, 93%, 94%, 95%, 96%, 97%, 99%, etc.

[0026] Preferably, the final volume concentration of alcohol after mixing the alcohol solution and the concentrate is 80-90%, such as 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, etc.

[0027] The final concentration of alcohol affects the alcohol precipitation effect to a certain extent. When the final volume concentration is 80-90%, it can balance the alcohol precipitation efficiency and the excellent skin barrier repair effect of the alcohol precipitation product.

[0028] Preferably, the alcohol solution is added to the concentrate dropwise while stirring is being applied.

[0029] Preferably, the alcohol is selected from ethanol, a combination of ethanol and butanediol, or a combination of ethanol, butanediol, and polyethylene glycol.

[0030] The alcohol used in the alcohol precipitation process of this invention can be ethanol alone, or a combination of ethanol and butylene glycol, or a combination of ethanol, butylene glycol, and polyethylene glycol. The choice of alcohol type affects the alcohol precipitation effect to a certain extent. When butylene glycol or polyethylene glycol is further added to ethanol, the skin barrier repair effect of the final product is further improved.

[0031] Preferably, the volume ratio of ethanol to butanediol is (1-4):1, for example, 1:1, 2:1, 3:1, 4:1, etc.

[0032] Preferably, the volume ratio of ethanol to butanediol to polyethylene glycol is (1-4):1:(0.05-0.2), wherein the specific values ​​in (0.05-0.2) can be selected from 0.05, 0.07, 0.1, 0.12, 0.14, 0.15, 0.2, etc.

[0033] Preferably, the number average molecular weight of the polyethylene glycol is 200-600, such as 200, 300, 400, 500, 600, etc.

[0034] Preferably, the filtration in step (4) is performed using a filter cloth with a pore size of 150-250 mesh (e.g., 150 mesh, 180 mesh, 200 mesh, 220 mesh, 240 mesh, 250 mesh, etc.).

[0035] Preferably, the drying method in step (4) includes freeze drying.

[0036] Other specific point values ​​within the range of all the values ​​mentioned above can be selected, but for the sake of brevity in the instruction manual, they will not be described in detail here.

[0037] In a second aspect, the present invention provides a *Prickly pear* fruit extract prepared according to the preparation method described in the first aspect.

[0038] Thirdly, the present invention provides the use of the extract of *Cynanchum paniculatum* according to the second aspect in the preparation of skin barrier repair products.

[0039] Preferably, the product includes skin care products, cosmetics, pharmaceuticals, or medical materials.

[0040] Preferably, the content of the sea buckthorn fruit extract in the product is 0.01-5%, such as 0.01%, 0.03%, 0.05%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0041] Compared with the prior art, the present invention has the following beneficial effects:

[0042] This invention creatively employs a combination of microwave treatment, vacuum reflux extraction, and alcohol precipitation to prepare *Prickly pear* fruit extract. This extract exhibits significant skin repair effects, specifically: it promotes increased epidermal thickness, reduces water loss from damaged skin, accelerates wound healing, and significantly promotes the expression of repair-related genes. Furthermore, its safety has been well-verified, making it safe for use in related products. Attached Figure Description

[0043] Figure 1 These are images showing the skin results of mice in each group during an acute mouse skin injury model experiment.

[0044] Figure 2 This is a graph showing the PPAR-β expression levels in each group during an acute mouse skin injury model experiment.

[0045] Figure 3 This is a graph showing the LOR expression levels in each group during an acute mouse skin injury model experiment.

[0046] Figure 4 This is a graph showing the Claudin-5 expression levels in each group during an acute mouse skin injury model experiment.

[0047] Figure 5 This is a graph showing the SPT expression levels in each group during an acute mouse skin injury model experiment. Detailed Implementation

[0048] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0049] Example 1

[0050] This embodiment provides a sea buckthorn fruit extract, the preparation method of which is as follows:

[0051] (1) Mix the seed meal powder of the Chinese prickly pear with water at a ratio of 1:10 g / mL and then microwave it. The microwave power is 250W and the microwave time is 60min. Then, the treated mixture is vacuum refluxed twice at a vacuum degree of 0.05MPa and a temperature of 40℃ for 90min each time. The two extracts are combined.

[0052] (2) Filter the extract and concentrate the filtrate under reduced pressure to 1 / 4 of the original volume to obtain the concentrate;

[0053] (3) Add 95% ethanol aqueous solution dropwise to the concentrate while stirring, and add it all within 30 minutes to make the final volume concentration of ethanol 85%.

[0054] (4) The alcohol precipitation solution was filtered through a filter cloth with a pore size of 200 mesh, the precipitate was freeze-dried, and the extract of the prickly pear fruit was obtained.

[0055] Example 2

[0056] This embodiment provides a sea buckthorn fruit extract, the preparation method of which is as follows:

[0057] (1) Mix the seed meal powder of the Chinese prickly pear with water at a ratio of 1:7 g / mL and then microwave it. The microwave power is 200W and the microwave time is 80min. Then, the treated mixture is vacuum refluxed twice at a vacuum degree of 0.03MPa and a temperature of 45℃ for 110min each time. The two extracts are combined.

[0058] (2) Filter the extract and concentrate the filtrate under reduced pressure to 1 / 6 of the original volume to obtain the concentrate;

[0059] (3) Add 95% ethanol aqueous solution dropwise to the concentrate while stirring, and add it all within 30 minutes to make the final volume concentration of ethanol 80%.

[0060] (4) The alcohol precipitation solution was filtered through a filter cloth with a pore size of 200 mesh, the precipitate was freeze-dried, and the extract of the prickly pear fruit was obtained.

[0061] Example 3

[0062] This embodiment provides a sea buckthorn fruit extract, the preparation method of which is as follows:

[0063] (1) Mix the seed meal powder of the Chinese prickly pear with water at a ratio of 1:15 g / mL and then microwave it. The microwave power is 350W and the microwave time is 40min. Then, the treated mixture is vacuum refluxed twice at a vacuum degree of 0.08MPa and a temperature of 35℃ for 60min each time. The two extracts are combined.

[0064] (2) Filter the extract and concentrate the filtrate under reduced pressure to 1 / 2 of the original volume to obtain the concentrate;

[0065] (3) Add 95% ethanol aqueous solution dropwise to the concentrate while stirring, and add it all within 30 minutes to make the final volume concentration of ethanol 90%.

[0066] (4) The alcohol precipitation solution was filtered through a filter cloth with a pore size of 200 mesh, the precipitate was freeze-dried, and the extract of the prickly pear fruit was obtained.

[0067] Example 4

[0068] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that: in step (2), the extract is filtered and the filtrate is concentrated under reduced pressure to 1 / 10 of the original volume to obtain a concentrated solution; all other steps remain unchanged.

[0069] Example 5

[0070] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that: in step (3), the 95% ethanol aqueous solution is replaced with the same concentration of butanediol aqueous solution, while other conditions remain unchanged.

[0071] Example 6

[0072] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that: in step (3), the 95% ethanol aqueous solution is replaced with a mixed alcohol aqueous solution of the same concentration (the volume ratio of ethanol and butanediol is 2:1), and all other conditions remain unchanged.

[0073] Example 7

[0074] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that: in step (3), the 95% ethanol aqueous solution is replaced with a mixed alcohol aqueous solution of the same concentration (the volume ratio of ethanol, butanediol and polyethylene glycol 300 is 2:1:0.1), and all other conditions remain unchanged.

[0075] Example 8

[0076] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that the microwave power of 250W and the microwave time of 60min in step (1) are replaced with a microwave power of 500W and a microwave time of 100min, while other conditions remain unchanged.

[0077] Example 9

[0078] This embodiment provides a Chinese prickly pear fruit extract, the preparation method of which differs from that of Example 1 only in that the microwave power of 250W and the microwave time of 60min in step (1) are replaced with a microwave power of 100W and a microwave time of 20min, while other conditions remain unchanged.

[0079] Comparative Example 1

[0080] This comparative example provides a *Prickly pear* fruit extract, the preparation method of which is as follows:

[0081] (1) Mix the seed meal powder of the Chinese prickly pear with water at a ratio of 1:10 g / mL, and then perform vacuum reflux extraction twice at a vacuum degree of 0.05 MPa and a temperature of 40℃, each time for 90 min, and combine the two extracts.

[0082] (2) Filter the extract and concentrate the filtrate under reduced pressure to 1 / 4 of the original volume to obtain the concentrate;

[0083] (3) Add 95% ethanol aqueous solution dropwise to the concentrate while stirring, and add it all within 30 minutes to make the final volume concentration of ethanol 85%.

[0084] (4) The alcohol precipitation solution was filtered through a filter cloth with a pore size of 200 mesh, the precipitate was freeze-dried, and the extract of the prickly pear fruit was obtained.

[0085] Comparative Example 2

[0086] This comparative example provides a *Prickly pear* fruit extract, the preparation method of which is as follows:

[0087] (1) The seed meal powder of the Chinese prickly pear fruit was mixed with water at a ratio of 1:10 g / mL and then subjected to microwave-enzyme combined treatment. The microwave power was 250W, the microwave time was 60min, the temperature was 40℃, the amount of cellulase added was 2%, and the pH was 5.0 to obtain the extract. The extract was then inactivated at 80℃ for 15min.

[0088] (2) Then filter the solution and concentrate the filtrate under reduced pressure to 1 / 4 of its original volume to obtain a concentrated solution.

[0089] (3) Add 95% ethanol aqueous solution dropwise to the concentrate while stirring, and add it all within 30 minutes to make the final volume concentration of ethanol 85%.

[0090] (4) The alcohol precipitation solution was filtered through a filter cloth with a pore size of 200 mesh, the precipitate was freeze-dried, and the extract of the prickly pear fruit was obtained.

[0091] Test Example 1

[0092] 3D Skin Epidermal Layer Thickness Change Evaluation Experiment:

[0093] Preparation of test sample solution: The extracts of *Cynanchum paniculatum* obtained in Examples 1-9 and Comparative Examples 1-2 were diluted with physiological saline solution to a concentration of 0.2%.

[0094] Experimental procedure: A commercially available 3D skin model-artificial epidermal detection kit (Jinan Pansheng Biotechnology Co., Ltd.) was purchased for the experiment. After adding the test sample solution and leaving it for 48 hours, if the epidermal layer thickness increased, it indicates that the test sample has a barrier repair effect. The blank control group was prepared by adding the same amount of physiological saline solution.

[0095] The experimental results are shown in Table 1.

[0096] Table 1

[0097]

[0098]

[0099] As shown in Table 1, the *Prickly pear* fruit extract obtained by the preparation method of this invention can significantly increase the thickness of the epidermis after use. However, relevant parameters in the preparation process, such as the concentration ratio of the extract, the solvent and its ratio for alcohol precipitation, and microwave process parameters, all affect the final increase in epidermal layer thickness to some extent. Furthermore, if one or more of the microwave extraction or vacuum reflux extraction steps are missing from the preparation process, or if one or more of these steps are replaced with other preparation processes, the ability of the *Prickly pear* fruit extract to promote the increase of epidermal layer thickness will be significantly reduced.

[0100] Test Example 2

[0101] Acute mouse skin injury model experiment:

[0102] Preparation of test sample solution: The extracts of *Cynanchum paniculatum* obtained in Examples 1-9 and Comparative Examples 1-2 were diluted with physiological saline solution to a concentration of 0.2%.

[0103] An acute mouse skin injury model was established using the acetone-ether method. Specifically, male SPF-grade BALB / c mice (16-18g) purchased after passing quarantine were housed in standard polycarbonate cages under a 12-hour light / dark cycle, with free access to food and water. All procedures were performed in accordance with relevant ethical requirements for laboratory animals. A total of three groups were established: a normal control group, a model group, and experimental groups 1-11, with six mice in each group.

[0104] Hair removal: On the first day of the experiment, the hair on the back of the animals was shaved off using a hair removal device, covering an area of ​​approximately 2×2cm. 2 If, during the experiment, animal hair growth becomes too large to interfere with sample administration and observation, shaving is necessary. The hair removal process should be gentle to avoid damaging the skin.

[0105] Sample loading: From day two to day seven of the experiment, medication was administered to mice after hair removal. Model group: Administered medication twice daily, at 9:00 AM and 5:00 PM. Each time, a cotton pad soaked in an equal volume mixture of acetone and ether was applied to the shaved area on the back of the mouse's neck for 15 seconds, followed by a cotton pad soaked in distilled water for another 15 seconds. This was repeated for 6 consecutive days. Normal control group: Compared to the model group, the acetone and ether mixture was replaced with physiological saline; all other parameters remained the same. Experimental groups 1-11: Compared to the model group, after modeling, distilled water was replaced with the experimental sample solution for each group; all other parameters remained the same.

[0106] Testing: On the eighth day of the experiment, the TEWL (transepidermal water loss) value of each group of mice was measured using an epidermal water loss meter. Compared with the model group, a smaller TEWL value indicates a better repair effect.

[0107] Skin images of mice in each group on day 8 of the experiment are shown below. Figure 1 As shown in Table 2.

[0108] Table 2

[0109]

[0110]

[0111] As shown in Table 2, the *Prickly pear* fruit extract obtained by the preparation method of this invention exhibits significant skin repair effects after use, with low transepidermal water loss rates. However, relevant parameters in the preparation process, such as the concentration ratio of the extract, the solvent and its ratio for alcohol precipitation, and microwave process parameters, all influence these repair effects to some extent. Furthermore, if one or more of the microwave extraction or vacuum reflux extraction steps are missing from the preparation process, or if one or more of these steps are replaced with other preparation processes, the repair effect of the final *Prickly pear* fruit extract will be significantly reduced.

[0112] Test Example 3

[0113] mRNA expression level assay for repair-related genes:

[0114] Experimental procedure: After the experiment in test example 2 was completed, all mice were euthanized by cervical dislocation, and the skin on the back of the mice after hair removal and drug administration was collected for relevant tests.

[0115] The mRNA expression levels of keratinocyte proliferation and differentiation-related protein (PPAR-β), keratinization mantle-related protein (LOR), tight junction protein (Claudin-5), and lipid synthase (SPT) in each group of mice are as follows: Figure 2 , Figure 3 , Figure 4 , Figure 5 As shown.

[0116] Depend on Figure 2-5 It is known that the *Prickly pear* fruit extract obtained by the preparation method involved in this invention has a good promoting effect on the mRNA expression level of repair-related genes. However, relevant parameters in the preparation process, such as the concentration ratio of the extract, the alcohol precipitation solvent and its ratio, and the microwave process parameters, all affect the above effect to a certain extent, that is, the promoting effect on the mRNA expression level of repair-related genes will be affected to some extent. In addition, if one or more of the microwave extraction treatment or vacuum reflux extraction steps are missing from the preparation process, or if one or more of the preparation steps are replaced with other preparation processes, the repair effect of the final *Prickly pear* fruit extract will be greatly reduced, that is, the mRNA expression level of repair-related genes will be significantly weakened.

[0117] Test Example 4

[0118] Rabbit wound healing experiment:

[0119] Preparation of test sample solution: The extracts of *Cynanchum paniculatum* obtained in Examples 1-9 and Comparative Examples 1-2 were diluted with physiological saline solution to a concentration of 0.2%.

[0120] A rabbit wound healing model was constructed as follows: Healthy rabbits purchased after passing quarantine were acclimatized for one week before the experiment, with free access to food and water, and kept at a room temperature of 23±2℃, humidity of 50-70%, and natural light. After one week, the rabbits were anesthetized by intravenous injection of 30 mg / kg sodium pentobarbital. After hair removal and disinfection of the back, a 1cm×1cm incision was made on the back with a scalpel, initially reaching the full thickness of the skin, and then carefully dissected down to the fascia layer, avoiding large blood vessels in the subcutaneous tissue. The wound was mainly characterized by oozing. A total of 6 rabbits were set up: a negative control group, a model group, and experimental groups 1-11.

[0121] Sample loading: On day 1 of the experiment, administration began after wounds were created in rabbits. Negative control group: Administered twice daily, at 9:00 AM and 5:00 PM, each time by applying a cotton pad soaked in physiological saline to the wound site for 15 seconds, for 14 consecutive days. Model group: Normal feeding after wound creation, without any other treatment. Experimental groups 1-11: The difference from the negative control group was that the physiological saline was replaced with the experimental sample solution of each group; all other aspects remained the same.

[0122] Detection: Images were analyzed using Photoshop 2023 software on days 4, 7, and 14, the wound area was measured, and the healing rate was calculated. A higher healing rate (%) indicates a better repair effect.

[0123] The wound healing rates of each group of rabbits on days 4, 7, and 14 are shown in Table 3.

[0124] Table 3

[0125] Example 1 31.65％ 59.26％ 96.83％ Example 2 31.53％ 59.14％ 96.48％ Example 3 31.40％ 59.03％ 96.12％ Example 4 30.07％ 57.77％ 92.32％ Example 5 24.18％ 52.20％ 75.52％ Example 6 32.21％ 59.79％ 98.42％ Example 7 38.87％ 66.08％ 99.97％ Example 8 29.17％ 56.92％ 89.76％ Example 9 29.67％ 57.39％ 91.17％ Comparative Example 1 21.85％ 50.01％ 68.89％ Comparative Example 2 22.76％ 50.93％ 71.49％ negative control group 18.15％ 46.57％ 58.09％ Model group 17.98％ 46.35％ 57.84％

[0126] As shown in Table 3, the *Prickly pear* fruit extract obtained by the preparation method of this invention has a good repair effect after use, and can basically completely heal rabbit wounds within 14 days. However, relevant parameters in the preparation process, such as the concentration ratio of the extract, the alcohol precipitation solvent and its ratio, and the microwave process parameters, all affect the above-mentioned repair effect to a certain extent, and thus significantly affect the wound healing rate of rabbits. In addition, if one or more of the microwave extraction process or vacuum reflux extraction steps are missing, or if one or more of the preparation steps are replaced with other preparation processes, the repair effect of the final *Prickly pear* fruit extract will be greatly reduced, that is, the wound healing rate of rabbits will be poor.

[0127] Test Example 5

[0128] Safety testing:

[0129] According to the "Cosmetic Safety Technical Specifications" (2015 edition), acute eye irritation test, cell sensitization test, and cell phototoxicity test were conducted on Examples 1-9 respectively; the safety of the products obtained in the examples was evaluated together. The results are shown in Table 4.

[0130] Table 4

[0131]

[0132] Therefore, it can be seen that the sea buckthorn fruit extract product involved in this invention is safe and non-irritating.

[0133] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0134] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0135] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. The application of a *Prickly pear* fruit extract in the preparation of skin barrier repair products, characterized in that, The preparation method of the sea buckthorn fruit extract includes the following steps: (1) Mix the raw material of the prickly pear fruit with water and then microwave it. Then, vacuum decompression reflux extraction is performed on the treated mixture to obtain the extract. The conditions for microwave processing include: microwave power of 150-400 W and microwave time of 30-90 min. The vacuum degree of the vacuum decompression reflux extraction is 0.03-0.08 MPa, and the temperature is 35-45℃; The raw material of the prickly pear fruit is prickly pear seed meal powder, which is the residue powder left after pressing the prickly pear kernels for oil. (2) Filter the extract and concentrate the filtrate to 1 / 6-1 / 2 of the filtrate volume to obtain the concentrate; (3) Mix the concentrate with the alcohol solution for alcohol precipitation; the alcohol is a combination of ethanol, butanediol and polyethylene glycol in a volume ratio of (1-4):1:(0.05-0.2), and the number average molecular weight of the polyethylene glycol is 200-600. (4) Filter the alcohol precipitation solution, dry the precipitate, and obtain the prickly pear fruit extract; The skin barrier repair is manifested in: promoting an increase in epidermal thickness, reducing the water loss rate of damaged skin, and accelerating wound healing.

2. The application according to claim 1, characterized in that, The ratio of raw prickly ash fruit to water is 1:(5-20)g / mL.

3. The application according to claim 1, characterized in that, The vacuum decompression reflux extraction process is performed 1-3 times, with each process lasting 60-120 minutes independently.

4. The application according to claim 1, characterized in that, The alcohol solution is an anhydrous alcohol solution or an aqueous alcohol solution, and the alcohol concentration of the aqueous alcohol solution is 90-99%.

5. The application according to claim 1, characterized in that, The final volume concentration of alcohol after mixing the alcohol solution and the concentrate is 80-90%.

6. The application according to claim 1, characterized in that, The alcohol solution is added to the concentrate dropwise while being stirred.

7. The application according to claim 1, characterized in that, The filtration in step (4) is performed using a filter cloth with a pore size of 150-250 mesh.

8. The application according to claim 1, characterized in that, The drying method described in step (4) includes freeze drying.

9. The application according to claim 1, characterized in that, The products include cosmetics, pharmaceuticals, or medical materials.

10. The application according to claim 1, characterized in that, The content of the sea buckthorn fruit extract in the product is 0.01%-5%.

Images