A cell wall polysaccharide of citrus sinensis peel and a preparation method and application thereof

Abstract

This application discloses a polysaccharide for the cell wall of Citrus reticulata peel, its preparation method, and its application. The preparation method of the polysaccharide includes: weighing and drying Citrus reticulata peel, pulverizing, and sieving; extracting the Citrus reticulata peel powder by reflux in an organic solvent, washing the filter residue to remove fat, and then removing starch; washing and drying the filter residue after starch removal to obtain Citrus reticulata peel cell wall material; extracting the Citrus reticulata peel cell wall material with an extraction solution, adjusting the pH of the extraction solution to neutral, concentrating, dialyzing, and freeze-drying to obtain the polysaccharide. The Citrus reticulata peel cell wall polysaccharide is prepared by this method. The application of the Citrus reticulata peel cell wall polysaccharide is its use in the preparation of skin care products or pharmaceuticals that have antioxidant properties and promote skin damage repair.

Description

Technical Field

[0001] This application relates to the field of cell wall polysaccharide preparation, and more particularly to a tea branch and citrus peel cell wall polysaccharide, its preparation method, and its application. Background Technology

[0002] *Citrus reticulata* cv. Chachiensis is a cultivated variety of citrus fruit belonging to the Rutaceae family and the *Citrus* genus. The peel of *Citrus reticulata* can be categorized based on its harvest time and maturity into several types: *Citrus reticulata* fetal peel, *Citrus reticulata* green peel, *Citrus reticulata* slightly red peel, and *Citrus reticulata* fully red peel. It also includes the dried, mature peel after aging, known as Guangchenpi (Guangdong aged tangerine peel). Guangchenpi is a major source of authentic Guangdong medicinal material and is recognized by the Ministry of Health as a dual-use medicinal and edible Chinese medicine. *Citrus reticulata* peel contains various active ingredients, with significant research and development currently focusing on its volatile oils, flavonoids, and trace elements. However, in recent years, the plant cell wall structure, traditionally considered inactive, has received increasing attention from researchers. As one of the main differences between plant and animal cells, the cell wall plays diverse roles in cell metabolism, intercellular transport, signal sensing and transduction, cell fate determination, and responses to pathogens and stress. It also influences cell differentiation, defense, and plant growth and development in these processes. As cell wall polysaccharides are the main chemical components of cell walls, the study of their activity is receiving increasing attention.

[0003] Cell wall polysaccharides can be broadly classified into pectin, hemicellulose, and cellulose. These polysaccharides collectively constitute the main structure of the cell wall, ensuring plant integrity and supporting and regulating life processes. Cell wall polysaccharides are also a major component of dietary fiber, playing an indispensable role in modern diets and considered a promising raw material for biomedicine and the food industry. In recent years, they have attracted considerable attention due to their antioxidant, antibacterial, anticoagulant, and antidiabetic biological activities. Furthermore, they possess adhesive properties, biodegradability, non-toxicity, and diverse physicochemical properties.

[0004] Skin, the largest organ in the human body, primarily protects the body from potential damage such as microorganisms, radiation, and mechanical injury; therefore, it is highly susceptible to damage. Rapid healing is crucial after skin injury to rebuild the protective barrier, making wound healing a significant clinical issue. However, in modern medicine, the number of drugs capable of stimulating wound healing remains limited. The functions of cell walls in cell differentiation, defense, and metabolism during plant growth and development provide a basis for further research into new natural compounds with antioxidant, skin repair, and wound-healing activities. In fact, existing literature reports that plant polysaccharides can accelerate healing, regulate the inflammatory phase, scavenge free radicals, and stimulate the proliferation of dermal fibroblasts and keratinocytes. However, to date, no research reports have been published on polysaccharides extracted from the cell walls of citrus peel with antioxidant and wound-healing activities. Summary of the Invention

[0005] This application provides a polysaccharide from the cell wall of citrus peel of tea branches, its preparation method, and its application, in order to solve the problems existing in related technologies. The technical solution is as follows:

[0006] In a first aspect, embodiments of this application provide a method for preparing polysaccharides from the cell walls of tea twig and citrus peel, comprising:

[0007] Weigh out tea branches and tangerine peel, dry them, crush them, and sieve them.

[0008] Tea branch and citrus peel powder was extracted by reflux in an organic solvent, and the filter residue was washed and defatted, and then the starch was removed.

[0009] The filter residue after removing starch was rinsed and dried to obtain the cell wall material of tea branch and citrus peel.

[0010] The cell wall material of tea branch and citrus peel was extracted with an extraction solution. The pH of the extraction solution was adjusted to neutral, and the mixture was concentrated, dialyzed, and freeze-dried to obtain polysaccharides from the cell wall of tea branch and citrus peel.

[0011] In one embodiment, tea branches and tangerine peel are weighed, dried, pulverized, and passed through a 40-60 mesh sieve; wherein the drying temperature is 50-60℃ and the drying time is 12-24 hours.

[0012] In one embodiment, the organic solvent is ethanol, with an ethanol concentration of 80-95% and a material-to-liquid ratio of 1:15-25.

[0013] The reflux extraction temperature is 60-90℃;

[0014] The degreasing conditions are as follows: degreasing at 4-10℃ for 12-15 hours, and the degreasing solvent is a solution of chloroform and methanol mixed in a mass ratio of 2:1, with a material-to-liquid ratio of 1:10-20.

[0015] Starch is removed by treatment with dimethyl sulfoxide (DMSO) at a concentration of 80-95% for 8-10 hours at a material-to-liquid ratio of 1:10-20.

[0016] In one embodiment, tea branch and citrus peel powder is refluxed in an organic solvent for 2-5 times, with each extraction time being 1-3 hours. The filter residue is then washed 3-5 times to remove fat, followed by treatment with dimethyl sulfoxide to remove starch.

[0017] Rinse the filter residue after removing starch 3-5 times.

[0018] In one embodiment, the solvent used for rinsing the filter residue after degreasing and for rinsing the filter residue after removing starch are both acetone or ethanol with a concentration of 80% or higher.

[0019] The extraction solution is a sodium carbonate solution containing sodium borohydride, wherein the sodium carbonate concentration is 0.05 mol / L and the sodium borohydride concentration is 0.02 mol / L.

[0020] In one embodiment, extraction is performed using an extraction solution at a temperature of 20-30°C for 2-4 hours, with a material-to-liquid ratio of 1:15-25.

[0021] Dialysis was performed using a 6-8 kDa dialysis bag;

[0022] The drying process is either vacuum drying or freeze drying.

[0023] In one embodiment, the source of the tea branch tangerine peel is the tangerine peel embryo, or green peel, or red peel, or second red peel, or aged peel from Guangchen.

[0024] Secondly, embodiments of this application provide a polysaccharide for the cell wall of citrus peel from tea branches, prepared by any of the above-mentioned methods for preparing polysaccharides for the cell wall of citrus peel from tea branches.

[0025] Thirdly, this application provides an application of tea branch and citrus peel cell wall polysaccharide, which is used in the preparation of skin care products or pharmaceuticals that have antioxidant properties and promote skin damage repair.

[0026] In one embodiment, the skincare product includes a toner, essence, hydrogel, serum, cream, lotion, or mask; the concentration of polysaccharides from the cell walls of Citrus reticulata peel in the skincare product is 2-15%.

[0027] The dosage forms of the drug include solutions, aerosols, sprays, ointments, and films.

[0028] The advantages or beneficial effects of the above technical solutions include at least the following:

[0029] The tea branch and citrus peel cell wall polysaccharide provided by this invention innovates and optimizes the extraction method of plant peel cell wall polysaccharides. Compared with traditional methods for preparing peel cell wall polysaccharides, it can more specifically provide the market with a polysaccharide raw material with better antioxidant and wound-healing activities. This invention greatly promotes the comprehensive development and utilization of tea branch and citrus peel, and broadens its application prospects in skin care products and pharmaceuticals.

[0030] The method for preparing polysaccharides from the cell walls of tea twig and citrus peel provided by this invention is simple and quick. Experimental studies have shown that these cell wall polysaccharides have antioxidant and wound-healing effects, and can therefore be applied as biomedical excipients for skin anti-oxidation and wound healing.

[0031] The polysaccharide from the cell wall of Citrus reticulata peel prepared by the above method has a sugar content of 60-85%, a protein content of 2.0-8.5%, a total phenol content of 0.3-1.5%, and a uronic acid content of 40-62%. It exhibits good hydroxyl radical scavenging ability in in vitro antioxidant activity tests, thus it can be used as a raw material for skincare products or pharmaceuticals with antioxidant effects. Furthermore, in experiments on the proliferation, migration, and wound healing of human keratinocytes, the polysaccharide from the cell wall of Citrus reticulata peel significantly promotes the proliferation and migration of human keratinocytes, accelerates wound healing and epithelialization, and is suitable for preparing skincare products or pharmaceuticals that promote skin renewal and repair, especially wound healing.

[0032] The above overview is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features of this application will become readily apparent from the accompanying drawings and the following detailed description. Attached Figure Description

[0033] In the accompanying drawings, unless otherwise specified, the same reference numerals throughout the various drawings denote the same or similar parts or elements. These drawings are not necessarily drawn to scale. It should be understood that these drawings depict only some embodiments disclosed in this application and should not be construed as limiting the scope of this application.

[0034] Figure 1 This is a diagram showing the hydroxyl radical scavenging capacity of cell wall polysaccharides.

[0035] Figure 2 The figure shows the results of the CCK8 assay for detecting the effect of cell wall polysaccharides on HaCaT cell proliferation.

[0036] Figure 3 Microscopic images of wound healing in HaCaT cells treated with cell wall polysaccharides during a cell scratch assay.

[0037] Figure 4This is a graph comparing the wound healing rates of cell wall polysaccharides and the control group in a scratch assay.

[0038] Figure 5 This is a graph comparing the degree of wound healing between the cell wall polysaccharide and control groups in a wound healing experiment. Detailed Implementation

[0039] In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of this application. Therefore, the drawings and description are considered to be exemplary in nature and not restrictive.

[0040] In a first aspect, embodiments of this application provide a method for preparing polysaccharides from the cell walls of tea twig and citrus peel, comprising:

[0041] Step 101: Weigh out tea branches and tangerine peel, dry them, crush them, and sieve them.

[0042] In one embodiment, the source of the peel of the tea branch mandarin orange is the peel of the tea branch mandarin orange embryo, or green peel, or red peel, or second red peel, or aged peel of Guangchen.

[0043] In one embodiment, tea branches and tangerine peel are weighed, dried, pulverized, and passed through a 40-60 mesh sieve; wherein the drying temperature is 50-60℃ and the drying time is 12-24 hours.

[0044] Step 102: Extract the tea branch and citrus peel powder by reflux in an organic solvent, and then degrease the filter residue by rinsing, followed by the removal of starch.

[0045] In one embodiment, the organic solvent is ethanol, the ethanol concentration is 80-95%, and the material-to-liquid ratio is 1:15-25.

[0046] In one embodiment, the reflux extraction temperature is 60–90°C.

[0047] In one embodiment, the degreasing conditions are 4-10°C for 12-15 hours, the degreasing solvent is a solution of chloroform and methanol mixed in a mass ratio of 2:1, and the material-to-liquid ratio is 1:10-20.

[0048] In one embodiment, starch is removed by treatment with dimethyl sulfoxide (DMSO), wherein the DMSO concentration is 80-95%, preferably 90%, the treatment time is 8-10 hours, and the material-to-liquid ratio is 1:10-20.

[0049] In one embodiment, tea branch and citrus peel powder is refluxed in an organic solvent for 2-5 times, with each extraction time being 1-3 hours. The filter residue is then washed 3-5 times to remove fat, followed by treatment with dimethyl sulfoxide to remove starch.

[0050] In one embodiment, the filter residue after starch removal is rinsed 3-5 times.

[0051] Step 103: Take the filter residue after removing starch, rinse and dry it to obtain the cell wall material of tea branch citrus peel.

[0052] In one embodiment, the solvents used for rinsing the filter residue in step 102 (after rinsing and degreasing) and for rinsing the filter residue after removing starch in step 103 are acetone or ethanol with a concentration of 80% or higher.

[0053] Step 104: Take the cell wall material of tea branch and citrus peel, extract it with the extraction solution, adjust the pH of the extraction solution to neutral, concentrate, dialyze, and freeze dry to obtain polysaccharide of tea branch and citrus peel cell wall.

[0054] In one embodiment, the extraction solution is a sodium carbonate solution containing sodium borohydride, wherein the sodium carbonate concentration is 0.05 mol / L and the sodium borohydride concentration is 0.02 mol / L.

[0055] In one embodiment, extraction is performed using an extraction solution at a temperature of 20-30°C for 2-4 hours, with a material-to-liquid ratio of 1:15-25.

[0056] In one embodiment, the drying is vacuum drying or freeze drying, preferably freeze drying.

[0057] Secondly, this application provides a polysaccharide for the cell wall of citrus peel from tea branches, prepared by a method for preparing polysaccharides for the cell wall of citrus peel from tea branches.

[0058] According to the test results, the polysaccharide in the cell wall of the tea branch and citrus peel prepared in this application has a sugar content of 60-85%, a protein content of 2.0-8.5%, a total phenol content of 0.3-1.5%, and a uronic acid content of 40-62%.

[0059] The polysaccharide in the cell wall of tea branch and citrus peel has a good ability to scavenge hydroxyl free radicals; at the same time, it has a significant promoting effect on the proliferation, migration and wound healing of human keratinocytes.

[0060] Thirdly, this application provides an application of tea branch and citrus peel cell wall polysaccharide, which is used in the preparation of skin care products or pharmaceuticals that have antioxidant properties and promote skin damage repair.

[0061] In one embodiment, the skincare product includes a toner, essence, hydrogel, serum, cream, lotion, or mask; the concentration of polysaccharides from the cell walls of Citrus reticulata peel in the skincare product is 2-15%.

[0062] The dosage forms of drugs include solutions, aerosols, sprays, ointments, and films.

[0063] Taking hydrogel as an example, hydrogel is used as a topical preparation for skin wound healing. The hydrogel includes polysaccharides from the cell walls of Citrus reticulata peel obtained by the above method. The preparation method of hydrogel is as follows:

[0064] Weigh out the polysaccharide from the cell wall of tea twig and citrus peel, dissolve it in a sterile solvent to prepare a polysaccharide aqueous solution, add a hydrophilic polymer and mix well, use a magnetic stirrer to help dissolve, after complete dissolution, add calcium chloride or ferric chloride solution and continue stirring the reaction, while adjusting the pH of the solution to 7.4, until the hydrogel changes from a flowing state to a non-flowing state, thus obtaining the hydrogel.

[0065] Preferably, the concentration of polysaccharides in the cell walls of tea twig and citrus peel in the hydrogel is 2-15%.

[0066] Preferably, the sterilizing solvent includes, but is not limited to, pure water or PBS.

[0067] Preferably, the hydrophilic polymer includes, but is not limited to, one or more polymers selected from sodium alginate, hyaluronic acid, chitosan and its derivatives, and cellulose and its derivatives.

[0068] Preferably, the concentration of calcium chloride or ferric chloride solution is 0.5-3%.

[0069] The above-mentioned method for preparing polysaccharide hydrogels for wound healing is simple, fast, convenient, and environmentally friendly.

[0070] Example 1: Preparation of polysaccharides from the cell walls of tea twig and citrus peel

[0071] Dry mature tea branch and citrus peel at 50℃, pulverize, and pass through a 40-mesh sieve. Mix 100g of the tea branch and citrus peel powder with 95% ethanol at a ratio of 1:20, and reflux extract three times at 80℃, 2 hours each time. After cooling, filter, wash the residue three times with 80% ethanol, then mix thoroughly with a chloroform / methanol (2:1) solution at a ratio of 1:10, and pretreat at 4℃ for 15 hours. Treat the resulting residue with 90% dimethyl sulfoxide solution at a ratio of 1:10 for 8 hours. Wash the residue three times with acetone, and freeze-dry to obtain the tea branch and citrus peel cell wall material.

[0072] The obtained tea branch and citrus peel cell wall material was extracted with 0.05 mol / L sodium carbonate (containing 0.02 mol / L sodium borohydride) solution at a material-to-liquid ratio of 1:20 at 25℃ for 2 hours. The pH of the filtrate was adjusted to neutral and concentrated. The filtrate was dialyzed using a 6-8 kDa dialysis bag and freeze-dried to obtain tea branch and citrus peel cell wall polysaccharide.

[0073] The polysaccharide obtained from the cell wall of tea twig and citrus peel prepared by the above method was found to have a sugar content of 62.8%, a protein content of 3.5%, a total phenol content of 0.4%, and a uronic acid content of 46.7% by phenol-sulfuric acid method, BCA protein method, gallic acid colorimetric method, and gas chromatography, respectively.

[0074] Comparative Example 1

[0075] This comparative embodiment provides a citrus peel cell wall polysaccharide, the purpose of which is to facilitate a comparison of the cell wall polysaccharide extracted and prepared by conventional methods with the tea branch citrus peel cell wall polysaccharide of the present invention in the following experiments. The preparation method refers to the citrus polysaccharide extraction method disclosed in Chinese Patent Document CN107033256A.

[0076] The preparation steps are as follows: Wash citrus peels, cut them into small pieces, and dry them in a 50℃ forced-air constant-temperature desiccator for 12 hours. Then, pulverize the dried citrus peels through a 40-mesh sieve to obtain citrus peel powder. Next, take the citrus peel powder and mix it thoroughly with petroleum ether at a material-to-liquid ratio of 1:15. Degrease and decolorize the mixture at 40℃, then discard the petroleum ether, evaporate to dryness, and soak in 20 times the amount of distilled water for 10 minutes. Finally, extract the citrus peel cell wall polysaccharide under ultrasonic conditions of 60℃, 220W, and 20KHZ for 10 minutes.

[0077] Example 2: Scavenging experiment of hydroxyl radicals from polysaccharides in the cell walls of tea branches and citrus peel

[0078] The polysaccharide samples from Example 1 and Comparative Example 1 were prepared into a series of sample solutions with concentration gradients (0.1, 0.2, 0.4, 0.8, 1.2, and 1.6 mg / mL), and control solutions (Vc) with the same concentration gradient were prepared simultaneously. 1 mL of each concentration of the test solution was added sequentially to 1 mL of ferrous sulfate solution (9 mmol / L), 1 mL of salicylic acid-ethanol solution (9 mmol / L), and 1 mL of hydrogen peroxide solution (9 mmol / L). After mixing, the mixture was incubated at 37°C for 30 minutes, centrifuged at 4000 rpm for 10 minutes, and the supernatant was measured at 510 nm. Deionized water was used as a blank control. The hydroxyl radical scavenging rate (SR) was calculated using the formula: SR(%) = (A0 - A) / A0 × 100. Where: A0 is the absorbance of the blank control, and A is the absorbance of the sample.

[0079] The above results are as follows Figure 1 As shown, the polysaccharide from the tea branch and citrus peel cell wall of Example 1 has a better hydroxyl radical scavenging ability within the experimental concentration range compared with the citrus peel cell wall polysaccharide prepared by the traditional method, and the scavenging rate is directly proportional to the polysaccharide concentration.

[0080] Example 3: Effects of tea branch and citrus peel cell wall polysaccharides on the proliferation of keratinocytes in the skin

[0081] Human immortalized keratinocytes (HaCaT cells) are an immortalized cell line that spontaneously transforms from adult epidermal cells. They participate in the formation of the skin's physical barrier, playing a protective role in the skin, and are widely used in research on skin damage repair, photoaging, and a range of skin diseases.

[0082] This experiment used the CCK8 assay to detect the effect of tea branch and citrus peel cell wall polysaccharides from Example 1 on keratinocyte proliferation. First, HaCaT cells were cultured at 4 × 10⁻⁶ cells / year. 3 Cells / well were seeded at a density of 4 × 10⁶ cells / well in 96-well plates, with 100 μL of cell suspension added to each well. 4 After incubation for 24 hours, the supernatant was discarded, and the samples were rinsed. Then, 100 μL of different concentrations of polysaccharide samples from Example 1 and Comparative Example 1 (25, 50, 100, 200 μg / mL) were added to each well, with three replicates per concentration. After incubation for 24 hours, 10 μL of CCK8 solution was added to each well, mixed, and incubated for another 2 hours. The absorbance was measured at 450 nm using a microplate reader.

[0083] Experimental results are as follows Figure 2 As shown, compared to the citrus peel cell wall polysaccharide in Comparative Example 1, the tea branch citrus peel cell wall polysaccharide in Example 1 exhibited a stronger promoting effect on keratinocyte proliferation within the tested concentration range, and the cell proliferation activity first increased and then decreased with increasing concentration. At the optimal proliferation concentration of 100 μg / mL, the tea branch citrus peel cell wall polysaccharide showed the most significant proliferative effect compared to the blank control group, with a cell proliferation activity reaching 160.9%.

[0084] Example 4: Wound healing activity of cell wall polysaccharides from tea branch and citrus peel

[0085] The cell scratch assay is an experimental method for measuring cell migration and repair capabilities. It establishes an in vitro wound healing model to study the mechanism of skin wound healing. This experiment used this method to obtain HaCaT cells at a concentration of 4 × 10⁻⁶ cells. 5 Cells / well were seeded at a density of 12-well plates, with 1 ml of cell suspension added to each well. After incubation for 24 h, vertical scratches were made in each well using a 10 μL pipette tip. The culture medium was discarded, and the cells were washed three times with PBS. The experimental group was treated with serum-free medium containing 100 μg / mL of tea twig and citrus peel cell wall polysaccharide, while the control group was treated with blank medium. Cell growth was determined by microscopic photography, and the scratch area was compared at 0, 24, and 48 h. The wound healing rate was calculated using ImageJ software.

[0086] Cell scratch assay results are as follows Figure 3 and Figure 4As shown, compared to citrus peel cell wall polysaccharides, the tea branch citrus peel cell wall polysaccharides of Example 1 exhibited higher wound healing rates at 24 and 48 hours after administration, and the healing rate at 48 hours was 13% higher than that of the control group. This experiment demonstrates that the tea branch citrus peel cell wall polysaccharides of the present invention have a more significant promoting effect on keratinocyte proliferation and migration compared to traditionally prepared citrus peel cell wall polysaccharides, and also show better skin repair and wound healing activity.

[0087] Example 5: Preparation and evaluation of the wound healing efficacy of tea branch and citrus peel cell wall polysaccharide hydrogel

[0088] Weigh 50 mg of the sample polysaccharide from Example 1 and Comparative Example 1 respectively, dissolve it in 1 mL of sterile PBS (pH 7.4) to prepare a polysaccharide aqueous solution, add 4% sodium alginate aqueous solution and mix well, use a magnetic stirrer to help dissolve, after complete dissolution, add 2% calcium chloride solution and continue stirring until the hydrogel changes from a flowing state to a non-flowing state, thus obtaining the hydrogel.

[0089] Subjects: Thirty participants aged 20-30 years with similar-sized skin wounds on their hands or arms were selected. They were randomly divided into three groups of 10 each: two experimental groups and one control group. Each participant in the experimental group applied an equal amount of the aforementioned hydrogel preparation to the wound site twice daily (morning and evening), while the control group applied an equal amount of the polysaccharide-free hydrogel preparation to the wound site. Wound healing was compared between the groups after one week (no other wound-healing medications or reagents were administered during the trial).

[0090] The results of the test are as follows Figure 5 As shown, the results indicate that the experimental group of subjects had better skin wound healing than the control group, and the hydrogel containing tea branch citrus peel cell wall polysaccharide had a better skin wound healing effect than the citrus peel cell wall polysaccharide hydrogel.

[0091] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of those different embodiments or examples.

[0092] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "a plurality of" means two or more, unless otherwise explicitly specified.

[0093] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in this application, and these should all be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. Use of a cell wall polysaccharide of Citrus sinensis peel in the preparation of a skin care product or a pharmaceutical product for promoting the repair of skin damage, characterized in that, The tea-citrus peel cell wall polysaccharide is prepared by a preparation method comprising the following steps: The tea-citrus peel is weighed, dried, crushed, and sieved; The tea-citrus peel powder is extracted by refluxing in an ethanol solvent with a concentration of 80-95%, and the filtrate is rinsed and then degreased using a solution of chloroform and methanol mixed at a mass ratio of 2:1, followed by treatment with dimethyl sulfoxide with a concentration of 80-95% to remove starch; The filtrate after removing starch is rinsed and dried to obtain tea-citrus peel cell wall material; The tea-citrus peel cell wall material is extracted using an extraction solution containing sodium borohydride and sodium carbonate, wherein the concentration of sodium carbonate in the extraction solution is 0.05 mol / L, and the concentration of sodium borohydride is 0.02 mol / L; the extraction solution is adjusted to neutral pH, concentrated, dialyzed, and freeze-dried to obtain tea-citrus peel cell wall polysaccharide.

2. Use according to claim 1, characterized in that, The tea-citrus peel is weighed, dried, crushed, and sieved through a 40-60 mesh sieve; wherein the drying temperature is 50-60°C, and the drying time is 12-24 hours.

3. Use according to claim 1, characterized in that, The ethanol reflux extraction liquid ratio is 1:15-25; The reflux extraction temperature is 60-90°C; The degreasing conditions are degreasing at 4-10°C for 12-15 hours, and the degreasing solvent degreasing liquid ratio is 1:10-20; The dimethyl sulfoxide treatment time is 8-10 hours, and the liquid ratio is 1:10-20.

4. Use according to claim 3, characterized in that, The tea-citrus peel powder is extracted by refluxing in an organic solvent for 2-5 times, each time for 1-3 hours, and the filtrate is rinsed 3-5 times before degreasing, followed by treatment with dimethyl sulfoxide to remove starch; The filtrate after removing starch is rinsed 3-5 times.

5. The use according to claim 1, characterized in that, The solvent used for rinsing the filtrate after degreasing the filtrate, and the solvent used for rinsing the filtrate after removing starch, are both acetone or ethanol with a concentration of 80% or above.

6. The use according to claim 1, characterized in that, The extraction solution is used for extraction at an extraction temperature of 20-30°C for 2-4 hours, and the liquid ratio is 1:15-25; Dialysis is performed using a 6-8 kDa dialysis bag; The drying is vacuum drying or freeze-drying.

7. The use according to any one of claims 1 to 6, characterized in that, The tea-citrus peel is derived from tea-citrus peel, or green peel, or red peel, or double red peel, or multi-year broad peel.

8. Use according to any one of claims 1 to 6, characterized in that, The skin care product includes cosmetic water, or essence water, or water gel, or essence liquid, or face cream, or emulsion, or face mask; the concentration of tea-citrus peel cell wall polysaccharide in the skin care product is 2-15%; The dosage form of the pharmaceutical product includes solution, or aerosol, or spray, or ointment, or film.

Images