Aromatic thread extract and its preparation process and application

Abstract

The present application proposes a roxburghii extract and its preparation process and application. The preparation process of roxburghii extract comprises: crushing the whole plant of roxburghii with a grinder and then sieving to obtain dry powder; adding the dry powder to an ethanol solvent and performing ultrasonic extraction at room temperature to obtain a crude extract of roxburghii; filtering the crude extract of roxburghii to remove the residue to obtain a filtrate; performing membrane separation and / or reduced pressure concentration on the obtained filtrate to remove ethanol, and fixing the volume to obtain a concentrated solution; adding a cosolvent and a stabilizer to the obtained concentrated solution, and then decolorizing to obtain a decolorized roxburghii solution; filtering and sterilizing the decolorized roxburghii solution; adding a preservative to the sterilized solution to obtain a roxburghii extract. The roxburghii extract is prepared by the preparation process of the roxburghii extract. The application of the roxburghii extract is the application of the roxburghii extract in skin care products for soothing, anti-inflammatory, anti-wrinkle, or repair.

Description

Technical Field

[0001] The present application relates to the technical field of skin care, and in particular to a roxburghii extract and a preparation process and application thereof. Background Art

[0002] Golden thread vine (Anoectochilus roxburghii (Wall.) Lindl.), also known as golden thread orchid, golden silk grass, bird ginseng, golden thread tiger head banana, golden thread into the bone, etc., is a rare perennial Chinese herbal medicine of the genus Anoectochilus in the Orchidaceae family. It is mainly distributed in southern provinces of my country such as Fujian, Guangdong, and Zhejiang. Fujian is the authentic production area of ​​​​Golden thread vine. The whole herb of Anoectochilus roxburghii can be used as medicine. The Standard of Traditional Chinese Medicine in Fujian Province (2006 edition) records Anoectochilus roxburghii: "This product is a rare folk herbal medicine in Fujian, Taiwan and other provinces. Its nature and flavor are flat and sweet; it can clear away heat and cool blood, dispel wind and dampness; it is used for nephritis, bronchitis, cystitis, diabetes, rheumatoid arthritis, acute convulsions in children, snake bites and other symptoms. Anoectochilus roxburghii is rich in flavonoids, polysaccharides and glycosides, amino acids, triterpenes, esters, sterols and trace elements. It has rich biological activity and can effectively resist inflammation, scavenge free radicals, and resist aging. At present, the development of Anoectochilus roxburghii products is mainly concentrated on fresh products, dry teas, health teas and other products. The structure is single and the innovation ability is weak. However, the application and research in skin care are rare and do not meet the market demand.

[0003] The currently disclosed preparation process of Anoectochilus roxburghii is relatively complicated and is not suitable for large-scale production. In addition, some preparation processes have low content of effective ingredients or low biological activity, instability of active ingredients, etc., which will cause waste of Anoectochilus roxburghii resources to a certain extent. Anoectochilus roxburghii is known as the "king of medicine" and "golden grass" and is one of the unique Chinese herbal medicines in my country. Therefore, optimizing the preparation process of Anoectochilus roxburghii extract and improving its application value are of great significance to fully develop Anoectochilus roxburghii resources.

[0004] Application Contents

[0005] The present application provides a roxburghii extract and its preparation process and application to solve the problems existing in the related technology. The technical solution is as follows:

[0006] In a first aspect, the present application provides a process for preparing a roxburghii roxburghii extract, comprising:

[0007] The dried whole plant of Anoectochilus roxburghii is crushed by a crusher and then sieved to obtain dry powder;

[0008] The dry powder is added into an ethanol solvent, and ultrasonic extraction is performed at room temperature to obtain a crude extract of Anoectochilus roxburghii;

[0009] Filtering the crude extract of Anoectochilus roxburghii to remove residues and obtain a filtrate;

[0010] The obtained filtrate is subjected to membrane separation and / or reduced pressure concentration to remove ethanol, and the volume is fixed to obtain a concentrated solution;

[0011] Adding a co-solvent and a stabilizer to the obtained concentrated solution, followed by decolorization to obtain a decolorized rosa canina liquid;

[0012] Filter and sterilize the decolorized solution of Anoectochilus roxburghii;

[0013] A preservative is added to the sterilized solution to obtain a golden thread vine extract.

[0014] In one embodiment, the dry powder is added to an ethanol solvent at a material-liquid ratio of 1:4-1:40, and ultrasonic extraction is performed at room temperature; wherein the volume fraction of the ethanol solvent is 5%-70%, the ultrasonic frequency is 480W, the ultrasonic extraction time is 10-60min, and the extraction times are 1-2 times.

[0015] In one embodiment, the volume is fixed to 10-20 times the mass of the dry powder of Anoectochilus roxburghii to obtain a concentrated solution.

[0016] In one embodiment, 20%-50% w / w of a co-solvent and 0.1%-5% w / w of a stabilizer are added to the obtained concentrate.

[0017] In one embodiment, the cosolvent is one or more of glycerol, 1,3-propylene glycol, and 1,3-butylene glycol;

[0018] The stabilizer is one or more of 1,2-hexanediol, 1,2-pentanediol and p-hydroxyacetophenone.

[0019] In one embodiment, a macroporous adsorption resin is used for adsorption decolorization, and the resin usage is 4%-40% w / v.

[0020] In one embodiment, the macroporous adsorption resin includes one or more of D101, AB-8, XAD-2, HDP-100, HP-10, and LSD-20;

[0021] The adsorption method is static adsorption, and the adsorption time is 0.5-8h.

[0022] In one embodiment, 0.1%-0.5% w / w of preservative is added to the sterilized solution; the preservative is one or more of sodium metabisulfite, phenoxyethanol, benzyl alcohol, potassium sorbate, and sodium sorbate.

[0023] In a second aspect, an embodiment of the present application provides a roxburghii roxburghii extract, which is prepared by any of the roxburghii roxburghii extract preparation processes described above.

[0024] In a third aspect, an embodiment of the present application provides an application of a roxburghii roxburghii extract, which is an application of the roxburghii roxburghii extract in skin care products for soothing, anti-inflammatory, anti-wrinkle, or repairing effects.

[0025] The advantages or beneficial effects of the above technical solution include at least:

[0026] (1) The present application uses an optimized extraction process, including the extraction material-liquid ratio, extraction solvent, extraction time, decolorization process, and adding a stabilizer, to extract the golden thread vine to obtain the golden thread vine extract. The extraction process is simple and efficient and is suitable for large-scale production.

[0027] (2) The ultrasonic extraction method is used to extract the golden thread vine plant, which makes the plant cell wall and the entire organism rupture in an instant, shortening the crushing time. At the same time, the vibration generated by the ultrasound enhances the release, diffusion and dissolution of the substances in the golden thread vine cells, thereby significantly improving the extraction efficiency and obtaining a golden thread vine extract with a high content of effective ingredients and stable properties.

[0028] (3) The roxburghii extract obtained by the present application is enriched with a variety of effective ingredients such as roxburghii glycosides, roxburghii polysaccharides, etc. The extract has stable properties, high content of effective ingredient polysaccharides, good activity, and significant soothing, anti-inflammatory, anti-wrinkle, and repair effects. It can be used as a skin protectant in skin care products, thereby realizing the high-value utilization of roxburghii.

[0029] The above summary 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 the present application will be readily apparent by reference to the accompanying drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In the accompanying drawings, unless otherwise specified, the same reference numerals throughout the multiple drawings represent the same or similar parts or elements. These drawings are not necessarily drawn to scale. It should be understood that these drawings only depict some embodiments disclosed in the present application and should not be regarded as limiting the scope of the present application.

[0031] Figure 1 The figure shows the result that the extract of Anoectochilus roxburghii of Example 1 reduces the expression level of inflammatory factor TNF-α in RAW264.7 cells.

[0032] Figure 2 The figure shows the result that the roxburghii extract of Example 1 reduces the expression level of the inflammatory factor IL-1α in RAW264.7 cells.

[0033] Figure 3This graph shows the results of the roxburghii extract of Example 1 promoting the synthesis of type I collagen.

[0034] Figure 4 The graph shows the results of the HaCaT cell proliferation experiment using the roxburghii extract of Example 1.

[0035] Figure 5 The imageJ quantitative analysis graph of the HaCaT cell migration experiment of the roxburghii extract in Example 1 is shown.

[0036] Figure 6 A bar graph showing the migration mobility of HaCaT cells of the roxburghii extract of Example 1 is shown. DETAILED DESCRIPTION

[0037] Hereinafter, only some exemplary embodiments are briefly described. As those skilled in the art will appreciate, the described embodiments may be modified in various ways without departing from the spirit or scope of the present application. Therefore, the drawings and descriptions are considered to be exemplary and non-restrictive in nature. The test methods used in the following examples are conventional methods unless otherwise specified, and the raw materials, reagents, etc. used, unless otherwise specified, can be obtained from conventional commercial sources such as commercial purchase.

[0038] In a first aspect, the present application provides a process for preparing a roxburghii roxburghii extract, comprising:

[0039] Step 101, crushing.

[0040] The dried whole plant of Anoectochilus roxburghii is crushed with a crusher and then sieved to obtain dry powder.

[0041] Step 102, extraction.

[0042] The dry powder was added into an ethanol solvent and ultrasonic extraction was performed at room temperature to obtain a crude extract of Anoectochilus roxburghii.

[0043] In one embodiment, the dry powder is added to an ethanol solvent at a material-liquid ratio of 1:4-1:40, and ultrasonic extraction is performed at room temperature; wherein the volume fraction of the ethanol solvent is 5%-70%, the ultrasonic frequency is 480W, the ultrasonic extraction time is 10-60min, and the extraction times are 1-2 times.

[0044] The main driving force of ultrasound in extracting comes from the ultrasonic cavitation effect. The ultrasonic cavitation process is a process of concentrating the energy of the sound field and releasing it immediately. It usually refers to a series of dynamic processes such as oscillation, growth, contraction, and collapse of tiny bubbles in the liquid that are activated by ultrasound. Ultrasonic cavitation causes turbulence effect, perturbation effect, interface effect, and energy-gathering effect. The turbulence effect thins the boundary layer and increases the mass transfer rate; the perturbation effect strengthens the microporous diffusion; the interface effect increases the mass transfer surface area; the energy-gathering effect activates the molecules of the separated substances; thus, the mass transfer rate and effect of the separation and purification process are strengthened as a whole. Ultrasonic extraction of Chinese medicinal materials is not limited by the polarity and molecular weight of the ingredients, and is suitable for the extraction of most types of Chinese medicinal materials and various ingredients; the effective ingredients are easy to separate and purify during the extraction of the medicinal solution; there is no chemical reaction during the entire extraction process, and the activity of the active ingredients is little affected. Ultrasonic extraction can shorten the extraction time and improve the extraction rate. Compared with conventional methods, ultrasonic extraction has the advantages of simple operation, short extraction time, high extraction efficiency, and no need for heating.

[0045] The ultrasonic extraction method is used to extract the golden thread vine plant, which makes the plant cell wall and the entire organism rupture in an instant, shortening the crushing time. At the same time, the vibration generated by the ultrasound enhances the release, diffusion and dissolution of the golden thread vine intracellular substances, thereby significantly improving the extraction efficiency and obtaining a golden thread vine extract with a high content of effective ingredients and stable properties.

[0046] Step 103, filtering.

[0047] The crude extract of Anoectochilus roxburghii is filtered to remove residues and obtain a filtrate.

[0048] Step 104, concentrating and fixing the volume.

[0049] The obtained filtrate is subjected to membrane separation and / or concentrated under reduced pressure to remove ethanol, and the volume is fixed to obtain a concentrated solution.

[0050] In one embodiment, the volume is fixed to 10-20 times the mass of the dry powder of Anoectochilus roxburghii to obtain a concentrated solution.

[0051] Step 105, dissolving and decolorizing.

[0052] A co-solvent and a stabilizer are added to the obtained concentrated solution, followed by decolorization to obtain a decolorized roxburghii liquid.

[0053] Since a small amount of water-insoluble precipitate may be precipitated from the extract in step 104, adding a co-solvent can dissolve the precipitate, and adding a stabilizer can stabilize the heating and concentration process, resulting in physical changes in the extract such as color. This step can improve the stability of the extract, improve the quality such as color, and improve solubility.

[0054] In one embodiment, 20% to 50% w / w of a co-solvent and

[0055] 0.1% - 5% w / w stabilizer.

[0056] In one embodiment, the co-solvent is one or more of glycerol, 1,3-propylene glycol, and 1,3-butylene glycol.

[0057] In one embodiment, the stabilizer is one or more of 1,2-hexanediol, 1,2-pentanediol, and p-hydroxyacetophenone.

[0058] In one embodiment, a macroporous adsorption resin is used for adsorption decolorization, and the resin usage is 4%-40% w / v.

[0059] In one embodiment, the macroporous adsorption resin includes one or more of D101, AB-8, XAD-2, HDP-100, HP-10, and LSD-20.

[0060] In one embodiment, the adsorption method is static adsorption, and the adsorption time is 0.5-8 hours.

[0061] Step 106, sterilization.

[0062] Filter and sterilize the decolorized solution of Anoectochilus roxburghii.

[0063] Step 107, adding preservatives.

[0064] A preservative is added to the sterilized solution to obtain a golden thread vine extract.

[0065] In one embodiment, 0.1%-0.5% w / w of preservative is added to the sterilized solution; the preservative is one or more of sodium metabisulfite, phenoxyethanol, benzyl alcohol, potassium sorbate, and sodium sorbate.

[0066] In a second aspect, an embodiment of the present application provides a roxburghii roxburghii extract, which is prepared by a roxburghii roxburghii extract preparation process.

[0067] In a third aspect, an embodiment of the present application provides an application of a roxburghii roxburghii extract, which is an application of the roxburghii roxburghii extract in skin care products for soothing, anti-inflammatory, anti-wrinkle, or repairing effects.

[0068] Example 1

[0069] S1, crushing: taking the whole plant of dried roxburghii roxburghii, crushing it with a crusher and passing it through a No. 1 sieve to obtain 20 g of roxburghii roxburghii dry powder;

[0070] S2, extraction: the dry powder obtained in S1 was added with 30% ethanol solvent at a solid-liquid ratio of 1:15, and ultrasonic extraction was performed once, the ultrasonic frequency was 480W, and the ultrasonic extraction time was 30min, to obtain a crude extract of Anoectochilus roxburghii;

[0071] S3, filtering: filtering the crude extract of Anoectochilus roxburghii obtained in S2 to remove residues;

[0072] S4, concentration: the filtrate obtained in S3 is concentrated under reduced pressure to remove ethanol, and the volume is fixed with water to 10 times the mass of the dry powder of Anoectochilus roxburghii;

[0073] S5, dissolving: adding glycerol, 1,2-hexanediol and p-hydroxyacetophenone to the concentrated solution obtained in S4, the final solution having a mass ratio of crude drug: solution = 1:20, and a composition of 48.6% w / w concentrated solution, 50% w / w glycerol, 0.7% w / w 1,2-hexanediol and 0.7% w / w p-hydroxyacetophenone;

[0074] S6, decolorization: the solution obtained in S5 is subjected to adsorption decolorization for 2 hours using 20% ​​w / w XAD-2 macroporous adsorption resin to obtain a decolorized solution of Anoectochilus roxburghii;

[0075] S7, sterilization: filtering and sterilizing the decolorizing solution described in S6;

[0076] S8. Preservative compounding: Add 0.1% w / w sodium pyrosulfite to the sterilized solution in S7 to obtain the roxburghii extract.

[0077] The auxiliary solvents are glycerol, 1,3-propylene glycol and 1,3-butylene glycol, all of which are miscible with water or ethanol.

[0078] In the preparation process of the golden thread vine extract, after the ethanol is concentrated and removed, active substances insoluble in water are precipitated, which can be dissolved by adding glycerol, 1,3-propylene glycol and 1,3-butylene glycol as auxiliary solvents.

[0079] 1,2-Hexanediol and 1,2-pentanediol have two hydroxyl hydrophilic groups and a linear alkyl hydrophobic group in their molecular structure, so they have oil-water amphiphilicity. During the preparation of the golden thread vine extract, a trace amount of water-insoluble precipitate was precipitated after the ethanol was concentrated and removed. Adding 1,2-hexanediol and 1,2-pentanediol as co-solvents can dissolve it. The verification experiment results are as follows:

[0080] Table 1 Effect of the addition amount of 1,2-hexanediol and 1,2-pentanediol on the clarity of Anoectochilus roxburghii extract

[0081]

[0082] In this embodiment, p-hydroxyacetophenone is selected as a stabilizer. p-hydroxyacetophenone is an antioxidant that can inhibit the oxidative discoloration of phenolic pigments in plant extracts. During the preparation of the golden thread vine extract, the addition of p-hydroxyacetophenone can inhibit the discoloration of the golden thread vine extract. The results of the verification experiment are as follows:

[0083] Table 2 Effect of the amount of p-hydroxyacetophenone added on the color of Anoectochilus roxburghii extract

[0084]

[0085] Example 2

[0086] S1, crushing: taking the whole plant of dried roxburghii roxburghii, crushing it with a crusher and passing it through a No. 4 sieve to obtain 50g of roxburghii roxburghii dry powder;

[0087] S2, extraction: the dry powder obtained in S1 was added with 10% ethanol solvent at a solid-liquid ratio of 1:20 for ultrasonic extraction twice, the ultrasonic frequency was 480W, the ultrasonic extraction time was 30min, and the crude extract of Anoectochilus roxburghii was combined;

[0088] S3, filtering: filtering the crude extract of Anoectochilus roxburghii obtained in S2 to remove residues;

[0089] S4, concentration: the filtrate obtained in S3 is concentrated under reduced pressure to remove ethanol, and the volume is fixed with water to 15 times the mass of the dry powder of Anoectochilus roxburghii;

[0090] S5, dissolving: adding 1,3-propylene glycol, 1,2-pentanediol and p-hydroxyacetophenone to the concentrated solution obtained in S4, the final solution having a mass ratio of crude drug:solution = 1:20, and a composition of 68.6% w / w concentrated solution, 30% w / w 1,3-propylene glycol, 0.7% w / w 1,2-pentanediol and 0.7% w / w p-hydroxyacetophenone;

[0091] S6, decolorization: The solution obtained in S5 was adsorbed and decolorized with 20% w / w D101 macroporous adsorption resin for 4 hours to obtain a decolorized solution of Anoectochilus roxburghii;

[0092] S7, sterilization: filtering and sterilizing the decolorizing solution described in S6;

[0093] S8. Preservative compounding: Add 0.5% w / w potassium sorbate to the sterilized solution in S7 to obtain the roxburghii extract.

[0094] Example 3

[0095] S1, crushing: taking the whole plant of dried roxburghii roxburghii, crushing it with a crusher and passing it through a No. 2 sieve to obtain 100 g of roxburghii roxburghii dry powder;

[0096] S2, extraction: the dry powder obtained in S1 was added to 50% ethanol solvent at a material-liquid ratio of 1:10 for ultrasonic extraction twice, the ultrasonic frequency was 480W, the ultrasonic extraction time was 10min, and the crude extract of Anoectochilus roxburghii was combined;

[0097] S3, filtering: filtering the crude extract of Anoectochilus roxburghii obtained in S2 to remove residues;

[0098] S4, concentration: the filtrate obtained in S3 is concentrated under reduced pressure to remove ethanol, and the volume is fixed with water to 10 times the mass of the dry powder of Anoectochilus roxburghii;

[0099] S5, dissolving: adding 1,3-butanediol, 1,2-hexanediol and p-hydroxyacetophenone to the concentrated solution obtained in S4, the final solution is crude drug: solution = 1:20 by mass ratio, and the composition is 68.6% w / w concentrated solution, 30% w / w 1,3-butanediol, 0.7% w / w 1,2-hexanediol and 0.7% w / w p-hydroxyacetophenone;

[0100] S6, decolorization: the solution obtained in S5 is subjected to adsorption decolorization for 2 hours using 10% w / w AB-8 and HP-10 macroporous adsorption resins to obtain a decolorized solution of Anoectochilus roxburghii;

[0101] S7, sterilization: filtering and sterilizing the decolorizing solution described in S6;

[0102] S8. Preservative compounding: Add 0.5% w / w benzoic acid to the sterilized solution of S7 to obtain the roxburghii extract.

[0103] Comparison example

[0104] This comparative example is the same as Example 1 except that the ultrasonic extraction in S2 and the addition of a co-solvent and a stabilizer in S5 are different from those in Example 1.

[0105] S1, crushing: taking the whole plant of dried roxburghii roxburghii, crushing it with a crusher and passing it through a No. 1 sieve to obtain 20 g of roxburghii roxburghii dry powder;

[0106] S2, extraction: the dry powder obtained in S1 is added to 30% ethanol solvent at a solid-liquid ratio of 1:15, and the mixture is soaked and extracted once to obtain a crude extract of Anoectochilus roxburghii;

[0107] S3, filtering: filtering the crude extract of Anoectochilus roxburghii obtained in S2 to remove residues;

[0108] S4, concentration: the filtrate obtained in S3 is concentrated under reduced pressure to remove ethanol, and the volume is fixed with water to 10 times the mass of the dry powder of Anoectochilus roxburghii;

[0109] S5, dissolving: adding pure water to the concentrated solution obtained in S4, the final solution is a crude drug: solution = 1:20 mass ratio, the composition is 50% w / w concentrated solution, 50% w / w water;

[0110] S6, decolorization: the solution obtained in S5 is subjected to adsorption decolorization for 2 hours using 20% ​​w / w XAD-2 macroporous adsorption resin to obtain a decolorized solution of Anoectochilus roxburghii;

[0111] S7, sterilization: filtering and sterilizing the decolorizing solution in S6 to obtain a roxburghii extract;

[0112] S8. Preservative compounding: Add 0.1% w / w sodium pyrosulfite to the sterilized solution in S7 to obtain the roxburghii extract.

[0113] Comparative experiment on polysaccharide content of Anoectochilus roxburghii extracts in Examples 1-3 and Control Example

[0114] Table 3 Comparison of polysaccharide content in extracts of Anoectochilus roxburghii of Examples 1-3 and Control Example

[0115]

[0116] As shown in Table 3, the polysaccharide content of Anoectochilus roxburghii extracted by ultrasonic method is relatively high.

[0117] Stability test of the extract of Anoectochilus roxburghii in Examples 1-3 and Control Example

[0118] The stability test of the golden thread extract obtained by the process of this application was carried out in accordance with TSHFCA002-2021 Cosmetic Stability Test Guidelines. The test was carried out for 6 months at 45℃±2℃ and relative humidity 45%RH±5%RH. The inspection items were appearance, odor, pH value, and golden thread polysaccharide content. The test standards are shown in Table 4 below:

[0119] Table 4 Stability test of roxburghii extract

[0120]

[0121]

[0122] According to the designed stability test plan, an accelerated test was conducted, and the results are the results of a 6-month accelerated test (Table 5).

[0123] Table 5 Stability test results of Anoectochilus roxburghii extract

[0124]

[0125]

[0126]

[0127]

[0128] As can be seen from Table 5, the Anoectochilus roxburghii extract obtained by the process of the present application meets the standard requirements in terms of quality control indicators such as raw material appearance, odor, pH value, relative density, microorganisms and Anoectochilus roxburghii polysaccharide content. Compared with the control example, its physical and chemical properties and active ingredient content will not change significantly over time, thereby ensuring the activity of the Anoectochilus roxburghii extract itself.

[0129] Test on the soothing and anti-inflammatory effects of Anoectochilus roxburghii extract

[0130] The inflammatory model of macrophage RAW264.7 cells was induced by lipopolysaccharide (LPS), and the levels of inflammatory factors TNF-α and IL-1α in RAW264.7 cells were detected to observe the anti-inflammatory effect of Anoectochilus roxburghii extract. 5 / well, the cultured cells were inoculated in a 6-well plate, 2 mL per well, placed in a 5% CO2 incubator, and cultured at 37°C for 24 hours; the supernatant was removed and 2 mL was administered, model group; 5 μg / mL lipopolysaccharide (LPS); sample group: 5 μg / mL lipopolysaccharide (LPS) + the golden thread lotus extract of the present application; blank control group: serum-free culture medium was added; placed in a CO2 incubator (37°C, 5% CO2) and continued to culture for 24 hours; the supernatant was aspirated and the operation was performed according to the instructions of the kit.

[0131] like Figure 1 and Figure 2 The results show that Control: blank control group; Model: 5μg / mL LPS-induced model group; Positive control group: 100μg / mL DXMS (dexamethasone); Experimental group: high, medium and low doses of Anoectochilus roxburghii extract, JXL-H, 5μg / mL LPS + 10% Anoectochilus roxburghii extract; JXL-M, 5μg / mL LPS + 5% Anoectochilus roxburghii extract; JXL-L, 5μg / mL LPS + 2.5% Anoectochilus roxburghii extract. # indicates a significant difference compared with Control (P < 0.05); * indicates a significant difference compared with Model (P < 0.05).

[0132] Control: blank control group; Model: 5μg / mL LPS induced group; Positive control group: 100μg / mL DXMS (dexamethasone); Experimental group: high, medium and low doses of Anoectochilus roxburghii extract, JXL-H, 5μg / mL LPS + 1% Anoectochilus roxburghii extract; JXL-M, 5μg / mL LPS + 0.5% Anoectochilus roxburghii extract; JXL-L, 5μg / mL LPS + 0.1% Anoectochilus roxburghii extract. # indicates significant difference compared with Control (P < 0.05); * indicates significant difference compared with Model (P < 0.05).

[0133] Compared with the blank control group (Control), the levels of TNF-α and IL-1α in the model group (Model) were significantly increased, indicating that the inflammatory cell model was successfully established. The roxburghii extract has a significant inhibitory effect on the TNF-α secretion of RAW264.7 cells induced by LPS; at the same time, the roxburghii extract also has a significant inhibitory effect on the IL-1α secretion of RAW264.7 cells induced by LPS, and is dose-dependent. This shows that the roxburghii extract of the present application has a good anti-inflammatory effect and is suitable for soothing skin care products.

[0134] Anti-wrinkle effect test of golden thread lotus extract

[0135] Human skin fibroblasts (HSF) in the logarithmic growth phase were resuspended and counted at 2.5×10 5 / well were inoculated into a 6-well cell culture plate, placed in a CO2 incubator, and cultured at 37°C for 24 hours; after the cells adhered to the wall, the original culture medium was removed and 2 mL of the drug was added to each well; the experimental group design was Control (blank control group), and a culture medium containing 1% serum was added without any treatment; the experimental group was added with the golden thread lotus extract of the present application diluted with a DMEM culture medium containing 1% serum; placed in a CO2 incubator, and cultured at 37°C for 24 hours; then the cell supernatant was collected, centrifuged at 8000 rpm for 10 minutes, and the supernatant was taken and operated according to the instructions of the Human COL1α1 (Collagen TypeⅠAlpha 1) ELISA Kit.

[0136] The experimental results are as follows Figure 3 As shown in the figure, Control: blank control group; Experimental groups: high, medium and low doses of Anoectochilus roxburghii extract, JXL-H, 1% (v / v) Anoectochilus roxburghii extract; JXL-M, 0.5% (v / v) Anoectochilus roxburghii extract; JXL-L, 0.25% (v / v) Anoectochilus roxburghii extract. * indicates significant difference compared with Control (P < 0.05).

[0137] The 1% and 0.5% concentrations of the roxburghii extract of the present application significantly promoted the synthesis of type I collagen by human skin fibroblasts (HSF), and compared with the blank control group, the synthesis of type I collagen increased by more than 20%. This shows that the roxburghii extract of the present application is suitable for anti-wrinkle skin care products.

[0138] Test on the repairing effect of golden thread lotus extract

[0139] (1) Cell proliferation assay

[0140] Human keratinocytes (HaCaT) were cultured at 5.0×10 3 / well was inoculated into a 96-well plate, 100uL per well, and placed in a CO2 incubator for 12h; the supernatant was discarded, and the test drug was added. The blank control group (Control) was added with 1% serum culture medium without any treatment. The competitor control group was 0.1mg / mL NMN, and the sample group was the golden thread lotus extract of the present application (concentrations were 1%, 0.75%, 0.5%, and 0.25%); after culturing for 24h, the supernatant was discarded, 100uL of complete culture solution containing 10% CCK-8 reagent was added to each well, incubated for 1.5h, and detected at 450nm with an enzyme reader.

[0141] The experimental results are as follows Figure 4 The results showed that compared with the blank control group, both the competitor control group and the sample group had a certain effect in promoting the proliferation of HaCaT cells, but the 1% roxburghii extract had a stronger effect in promoting the proliferation of HaCaT cells.

[0142] (2) Cell migration assay

[0143] Human keratinocytes (HaCaT) were cultured at 3×10 5 / well were inoculated into 6-well plates, 2 mL per well, and cultured in a CO2 incubator for 24 h; each well was scratched with a 200 μL pipette tip to remove the culture solution, and the excess cells were washed with PBS to remove the solution; the test drug was added, the blank control group (Control) was added with 1% serum culture medium without any treatment, the positive control group was added with 10 ng / mL EGF, the sample group was added with the golden thread lotus extract of the present application (concentrations of 1% and 0.25%), and the plates were cultured in a CO2 incubator, and photographed and recorded at 0 h, 12 h, 24 h, and 48 h, respectively; imageJ quantitative analysis was performed to obtain the migration area.

[0144] like Figure 5 -6, the migration rate of HaCaT cells is positively correlated with time. At 48 hours after drug addition, the migration rate of the positive control group was 30.5%; while in the sample group, the migration rate of HaCaT cells treated with 1% roxburghii extract was 23.9%, and the migration rate of HaCaT cells treated with 0.5% roxburghii extract was 16.6%; the migration rate of the blank control group (Control) was 14.7%. Compared with the blank control group (Control), the roxburghii extract promoted the migration of HaCaT cells, which shows that the roxburghii extract of the present application has a certain repair effect and is suitable for use in repair skin care products.

[0145] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine different embodiments or examples described in this specification and the features of different embodiments or examples, unless they are contradictory.

[0146] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the features. In the description of this application, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0147] The above is only a specific implementation of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can easily think of various changes or substitutions within the technical scope disclosed in the present application, which should be included in the protection scope of the present application. Therefore, the protection scope of the present application should be based on the protection scope of the claims.

Claims

1. A process for preparing a roxburghii extract, characterized in that: include: The dried whole plant of Anoectochilus roxburghii is crushed by a crusher and then sieved to obtain dry powder; The dry powder is added to an ethanol solvent at a material-liquid ratio of 1:10-1:20, and ultrasonic extraction is performed at room temperature; wherein the volume fraction of the ethanol solvent is 10%-50%, the ultrasonic frequency is 480W, the ultrasonic extraction time is 10-30min, and the extraction times are 1-2 times; and a crude extract of Anoectochilus roxburghii is obtained; Filtering the crude extract of Anoectochilus roxburghii to remove residues and obtain a filtrate; The obtained filtrate is subjected to membrane separation and / or reduced pressure concentration to remove ethanol, and the volume is fixed to 10-15 times the mass of the dry powder of Anoectochilus roxburghii to obtain a concentrated solution; 30%-50% w / w of a cosolvent and 1.4%-5% w / w of a stabilizer are added to the obtained concentrated solution, and then adsorption and decolorization are performed using a macroporous adsorption resin to obtain a decolorized solution of Anoectochilus roxburghii; the cosolvent is one or more of glycerol, 1,3-propylene glycol, and 1,3-butanediol; the stabilizer is one or more of 1,2-hexanediol, 1,2-pentanediol, and p-hydroxyacetophenone; the resin usage is 10%-20% w / v; the macroporous adsorption resin includes one or more of D101, AB-8, XAD-2, and HP-10; the adsorption method is static adsorption, and the adsorption time is 2-4 hours; Filter and sterilize the decolorized solution of Anoectochilus roxburghii; Adding 0.1%-0.5% w / w of preservative to the sterilized solution to obtain a roxburghii extract; The preservative is one or more of sodium pyrosulfite, phenoxyethanol, benzyl alcohol, potassium sorbate and sodium sorbate.

2. A roxburghii extract, characterized in that The roxburghii roxburghii extract is prepared by the roxburghii roxburghii extract preparation process described in claim 1.

3. An application of a roxburghii extract, characterized in that: The application is the use of the golden thread vine extract described in claim 2 in skin care products for soothing, anti-inflammatory, or repairing.

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