Preparation process and application of fructus chebulae-containing extract

Abstract

The invention provides a preparation process and application of an extract containing fructus chebulae. Specifically, the invention provides the medicine terminalia fruit wrapping liquid, and the medicine terminalia fruit wrapping liquid comprises medicine terminalia fruit extract, lecithin, caprylic / capric triglyceride, organic alcohol, p-hydroxyacetophenone, polysorbate-80 and a humectant, so that the medicine terminalia fruit wrapping liquid is more stable in active matter, color and state and smaller in smell. And the medicine terminalia fruit coating liquid is safer, has stronger soothing property, and has better anti-oxidation, anti-aging and nerve analgesic effects.

Description

Technical Field

[0001] The present invention relates to the field of phytochemistry, and particularly relates to a preparation process and application of an extract containing Terminalia chebula Retz.. Background Art

[0002] Terminalia chebula Retz. Terminalia chebula Retz. , also known as Terminalia chebula, Terminalia fruit, is a deciduous tree belonging to the genus Terminalia ( Combretaceae ), Combretaceae ( Terminalia L. ). Its fruit is a traditional medicine included in the Chinese Pharmacopoeia. Terminalia chebula Retz. is native to India, Myanmar and other places. At present, its wild populations are mainly distributed in the low-heat river valley areas with an altitude of 800-1100 m in the Nujiang River Basin in southwestern Yunnan, China.

[0003] In various traditional medicine systems, Terminalia chebula Retz. has been widely used as a crude drug. So far, the chemical constituents reported for Terminalia chebula Retz. include various bioactive compounds such as tannins, phenolic acids, triterpenoids, flavonoids, aliphatics, etc. In recent years, with the continuous in-depth research on Terminalia chebula Retz., it has been found that Terminalia chebula Retz. has pharmacological effects such as antioxidant, anti-tumor, antiviral, antibacterial and neuroprotective effects.

[0004] In the prior art, the preparation process of the extract of Terminalia chebula Retz. is mainly simple extraction and purification. For example, CN102526136B discloses an antioxidant of Terminalia chebula Retz. and its preparation method. The antioxidant is extracted by a solvent, then extracted with an organic solvent, freeze-dried after purification, and applied to oral foods, health products and drugs. At present, there is an urgent need to develop a preparation process of an extract containing Terminalia chebula Retz. that is simple, non-toxic and has improved activity. Summary of the Invention

[0005] The object of the present invention is to provide a preparation process of an extract containing Terminalia chebula Retz. that is simple, non-toxic and has improved activity.

[0006] In the first aspect of the present invention, a Terminalia chebula Retz. wrapping solution is provided, and the Terminalia chebula Retz. wrapping solution includes: 1-25 wt% of Terminalia chebula Retz. extract; 2-4 wt% of lecithin; 5-15 wt% of caprylic / capric triglyceride; 1-5 wt% of organic polyol; 0.1-1 wt% of p-hydroxyacetophenone; 10-30 wt% of polysorbate-80; 2-10 wt% of a humectant, based on the total mass of the wrapping material; the balance is made up with water.

[0007] In another preferred example, the content of lecithin in the Terminalia chebula Retz. wrapping solution is 2.8-3.2 wt%.

[0008] In another preferred example, the content of caprylic / capric triglyceride in the Terminalia chebula encapsulating liquid is 8 - 12 wt%.

[0009] In another preferred example, the Terminalia chebula encapsulating liquid further comprises 10 - 80 wt% of water, preferably 30 - 50 wt% of water.

[0010] In another preferred example, the organic polyol is selected from the group consisting of: hexylene glycol, pentylene glycol, or a combination thereof.

[0011] In another preferred example, the humectant is selected from the group consisting of: glycerol, butanediol, or a combination thereof.

[0012] In another preferred example, the Terminalia chebula extract in the Terminalia chebula encapsulating liquid is an extract, an extract paste, or a freeze-dried powder.

[0013] In another preferred example, the Terminalia chebula extract is a freeze-dried powder.

[0014] In another preferred example, the Terminalia chebula extract is obtained by extraction with 50 - 70 (v / v)% ethanol.

[0015] In another preferred example, the method for preparing the Terminalia chebula extract comprises the steps of: Weigh an appropriate amount of Terminalia chebula, add 50 - 70 (v / v)% ethanol; reflux and extract at 70 - 80 °C; filter the obtained extract to obtain a filtrate; for the remaining filter residue, add 50 - 70 (v / v)% ethanol and reflux and extract at 70 - 80 °C. After combining the filtrates, concentrate under reduced pressure to obtain an extract paste, and obtain the Terminalia chebula extract.

[0016] In another preferred example, the method for preparing the Terminalia chebula extract comprises the steps of: weigh an appropriate amount of Terminalia chebula, add 5 - 12 times the amount of raw material of 50 - 70 (v / v)% ethanol; reflux and extract at 70 - 80 °C for 1.0 - 3.0 hours; filter the obtained extract to obtain a filtrate; for the remaining filter residue, add 5 - 12 times the amount of raw material of 50 - 70 (v / v)% ethanol and reflux and extract at 70 - 80 °C for 1.0 - 3.0 hours. After combining the filtrates, concentrate under reduced pressure to obtain an extract paste, and concentrate to 0.5 - 2.2 times the amount of the charged material. Filter the above concentrated solution to obtain a filtrate, which is the Terminalia chebula extract.

[0017] In the second aspect of this aspect, there is provided a method for preparing the Terminalia chebula encapsulating liquid as described in the first aspect of the present invention, and the method comprises the steps of: (1) Take the Terminalia chebula extract, lecithin and caprylic / capric triglyceride and mix them to form an encapsulating mixture, and stir under heating conditions at 50 - 90 °C to form phase A; (2) Mix water, organic polyol and p - hydroxyacetophenone to form phase B; (3) Mix polysorbate - 80 with glycerol to form Phase C; (4) Add Phase A to Phase C to obtain Phase A + C; (5) Add Phase B to the above - mentioned Phase A + C, and stir evenly to obtain the Terminalia chebula encapsulated liquid.

[0018] In another preferred example, step (5) further includes: adding Phase B to the above - mentioned Phase A + C, pre - emulsifying first, then performing high - pressure homogenization stirring and then converting to mechanical stirring, and standing for aging for 12 - 48 hours to obtain the Terminalia chebula encapsulated liquid.

[0019] In another preferred example, the pressure of the high - pressure homogenization stirring is 100 - 2000 Pa.

[0020] In another preferred example, the pressure of the high - pressure homogenization stirring is 400 - 1800 Pa, preferably 1200 - 1800 Pa.

[0021] In another preferred example, the Terminalia chebula encapsulated liquid further has one or more characteristics selected from the following group: (1) Increase the content of nuclear factor E2 - related factor 2 (Nrf2); (2) Inhibit the expression of TRPV1 content; (3) Inhibit the decrease of type I collagen.

[0022] In another preferred example, the Terminalia chebula encapsulated liquid further has one or more characteristics selected from the following group: (1) No skin irritation; (2) Antioxidant and anti - inflammatory; (3) Anti - allergic, soothing and analgesic; (4) Anti - aging.

[0023] In the third aspect of the present invention, there is provided the use of the Terminalia chebula encapsulated liquid as described in the first aspect of the present invention, which is used for preparing daily products or daily chemical products with anti - inflammatory, antioxidant, anti - allergic, anti - glycation, anti - aging and / or nerve - type analgesic effects.

[0024] In another preferred example, the dosage of the Terminalia chebula encapsulated liquid in the product is 0.05 - 99 wt%.

[0025] It should be understood that within the scope of the present invention, the above - mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. Description of the Drawings

[0026] Figure 1 It is the liquid phase diagram of the Terminalia chebula encapsulated liquid.

[0027] Figure 2Results of cell viability tests for Terminalia chebula Retz. extracts and Terminalia chebula Retz. encapsulation solutions.

[0028] Figure 3 Results of experiments on the effects of Terminalia chebula Retz. extracts and Terminalia chebula Retz. encapsulation solutions on the content of Nrf2.

[0029] Figure 4 Results of experiments on the effects of Terminalia chebula Retz. extracts and Terminalia chebula Retz. encapsulation solutions on type I collagen. Detailed implementation manners

[0030] Through extensive and in - depth research, and through a large number of screenings and tests, a preparation process of Terminalia chebula Retz. extract and its application are provided. The preparation process is an encapsulation process. Terminalia chebula Retz. extract is provided, and then a variety of excipients such as lecithin, caprylic / capric triglyceride, organic polyols, p - hydroxyacetophenone, polysorbate - 80 and humectants are compounded, so that the active substances, color, state of the Terminalia chebula Retz. encapsulation solution are more stable and have a smaller odor. And the Terminalia chebula Retz. encapsulation solution is safer, has stronger soothing properties, better anti - glycation, anti - aging and neuro - analgesic effects. The Terminalia chebula Retz. extract used in the encapsulation process is extracted with 65% ethanol, and the content of active ingredients such as galloyl glucoside compounds is higher. On this basis, the present invention is completed.

[0031] Terms Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0032] As used herein, when referring to a specifically recited numerical value, the term "about" means that the value can vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0033] As used herein, the term "comprising" or "including" can be open - ended, semi - closed and closed. In other words, the term also includes "consisting essentially of...", or "consisting of...".

[0034] As used herein, the term "room temperature" or "normal temperature" refers to a temperature of 4 - 40 o °C, preferably, 25 ± 5 o °C.

[0035] Preparation of Terminalia chebula Retz. extract The Terminalia chebula Retz. extract described in the present invention can be prepared by conventional methods. Preferably, the method includes the steps: Weigh an appropriate amount of Terminalia chebula, add 5 - 12 times the amount of raw materials of 50 - 70 (v / v)% ethanol; reflux and extract at 70 - 80 °C for 1.0 - 3.0 hours; naturally filter the obtained extract through a 300 - mesh sieve to obtain a filtrate; for the remaining filter residue, add 5 - 12 times the amount of raw materials of 50 - 70 (v / v)% ethanol, and reflux and extract at 70 - 80 °C for 1.0 - 3.0 hours. After combining the filtrates, concentrate under reduced pressure (50 °C - 70 °C, vacuum pressure 0.060~0.08 Mpa) into an extract, concentrating to 0.5 - 2.2 times the feeding amount. Filter the above - concentrated solution through a 0.45 - um membrane to obtain a filtrate.

[0036] Freeze - dry the above filtrate (pre - freezing: temperature - 40 °C~ - 45 °C, time 60 - 90 min; sublimation drying: temperature - 30 °C~ - 10 °C, vacuum degree 5 Pa~20 Pa, time 8 - 12 h; desorption drying: temperature 35 °C~40 °C, vacuum degree < 5 Pa, time 12 - 20 h; cold trap temperature: - 60 °C~ - 50 °C). After pulverizing with a pulverizer and passing through an 80 - mesh sieve, obtain the freeze - dried powder of Terminalia chebula extract, or dry it into a powder to obtain the powder of Terminalia chebula extract.

[0037] Preparation process of Terminalia chebula extract The preparation process of the Terminalia chebula extract - containing composition of the present invention is preferably encapsulation, and the encapsulation includes: (1) Mix 3% - 25% of the powder of Terminalia chebula extract or the liquid of Terminalia chebula extract, 2.5 - 3.8% of lecithin and 10 - 15% of caprylic / capric triglyceride, and stir to dissolve them thoroughly under heating conditions at 50 - 80 °C to prepare phase A; At room temperature, mix 40% - 68% of water, 1% - 5% of hexylene glycol or pentylene glycol and 0.1% - 1% of p - hydroxyacetophenone, and stir to dissolve them thoroughly to prepare phase B; At room temperature, mix 15% - 25% of polysorbate - 80 and 1% - 5% of glycerol or butanediol, and stir evenly to prepare phase C; Under slow stirring, slowly add phase A to phase C to form a homogeneous state to obtain phase A + C; under slow stirring, slowly add phase B to the above - mentioned phase A + C, and make up the rest with water. First, pre - emulsify, then homogenize at high pressure (1000 bar - 1500 bar) for 10 - 30 min and then switch to mechanical stirring; cool to room temperature, degas, stir evenly and then stand for aging for 24 hours.

[0038] Nrf2 Nrf2, short for "Nuclear factor erythroid 2-related factor 2", is a transcription factor that plays an important role in cells. Nrf2 plays a key role in the cellular oxidative stress response. By interacting with the antioxidant response element (ARE), it regulates the expression of antioxidant proteins and phase II detoxifying enzymes, thereby protecting cells from oxidative stress damage.

[0039] Nrf2 plays multiple important roles in biology. First, it is a key regulator in the cellular oxidative stress response. Under oxidative stress conditions, Nrf2 is activated and enters the nucleus to bind to ARE, initiating the expression of a series of anti-oxidation genes and phase I detoxifying enzymes, thus enhancing the antioxidant defense ability of cells. Second, Nrf2 is also involved in regulating cell proliferation, inflammatory responses, and various metabolic processes. For example, in cancer research, it has been found that the activity of Nrf2 usually increases, providing a growth advantage for cells and promoting the progression and metastasis of cancer. In addition, Nrf2 is also closely related to the occurrence and development of various diseases such as reproductive system diseases and cardiovascular diseases.

[0040] Transient receptor potential vanilloid 1 (Trpv1) External stimuli or endogenous substances including bradykinin, nerve growth factor, etc. can activate TRPV1, thereby causing an influx of Ca 2+ ions, resulting in increased skin permeability and a stinging and burning sensation. At the same time, TRPV1 also promotes the release of neurogenic inflammatory factors including substance P, further inducing the secretion of inflammatory factors and causing local capillary dilation. Currently, most of the cosmetic raw materials developed for neurogenic skin sensitivity are TRPV1 expression inhibitors or functional antagonists. By detecting a decrease in its expression level, it shows that the active substance has anti-allergic and soothing effects in relieving facial discomfort.

[0041] The main advantages of the present invention include: 1. The preparation method of the Terminalia chebula wrapped solution provided by the present invention is simple, without using strongly irritating organic solvents, and has no irritation when used transdermally, and the cell viability of tissues is higher.

[0042] 2. The Terminalia chebula wrapped solution provided by the present invention improves the activity of the Terminalia chebula extract, especially in terms of increasing the content of Nrf2 and inhibiting the expression of the content of TRPV1. The Terminalia chebula wrapped solution shows an equivalent effect to the Terminalia chebula extract at a lower concentration.

[0043] 3. The Terminalia chebula wrapped solution provided by the present invention has better antioxidant and anti-aging effects on transdermal products.

[0044] The present invention will be further described below in connection with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. For the experimental methods without specific conditions noted in the following embodiments, they are generally carried out under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0045] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to persons skilled in the art. In addition, any methods and materials similar or equivalent to those described may be applied in the method of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0046] Example 1 Weigh 10 g of Terminalia chebula Retz., add 100 ml of 65% ethanol; reflux and extract at 80 °C for 2.0 hours; naturally filter the obtained extract through a 300-mesh sieve to obtain a filtrate; for the remaining filter residue, add 100 ml of 65% ethanol and reflux and extract at 80 °C for 2.0 hours. Combine the filtrates and concentrate under reduced pressure (50 °C - 70 °C, vacuum pressure 0.060 - 0.08 Mpa) to obtain an extract, and concentrate to 10 g. Filter the above concentrate through a 0.45-μm membrane to obtain a filtrate. Lyophilize the above filtrate, use gallic acid as a reference substance, and test the content of galloyl glucoside components to be 75.12% (calculated based on the mass of the freeze-dried powder), and the extraction rate is 90.61%.

[0047] Example 2 Weigh 10 g of Terminalia chebula Retz., add 100 mL of water; extract at 80 °C for 2.0 hours; naturally filter the obtained extract through a 300-mesh sieve to obtain a filtrate; for the remaining filter residue, add 100 ml of water and reflux and extract at 80 °C for 2.0 hours. Combine the filtrates and concentrate under reduced pressure (50 °C - 70 °C, vacuum pressure 0.060 - 0.08 Mpa) to obtain an extract, and concentrate to 10 g. Filter the above concentrate through a 0.45-μm membrane to obtain a filtrate. Lyophilize the above filtrate, use gallic acid as a reference substance, and test the content of galloyl glucoside components to be 46.98%, and the extraction rate is 66.82%.

[0048] Example 3 Weigh 10 g of Terminalia chebula Retz., add 100 mL of 75% ethanol; extract at 80 °C for 2.0 hours; naturally filter the obtained extract through a 300-mesh sieve to obtain a filtrate; for the remaining filter residue, add 100 mL of 75% ethanol, and reflux extract at 80 °C for 2.0 hours. Combine the filtrates and concentrate under reduced pressure (50 °C - 70 °C, vacuum pressure 0.060~0.08 Mpa) to obtain an extract, and concentrate it to 10 g. Filter the above concentrated solution through a 0.45-μm membrane to obtain a filtrate. Freeze-dry the above filtrate. Using gallic acid as a reference substance, test the content of galloylglucosides to be 50.12%, and the extraction rate is 88.76%.

[0049] As can be seen from the results of Examples 1-3, in the heating reflux extraction method, 65% ethanol is the optimal extraction solvent, and the highest content of galloylglucosides in the extract is 75.12%. When the ethanol concentration increases, the content of galloylglucosides in the extract will instead decrease to 50.12%. Increasing the polarity of the extraction solvent will also reduce the yield of galloylglucosides. For example, when water is used as the extraction solvent, the content of galloylglucosides in the extract decreases to 46.98%.

[0050] Example 4 Take 3% of the Terminalia chebula Retz. extract (prepared in Example 1, calculated by the mass of the freeze-dried powder), 3.0% lecithin and 10% caprylic / capric triglyceride, mix them, and stir to dissolve them fully under heating conditions at 70 °C to prepare Phase A (the percentage is by mass ratio and based on the total amount of the whole encapsulation); at room temperature, mix 40% water, 3% hexylene glycol and 0.5% p-hydroxyacetophenone, and stir to dissolve them fully to prepare Phase B; at room temperature, mix 20% polysorbate-80 and 5% glycerol, and stir evenly to prepare Phase C; under slow stirring, slowly add Phase A to Phase C to form a homogeneous state to obtain Phase A+C; under slow stirring, slowly add Phase B to the above Phase A+C, and make up the rest with water to 100%. First, pre-emulsify, then homogenize at 1500 bar for 30 min and then switch to mechanical stirring; cool to room temperature, degas, stir evenly and then let it stand and age for 24 hours. Obtain the Terminalia chebula Retz. encapsulation liquid, which naturally settles at room temperature and does not precipitate within 2 months, and the solution is homogeneous. Centrifuge for 30 minutes (10000 r / min) without precipitation, and the solution is homogeneous. The color chromaticity value of the 10% aqueous solution is 5.04.

[0051] Example 5 Take 3% of the Terminalia chebula extract (prepared in Example 1, calculated by the mass of the freeze-dried powder), 3.0% lecithin and 10% caprylic / capric triglyceride, mix them, and stir to dissolve them completely under heating conditions at 70 °C to prepare Phase A; at room temperature, mix 40% water, 3% hexylene glycol and 0.5% p-hydroxyacetophenone, and stir to dissolve them completely to prepare Phase B; at room temperature, mix 20% polysorbate-80 and 5% glycerol, stir evenly to prepare Phase C; under slow stirring, slowly add Phase A to Phase C to form a homogeneous state to obtain Phase A+C; under slow stirring, slowly add Phase B to the above Phase A+C, and make up the rest with water to 100%. First, pre-emulsify, then homogenize at 500 bar for 30 min and then switch to mechanical stirring; cool to room temperature, degas, stir evenly and then let it stand for aging for 24 hours. Obtain the Terminalia chebula encapsulation liquid, let it settle naturally at room temperature, and precipitate within 1 month. Centrifuge for 30 minutes (10,000 r / min) without precipitation, and the solution is homogeneous.

[0052] Example 6 Take 3% (prepared in Example 1, calculated by the mass of the freeze-dried powder) of the Terminalia chebula extract, 1.0% lecithin and 8% caprylic / capric triglyceride, mix them, and stir to dissolve them completely under heating conditions at 70 °C to prepare Phase A; at room temperature, mix 40% water, 3% hexylene glycol and 0.5% p-hydroxyacetophenone, and stir to dissolve them completely to prepare Phase B; at room temperature, mix 20% polysorbate-80 and 2% glycerol, stir evenly to prepare Phase C; under slow stirring, slowly add Phase A to Phase C to form a homogeneous state to obtain Phase A+C; under slow stirring, slowly add Phase B to the above Phase A+C, and make up the rest with water to 100%. First, pre-emulsify, then homogenize at 1500 bar for 30 min and then switch to mechanical stirring; cool to room temperature, degas, stir evenly and then let it stand for aging for 24 hours. Obtain the Terminalia chebula encapsulation liquid, let it settle naturally at room temperature, and precipitate within 1 month. Centrifuge for 30 minutes (10,000 r / min) without precipitation, and the solution is homogeneous.

[0053] From the results of Example 4 and Example 6, it can be seen that the concentrations of lecithin and caprylic / capric triglyceride in Phase A will affect the stability of the Terminalia chebula encapsulation liquid. The optimal concentration of lecithin is 10%, and the optimal concentration of caprylic / capric triglyceride is 10%. After the contents of lecithin and caprylic / capric triglyceride decrease, the stability of the Terminalia chebula encapsulation liquid decreases, and precipitation occurs within 1 month.

[0054] From the results of Example 5 and Example 6, it can be seen that the pressure of high-pressure homogenization during the stirring process will affect the stability of the Terminalia chebula encapsulation liquid. The higher the pressure, the better the stability effect, and the optimal pressure condition is 1500 bar. When the pressure is reduced to 500 bar, the stability of the Terminalia chebula encapsulation liquid decreases, and precipitation occurs within 1 month.

[0055] Safety evaluation Example 1 of safety evaluation - Skin irritation evaluation The irritations of Terminalia chebula Retz. extract and Terminalia chebula Retz. encapsulation solution to the skin were evaluated respectively, referring to OECD TG439 "In vitro skin irritation: Reconstructed human epidermis test method". Experimental samples: 4% Terminalia chebula Retz. extract (calculated based on the total weight of the freeze-dried powder); 100% Terminalia chebula Retz. encapsulation solution (calculated based on the total weight of the encapsulation liquid). Preparation of 4% Terminalia chebula Retz. extract: Weigh 0.0220 g of Terminalia chebula Retz. extract freeze-dried powder, add 0.528 ml of DPBS buffer solution, mix well, and prepare a 4% concentration. 100% Terminalia chebula Retz. encapsulation solution: Test the stock solution.

[0056] 1.1 Skin model preparation: Add the pre-warmed maintenance culture medium into a 12-well culture plate at 2 mL / well, transfer the skin model to the 12-well plate containing the maintenance culture medium, and place it in the incubator for overnight culture. 1.2 Reaction of the test sample with MTT Add 2 mL of 0.3 mg / mL MTT solution and 10 μL of the test sample to the 12-well plate, mix well, and incubate in the incubator for 3 h. 1.3 Sample addition: Take 10 μL of the test sample (including NC and PC), evenly apply it on the surface of the skin model, after standing at room temperature for 15 min, thoroughly rinse the sample with DPBS until there is no residue, and then place it in a well containing fresh maintenance culture medium for incubation for 42 h. Set 3 parallels for each group. 1.4 MTT reaction: Transfer the skin model to a well containing 2 mL of 0.3 mg / mL MTT solution, and place it in the incubator for 3 h. After the MTT reaction is completed, use a punch to remove the epidermal tissue, transfer the tissue to a 2 mL centrifuge tube, add 500 mL of isopropyl alcohol hydrochloride to each tube, mix well, and place it in the dark at room temperature overnight. 1.5 Detection: Take 200 μL of the solution from each tube and place it in a 96-well plate. Use isopropyl alcohol hydrochloride as the blank control, and measure the absorbance value at a wavelength of 570 nm.

[0057] 1.6 Data collation and analysis Using isopropyl alcohol hydrochloride as the blank control (Blank), the tissue activity of the negative control group (NC) is 100%, and calculate the relative tissue activities of the experimental group and the positive control group (PC): Relative tissue activity (%) = [((OD 实验 / PC - OD Blank ) / (OD NC - OD Blank )] * 100% OD experiment: OD of the test sample ODPC : OD value of the positive control group NC : OD value of the negative control group OD Blank : OD value of the blank control (isopropyl alcohol hydrochloride) Classify skin irritation according to Table 1: Table 1: Classification of skin irritation

[0058] 1.7. Experimental results: Table 2: Experimental results

[0059] The experimental results are as Figure 2 shown. Under the conditions of this experiment, the test samples of 4% Terminalia chebula extract and 100% Terminalia chebula encapsulation solution are non-irritating, but the cell viability of 4% Terminalia chebula extract is 14% lower than that of 100% Terminalia chebula encapsulation solution, proving that the Terminalia chebula encapsulation solution has higher safety and lower irritation.

[0060] Efficacy test Effect of Efficacy Example 1 on nuclear factor E2-related factor (Nrf2) 1.1 Experimental method 1.1.1 Administration: According to the test grouping, transfer the model to a 6-well plate (add 0.9 mL of EpiGrowth culture medium in advance), and mark the test group number on the 6-well plate. Add 0.9 mL of model culture medium to each well in the BC group and NC group, and add 0.9 mL of model culture medium containing vitamin E (VE) to each well in the PC group. The sample group is administered by surface application, with a dosing volume of 25 μL / model. When administering, apply it evenly in a gentle circular motion to ensure full absorption of the sample.

[0061] Positive control (PC): Weigh 7.0 mg of VE precisely, dissolve it in PBS buffer solution and make up the volume to 100 ml, shake well, precisely take 1.0 ml and transfer it to a 10-ml volumetric flask, and make up the volume with PBS buffer solution.

[0062] Experimental sample: 0.1% Terminalia chebula extract: Weigh 50 mg of Terminalia chebula extract precisely, dissolve it with PBS buffer solution, and make up the volume to 50 ml, shake well.

[0063] 0.03% Terminalia chebula encapsulation solution (calculated based on the amount of Terminalia chebula extract in the Terminalia chebula encapsulation solution): Precisely take 0.1 ml of Terminalia chebula encapsulation solution and transfer it to a 10-ml volumetric flask, and make up the volume with PBS buffer solution, shake well.

[0064] 1.1.2 Irradiation: After the administration, all groups except the blank control group were irradiated with UVB, and the irradiation dose was 600 mJ / cm 2 .

[0065] 1.1.3 Model incubation: After irradiation, the 6-well plates were placed in a CO2 incubator (37 °C, 5% CO2) for 24 h of incubation.

[0066] 1.1.4 Washing: After incubation, the remaining test substances were washed with sterile PBS, and the remaining liquid was wiped off with a sterile cotton swab.

[0067] 1.1.5 Immunofluorescence detection: The models used for detection were fixed with 4% paraformaldehyde. After 24 h of fixation, immunofluorescence detection was performed, photographed and observed under a microscope, and the pictures were collected and analyzed.

[0068] 1.1.6 Calculation of promotion rate: Promotion rate (%) = (sample group - negative control group) / negative control group × 100%.

[0069] 1.1.7 Result statistical analysis: GraphPad Prism was used for plotting, and the results were expressed as Mean ± SD. The t-test statistical analysis was used for comparison between groups. All statistical results were two-tailed. P < 0.05 was considered to have a significant difference, and P < 0.01 was considered to have a highly significant difference.

[0070] 1.2 Experimental results Table 3 Summary of the results of Nrf2 immunofluorescence analysis

[0071] Note: Integrated optical density (IOD), the value of which reflects the content of Nrf2. When using the t-test method for statistical analysis, compared with the BC group, the significance is indicated by #, P-value < 0.05 is indicated by #, and P-value < 0.01 is indicated by ##; compared with the NC group, the significance is indicated by *, P-value < 0.05 is indicated by *, and P-value < 0.01 is indicated by **.

[0072] The experimental results are as Figure 3 shown in and Table 3. It can be seen that both the Terminalia chebula extract and the Terminalia chebula encapsulation solution have the active effect of increasing the content of Nrf2, and the active effect of the Terminalia chebula encapsulation solution in increasing the content of Nrf2 is greater than that of the unencapsulated Terminalia chebula extract at a lower concentration, increasing from 64.81% to 118.52%, which is equivalent to the effect of vitamin E.

[0073] Effect of Efficacy Example 2 on transient receptor potential vanilloid 1 (Trpv1) 2.1 Experimental method: 2.1.1 Cell viability assay: Cells in the logarithmic growth phase were collected, and the cell density was adjusted and inoculated into a 96-well plate, with 100 μL of culture medium in each well. After culturing in a CO2 cell incubator for 24 h, the culture medium was aspirated, and the cells were washed twice with PBS buffer. The samples were prepared according to the usage method of the samples, and then the samples were grouped and administered according to the final concentrations of the sample preparations in the following table.

[0074] Table 4 Concentration grouping

[0075] 2.1.2 Administration: Discard the culture medium in each well of the plate. Add normal culture medium to the blank well and negative control, add capsazepine to the positive control, and add samples with different concentrations in Table 4 to the sample groups. After adding the drugs, continue to culture in a CO2 cell incubator for 24 h, and then perform the MTT cell toxicity assay. Read the absorbance OD value at 490 nm. Calculate the cell survival rate. Only culture medium is added to the blank well. Use the sample concentration with a cell viability ≥ 90% for subsequent efficacy detection.

[0076] 2.1.3 Detection: Adjust cells at a certain concentration (see Table 5 specifically) and re-seed them into a 96-well plate. After 24 h ± 1 h, when the cells adhere to the wall, perform grouped administration and proceed to the next experiment. Set 3 replicate wells for each group.

[0077] Table 5 Experimental grouping

[0078] 2.1.4 Sample collection: At the end of the 24-h culture, collect the supernatant for ELISA detection. Collect 150 μL of supernatant from each well into a 1.5-mL centrifuge tube, centrifuge at 10,000 rPm for 10 min before ELISA detection, and collect the supernatant for the experiment.

[0079] 2.1.5 ELISA detection: TRPV1 detection was selected for this experiment. Perform the experiment according to the kit instructions.

[0080] 2.2 Data processing 2.2.1 Cell viability (%) = OD of the sample group 490 / OD of the negative control group 490 * 100% Note: The blank well is deducted from each group of data. 2.2.2 Inhibition rate (%) = (1 - sample group / model group) * 100% 2.3 Experimental results Table 6 Results of Trpv1 inhibition rate

[0081] As can be seen from the above experimental results, the inhibition rates of Terminalia chebula Retz. extract at concentrations of 0.078 mg / mL, 0.039 mg / mL, and 0.020 mg / mL on Trpv1 were 50.35% ± 1.65%, 34.76% ± 1.51%, and 21.05% ± 0.47% respectively. Compared with the model group, P < 0.05, showing significant differences. The inhibition rates of the Terminalia chebula Retz. encapsulated liquid at concentrations of 0.00234%, 0.00117%, and 0.0006% on TRPV1 were 45.02% ± 3.77%, 31.94% ± 0.49%, and 25.07% ± 0.96% respectively. Compared with the model group, P < 0.05, showing significant differences. It indicates that both the Terminalia chebula Retz. encapsulated liquid and the Terminalia chebula Retz. extract have the effect of inhibiting the expression of TRPV1 content, and their inhibition rates on Trpv1 are 25 - 55%, showing anti-allergic, soothing, and analgesic effects.

[0082] In addition, the inhibitory effect of the Terminalia chebula Retz. encapsulated liquid is better than that of the unencapsulated Terminalia chebula Retz. extract. When the inhibitory effects are equivalent, the dosage of the Terminalia chebula Retz. encapsulated liquid is 1 / 33 of that of the unencapsulated Terminalia chebula Retz. extract, with a lower dosage, suggesting that the Terminalia chebula Retz. encapsulated liquid of the present invention can achieve improved administration effects.

[0083] Effect Example 3 Influence on Type I Collagen 3.1 Experimental Method 3.1.1 Seeding: When the cell density ≥ 70%, collect the cells and count them, adjust the cell density to a certain concentration, and seed the cells into a 6-well plate at a volume of 2000 uL per well, then put it back into the incubator.

[0084] 3.1.2 Experimental Grouping: Plate according to 2 replicate wells for each concentration in each group in Table 7; among them, the Terminalia chebula Retz. extract is 6.25 μg / mL (calculated based on the weight of the freeze-dried powder), and the Terminalia chebula Retz. encapsulated liquid is 4.5 μg / mL (calculated based on the Terminalia chebula Retz. extract).

[0085] 3.1.3 Solution Preparation: According to the obtained cell viability test results, select an appropriate sample concentration as the test concentration and prepare the sample solution to be tested: Use vitamin E as the positive control for the experiment.

[0086] Table 7 Experimental Grouping

[0087] 3.1.4 Administration: Take out the 6-well plate after 24 h, discard the old culture medium, add samples according to 2 replicate wells for each concentration in each group. Add 2000 uL of basal medium to each well in the blank control group (BC) and the negative control group (NC), add 2000 uL of the positive control sample to each well in the positive control group, and add 2000 uL of the corresponding concentration sample to each of the 2 replicate wells for each concentration in the experimental group, then put it back into the incubator for culture (37 °C, 5% CO2); 3.1.5 UVA irradiation: Take out the 24-well plates after 1 h. According to the experimental grouping, perform UVA irradiation at 20 J / cm 2 on the negative control group (NC), positive control group (PC) and experimental group, and the blank control group (BC) is not irradiated. Incubate overnight in an incubator.

[0088] 3.1.6 Fluorescent staining of type I collagen 3.1.7 Data processing and analysis: Analyze the fluorescent expression intensity of type I collagen in each group using Image J, plot graphs using GraphPad Prism software, and perform statistical analysis using T-test between groups. # P < 0.05 indicates significant difference, ## P < 0.01 indicates extremely significant difference, ### P < 0.001 indicates highly significant difference.

[0089] 3.2 Experimental results The experimental results are shown in Figure 4 . Compared with the UVA irradiation group, the Terminalia chebula extract and the Terminalia chebula encapsulation solution have an obvious inhibitory effect on the decrease of type I collagen in UVA-induced 3T3 fibroblasts. It shows that under the experimental conditions, the Terminalia chebula extract and the Terminalia chebula encapsulation solution have an inhibitory effect on the loss of type I collagen in the photoaging model of UVA-induced 3T3 fibroblasts and have an anti-wrinkle effect. When the fluorescent expression intensity of type I collagen is 39 AU, the dosage of the Terminalia chebula extract is 1.38 times that of the Terminalia chebula encapsulation solution, proving that the Terminalia chebula encapsulation solution has a better effect and a better inhibitory effect on the decrease of type I collagen.

[0090] All the documents mentioned in the present invention are cited in this application as references, just as if each document was cited separately as a reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. A terminalia chebula coating liquid, characterized in that: The terminalia chebula packaging liquid comprises: 1-25wt% Terminalia chebula extract; 2-4 wt % lecithin; 5-15 wt % caprylic / capric triglyceride; 1-5 wt % of an organic polyol; 0.1-1 wt % p-hydroxyacetophenone; 10-30 wt % polysorbate-80; 2-10 wt % of the moisturizing agent, based on the total mass of the package; the balance is made up with water.

2. The terminalia chebula packaging liquid according to claim 1, characterized in that The content of lecithin in the terminalia chebula coating liquid is 2.8-3.2 wt %.

3. The terminalia chebula packaging liquid according to claim 1, characterized in that The content of caprylic / capric triglyceride in the terminalia chebula packaging liquid is 8-12 wt %.

4. The terminalia chebula coating liquid according to claim 1, characterized in that The organic polyol is selected from the group consisting of hexanediol, pentanediol, or a combination thereof.

5. The terminalia chebula coating liquid according to claim 1, characterized in that The humectant is selected from the group consisting of glycerin, butylene glycol, or a combination thereof.

6. The method for preparing the terminalia chebula coating liquid according to claim 1, characterized in that: The method comprises the steps of: (1) mixing terminalia chebula extract, lecithin and caprylic / capric triglyceride to form a coating mixture, and stirring under heating conditions of 50-90° C. to form phase A; (2) mixing water, organic polyol and p-hydroxyacetophenone to form phase B; (3) mixing polysorbate 80 with glycerol to form phase C; (4) Add phase A to phase C to obtain phase A+C; (5) Add phase B to the above phases A+C and stir evenly to obtain the terminalia chebula coating liquid.

7. The preparation method according to claim 6, characterized in that: The step (5) further comprises: adding phase B into the above phase A+C, pre-emulsifying, homogenizing under high pressure and then switching to mechanical stirring, standing and aging for 12-48 hours to obtain the terminalia chebula coating liquid.

8. The preparation method according to claim 6, characterized in that: The pressure of the high-pressure homogenization stirring is 100-2000Pa.

9. The terminalia chebula coating liquid according to claim 1, characterized in that The terminalia chebula packaging liquid also has one or more characteristics selected from the following group: (1) Increase the content of nuclear factor E2-related factor 2 (Nrf2); (2) Inhibit the expression of TRPV1 content; (3) Inhibit the decline of type I collagen.

10. The use of the terminalia chebula packaging liquid as claimed in claim 1, characterized in that: Used to prepare daily products or daily chemical products with anti-inflammatory, antioxidant, anti-allergic and soothing, anti-glycation, anti-aging and / or neuro-analgesic effects.

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