A composition with moisturizing and anti-allergic effects and its application

Abstract

The present invention relates to a composition with moisturizing and anti-allergic effects and its application. The composition includes tea tree flower extract, houttuynia cordata extract and elsholtzia ciliata extract. The present invention prepares a natural plant composition using water as an extraction solvent. The obtained composition is safe and reliable, with clear biological activity, and can synergistically improve skin moisture, allergies and inflammation, and has the effect of promoting skin health. The composition can be applied to the fields of food, health food, functional food or cosmetics.

Description

Technical Field

[0001] The present invention belongs to the technical field of substance extraction, and relates to a composition with moisturizing and anti-allergic effects and its application. Background Art

[0002] The skin is the largest organ of the human body, undertaking various important functions such as barrier protection, body temperature regulation, and immune defense. The water retention and anti-allergic ability of the skin are directly related to its health status and protection ability. However, various factors in daily life, such as environmental pollution, ultraviolet exposure, bad eating habits, and mental stress, can lead to damage to the skin barrier function, thereby causing skin dryness, allergic reactions, and other skin problems.

[0003] With the continuous improvement of people's pursuit of beauty and health, many oral beauty products have emerged on the market, aiming to improve skin moisture through internal regulation. A Chinese patent with the application number CN 105582527 B discloses a composition for improving skin moisture and its preparation method. The composition contains okra, boletus edulis, roselle, collagen, hyaluronic acid, sodium erythorbate, and vitamin E, and has a good effect on improving skin moisture. A Chinese patent with the application number CN 117694549 B discloses a composition, preparation method, and application for moisturizing, removing spots, and whitening the skin. The composition contains elastin peptide, tremella, aronia melanocarpa fruit powder, prunus serrulata lindl. var. lannesiana Wils. powder, and lactobacillus plantarum S58, and has the effects of moisturizing and improving skin elasticity.

[0004] The prior art has explored the improvement of skin moisture by oral products, but many product formulas contain too many raw materials, which may lead to mutual interference between components, increase the complexity of the formula and production cost, and increase the risk of side effects or allergic reactions. In addition, improving skin moisture is a complex process that requires a scientific and reasonable combination of ingredients and comprehensive conditioning from multiple perspectives to achieve the best effect. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a composition with moisturizing and anti-allergic effects and its application.

[0006] To achieve the purpose of this invention, the present invention adopts the following technical solutions:

[0007] In the first aspect, the present invention provides a composition with moisturizing and anti-allergic effects, and the composition includes tea tree flower extract, houttuynia cordata extract, and elsholtzia ciliata extract.

[0008] The present invention provides a plant composition for synergistically improving skin moisture and anti-allergy, and a preparation method thereof. Three natural plants, namely tea tree flower, houttuynia cordata, and elsholtzia ciliata, are scientifically selected and combined. By inhibiting the activity of hyaluronidase, promoting the expression of aquaporin AQP3, reducing the secretion of inflammatory factors, reducing allergic reactions, and promoting skin repair, the purpose of comprehensively improving skin moisture and enhancing the skin barrier is achieved, fundamentally improving the skin's water retention ability and promoting the healthy resistance of the skin. In addition, the composition formula is designed simply and efficiently, which can not only effectively avoid the mutual interference between components, but also maximize the efficacy of each component. The polysaccharide content in the tea tree flower extract is not less than 2.5%, the flavonoid content in the houttuynia cordata extract is not less than 10%, the flavonoid content in the elsholtzia ciliata extract is not less than 35%, and the rosmarinic acid content is not less than 1.2%. The three extracts can cooperate with each other and have potential synergistic effects.

[0009] Preferably, the composition comprises 3-10 parts by mass of tea tree flower extract, 1-2 parts by mass of houttuynia cordata extract, and 1-5 parts by mass of elsholtzia ciliata extract.

[0010] The mass parts of the tea tree flower extract can be selected as 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, 5.5 parts, 6 parts, 6.5 parts, 7 parts, 7.5 parts, 8 parts, 8.5 parts, 9 parts, 9.5 parts, 10 parts, etc.; the mass parts of the houttuynia cordata extract can be selected as 1 part, 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, etc.; the mass parts of the elsholtzia ciliata extract can be selected as 1 part, 1.2 parts, 1.5 parts, 1.8 parts, 2 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3 parts, 3.2 parts, 3.5 parts, 3.8 parts, 4 parts, 4.2 parts, 4.5 parts, 4.8 parts, 5 parts, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated here one by one.

[0011] Preferably, the composition is prepared by the following method: physically mixing the preparation raw materials to obtain the composition.

[0012] Preferably, the tea tree flower extract is prepared by the following method:

[0013] (1) Mix the tea tree flower with water and perform vacuum reflux extraction to obtain a tea tree flower extract solution;

[0014] (2) Filter the tea tree flower extract solution through a ceramic membrane and perform freeze-drying to obtain the tea tree flower extract.

[0015] Preferably, the houttuynia cordata extract is prepared by the following method:

[0016] (1) Mix the houttuynia cordata with water and extract to obtain a houttuynia cordata extract solution;

[0017] (2) Concentrate the houttuynia cordata extract to obtain a concentrated solution, refrigerate and flocculate the concentrated solution, and then centrifuge to obtain the product.

[0018] Preferably, the elsholtzia ciliata extract is prepared by a method comprising the following steps:

[0019] (1) Mix elsholtzia ciliata with water, then mix with an acid, and perform ultrasonic extraction to obtain an elsholtzia ciliata extract;

[0020] (2) Filter the elsholtzia ciliata extract through a ceramic membrane and dry to obtain the product.

[0021] Preferably, the pore size of the ceramic membrane used in the extraction of camellia sinensis flowers is 200 - 400 nm, such as 200 nm, 220 nm, 240 nm, 260 nm, 280 nm, 300 nm, 320 nm, 340 nm, 360 nm, 380 nm, 400 nm, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0022] Preferably, the pore size of the ceramic membrane used in the extraction of elsholtzia ciliata is 50 - 100 nm, such as 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0023] Preferably, the mass ratio of camellia sinensis flowers to water is 1:(20 - 30).

[0024] Preferably, the mass ratio of houttuynia cordata to water is 1:(10 - 15).

[0025] Preferably, the mass ratio of elsholtzia ciliata to water is 1:(10 - 30).

[0026] Among them, the specific point values in (20 - 30) can be selected as 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, etc.; the specific point values in (10 - 15) can be selected as 10, 11, 12, 13, 14, 15, etc.; the specific point values in (10 - 30) can be selected as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0027] Preferably, the temperature of reduced-pressure reflux extraction is 60 - 70 °C, the time is 1 - 2 h, and the pressure is 0.02 - 0.08 MPa.

[0028] The temperature can be selected as 60°C, 62°C, 64°C, 66°C, 68°C, 70°C, etc., the time can be selected as 1 h, 1.1 h, 1.2 h, 1.3 h, 1.4 h, 1.5 h, 1.6 h, 1.7 h, 1.8 h, 1.9 h, 2 h, etc., and the pressure can be selected as 0.02 MPa, 0.025 MPa, 0.03 MPa, 0.035 MPa, 0.04 MPa, 0.045 MPa, 0.05 MPa, 0.055 MPa, 0.06 MPa, 0.065 MPa, 0.07 MPa, 0.075 MPa, 0.08 MPa, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0029] Preferably, the temperature for the extraction of Houttuynia cordata is 70 - 90°C and the time is 40 - 60 min.

[0030] The temperature can be selected as 70°C, 72°C, 74°C, 76°C, 78°C, 80°C, 82°C, 84°C, 86°C, 88°C, 90°C, etc., and the time can be selected as 40 min, 42 min, 44 min, 46 min, 48 min, 50 min, 52 min, 54 min, 56 min, 58 min, 60 min, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0031] Preferably, before the extraction, it also includes treating the mixed solution of Houttuynia cordata and water at 0.2 - 0.4 MPa and 120 - 140°C for 5 - 10 min.

[0032] The pressure can be selected as 0.2 MPa, 0.22 MPa, 0.24 MPa, 0.26 MPa, 0.28 MPa, 0.3 MPa, 0.32 MPa, 0.34 MPa, 0.36 MPa, 0.38 MPa, 0.4 MPa, etc., the temperature can be selected as 120°C, 122°C, 124°C, 126°C, 128°C, 130°C, 132°C, 134°C, 136°C, 138°C, 140°C, etc., and the time can be selected as 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0033] Preferably, the acid includes malic acid and / or citric acid.

[0034] Preferably, the time for mixing with the acid is 5 - 20 min, such as 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min, 20 min, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0035] Preferably, the solid content of the concentrated solution is 15 - 25%, the refrigeration temperature is 2 - 4°C, and the time is 4 - 12 h.

[0036] The solid content can be selected as 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, etc.; the temperature can be selected as 2°C, 2.2°C, 2.4°C, 2.6°C, 2.8°C, 3°C, 3.2°C, 3.4°C, 3.6°C, 3.8°C, 4°C, etc.; the time can be selected as 4 h, 4.5 h, 5 h, 5.5 h, 6 h, 6.5 h, 7 h, 7.5 h, 8 h, 8.5 h, 9 h, 9.5 h, 10 h, 10.5 h, 11 h, 11.5 h, 12 h, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0037] Preferably, the rotation speed of centrifugation is 6000 - 10000 rpm, and the time is 5 - 10 min.

[0038] The rotation speed can be selected as 6000 rpm, 6500 rpm, 7000 rpm, 7500 rpm, 8000 rpm, 8500 rpm, 9000 rpm, 9500 rpm, 10000 rpm, etc.; the time can be selected as 5 min, 6 min, 7 min, 8 min, 9 min, 10 min, etc. Other specific point values within the above numerical range can be selected and will not be elaborated one by one here.

[0039] Preferably, the power of ultrasonic extraction is 200 - 500 W, the frequency is 20 - 50 kHz, the temperature is 65 - 80°C, the time is 40 - 60 min, and the pH is 3 - 4.5.

[0040] The pH can be selected from 3, 3.2, 3.4, 3.6, 3.8, 4, 4.1, 4.2, 4.3, 4.4, 4.5, etc.; the power can be selected from 200W, 220W, 240W, 260W, 280W, 300W, 320W, 340W, 360W, 380W, 400W, 420W, 440W, 460W, 480W, 500W, etc.; the frequency can be selected from 20kHz, 22kHz, 24kHz, 26kHz, 28kHz, 30kHz, 32kHz, 34kHz, 36kHz, 38kHz, 40kHz, 42kHz, 44kHz, 46kHz, 48kHz, 50kHz, etc.; the temperature can be selected from 65°C, 66°C, 67°C, 68°C, 69°C, 70°C, 71°C, 72°C, 73°C, 74°C, 75°C, 76°C, 77°C, 78°C, 79°C, 80°C, etc.; the time can be selected from 40min, 42min, 44min, 46min, 48min, 50min, 52min, 54min, 56min, 58min, 60min, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0041] In a second aspect, the present invention provides an application of the composition having moisturizing and anti-allergic effects according to the first aspect in the preparation of a product having moisturizing, anti-allergic, and anti-inflammatory effects.

[0042] Preferably, the product includes food, cosmetics, or health products.

[0043] Preferably, the addition amount of the composition having moisturizing and anti-allergic effects in the product is 0.1 - 50%, such as 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc. Other specific point values within the above numerical ranges can be selected and will not be elaborated one by one here.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] (1) The present invention uses water as the extraction solvent, which can not only efficiently obtain the active ingredients in the three natural herbal plants, but also does not use organic reagents throughout the process, making it safer, more reliable, and having a wide range of applications.

[0046] (2) The composition prepared by the present invention can significantly inhibit the activity of hyaluronidase, promote the expression of AQP-3, inhibit the secretion of NO and the expression of TNF-α and tryptase, reduce the number of neutrophils in the zebrafish inflammation model, and promote the repair of HaCat cells. It has the effects of improving skin moisture, anti-allergy, anti-inflammation and repair, and promotes skin health and water retention ability in multiple aspects.

[0047] (3) The composition prepared by the present invention has synergistic effects in improving skin moisture, anti-allergy, anti-inflammation and repair effects. Description of the Drawings

[0048] Figure 1 It is a graph showing the results of the expression level of zebrafish tryptase. indicates P < 0.001 compared with the control group; * indicates P < 0.05 compared with the model group; *** indicates P < 0.001 compared with the model group.

[0049] Figure 2 It is a fluorescence staining map of zebrafish neutrophils.

[0050] Figure 3 It is a graph showing the results of the number of zebrafish neutrophils. indicates P < 0.001 compared with the control group; *** indicates P < 0.001 compared with the model group.

[0051] Figure 4 It is a microscopic field-of-view result map of the scratch repair test of HaCat cells.

[0052] Figure 5 It is a graph showing the results of the healing rate of the scratch repair test of HaCat cells. ** indicates P < 0.01 compared with the control group; *** indicates P < 0.001 compared with the control group. Detailed Embodiments

[0053] The technical solutions of the present invention will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations of the present invention.

[0054] Preparation Example 1-1

[0055] This preparation example provides a preparation method of tea tree flower extract, and the preparation method includes:

[0056] (1) After crushing the tea tree flowers, mix them with water. The mass ratio of tea tree flowers to water is 1:25, and reflux extraction is carried out at 65 °C and 0.05 MPa for 1.5 h;

[0057] (2) Filter the extract through a 600-mesh sieve and a 300-nm ceramic membrane in sequence;

[0058] (3) Sterilize the filtrate and freeze-dry it to obtain the product.

[0059] Preparation Example 1-2

[0060] This preparation example provides a method for preparing a tea tree flower extract, and the preparation method includes:

[0061] (1) Crush the tea tree flowers and mix them with water. The mass ratio of tea tree flowers to water is 1:20, and reflux extraction is carried out at 60 °C and 0.02 MPa for 2 h;

[0062] (2) Filter the extract through a 600-mesh sieve and a 200-nm ceramic membrane in sequence;

[0063] (3) Centrifuge, sterilize, and freeze-dry the filtrate to obtain the product.

[0064] Preparation Example 1-3

[0065] This preparation example provides a method for preparing a tea tree flower extract, and the preparation method includes:

[0066] (1) Crush the tea tree flowers and mix them with water. The mass ratio of tea tree flowers to water is 1:30, and reflux extraction is carried out at 70 °C and 0.08 MPa for 1 h;

[0067] (2) Filter the extract through a 600-mesh sieve and a 400-nm ceramic membrane in sequence;

[0068] (3) Centrifuge, sterilize, and freeze-dry the filtrate to obtain the product.

[0069] Preparation Example 2-1

[0070] This preparation example provides a method for preparing a houttuynia cordata extract, and the preparation method includes:

[0071] (1) Crush the houttuynia cordata and mix it with water. The mass ratio of houttuynia cordata to water is 1:12, and it is treated at 0.3 MPa and 130 °C for 8 min, and extracted at 80 °C for 50 min to obtain an extract;

[0072] (2) Concentrate the extract to a solid content of 20%, refrigerate the concentrated solution at 3 °C for 8 h, centrifuge at 8000 rpm for 8 min, sterilize and dry the supernatant to obtain the product.

[0073] Preparation Example 2-2

[0074] This preparation example provides a method for preparing a houttuynia cordata extract, and the preparation method includes:

[0075] (1) Crush the houttuynia cordata and mix it with water. The mass ratio of houttuynia cordata to water is 1:15, and it is treated at 0.2 MPa and 120 °C for 10 min, and extracted at 70 °C for 60 min to obtain an extract;

[0076] (2) Concentrate the extract to a solid content of 15%, refrigerate the concentrated solution at 2°C for 4 h, centrifuge at 6000 rpm for 10 min, sterilize the supernatant, and dry it to obtain the product.

[0077] Preparation Example 2-3

[0078] This preparation example provides a method for preparing Houttuynia cordata extract, and the preparation method includes:

[0079] (1) Crush Houttuynia cordata and mix it with water. The mass ratio of Houttuynia cordata to water is 1:10. Treat it at 0.4 MPa and 140°C for 5 min, and extract at 90°C for 40 min to obtain an extract.

[0080] (2) Concentrate the extract to a solid content of 25%, refrigerate the concentrated solution at 4°C for 12 h, centrifuge at 10000 rpm for 5 min, sterilize the supernatant, and dry it to obtain the product.

[0081] Preparation Example 2-4

[0082] This preparation example provides a method for preparing Houttuynia cordata extract, and the difference from Preparation Example 2-1 is only that step (1) is "Crush Houttuynia cordata and mix it with water. The mass ratio of Houttuynia cordata to water is 1:12. Extract at 80°C for 50 min to obtain an extract", and other operations remain unchanged.

[0083] Preparation Example 2-5

[0084] This preparation example provides a method for preparing Houttuynia cordata extract, and the difference from Preparation Example 2-1 is only that step (2) is "Concentrate the extract to a solid content of 10%, refrigerate the concentrated solution at 3°C for 8 h, centrifuge at 8000 rpm for 8 min, sterilize the supernatant, and dry it to obtain the product", and other operations remain unchanged.

[0085] Preparation Example 2-6

[0086] This preparation example provides a method for preparing Houttuynia cordata extract, and the difference from Preparation Example 2-1 is only that step (2) is "Concentrate the extract to a solid content of 30%, refrigerate the concentrated solution at 3°C for 8 h, centrifuge at 8000 rpm for 8 min, sterilize the supernatant, and dry it to obtain the product", and other operations remain unchanged.

[0087] Preparation Example 2-7

[0088] This preparation example provides a method for preparing Houttuynia cordata extract, and the difference from Preparation Example 2-1 is only that step (2) is "Concentrate the extract to a solid content of 20%, centrifuge at 8000 rpm for 8 min, sterilize the supernatant, and dry it to obtain the product", and other operations remain unchanged.

[0089] Preparation Example 2-8

[0090] This preparation example provides a method for preparing Houttuynia cordata Thunb. extract. The difference from Preparation Example 2-1 is only that in step (2), it is "concentrate the extract to a solid content of 20%, refrigerate the concentrated solution at 3°C for 8 h, pass the refrigerated concentrated solution through a 400-mesh sieve, sterilize the supernatant, and dry it to obtain the product", and other operations remain unchanged.

[0091] Preparation Example 3-1

[0092] This preparation example provides a method for preparing Elsholtzia ciliata (Thunb.) Hyland. extract. The preparation method includes:

[0093] (1) After pulverizing Elsholtzia ciliata (Thunb.) Hyland., mix it with water at a mass ratio of Elsholtzia ciliata (Thunb.) Hyland. to water of 1:20, then mix it with citric acid for 5 min, and adjust the pH to 4 to obtain a mixed solution;

[0094] (2) Perform ultrasonic extraction on the extract. The power of ultrasonic extraction is 300 W, the frequency is 35 kHz, the temperature is 70°C, and the time is 50 min. Filter it through an 80-nm ceramic membrane, concentrate the filtrate, sterilize it, and dry it to obtain the product.

[0095] Preparation Example 3-2

[0096] This preparation example provides a method for preparing Elsholtzia ciliata (Thunb.) Hyland. extract. The preparation method includes:

[0097] (1) After pulverizing Elsholtzia ciliata (Thunb.) Hyland., mix it with water at a mass ratio of Elsholtzia ciliata (Thunb.) Hyland. to water of 1:10, then mix it with citric acid for 10 min, and adjust the pH to 3.5 to obtain a mixed solution;

[0098] (2) Perform ultrasonic extraction on the extract. The power of ultrasonic extraction is 200 W, the frequency is 20 kHz, the temperature is 65°C, and the time is 60 min. Filter it through a 100-nm ceramic membrane, concentrate the filtrate, sterilize it, and dry it to obtain the product.

[0099] Preparation Example 3-3

[0100] This preparation example provides a method for preparing Elsholtzia ciliata (Thunb.) Hyland. extract. The preparation method includes:

[0101] (1) After pulverizing Elsholtzia ciliata (Thunb.) Hyland., mix it with water at a mass ratio of Elsholtzia ciliata (Thunb.) Hyland. to water of 1:30, then mix it with citric acid for 15 min, and adjust the pH to 4.5 to obtain a mixed solution;

[0102] (2) Perform ultrasonic extraction on the extract. The power of ultrasonic extraction is 500 W, the frequency is 50 kHz, the temperature is 80°C, and the time is 40 min. Filter it through a 50-nm ceramic membrane, concentrate the filtrate, sterilize it, and dry it to obtain the product.

[0103] Preparation Example 3-4

[0104] This Preparation Example provides a method for preparing elsholtzia extract, which is only different from Preparation Example 3-1 in that step (1) is "After elsholtzia is crushed, it is mixed with water, and the mass ratio of elsholtzia to water is 1:20 to obtain a mixed solution", and other operations remain unchanged.

[0105] Preparation Example 3-5

[0106] This Preparation Example provides a method for preparing elsholtzia extract, which is only different from Preparation Example 3-1 in that step (2) is "The mixed solution is extracted at 70°C for 50 min, filtered through an 80-nm ceramic membrane, the filtrate is concentrated, sterilized, and dried to obtain the product", and other operations remain unchanged.

[0107] Preparation Example 3-6

[0108] This Preparation Example provides a method for preparing elsholtzia extract, which is only different from Preparation Example 3-1 in that step (2) is "The extract is subjected to heating reflux extraction for 50 min, filtered through an 80-nm ceramic membrane, the filtrate is concentrated, sterilized, and dried to obtain the product.

[0109] Example 1

[0110] This Example provides a composition with moisturizing and anti-allergic effects. The composition includes 5 parts of the tea tree flower extract described in Preparation Example 1-1, 1.5 parts of the houttuynia cordata extract described in Preparation Example 2-1, and 3 parts of the elsholtzia extract described in Preparation Example 3-1 by mass.

[0111] Its preparation method is: physically mix the preparation raw materials to obtain the product.

[0112] Example 2

[0113] This Example provides a composition with moisturizing and anti-allergic effects. The composition includes 10 parts of the tea tree flower extract described in Preparation Example 1-2, 1 part of the houttuynia cordata extract described in Preparation Example 2-2, and 1 part of the elsholtzia extract described in Preparation Example 3-2 by mass.

[0114] The preparation method refers to Example 1.

[0115] Example 3

[0116] This Example provides a composition with moisturizing and anti-allergic effects. The composition includes 3 parts of the tea tree flower extract described in Preparation Example 1-3, 2 parts of the houttuynia cordata extract described in Preparation Example 2-3, and 5 parts of the elsholtzia extract described in Preparation Example 3-3 by mass.

[0117] The preparation method refers to Example 1.

[0118] Examples 4-8

[0119] This example provides 5 kinds of compositions with moisturizing and anti-allergic effects. The difference from Example 1 is only that the houttuynia cordata extract obtained from Preparation Example 2-1 is successively replaced with the houttuynia cordata extracts obtained from Preparation Examples 2-4, 2-5, 2-6, 2-7, and 2-8, and other components and their contents remain unchanged.

[0120] The preparation method refers to Example 1.

[0121] Examples 9-11

[0122] This example provides 3 kinds of compositions with moisturizing and anti-allergic effects. The difference from Example 1 is only that the elsholtzia ciliata extract obtained from Preparation Example 3-1 is successively replaced with the elsholtzia ciliata extracts obtained from Preparation Examples 3-4, 3-5, and 3-6, and other components and their contents remain unchanged.

[0123] The preparation method refers to Example 1.

[0124] Comparative Example 1

[0125] This comparative example provides a composition with moisturizing and anti-allergic effects. The difference from Example 1 is only that it does not contain the tea tree flower extract, and the reduced mass is proportionally distributed to the masses of the houttuynia cordata extract and the elsholtzia ciliata extract, and other aspects remain unchanged.

[0126] The preparation method refers to Example 1.

[0127] Comparative Example 2

[0128] This comparative example provides a composition with moisturizing and anti-allergic effects. The difference from Example 1 is only that it does not contain the houttuynia cordata extract, and the reduced mass is proportionally distributed to the masses of the tea tree flower extract and the elsholtzia ciliata extract, and other aspects remain unchanged.

[0129] The preparation method refers to Example 1.

[0130] Comparative Example 3

[0131] This comparative example provides a composition with moisturizing and anti-allergic effects. The difference from Example 1 is only that it does not contain the elsholtzia ciliata extract, and the reduced mass is proportionally distributed to the masses of the tea tree flower extract and the houttuynia cordata extract, and other aspects remain unchanged.

[0132] The preparation method refers to Example 1.

[0133] Test Example 1

[0134] Hyaluronidase inhibition effect test

[0135] Prepare a 0.25 mg / mL hyaluronidase (HAS) solution and a 1 mg / mL sodium hyaluronate solution with 0.1 mM acetate buffer. Prepare a 1 mg / mL tannic acid and sample solution with water as the solvent. The operations are shown in Table 1:

[0136] Table 1

[0137]

[0138]

[0139] A—Absorbance value of the sample solution B—Absorbance value of the sample blank C—Absorbance value of the control solution D—Absorbance value of the control blank

[0140] Table 2

[0141]

[0142]

[0143] It can be seen from the data in Table 2 that the composition described in this application has excellent hyaluronidase inhibitory effect, and the preparation methods of Houttuynia cordata extract and Elsholtzia ciliata extract will affect the efficacy of the composition. Camellia sinensis flower extract, Houttuynia cordata extract and Elsholtzia ciliata extract have a certain synergistic effect in the above efficacy, and the lack of any one of them will affect the efficacy of the composition.

[0144] Test Example 2

[0145] Determination of moisturizing efficacy

[0146] Aquaporin-3 (AQP-3) plays an important role in maintaining skin moisture and skin function. The experiment uses fluorescence quantitative PCR method to determine the effect of the sample on the expression level of aquaporin-3. Seed HaCaT cells in a 24-well plate. Set up a normal control group and a sample administration group (120 μg / mL) in the experiment. Incubate in an incubator (37 °C, 5% CO2) for 12 h. When the plating rate reaches 40% - 60%, conduct grouped administration. After the administration is completed, incubate in the incubator for 24 h. After the incubation is over, use a total RNA extraction kit to perform RNA determination, and fluorescence quantitative PCR to detect the expression of the AQP3 gene. The results are shown in Table 3.

[0147] Table 3

[0148]

[0149]

[0150] As can be seen from the data in Table 3, the composition described in the present application can significantly promote the expression of AQP-3, has excellent moisturizing efficacy, and the preparation methods of Houttuynia cordata extract and Elsholtzia ciliata extract will affect the efficacy of the composition. There is a certain synergistic effect among Camellia sinensis flower extract, Houttuynia cordata extract and Elsholtzia ciliata extract in the above-mentioned efficacy, and the lack of any one of them will affect the efficacy of the composition.

[0151] Test Example 3

[0152] Determination of anti-inflammatory efficacy

[0153] According to the results of the CCK-8 method, the test concentration of the sample was selected as 250 μg / mL for the determination of the anti-inflammatory efficacy of macrophage RAW264.7. The inhibitory effects of the sample on TNF-α and NO were detected by ELISA. The RAW264.7 cell suspension was prepared with complete medium to 5×10 4 cells / mL, and it was inoculated into a 24-well plate, 1000 μL per well, and incubated for 24 hours. A blank control group, a positive control group (5 μM dexamethasone acetate) and a sample group were set up. Add 250 μg / mL of the sample to be tested to each well. After incubating for 1 hour in an incubator at 37 °C and 5% CO2, except for the blank control group, add lipopolysaccharide (LPS) with a final concentration of 1 μg / mL to each well and continue to culture for 24 hours. After completion, collect the cell supernatant by low-temperature centrifugation, and use an ELISA kit and a nitric oxide detection kit to detect the contents of TNF-α and NO according to the instructions of the manual. The results are shown in Table 4.

[0154] Table 4

[0155] Group Relative expression level of TNF-α Relative expression level of NO Example 1 0.63±0.01 0.37±0.02 Example 2 0.66±0.02 0.40±0.03 Example 3 0.65±0.01 0.38±0.01 Example 4 0.73±0.04 0.47±0.06 Example 5 0.72±0.04 0.46±0.01 Example 6 0.83±0.02 0.52±0.03 Example 7 0.85±0.03 0.57±0.02 Example 8 0.82±0.02 0.63±0.07 Example 9 0.78±0.02 0.76±0.03 Example 10 0.80±0.06 0.69±0.01 Example 11 0.70±0.03 0.49±0.03 Control Example 1 0.79±0.03 0.59±0.01 Control Example 2 0.86±0.04 0.52±0.01 Control Example 3 0.96±0.05 0.71±0.02 Preparation Example 1-1 0.95±0.11 0.53±0.02 Preparation Example 2-1 0.88±0.06 0.82±0.01 Preparation Example 3-1 0.75±0.04 0.58±0.02 Positive control 0.56±0.01 0.85±0.04 Model group 0.96±0.08 1.01±0.03 Normal control group 0.02±0.00 0.13±0.03

[0156] As can be seen from the data in Table 4, the composition described in the present application can significantly inhibit the generation of NO and the expression of TNF-α, has excellent anti-inflammatory efficacy, and the preparation methods of Houttuynia cordata extract and Elsholtzia ciliata extract will affect the efficacy of the composition. There is a certain synergistic effect among Camellia sinensis flower extract, Houttuynia cordata extract and Elsholtzia ciliata extract in the above-mentioned efficacy, and the lack of any one of them will affect the efficacy of the composition.

[0157] Test Example 4

[0158] Determination of anti-allergic efficacy

[0159] Dinitrochlorobenzene induces a significant increase in the serum tryptase level. Tryptase complexes with the BAPNA substrate and has a maximum absorption peak at 405 nm, and the degree of allergy can be detected.

[0160] The zebrafish rearing method is carried out according to "The zebrafish book".

[0161] Randomly select 2 dpf wild-type AB strain zebrafish and place them in a 12-well cell culture plate, with 10 zebrafish treated in each well. Sample solutions (Example 1, Preparation Example 1-1, Preparation Example 2-1, Preparation Example 3-1) with a concentration of 100 μg / mL were administered by dissolving in water. Cromolyn sodium at a concentration of 1000 μg / mL was used as the positive control. At the same time, a normal control group and a model control group were set up, and the experiment was repeated in parallel 3 times. Except for the normal control group, a zebrafish allergy model was established by administering 3 μg / mL of dinitrochlorobenzene to the other experimental groups. Each experimental group was given 100 μg / mL of BAPNA and incubated in the dark at (28.5 °C ± 1.0 °C) in a biochemical incubator for 22 h. Then, the 12-well plate was taken out, shaken well, and 200 μL of the liquid was pipetted from each well into a 96-well plate. The absorbance was measured at a wavelength of 405 nm to determine the relative expression level of tryptase.

[0162]

[0163] Sample group: The average value of the absorbance of each sample group

[0164] Normal control group: The average value of the absorbance of the normal control

[0165] Sample blank group: The absorbance value of the sample with the same volume

[0166] Normal blank group: The absorbance value of the embryo culture medium with the same volume

[0167] The results are as Figure 1 shown. It can be Figure 1 seen that the composition described in this application has excellent anti-allergic effects, can significantly reduce the expression level of tryptase, and the effect is better than that of tea flower extract, elsholtzia extract, and houttuynia cordata extract.

[0168] Test Example 5

[0169] Determination of anti-inflammatory efficacy

[0170] Randomly select 3 dpf transgenic neutrophil red fluorescence MPX and place them in a 6-well plate, with 15 zebrafish treated in each well (experimental group). The sample of Example 1 was administered by dissolving in water (test concentration: 100 μg / mL), and dexamethasone acetate (concentration: 5 μg / mL) was used as the positive control. At the same time, a normal control group and a model control group were set up, and the volume of each well was 3 mL. Except for the normal control group, a zebrafish skin inflammation model was established by administering 1 μM of copper sulfate solution to the other experimental groups. After treatment at 28 °C for 2 h, 10 zebrafish were randomly selected from each group and photographed under a fluorescence microscope and the pictures were saved ( Figure 2), the data was analyzed and collected using NIS-Elements D 3.20 advanced image processing software to analyze the number of neutrophils in the skin of zebrafish. Neutrophils in zebrafish (Tg(mpx:dsRED)) are red-fluorescently labeled, so the behaviors such as the migration of neutrophils in live zebrafish can be directly observed under a stereomicroscope with fluorescence. After the incubation, zebrafish were randomly selected from those with normal phenotypes and behaviors, fixed with 3% methylcellulose, observed and photographed under a stereomicroscope, and the statistical results are as Figure 3 shown, and it can be seen from Figure 3 the results that the composition described in the present application can significantly reduce the number of neutrophils and has excellent anti-inflammatory effects.

[0171] Test Example 6

[0172] Determination of scratch repair activity

[0173] HaCaT cells were inoculated into 12-well plates and cultured in maintenance medium in an incubator at 37°C with 5% CO2 for 24 h. A cell scratch model was created by placing inserts in each well of the culture plate and cultured in the incubator for 24 h. A normal control group (DMEM medium without serum), a positive control group (EGF solution with a concentration of 10,000 IU / mL), and treatment groups with the test substances (Example 1, Preparation Example 1-1, Preparation Example 2-1, Preparation Example 3-1) at a concentration of 250 μg / mL were set. The cell growth status at the scratch was observed under a microscope for 24 hours and photographed respectively, as Figure 4 shown, and the healing area was calculated using ImageJ software. The formula for the healing rate is as follows:

[0174] Healing rate = (Initial scratch area - Scratch area at the time to be measured) / Initial scratch area.

[0175] The calculation results are as Figure 5 shown, and it can be seen from Figure 5 the results that the composition described in the present application can significantly increase the cell healing rate, and the effect is better than that of extracts of Camellia sinensis flower, Elsholtzia ciliata, and Houttuynia cordata.

[0176] The applicant declares that the present invention uses the above embodiments to illustrate a composition with moisturizing and anti-allergic effects and its application, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, and the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

[0177] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0178] In addition, it should be noted that, in the various specific technical features described in the above specific embodiments, they can be combined in any appropriate manner without contradiction. To avoid unnecessary repetition, the present invention will not separately describe various possible combination manners.

Claims

1. A composition with moisturizing and anti-allergic effects, characterized in that, The composition comprises 3 - 10 parts by mass of tea tree flower extract, 1 - 2 parts by mass of houttuynia cordata extract, and 1 - 5 parts by mass of elsholtzia ciliata extract; The tea tree flower extract is prepared by a method comprising the following steps: (1) Mix tea tree flowers with water and perform vacuum reflux extraction to obtain a tea tree flower extract solution; (2) Filter the tea tree flower extract solution through a ceramic membrane and freeze-dry it to obtain the tea tree flower extract; The houttuynia cordata extract is prepared by a method comprising the following steps: (1) Mix houttuynia cordata with water, and treat the mixed solution of houttuynia cordata and water at 0.2 - 0.4 MPa and 120 - 140 °C for 5 - 10 min, then extract to obtain a houttuynia cordata extract solution; (2) Concentrate the houttuynia cordata extract solution to obtain a concentrated solution, refrigerate the concentrated solution for flocculation, and centrifuge it to obtain the houttuynia cordata extract; The solid content of the concentrated solution is 15 - 25%, the refrigeration temperature is 2 - 4 °C, and the time is 4 - 12 h; The elsholtzia ciliata extract is prepared by a method comprising the following steps: (1) Mix elsholtzia ciliata with water, then mix with an acid, and perform ultrasonic extraction to obtain an elsholtzia ciliata extract solution; (2) Filter the elsholtzia ciliata extract solution through a ceramic membrane and dry it to obtain the elsholtzia ciliata extract; 2. The composition with moisturizing and anti-allergic effects according to claim 1, wherein The temperature of the vacuum reflux extraction is 60 - 70 °C, the time is 1 - 2 h, and the pressure is 0.02 - 0.08 MPa.

3. The composition with moisturizing and anti-allergic effects according to claim 1, characterized in that, The power of the ultrasonic extraction is 200 - 500 W, the frequency is 20 - 50 kHz, the temperature is 65 - 80 °C, the time is 40 - 60 min, and the pH is 3 - 4.

5.

4. Use of the composition with moisturizing and anti-allergic effects according to any one of claims 1 - 3 in the preparation of a product with moisturizing, anti-allergic or anti-inflammatory effects.

Images