Traditional Chinese medicine composition with anti-inflammatory and anti-oxidation effects as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention belongs to the field of traditional Chinese medicine extraction, and particularly relates to a traditional Chinese medicine composition with anti-inflammatory and anti-oxidation effects and a preparation method and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials and / or extracts of the following raw materials in parts by mass: 8 to 12 parts of black ants, 8 to 12 parts of cortex fraxini, 4 to 6 parts of yam rhizome, 2 to 4 parts of rhizoma polygoni cuspidati, 2 to 4 parts of fructus axilli, 2 to 4 parts of rhizoma chuanxiong, 1 to 3 parts of radix paeoniae rubra, 1 to 3 parts of cassia twig and 0.5 to 2 parts of liquorice root. The raw materials of the traditional Chinese medicine composition are dried, crushed, sieved, extracted and separated to obtain the traditional Chinese medicine composition with anti-inflammatory and anti-oxidation effects. The invention also discloses application of the traditional Chinese medicine composition in the health field. Compared with the prior art, the traditional Chinese medicine composition with the anti-inflammatory and anti-oxidation effects and the preparation method of the traditional Chinese medicine composition are provided, and the prepared traditional Chinese medicine composition extract has excellent oxidation resistance, anti-inflammatory performance, ultraviolet resistance and xanthine oxidase inhibition capacity.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine extraction, and specifically relates to a traditional Chinese medicine composition with anti-inflammatory and antioxidant effects, and a preparation method and application thereof. Background Art

[0002] The application scope of Chinese herbal medicine extracts is now very wide. In addition to the traditional use in Chinese medicine products, as people's trust and dependence on natural products gradually increase, a large part of Chinese herbal medicine extracts have been used in health care products and food ingredients. In addition, Chinese herbal medicine extracts have been widely used in cosmetics and other fields in recent years.

[0003] Black ants are a kind of Chinese medicinal material, which is often used in traditional Chinese medicine to treat joint pain, rheumatic diseases, etc. It has the effects of promoting blood circulation and removing blood stasis, reducing swelling and relieving pain. Qinpi is a common Chinese medicinal material, which is often used to clear away heat and detoxify, cool blood and stop bleeding. In traditional Chinese medicine, Qinpi can be used to treat bleeding symptoms, colds, etc. Dioscorea has the effects of clearing heat and detoxifying, cooling blood and stopping bleeding, and is often used in traditional Chinese medicine to treat symptoms such as fever, dysentery, and hemorrhoids. Polygonum cuspidatum is a Chinese medicinal material that has the effects of clearing heat and detoxifying, and diuresis and stranguria. Six axis has the effects of clearing heat and detoxifying, dispelling wind and dampness, and is often used in traditional Chinese medicine to treat eczema, eczematous skin diseases, etc. Chuanxiong is a Chinese medicinal material that has the effects of promoting blood circulation and removing blood stasis, stopping bleeding and dispersing blood stasis. In traditional Chinese medicine, Chuanxiong is often used to treat symptoms such as blood stasis, amenorrhea, and stagnation. Chiyao has the effects of promoting blood circulation and removing blood stasis, reducing swelling and relieving pain, and is often used in traditional Chinese medicine to treat blood stasis symptoms, sores, etc. Cinnamon twig is a commonly used Chinese herbal medicine with the effects of warming the meridians and dispelling cold, relieving the symptoms and inducing sweating. It is often used in traditional Chinese medicine to treat colds, numbness of limbs and other symptoms. Licorice can be used in combination with Chinese herbal medicines to reconcile the properties of the medicines and increase the efficacy. It also has certain effects of nourishing yin and relieving pain. This Chinese medicine composition combines the unique properties of various herbs and has excellent anti-inflammatory and repair functions. The black ant, Qinpi and red medicine in its ingredients have anti-inflammatory effects, the knotweed and six-axis seeds help to repair the skin, and the cinnamon twig and licorice have the effect of balancing and enhancing the effects of other ingredients.

[0004] Traditional Chinese medicine has a wide range of applications and a long history in traditional medicine. It is generally considered safe and mild. Its application in the field of big health can further reduce potential irritation and allergic reactions. Summary of the invention

[0005] The purpose of the present invention is to solve at least one of the above problems and to provide a Chinese medicine composition with anti-inflammatory and antioxidant effects and its preparation method and application, so as to solve the lack of a Chinese medicine composition with anti-inflammatory and repair effects in the prior art, and to obtain a Chinese medicine composition with excellent antioxidant, anti-inflammatory and anti-ultraviolet properties.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] One of the technical solutions of the present invention is to provide a traditional Chinese medicine composition with anti-inflammatory and antioxidant effects, which comprises the following raw materials and / or extracts of the following raw materials, measured by mass: 8-12 parts of black ants, 8-12 parts of Qinpi, 4-6 parts of Dioscorea, 2-4 parts of Polygonum cuspidatum, 2-4 parts of Hexagonal Fructus, 2-4 parts of Chuanxiong, 1-3 parts of Red Medicine, 1-3 parts of Cassia Twig, and 0.5-2 parts of Licorice.

[0008] Furthermore, the Chinese medicine composition with anti-inflammatory and antioxidant effects includes the following raw materials and / or extracts of the following raw materials, measured by mass: 10 parts of black ants, 10 parts of Qinpi, 5 parts of Dihuangyuan, 3 parts of Polygonum cuspidatum, 3 parts of Hexagonal Seed, 3 parts of Chuanxiong, 2 parts of Red Medicine, 2 parts of Cassia Twig, and 1 part of Licorice.

[0009] The second technical solution of the present invention is to provide a method for preparing the Chinese medicine composition with anti-inflammatory and antioxidant effects as described above, wherein the raw materials of the Chinese medicine composition, including black ant, Qinpi, Dihuangjie, Polygonum cuspidatum, Hexagonal fruit, Chuanxiong, Red Medicine, Cassia twig and Licorice, are dried, crushed, sieved, extracted and separated to obtain the Chinese medicine composition with anti-inflammatory and antioxidant effects.

[0010] Furthermore, the drying temperature of the raw materials of the traditional Chinese medicine composition is 46-50° C., preferably 48° C., and the drying time is 48 hours.

[0011] Furthermore, the raw materials of the Chinese medicine composition are crushed to a size not exceeding 80 meshes.

[0012] Furthermore, the sieved raw materials of the Chinese medicine composition are mixed with the organic solvent in proportion, and then ultrasonically treated to obtain the extract of the Chinese medicine composition. The ultrasonic time is 100 to 150 minutes and the temperature is 40 to 60°C.

[0013] Furthermore, the ultrasonic time is preferably 120 min, and the temperature is preferably 50°C.

[0014] Furthermore, the mixing ratio of the raw materials of the traditional Chinese medicine composition to the organic solvent is 1 g: (3-6) mL, preferably 1 g: 4 mL.

[0015] Furthermore, the organic solvent is an ethanol solution with a volume fraction of 30%.

[0016] Furthermore, the separation includes centrifugation, concentration and freeze-drying.

[0017] Furthermore, the centrifugal speed is 8000-12000 r / min, and the time is 8-12 min; the concentration is concentrated to 0.25-20% of the original volume; the freeze-drying temperature is -60--40°C, and the time is 24-48 h.

[0018] Furthermore, the centrifugal speed is preferably 10000 r / min, and the time is preferably 10 min; the pore size of the filter membrane after centrifugation is 40 μm, and the concentration is concentrated to 5% of the original volume; the freeze-drying temperature is -50°C, and the time is 48 h.

[0019] The third technical solution of the present invention is to provide an application of the above-mentioned Chinese medicine combination with anti-inflammatory and antioxidant effects in the field of cosmetics and health products.

[0020] Furthermore, the application includes application in anti-oxidation, anti-inflammatory and anti-ultraviolet light.

[0021] Reactive oxygen free radicals are some oxygen-containing groups in an excited state, which are toxic to the human body. The antioxidant components in traditional Chinese medicine can react with these free radicals to remove them; antioxidant enzymes in cells (such as superoxide dismutase, glutathione peroxidase, etc.) can catalyze the decomposition of free radicals; Chinese medicine compositions can enhance the activity of these enzymes, thereby improving the antioxidant capacity of cells; anti-inflammatory components (saponins, flavonoids) in Chinese medicine compositions may also play an anti-inflammatory role by regulating signal pathways related to inflammation (such as NF-κB, AP-1, IRF3, etc.). The Chinese medicine composition of the present invention contains active ingredients such as flavonoids, polyphenols, saponins, polysaccharides, etc., which can remove free radicals in the human body and reduce oxidized substances, thereby achieving the effect of protecting the body.

[0022] In addition, ultraviolet radiation can cause skin cell apoptosis, leading to thinning of the skin and increased wrinkles, while the antioxidant ingredients in Chinese herbal compositions (such as flavonoids, polyphenols, etc.) can scavenge free radicals produced by ultraviolet radiation, thereby reducing oxidative stress and protecting skin cells from damage. At the same time, certain ingredients in Chinese medicine (such as saponins, polysaccharides, etc.) can inhibit ultraviolet-induced cell apoptosis and protect the survival and proliferation of skin cells.

[0023] Furthermore, the application of the Chinese medicine composition in the field of cosmetics includes preparing the Chinese medicine composition into a preparation type that is easy to use, including any one of an aqueous product, an oil product, an emulsion product, a surfactant solvent product, a gel product, and a paste product.

[0024] Furthermore, the application of the Chinese medicine composition in the field of health care products includes preparing the Chinese medicine composition into a preparation type that is easy to use, including any one of a solid dosage form, a semi-solid dosage form, a liquid dosage form and a gaseous dosage form.

[0025] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0026] Compared with the prior art, the present invention has the following advantages:

[0027] 1. The Chinese medicine composition extract prepared by the present invention has excellent antioxidant properties. The scavenging rate of the Chinese medicine composition extract solution with a concentration of 1 mg / mL reaches 97.95% for DPPH free radicals and 99.85% for ABTS free radicals. Its antioxidant activity is equivalent to that of vitamin C. After the important components of the Chinese medicine composition, black ants, Qinpi and Dihuangjie, are compounded and synergistically, the scavenging ability of the scavenging rate of DPPH free radicals and ABTS free radicals is significantly improved. It has excellent antioxidant properties when used in cosmetic preparations or health care product preparations.

[0028] 2. The Chinese medicine composition extract prepared by the present invention has excellent anti-inflammatory properties. The Chinese medicine composition extract solution with a concentration of 1 mg / mL has no inhibitory effect on macrophages and has the effect of promoting cell proliferation. The Chinese medicine composition extract solution with a concentration of 1 mg / mL has a significant inhibitory effect on the NO concentration in macrophages, indicating that the Chinese medicine composition extract can inhibit the occurrence of inflammatory reactions in macrophages through the concentration of nitric oxide; and after the important components of the Chinese medicine composition, black ants, Qinpi and Dihuang are compounded and synergistically, the inhibitory ability of hyaluronidase is significantly improved, which is 10.71%, 17.36% and 25.93% higher than that of using black ants, Qinpi or Dihuang respectively. Therefore, the Chinese medicine composition extract prepared by the present invention will have excellent anti-inflammatory properties when used in cosmetic preparations or health care product preparations.

[0029] 3. The Chinese medicine composition extract prepared by the present invention has xanthine oxidase inhibitory activity. When the Chinese medicine composition extract is at a concentration gradient of 0.2 to 1 mg / L, the xanthine oxidase inhibition rate increases with the increase in concentration. At a concentration of 1 mg / mL, the xanthine oxidase inhibition rate of the Chinese medicine composition extract is close to that of allopurinol. When used in health care preparations, it has excellent xanthine oxidase inhibitory activity and thus has an excellent uric acid control effect.

[0030] 4. The Chinese medicine composition extract prepared by the present invention has excellent anti-ultraviolet performance. The absorbance of the Chinese medicine composition extract solution under ultraviolet light in the wavelength range of 280 to 400 nm decreases with the increase of the wave number, and the absorbance increases with the concentration of the Chinese medicine composition extract. In the UVB segment at a wavelength of 320 nm, the absorbance of 1 mg / mL of the Chinese medicine composition extract reaches 99.51%, and in the UVA segment at a wavelength of 400 nm, the absorbance of 1 mg / mL of the Chinese medicine composition extract reaches 65.15%, indicating that the Chinese medicine composition extract has excellent anti-ultraviolet ability, and has excellent anti-ultraviolet ability when used in cosmetic preparations or health care product preparations. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The graph shows the test results of DPPH free radical scavenging rate of different concentrations of Chinese herbal composition extract (FDC) and vitamin C (VC).

[0032] Figure 2 The test results of the synergistic scavenging rate of DPPH free radicals by the combination of black ants, Radix Dioscoreae and Cortex Fraxini.

[0033] Figure 3 The graph shows the test results of different concentrations of Chinese herbal composition extract (FDC) and vitamin C (VC) on the ABTS free radical scavenging rate.

[0034] Figure 4 This is a test result chart of the synergistic scavenging rate of ABTS free radicals by the combination of black ants, Radix Dioscoreae and Cortex Fraxini.

[0035] Figure 5 This is a test result chart of the synergistic inhibition rate of hyaluronidase by the combination of black ants, Radix Dioscoreae and Cortex Fraxini.

[0036] Figure 6 The graph shows the test results of different concentrations of Chinese herbal composition extract (FDC) on macrophage activity.

[0037] Figure 7 The graph shows the test results of the inhibition rate of nitric oxide by different concentrations of Chinese herbal composition extract (FDC).

[0038] Figure 8 This is a graph showing the test results of the xanthine oxidase inhibition rate of different concentrations of Chinese herbal composition extract (FDC).

[0039] Figure 9 The graph shows the test results of ultraviolet absorption rate of traditional Chinese medicine composition extract (FDC) with different concentrations. DETAILED DESCRIPTION

[0040] The present invention is described in detail below in conjunction with the accompanying drawings and specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and specific operation process, but the protection scope of the present invention is not limited to the following embodiments.

[0041] In the following examples, unless otherwise specified, the reagents used are conventional commercial products in the art, and the methods adopted are conventional means known in the art.

[0042] Example 1

[0043] This embodiment provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0044] S1: Take the raw materials of the Chinese medicine composition: 10 parts of black ants, 10 parts of Qinpi, 5 parts of Dihuangjie, 3 parts of Polygonum cuspidatum, 3 parts of Hexigastrointestinal Root, 3 parts of Chuanxiong, 2 parts of Chiyao, 2 parts of Cinnamomum cassia, and 1 part of Licorice, dry them at room temperature at 48±2℃ for 48h, and grind them into powder using a machine;

[0045] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0046] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0047] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0048] The FDC prepared in Example 1 was dissolved in deionized water and its performance was measured.

[0049] Comparative Example 1

[0050] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0051] S1: Take 5 portions of Radix Dioscoreae, a raw material of the Chinese medicine composition, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0052] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0053] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0054] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0055] The FDC prepared in Comparative Example 1 was dissolved in deionized water to measure its performance.

[0056] Comparative Example 2

[0057] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0058] S1: Take 10 portions of the raw material of the Chinese medicine composition, Cortex Fraxini, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0059] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0060] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0061] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0062] The FDC prepared in Comparative Example 2 was dissolved in deionized water to measure its performance.

[0063] Comparative Example 3

[0064] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0065] S1: Take 10 portions of the raw material of the Chinese medicine composition, black ants, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0066] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0067] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0068] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0069] The FDC prepared in Comparative Example 3 was dissolved in deionized water to measure its performance.

[0070] Comparative Example 4

[0071] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0072] S1: Take 10 parts of Fraxinus Fraxinus and 5 parts of Radix Dioscoreae, the raw materials of the Chinese medicine composition, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0073] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0074] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0075] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0076] The FDC prepared in Comparative Example 4 was dissolved in deionized water to measure its performance.

[0077] Comparative Example 5

[0078] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0079] S1: Take 10 parts of black ants and 5 parts of Radix Dioscoreae, the raw materials of the Chinese medicine composition, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0080] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0081] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0082] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0083] The FDC prepared in Comparative Example 5 was dissolved in deionized water to measure its performance.

[0084] Comparative Example 6

[0085] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0086] S1: Take 10 parts of Qinpi and 10 parts of black ants as raw materials of the Chinese medicine composition, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0087] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0088] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0089] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0090] The FDC prepared in Comparative Example 6 was dissolved in deionized water to measure its performance.

[0091] Comparative Example 7

[0092] This comparative example provides a Chinese medicine composition with anti-inflammatory and antioxidant effects and a preparation method thereof, comprising the following steps:

[0093] S1: Take 10 parts of black ants, 10 parts of Qinpi, and 5 parts of Radix Dioscoreae as raw materials of the Chinese medicine composition, dry them at room temperature at 48±2°C for 48h, and grind them into powder using a machine;

[0094] S2: passing the ground powder through an 80-mesh sieve to obtain the raw material powder of the traditional Chinese medicine composition;

[0095] S3: The raw material powder of the Chinese medicine composition and the 30% ethanol solution are uniformly mixed in a ratio of 1 g: 4 mL, and ultrasonicated in an ultrasonic extraction device for 120 min in a water bath at a temperature of 50° C. to obtain an extract of the Chinese medicine composition;

[0096] S4: The Chinese herbal composition extract was centrifuged at a speed of 10000 r / min for 10 min, and the upper liquid was filtered after centrifugation. The pore size of the filter membrane was 40 μm, and the obtained filtrate was concentrated to 5% of the original filtrate volume. It was freeze-dried at -50°C for 48 hours to obtain a Chinese herbal composition extract (FDC) with anti-inflammatory and antioxidant effects.

[0097] The FDC prepared in Comparative Example 7 was dissolved in deionized water to measure its performance.

[0098] The raw material composition ratios of Example 1 and Comparative Examples 1-5 are shown in Table 1:

[0099] Table 1 Raw material components and ratios of Example 1 and Comparative Examples 1-7

[0100] Black ant Chinese ash bark Rhizoma dioscoreae hypoglaucae Polygonum cuspidatum Aconitum carmichaelii var. wutaiense Szechuan lovage rhizome Red peony root Cassia twig Licorice root Example 1 10 10 5 3 3 3 2 2 1 Comparative Example 1 0 0 5 0 0 0 0 0 0 Comparative Example 2 0 10 0 0 0 0 0 0 0 Comparative Example 3 10 0 0 0 0 0 0 0 0 Comparative Example 4 0 10 5 0 0 0 0 0 0 Comparative Example 5 10 0 5 0 0 0 0 0 0 Comparative Example 6 10 10 0 0 0 0 0 0 0 Comparative Example 7 10 10 5 0 0 0 0 0 0

[0101] The following is a detailed description of the Chinese medicine composition extract (FDC) provided by the present invention and its application in antioxidant, anti-inflammatory, anti-ultraviolet and xanthine oxidase inhibition in combination with Example 1 and Comparative Examples 1-7, but they should not be construed as limiting the scope of protection of the present invention.

[0102] Test Example 1

[0103] This test case combines Figures 1-4 , the antioxidant properties of the Chinese medicine composition extract (FDC) prepared in Example 1 were determined.

[0104] 1.1 DPPH free radical scavenging rate determination

[0105] The FDC prepared in Example 1 was dissolved in deionized water to obtain FDC mixed solutions with concentrations of 0.1, 0.2, 0.4, 0.8 and 1 mg / mL, respectively; the FDC prepared in Comparative Examples 1-7 was dissolved in deionized water to obtain an FDC mixed solution with a concentration of 1 mg / mL; the above two batches of samples were respectively subjected to DPPH free radical scavenging rate determination.

[0106] a. Prepare DPPH solution (2.3 mg DPPH: 50 mL anhydrous ethanol) and store in dark place. The absorbance at 517 nm should be between 1.2 and 1.3. Set aside.

[0107] b. Take a clean centrifuge tube, add 2 mL of sample FDC mixed solution, and then add 2 mL of DPPH solution, record it as A1;

[0108] c. Take a clean centrifuge tube, add 2 mL of sample FDC mixed solution, and then add 2 mL of deionized water solution, record it as A2;

[0109] d. Take a clean centrifuge tube, add 2 mL of deionized water, and then add 2 mL of DPPH solution, record it as A3;

[0110] e. The DPPH free radical scavenging ability of the Chinese medicine composition solution is calculated by the following formula:

[0111]

[0112] In the formula,

[0113] A1: absorbance after mixing DPPH and sample;

[0114] A2: Absorbance after mixing sample solution and deionized water:

[0115] A3: absorbance after mixing DPPH and deionized water;

[0116] A0: absorbance of deionized water (only one group needs to be tested).

[0117] The results are as follows Figures 1-2 As shown, the DPPH free radical scavenging rate of the Chinese medicine composition extracts (FDC) of Example 1 and Comparative Examples 1-7 was measured at different concentrations. Figure 1 It can be seen that, with vitamin C (VC) as a positive control, the DPPH free radical scavenging rate of the Chinese herbal composition extract (FDC) increased with the increase of concentration at a concentration gradient of 0.1 to 1 mg / mL. The DPPH free radical scavenging rate of the Chinese herbal composition extract (FDC) at a concentration of 1 mg / mL reached 97.95%. Compared with the positive control VC, the DPPH free radical scavenging ability of the Chinese herbal composition extract (FDC) is basically equivalent to that of the positive control, indicating that the Chinese herbal composition has a relatively excellent antioxidant capacity. Figure 2 It can be seen that when black ants, Qinpi or Dichroa are used alone, the scavenging ability of Qinpi and Dichroa on DPPH free radicals is not as good as that of black ants; in addition, the synergistic effect of using Qinpi and Dichroa in combination did not significantly exceed the single effect of black ants; but after the black ants, Qinpi and Dichroa were combined and synergistic, the scavenging ability of DPPH free radicals was significantly improved, which was increased by 25.57%, 33.33% and 49.23% respectively compared with using black ants, Qinpi or Dichroa alone.

[0118] 1.2 ABTS free radical scavenging rate determination

[0119] The FDC prepared in Example 1 was dissolved in deionized water to obtain FDC mixed solutions with concentrations of 0.1, 0.2, 0.4, 0.8 and 1 mg / mL, respectively; the FDC prepared in Comparative Examples 1-7 was dissolved in deionized water to obtain an FDC mixed solution with a concentration of 1 mg / mL; the above two batches of samples were respectively subjected to ABTS free radical scavenging rate determination.

[0120] a. Mix 5 mL of 7.4 mmol / L ABTS stock solution with 88 μL of 2.6 mmol / L K2S2O8 and let stand for 12 to 16 hours to prepare ABTS working solution; take 0.4 mL of ABTS solution and dilute it with PBS solution; an absorbance of 734 nm is required at room temperature;

[0121] b. Take a clean centrifuge tube, add 0.5mL of sample FDC mixed solution, and then add 2mL of ABTS solution, record it as A1;

[0122] c. Take a clean centrifuge tube, add 0.5mL of sample FDC mixed solution, and then add 2mL of PBS solution, record it as A2;

[0123] d. Take a clean centrifuge tube, add 0.5mL PBS solution, then add 2mL ABTS solution, record it as A3;

[0124] e. The ABTS free radical scavenging capacity of the Chinese medicine composition solution was calculated by the following formula:

[0125]

[0126] In the formula,

[0127] A1: absorbance of ABTS containing sample;

[0128] A2: Absorbance of sample solution and distilled water:

[0129] A3: Absorbance of ABTS without sample;

[0130] A0: absorbance of distilled water.

[0131] The results are as follows Figures 3-4 As shown, the extracts of the Chinese medicine composition prepared in Example 1 and Comparative Examples 1-7 were divided into different concentrations to determine the ABTS free radical scavenging rate. Figure 3It can be seen that, with vitamin C (VC) as a positive control, the ABTS free radical scavenging rate of the Chinese herbal composition extract (FDC) increased with the increase of concentration at a concentration gradient of 0.1 to 1 mg / mL. At a concentration of 1 mg / mL, the ABTS free radical scavenging rate of the Chinese herbal composition extract (FDC) reached 99.85%. Compared with the positive control VC, the ABTS free radical scavenging ability of the Chinese herbal composition extract (FDC) is similar to that of VC, indicating that the Chinese herbal composition has excellent antioxidant capacity. Figure 4 It can be seen that when a certain component of the Chinese medicine composition extract is used alone, the ABTS free radical scavenging ability exhibited by black ants is stronger than that of Qinpi and Dixie. When Qinpi and Dixie are used in combination, their free radical scavenging ability is improved. However, the ABTS free radical scavenging ability of the combination of black ants, Qinpi and Dixie is significantly enhanced after synergistic use, which is 9%, 16.67% and 30.07% higher than that of using black ants, Qinpi or Dixie alone, respectively.

[0132] Test Example 2

[0133] This test case combines Figures 5-7 The anti-inflammatory properties of the Chinese medicine composition extracts (FDC) prepared in Example 1 and Comparative Examples 1-7 were determined.

[0134] 2.1 Determination of hyaluronidase inhibition rate

[0135] The FDC prepared in Comparative Examples 1-7 was dissolved in water to obtain an FDC mixed solution with a concentration of 1 mg / mL for determination of hyaluronidase inhibition rate.

[0136] a. Add 0.1 mL of calcium chloride solution, 0.05 mL of hyaluronidase, and 0.1 mL of FDC mixed solution, and keep warm at 37°C for 40 min; add 0.2 mL of sodium hyaluronate solution, and keep warm at 37°C for 60 min; add 0.1 mL of sodium hydroxide solution and 0.1 mL of acetylacetone solution, and boil in a water bath for 15 min, and immediately cool to room temperature; add 0.5 mL of p-dimethylaminobenzaldehyde solution, mix thoroughly, and keep warm at 37°C for 10 min; record as T;

[0137] b. Add 0.1 mL of calcium chloride solution, 0.05 mL of acetic acid buffer solution, and 0.1 mL of flavonoid extract solution from burdock root, and keep warm at 37°C for 40 min; add 0.2 mL of deionized water, and keep warm at 37°C for 60 min; add 0.1 mL of sodium hydroxide solution and 0.1 mL of acetylacetone solution, and boil in a boiling water bath for 15 min, and immediately cool to room temperature; add 0.5 mL of p-dimethylaminobenzaldehyde solution, mix thoroughly, and keep warm at 37°C for 10 min; record as T0;

[0138] c. Add 0.1 mL of calcium chloride solution, 0.05 mL of hyaluronidase, and 0.1 mL of 60% anhydrous ethanol, and keep warm at 37°C for 40 min; add 0.2 mL of sodium hyaluronate solution, and keep warm at 37°C for 60 min; add 0.1 mL of sodium hydroxide solution and 0.1 mL of acetylacetone solution, and boil in a boiling water bath for 15 min, and immediately cool to room temperature; add 0.5 mL of p-dimethylaminobenzaldehyde solution, mix well, and keep warm at 37°C for 10 min; record as C;

[0139] d. Add 0.1 mL of calcium chloride solution, 0.05 mL of acetate buffer solution, and 0.1 mL of 60% anhydrous ethanol, and keep warm at 37°C for 40 min; add 0.2 mL of sodium hyaluronate solution, and keep warm at 37°C for 60 min; add 0.1 mL of sodium hydroxide solution and 0.1 mL of acetylacetone solution, and boil in a boiling water bath for 15 min, and immediately cool to room temperature; add 0.5 mL of p-dimethylaminobenzaldehyde solution, mix well, and keep warm at 37°C for 10 min; record as C0;

[0140] e. The hyaluronidase inhibition rate of the black ant, Qinpi, and Dihuangjie compound synergistic solution was calculated by the following formula:

[0141]

[0142] In the formula,

[0143] T: absorbance value of sample group;

[0144] T0: absorbance value of sample control group;

[0145] C: absorbance value of negative control group;

[0146] C0: absorbance value of blank control group.

[0147] The results are as follows Figure 5 As shown in the results, when the combination of black ants, qinpi and dauerkraut was tested for synergistic hyaluronidase inhibition, the structure and antioxidant effect trends were similar. When black ants were added to the combination of qinpi and dauerkraut, the anti-inflammatory activity of qinpi and dauerkraut was significantly improved compared to when they were used as single substances, which was increased by 10.71%, 17.36% and 25.93% respectively compared to the use of black ants, qinpi or dauerkraut alone. This result further verified that black ants can produce a positive synergistic effect when combined with qinpi and dauerkraut, thereby enhancing the overall anti-inflammatory effect.

[0148] 2.2 Macrophage RAW264.7 cell viability assay

[0149] The FDC prepared in Example 1 was dissolved in deionized water to obtain FDC mixed solutions with concentrations of 0.1, 0.125, 0.25, 0.5, 1 and 2 mg / mL, respectively, for macrophage RAW264.7 viability assay.

[0150] a. RAW264.7 macrophages were plated with DMEM medium in 96-well plates and incubated in a carbon dioxide cell culture incubator for 24 h;

[0151] b. Discard the supernatant on the 96-well plate, dilute the FDC mixed solution sample step by step with DMEM, add the diluted sample to the 96-well plate, and incubate in a carbon dioxide cell culture incubator for 24 hours;

[0152] c. Discard the supernatant of the 96-well plate and use a CCK-8 detection kit to detect the viability of RAW264.7 macrophages (RAW264.7 macrophages + diluted Chinese medicine composition sample + CCK-8 are recorded as the OD value of the experimental well; RAW264.7 macrophages + CCK-8 are recorded as the OD value of the blank well; RAW264.7 macrophages + DMEM medium + CCK-8 are recorded as the OD value of the control well);

[0153] d. The activity of the Chinese medicine composition solution on macrophage RAW264.7 cells was calculated according to the following formula:

[0154] Cell survival rate = (OD value of experimental well - OD value of blank well) / (OD value of control well - OD value of blank well) × 100%.

[0155] The results are as follows Figure 6 As shown, after the FDC mixed solution was incubated with RAW264.7 macrophages at different concentrations for 24 hours, the activity of macrophages RAW264.7 was measured. When the concentration of the Chinese medicine composition extract (FDC) was between 0.125 and 1 mg / mL, there was no significant difference in cell viability compared with that without the Chinese medicine composition extract, and the cell viability remained above 100% and had the effect of promoting cell proliferation. When the concentration of the Chinese medicine composition extract (FDC) reached 2g / mL, there was a significant difference compared with that without the Chinese medicine composition extract (FDC), and the cell viability reached 80.5%, showing inhibitory effect on cells; In summary, 1 mg / mL was selected as the maximum concentration of the Chinese medicine composition extract to conduct a series of cell experiments.

[0156] 2.3 Nitric oxide concentration determination

[0157] The FDC prepared in Example 1 was dissolved in deionized water to obtain FDC mixed solutions with concentrations of 0.25, 0.5, 1 and 2 mg / mL, respectively, for determination of nitric oxide concentration.

[0158] NO is an important cell signaling molecule in the body. Although it is active and extremely unstable, it plays an important role in maintaining the homeostasis of the body's internal environment. NO can participate in many physiological functions of the body, including vasodilation, smooth muscle relaxation, and neurotransmission. It is also one of the important inflammatory immune mediators secreted by inflammatory macrophages. Therefore, NO concentration determination is used to evaluate the anti-inflammatory activity of the extracts of the Chinese medicine composition.

[0159] a. RAW264.7 macrophages and DMEM medium were plated in 96-well plates and incubated in a carbon dioxide cell culture incubator for 24 h;

[0160] b. Discard the supernatant of the 96-well plate, co-culture with 1 μg / mL lipopolysaccharide (LPS) and FDC mixed solution (25, 50, 100, 200 μg / mL), co-culture with 1 μg / mL lipopolysaccharide (LPS) and DMEM medium, and culture with only DMEM in a carbon dioxide-based cell culture incubator for 24 hours;

[0161] c. After incubation for 24 h, the supernatant of the 96-well plate was discarded, and the concentration of nitric oxide in RAW264.7 macrophages was detected using a nitric oxide detection kit;

[0162] The results are as follows Figure 7 As shown, the inflammatory model was successfully established by stimulating RAW264.7 macrophages with 1 μg / mL lipopolysaccharide (LPS). The concentration of nitric oxide in macrophages RAW264.7 was measured with different concentrations of the Chinese herbal composition extract (FDC). When the concentration of the Chinese herbal composition extract (FDC) was 250 μg / mL, the concentration of nitric oxide in macrophages was significantly reduced. As the concentration of the Chinese herbal composition extract (FDC) increased, the concentration of nitric oxide in macrophages also decreased significantly. When the concentration of the Chinese herbal composition extract (FDC) was 1 mg / mL, the concentration of nitric oxide in macrophages was extremely significantly reduced. This shows that the Chinese herbal composition extract can inhibit the occurrence of inflammatory reactions in macrophages through the concentration of nitric oxide.

[0163] Test Example 3

[0164] This test case combines Figure 8 The xanthine oxidase inhibitory activity of the traditional Chinese medicine composition extract (FDC) prepared in Example 1 was determined. The FDC prepared in Example 1 was dissolved in deionized water to obtain FDC mixed solutions with concentrations of 0.2, 0.4, 0.8 and 1 mg / mL for determination.

[0165] The extract of the Chinese medicine composition was tested for xanthine oxidase inhibition activity, using xanthine as a substrate and allopurinol as a positive control, and a 96-well plate was used for activity testing;

[0166] a. Sample group: 100 μL sample + 40 μL xanthine oxidase;

[0167] b. Blank group, take 140 μL of PBS;

[0168] c. Control group, allopurinol 100 μL + xanthine oxidase 40 μL;

[0169] After incubation at 37°C for 3 min, 60 μL of substrate was added and the absorbance was measured at 295 nm (every 30 s) for a total of 10 min. The inhibition rate was calculated as follows:

[0170] Inhibition rate (%) = [1-(△ sample / -△ blank group)] × 100%.

[0171] The results are as follows Figure 8 As shown, the xanthine oxidase inhibitory activity of the extract of the Chinese medicine composition was determined at different concentrations, and allopurinol was used as a positive control. The xanthine oxidase inhibition rate of the extract of the Chinese medicine composition increased with the increase of concentration at a concentration gradient of 0.2 to 1 mg / L. At a concentration of 1 mg / mL, the xanthine oxidase inhibition rate of the extract of the Chinese medicine composition was close to that of allopurinol, indicating that the Chinese medicine composition has a relatively excellent xanthine oxidase inhibitory activity.

[0172] Test Example 4

[0173] This test case combines Figure 9 The anti-ultraviolet performance of the traditional Chinese medicine composition extract (FDC) prepared in Example 1 was determined.

[0174] The transmittance of the extracts of the Chinese herbal composition at different concentrations within a wavelength range of 200-400 nm was measured by ultraviolet spectrophotometry to evaluate the sunscreen performance of the extracts of the Chinese herbal composition. The transmittance of the samples was measured at 280, 290, 300, 310 and 320 nm in the UVB region, and at 320, 330, 340, 350, 360, 370, 380, 390 and 400 nm in the UVA region.

[0175] UV absorption formula: Absorption = (1-transmittance) × 100%;

[0176] The results are as follows Figure 9As shown, the extract of the Chinese herbal composition is divided into different concentrations for anti-ultraviolet measurement. As the concentration of the extract of the Chinese herbal composition increases, its absorbance in the wavelength range of 280-400nm gradually increases, and the absorbance of the same concentration decreases as the wave number increases. In the UVB section at a wavelength of 320nm, the absorbance of 1mg / mL of the Chinese herbal composition extract reaches 99.51%. In the UVA section at a wavelength of 400nm, the absorbance of 1mg / mL of the Chinese herbal composition extract reaches 65.15%. From the results of the anti-ultraviolet measurement of the extract of the Chinese herbal composition, it is known that the extract of the Chinese herbal composition has excellent anti-ultraviolet ability.

[0177] In summary, the Chinese medicine composition extract (FDC) provided by the present invention has excellent antioxidant, anti-inflammatory, anti-ultraviolet and xanthine oxidase inhibitory abilities, and there is a synergistic effect between the important components in the Chinese medicine composition, resulting in stronger antioxidant and anti-inflammatory properties.

[0178] The above description of the embodiments is to facilitate the understanding and use of the invention by those skilled in the art. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative work. Therefore, the present invention is not limited to the above embodiments, and improvements and modifications made by those skilled in the art based on the disclosure of the present invention without departing from the scope of the present invention should be within the scope of protection of the present invention.

Claims

1. A Chinese medicine composition with anti-inflammatory and antioxidant effects, characterized in that: The invention comprises the following raw materials and / or extracts of the following raw materials, in parts by mass: 8-12 parts of black ants, 8-12 parts of Qinpi, 4-6 parts of Dihuangyuan, 2-4 parts of Polygonum cuspidatum, 2-4 parts of Hexagonal Fructus, 2-4 parts of Chuanxiong, 1-3 parts of Red Medicine, 1-3 parts of Cassia Twig, and 0.5-2 parts of Licorice.

2. A Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 1, characterized in that: The invention comprises the following raw materials and / or extracts of the following raw materials, in parts by mass: 10 parts of black ants, 10 parts of Qinpi, 5 parts of Dihuangyuan, 3 parts of Polygonum cuspidatum, 3 parts of Hexagonal Fructus, 3 parts of Chuanxiong, 2 parts of Chiyao, 2 parts of Cinnamomum cassia, and 1 part of Licorice.

3. A method for preparing the Chinese medicine composition with anti-inflammatory and antioxidant effects as claimed in claim 1, characterized in that: The raw materials of the Chinese medicine composition, including black ants, qinpi, baixie, polygonum cuspidatum, hexapod seeds, chuanxiong, red scutellaria, cassia twig and liquorice, are dried, crushed, sieved, extracted and separated to obtain a Chinese medicine composition with anti-inflammatory and antioxidant effects.

4. The method for preparing a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 3, characterized in that: The raw materials of the Chinese medicine composition are crushed to a size not exceeding 80 meshes.

5. The method for preparing a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 3, characterized in that: The sieved Chinese medicine composition raw material is mixed with the organic solvent in proportion, and then ultrasonicated to obtain the Chinese medicine composition extract. The ultrasonication time is 100-150 minutes and the temperature is 40-60°C.

6. The method for preparing a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 5, characterized in that: The mixing ratio of the raw materials of the traditional Chinese medicine composition and the organic solvent is 1g: (3-6)mL.

7. The method for preparing a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 3, characterized in that: The separation includes centrifugation, concentration and freeze-drying.

8. The method for preparing a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 7, characterized in that: The centrifugal speed is 8000-12000 r / min, and the time is 8-12 min; the concentration is concentrated to 0.25-20% of the original volume; the freeze-drying temperature is -60--40°C, and the time is 24-48 h.

9. Use of the Chinese medicine composition with anti-inflammatory and antioxidant effects as claimed in claim 1 in the fields of cosmetics and health products.

10. The use of a Chinese medicine composition with anti-inflammatory and antioxidant effects according to claim 9, characterized in that: The applications include anti-oxidation, anti-inflammatory and anti-ultraviolet applications.

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