A wormwood extract with antioxidant and hyaluronidase activity inhibition function and its preparation method and application

Abstract

The present invention relates to the technical field of plant extracts, and specifically discloses a mugwort extract having both antioxidant and hyaluronidase activity inhibition, wherein the mugwort extract is extracted by drying and crushing mugwort tips with ethanol / water as an extraction solvent. A preparation method of a mugwort extract having both antioxidant and hyaluronidase activity inhibition, wherein mugwort powder is mixed with an extraction solvent, wherein the extraction solvent is an ethanol solution of 30%-70% or deionized water, and the mixture is extracted at an extraction temperature of 50-80°C, and the mixture is centrifuged after extraction, and the supernatant obtained by centrifugation is the mugwort extract. An application of a mugwort extract having both antioxidant and hyaluronidase activity inhibition, wherein the mugwort extract is used to be added to cosmetics. This scheme is used to solve the problem that the current mugwort extract has low hyaluronidase inhibition rate and low antioxidant activity to DPPH, while taking into account the high efficiency and simplicity of preparation.

Description

Technical Field

[0001] The present invention relates to the field of plant extracts, and in particular to an wormwood extract having the functions of anti-oxidation and inhibition of hyaluronidase activity, and a preparation method and application thereof. Background Art

[0002] With the advancement of modern technology and people's pursuit of health and beauty, the application of natural plant extracts in the cosmetics industry is becoming more and more extensive. As a traditional Chinese medicinal material, mugwort has rich medicinal value, and its application in the cosmetics field has also attracted much attention in recent years.

[0003] Mugwort, also known as Artemisia argyi, is a common Chinese herbal medicine with multiple biological activities such as antibacterial, antiviral, and antioxidant. Traditionally, mugwort has been widely used in the field of traditional Chinese medicine to treat various diseases. In recent years, with the rapid development of the cosmetics industry, mugwort extracts have gradually been favored by the cosmetics industry due to their natural, safe, and effective characteristics. At present, there are few applications of mugwort extracts in cosmetics on the market. In patent data, some studies on the use of mugwort extracts in cosmetics have been reflected.

[0004] For example, the patent publication number CN113368164A discloses a Chinese herbal formula fermentation product for anti-allergic purposes and a preparation method thereof and its application in cosmetics. In this technology, it is mentioned that a Chinese herbal formula fermentation product is prepared by combining Centella asiatica, Artemisia argyi and Gardenia jasminoides, which can achieve a hyaluronidase inhibition rate and a DPPH scavenging ability of more than 90%. However, the current preparation method of the Chinese herbal formula fermentation product is very complicated, and it is necessary to prepare a fermentation medium obtained by combining a Chinese herbal medicine composition with skimmed milk powder, honey and glucose, and to prepare a specific yeast liquid, and then to inoculate the yeast liquid into the fermentation medium. After 3-4 days, the Chinese herbal formula fermentation filtrate is obtained. The process is extremely complicated, and the hyaluronidase inhibition rate and DPPH scavenging ability of the Artemisia argyi fermentation liquid alone are both less than 60%.

[0005] In other disclosed patent technologies, for example, the patent announcement number CN115737754B is a Chinese medicine composition with dual anti-allergic and anti-inflammatory effects and its use in skin care products. This technology also makes use of mugwort. The Chinese medicine composition contains Ligustrum lucidum extract, Dendrobium nobile extract, mugwort extract and tea extract. The hyaluronidase inhibition rate of the Chinese medicine composition can reach up to about 95%, while the hyaluronidase inhibition rate of the mugwort extract after treatment using only 75% ethanol as the extraction solvent is less than 60%.

[0006] Artemisia argyi is also used in the technology of "A compound anti-acne composition and its application" with patent publication number CN118477025A, and it is pointed out that the chemical components of artemisia argyi extract mainly include volatile oil, flavonoids, eucalyptone and triterpenoid components. However, the current artemisia argyi extract in this composition has a certain inhibitory effect on hyaluronidase, but the hyaluronidase inhibition rate is less than 60%, and the hyaluronidase inhibition rate of the artemisia argyi extract disclosed in this technology is less than 20%.

[0007] It can be seen that in current applications, the hyaluronidase inhibition rate and antioxidant activity against DPPH of Artemisia argyi extract are both low. It can only be combined with other traditional Chinese medicines and a complex processing process to achieve a balance between hyaluronidase inhibition rate and antioxidant activity against DPPH. Summary of the invention

[0008] The present invention aims to provide an wormwood extract having both antioxidant and hyaluronidase inhibitory properties and a preparation method, so as to solve the problem that the current wormwood extract has low hyaluronidase inhibition rate and low antioxidant activity against DPPH, while taking into account the high efficiency and simplicity of preparation.

[0009] In order to achieve the above object, the present invention adopts the following technical scheme:

[0010] The invention discloses an wormwood extract having the functions of anti-oxidation and inhibition of hyaluronidase activity. The wormwood extract is obtained by drying and crushing the wormwood tips and then extracting them using ethanol / water as extraction solvent.

[0011] The present invention also provides a method for preparing an wormwood extract having both antioxidant and hyaluronidase activity inhibition properties. The wormwood powder made from wormwood tips is mixed with an extraction solvent, wherein the extraction solvent is a 30%-70% ethanol solution or deionized water. After mixing, extraction is performed at an extraction temperature of 50-80°C, and centrifugation is performed after extraction. The supernatant obtained by centrifugation is the wormwood extract.

[0012] Preferably, as an improvement, the solid-liquid ratio of wormwood powder to extraction solvent is 1:20.

[0013] Preferably, as an improvement, the preparation of mugwort powder is: take the tips of mugwort ten days before and after Qingming Festival, dry them at 50°C, and then grind them into mugwort powder.

[0014] Beneficial effects: In traditional applications, mugwort extracts are used in cosmetics and other products. These mugwort leaves are usually taken from long-term stored mugwort leaves. Since old mugwort is not restricted by time or season, and its smell is mild and mellow, and its medicinal properties are stable, researchers prefer to use mugwort leaves from old mugwort as the research object. In addition, according to traditional Chinese medicine theory, "seven years of illness, three years of mugwort", it is believed that mugwort stored for more than three years has the best effect, which further consolidates the status of old mugwort in traditional applications.

[0015] However, the present invention breaks this inherent thinking and uses the tips of wormwood before and after Qingming as the raw material of the extract for the first time; on the one hand, the tips of wormwood before and after Qingming belong to fresh wormwood, and its active ingredients such as flavonoids, polysaccharides and polyphenols are significantly higher. The flavonoid content in the Qingming wormwood of the present invention is 65-75mg / g, the polysaccharide content is 60-70mg / g, and the polyphenol content is 10-20mg / g, and these components will slowly decrease with the extension of growth time, and essential oils and volatile components will increase; in terms of specific component proportions, wormwood varieties of different varieties in different regions will also be different. The present invention uses these wormwood tips rich in active substances to greatly improve the hyaluronidase inhibition rate and the antioxidant activity against DPPH free radicals in the obtained wormwood extract.

[0016] On the other hand, pinching the tips of mugwort around the Qingming Festival will not affect the growth of mugwort in the later period and the yield of old mugwort, but will promote the rapid growth of mugwort. For farmers who grow mugwort, this is equivalent to one more harvesting opportunity in a growing period, greatly increasing economic income. Preferably, as an improvement, after the mugwort powder is mixed with the extraction solution, the container containing the mixture is sealed to avoid evaporation.

[0017] Preferably, as an improvement, the prepared wormwood extract is vacuum freeze-dried for preservation, and then diluted to the required concentration with an extraction solvent of corresponding concentration before use.

[0018] Preferably, as an improvement, when an ethanol solution is used as the extraction solvent, when the ethanol concentration is 40%, the extraction temperature is 60°C, and the inhibition rate of the obtained wormwood extract on hyaluronidase is 87.2%±3.66.

[0019] Preferably, as an improvement, when an ethanol solution is used as the extraction solvent, when the ethanol concentration is 60%, the extraction temperature is 60° C., the DPPH oxidation resistance of the obtained wormwood extract is 95.94%±3.52.

[0020] Preferably, as an improvement, when deionized water is used as the extraction solvent, the extraction temperature is 60-70° C., and the inhibition rate of the obtained wormwood extract on hyaluronidase is not less than 90%.

[0021] Preferably, as an improvement, when deionized water is used as the extraction solvent, the extraction temperature is 50-60° C., and the DPPH oxidation resistance of the obtained wormwood extract is not less than 85%.

[0022] The present invention also provides an application of an wormwood extract having both antioxidant and hyaluronidase activity inhibition properties, wherein the wormwood extract is added to cosmetics.

[0023] The present invention uses two processes of extracting mugwort with ethanol and extracting mugwort with deionized water, effectively retaining the active ingredients in mugwort and giving it anti-allergic and antioxidant effects. The mugwort extract of the present invention can take into account both the hyaluronidase inhibition rate and the antioxidant activity against DPPH, and the ethanol extract of mugwort is very significant in antioxidant performance. When the extraction temperature is 60°C and the ethanol concentration used during extraction is 60%, the anti-DPPH oxidation capacity can be as high as 95%. The mugwort water extract obtained by extraction with deionized water also has very good antioxidant performance, and is most significant in the inhibition rate of hyaluronic acid plum, which can be as high as 94%.

[0024] The present invention breaks the traditional method of extracting wormwood, greatly improving the anti-allergic and antioxidant properties of the wormwood extract. In addition, the entire production method has low material costs, and the production process is environmentally friendly, efficient, simple and convenient. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a bar graph of the in vitro anti-allergic effects of ethanol-wormwood extracts of different concentrations at an extraction temperature of 60° C. in the first test group of the embodiment of the present invention.

[0026] Figure 2 This is a bar graph of the in vitro anti-allergic effects of the wormwood extract of the second test group of the embodiment of the present invention at different extraction temperatures in 40% ethanol.

[0027] Figure 3 This is a bar graph showing the anti-DPPH oxidation capacity of ethanol-wormwood extracts of different concentrations at an extraction temperature of 60° C. in the third test group of the invention example.

[0028] Figure 4 This is a bar graph of the in vitro anti-allergic effect of the wormwood water extract in the fourth test group of the invention embodiment at different extraction temperatures.

[0029] Figure 5 This is a bar chart of the anti-DPPH oxidation ability of the wormwood water extract at different extraction temperatures in the fourth test group of the invention example. DETAILED DESCRIPTION

[0030] The following is further described in detail through specific implementation methods:

[0031] Example

[0032] The invention discloses an wormwood extract having antioxidant and hyaluronidase inhibitory effects. The wormwood tips collected ten days before and after the Qingming Festival are dried and crushed, and then extracted in a water bath heating environment using ethanol / water as the extraction solvent.

[0033] A specific method for preparing an wormwood extract having both antioxidant and hyaluronidase activity inhibition properties comprises the following steps:

[0034] S1. Drying and crushing of mugwort: pinch the tips of mugwort ten days before and after Qingming Festival, dry them at 50℃ for 10h~12h, and then grind them into mugwort powder with a Chinese medicine grinder for later use. The mugwort variety is a local variety cultivated by Chongqing Yuanxiang Xiaoai Technology Co., Ltd. at the company's planting base in Tongliang District, Chongqing; the flavonoid content of mugwort ten days before and after Qingming Festival is 65-75mg / g, the polysaccharide content is 60-70mg / g, and the polyphenol content is 10-20mg / g.

[0035] S2, ethanol extraction: Take the wormwood powder of S1 and add 30%-70% ethanol solution in a conical flask according to the solid-liquid ratio of 1:20, stir evenly and seal to avoid evaporation, then heat it in a constant temperature water bath to make the extraction temperature at 50-80°C, centrifuge after extraction, take the supernatant and centrifuge it again under the same conditions to further remove the insoluble matter, and the supernatant is the ethanol extract of wormwood. The ethanol extract of wormwood is vacuum freeze-dried for preservation, and diluted to the required concentration with ethanol of the corresponding concentration before use.

[0036] S3, water extraction: Take the wormwood powder of S1 and add an appropriate amount of deionized water in a triangular flask according to the solid-liquid ratio of 1:20, stir evenly and seal to avoid evaporation, then heat it in a constant temperature water bath to make the extraction temperature at 50-80°C, centrifuge after extraction, take the supernatant and centrifuge it again under the same conditions, and the supernatant is the water extract of wormwood. The water extract of wormwood is vacuum freeze-dried for storage, and diluted to the required concentration with deionized water of corresponding concentration before use.

[0037] In order to verify the hyaluronidase inhibition rate and antioxidant activity of the ethanol extract of wormwood at different concentrations and temperatures against DPPH, the following first test group was designed. In the first test group, in order to verify the in vitro anti-allergic activity of wormwood extracted with ethanol at different concentrations (hyaluronidase is a hydrolase that degrades hyaluronic acid and is the main component of the extracellular matrix of the body's connective tissue. It is related to most IgE-mediated type I and T-cell-mediated type IV allergic reactions. Therefore, the in vitro inhibition experiment of hyaluronidase is an effective method for rapid in vitro screening of anti-allergic drugs), in the first test group, the ethanol solution is set to 30%, 40%, 50%, 60%, and 70% five gradients (the percentages in this embodiment are all volume percentages), extracted at an extraction temperature of 70°C in a constant temperature water bath for 1h, and then centrifuged at 4000rpm for 3 minutes, the supernatant was taken and centrifuged again under the same conditions, and the supernatant was obtained. The ethanol extract of wormwood obtained in the first test group has an inhibition rate of hyaluronidase as shown in Tables 1 and Figure 1 shown.

[0038] In the second test group, 40% ethanol solution was used for extraction under different temperature gradients. The difference between the second test group and the first test group was that the ethanol solution used during extraction was limited to a fixed value of 40%, while the extraction temperature was varied. Three different test cases of 50°C, 60°C, and 70°C were set respectively. The hyaluronidase inhibition rate test was also carried out. The test results are shown in Tables 2 and 3. Figure 2 shown.

[0039] Table 1 In vitro anti-allergic effects of 70°C extraction temperature-different concentrations of ethanol-wormwood extracts in the first test group

[0040] Ethanol concentration (%) Mean inhibition rate % 30 76.77±6.36 40 80.9±3.18 50 71.1±3.11 60 65.2±3.02 70 47.5±7.81

[0041] Table 2 In vitro anti-allergic effects of wormwood extracts at different extraction temperatures with 40% ethanol in the second test group

[0042] Extraction temperature ℃ Inhibition rate % 50 74±7.72 60 87.2±3.66 70 80.1±7.28

[0043] From Table 1 and Figure 1 It can be seen that the extract obtained by extracting mugwort with 40% ethanol at a constant temperature of 70°C for 1 hour has the ability to inhibit hyaluronidase activity by 80.9±3.18%. The other concentrations of mugwort ethanol extracts have different degrees of inhibition on hyaluronidase, ranging from 47.5±7.81% to 76.77±6.36%.

[0044] From Table 2 and Figure 2 Among the extracts obtained by extracting mugwort with 40% ethanol at different temperatures for 1h, the 60°C extract had an inhibition degree of 87.2±3.66% on hyaluronidase. It can be seen that 40% ethanol concentration as a solvent and 60°C extraction is conducive to the extraction of anti-allergic substances, and increasing or decreasing the temperature has different effects on the extraction degree of anti-allergic substances in mugwort. In the test example in Table 2, the flavonoids concentration in the ethanol extract of mugwort using 40% ethanol at an extraction temperature of 60°C is 19.4 mg / g, the polyphenol concentration is 8.8 mg / g, the polysaccharide concentration is 50.4 mg / g, and the terpenoid concentration is 1612 mg / g.

[0045] In order to test the anti-DPPH oxidation ability of ethanol extract of wormwood, a third test group was designed. The difference between this test group and the first test group was that the extraction temperature was 70°C. The ethanol concentrations used in the sample extraction of the test group were different, namely 30%, 40%, 50%, 60%, and 70%. The anti-DPPH oxidation ability of wormwood extracts with different ethanol concentrations was tested and recorded, and the results were obtained in Tables 3 and Figure 3 .

[0046] Table 3 The anti-DPPH oxidation ability of ethanol-wormwood extracts of different concentrations at 60℃ extraction temperature in the third test group

[0047] Ethanol concentration% Antioxidant % 30 82.76±2.85 40 78.45±6.73 50 85.75±5.56 60 95.94±3.52 70 91.20±3.36

[0048] From Table 3 and Figure 3 It can be seen that the extracts of mugwort extracted at 60℃ for 1h with different ethanol concentrations showed strong resistance to DPPH oxidation, reaching 78.45±6.73% to 95.94±3.52%, among which the 60% ethanol extract had the strongest antioxidant capacity against DPPH.

[0049] In order to verify the hyaluronidase inhibition ability and anti-DPPH oxidation ability of the wormwood water extract under different extraction temperatures under the water extraction method, the following fourth test group was set up. In the fourth test group, the extraction temperature was changed, and four different test cases of 50°C, 60°C, 70°C, and 80°C were set respectively. The four test cases were extracted in a constant temperature water bath at the corresponding temperature for 1h, centrifuged at 4000rpm for 3 minutes, and the supernatant was taken and centrifuged again under the same conditions to obtain the wormwood water extract. The inhibition rate of the wormwood water extract obtained under the fourth test group on hyaluronidase is shown in Tables 4 and Figure 4 As shown in Table 5 and Figure 5 shown.

[0050] Table 4 In vitro anti-allergic effects of wormwood water extract at different extraction temperatures in the fourth experimental group

[0051]

[0052]

[0053] Combining Table 4 and Figure 4 It can be seen that the water extracts obtained by extracting mugwort with deionized water at different temperatures showed good in vitro anti-allergic activity, and the ability to inhibit hyaluronidase ranged from 79.4±3.87% to 94.67±2.24%, among which the inhibition rates at 60°C and 70°C exceeded 90%.

[0054] Table 5 The anti-DPPH oxidation ability of the water extract of Artemisia argyi at different extraction temperatures in the fourth experimental group

[0055] Extraction temperature ℃ Antioxidant % 50 88.56±2.07 60 87.85±2.88 70 74.98±5.68 80 72.46±3.02

[0056] Among the test examples in Table 4 and Table 5, at the extraction temperature of 60°C, the flavonoids concentration in the obtained mugwort water extract was 17.8 mg / g, the polyphenol concentration was 8.0 mg / g, the polysaccharide concentration was 39.4 mg / g, and the terpenoid concentration was 1223.5 mg / g.

[0057] From Table 5 and Figure 5It can be seen that the antioxidant capacity of the mugwort water extract decreases with the increase of temperature after the temperature exceeds 50°C, but the overall anti-DPPH oxidation capacity remains in the range of 72.46±3.02% to 88.56±2.07%.

[0058] It can be seen from the above test group that whether the present invention uses ethanol or deionized water to extract wormwood, the obtained extract has a strong hyaluronidase inhibition rate and antioxidant activity, achieving the effect of both parties. The ethanol extract of wormwood is very significant in antioxidant performance, especially at an extraction temperature of 60°C and an ethanol concentration of 60% during extraction, the anti-DPPH oxidation ability can be as high as 95%; when the ethanol concentration used in the extraction is 50% and the extraction temperature is controlled at about 60°C, the obtained wormwood ethanol extract has a hyaluronic acid plum The inhibition rate can exceed 71%, and the antioxidant performance can be around 85%. The extract obtained by extracting at an ethanol concentration of 40% and an extraction temperature of 60°C can have a hyaluronidase inhibition rate of up to 87.2±3.66%, and an anti-DPPH oxidation capacity of up to 78.45%±6.73%. That is, under the selection of this ethanol concentration and extraction temperature, the hyaluronidase inhibition rate and the anti-DPPH oxidation capacity are both around 80%, achieving a balance between the two effects. Compared with the prior art wormwood extract in which both effects are less than 60%, a substantial improvement has been achieved.

[0059] The anti-oxidant property of the wormwood water extract obtained by deionized water extraction is also very good, with the most significant inhibition rate of hyaluronic acid plum reaching 94%. The wormwood water extract obtained at an extraction temperature of 60°C can achieve a hyaluronidase inhibition rate of about 93%, and an anti-oxidant property of about 87%.

[0060] The present invention uses deionized water or ethanol to extract wormwood, thereby retaining the active ingredients in the wormwood, not only obtaining extremely high antioxidant and anti-allergic properties, but also having low raw material costs and high extraction efficiency, and the production process is also efficient, simple, convenient and environmentally friendly, and has extremely important practical significance and application value.

[0061] To ensure the performance of the present invention, the wormwood extract was also subjected to safety evaluations such as acute toxicity and skin irritation. The results showed that the wormwood extract obtained by the present invention had no obvious toxicity, no irritation to the skin, and was safe and reliable.

[0062] The present invention also provides an application of an wormwood extract that has both antioxidant and hyaluronidase activity inhibition properties. The wormwood ethanol extract obtained by the above-mentioned ethanol extraction method and / or the wormwood water extract obtained by the deionized water extraction method are added to cosmetics. When adding, the wormwood ethanol extract and the wormwood water extract are mixed in the required proportion according to needs.

[0063] The above is only an embodiment of the present invention, and the common knowledge such as the known specific technical solutions and / or characteristics in the solution is not described in detail here. It should be pointed out that for those skilled in the art, without departing from the technical solution of the present invention, several modifications and improvements can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A mugwort extract having both antioxidant and hyaluronidase inhibitory properties, characterized in that: The wormwood extract is obtained by drying and crushing the wormwood tips and then extracting them using ethanol / water as an extraction solvent.

2. A method for preparing an wormwood extract having both antioxidant and hyaluronidase inhibitory properties, characterized in that: The wormwood powder made from wormwood tips is mixed with an extraction solvent, which is a 30%-70% ethanol solution or deionized water. After mixing, extraction is performed at an extraction temperature of 50-80°C, and centrifugation is performed after extraction. The supernatant obtained by centrifugation is the wormwood extract.

3. The method for preparing a wormwood extract having both antioxidant and hyaluronidase inhibitory properties according to claim 2, characterized in that: The solid-liquid ratio of wormwood powder to extraction solvent is 1:

20.

4. The method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to claim 2, characterized in that: The preparation method of mugwort powder is as follows: take mugwort tips ten days before and after Qingming Festival, dry them at 50°C, and then grind them into mugwort powder.

5. The method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to claim 2, characterized in that: The prepared wormwood extract is vacuum freeze-dried for preservation, and is diluted to a required concentration with an extraction solvent of corresponding concentration before use.

6. A method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to any one of claims 2 to 5, characterized in that: When ethanol solution is used as the extraction solvent, the ethanol concentration is 40%, and the extraction temperature is 60° C., the inhibition rate of the obtained wormwood extract on hyaluronidase is 87.2%±3.

66.

7. A method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to any one of claims 2 to 5, characterized in that: When ethanol solution is used as the extraction solvent, the ethanol concentration is 60%, and the extraction temperature is 60°C, the DPPH oxidation resistance of the obtained wormwood extract is 95.94%±3.

52.

8. A method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to any one of claims 2 to 5, characterized in that: When deionized water is used as the extraction solvent, the extraction temperature is 60-70° C., and the inhibition rate of the obtained wormwood extract on hyaluronidase is not less than 90%.

9. A method for preparing a mugwort extract having both antioxidant and hyaluronidase inhibitory properties according to any one of claims 2 to 5, characterized in that: When deionized water is used as the extraction solvent, the extraction temperature is 50-60° C., and the DPPH oxidation resistance of the obtained wormwood extract is not less than 85%.

10. An application of an wormwood extract having both antioxidant and hyaluronidase activity inhibition properties, characterized in that: The wormwood extract according to any one of claims 1 to 9 is used for adding into cosmetics.

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