Preparation method of pine mushroom extract with soothing and repairing effects, product and application thereof

Abstract

The present invention relates to a preparation method of a pine mushroom extract with soothing and repairing effects, and its products and applications. The preparation method comprises the following steps: mixing pine mushroom superfine powder with a low eutectic solvent and a complex enzyme for extraction to obtain an extraction product; centrifuging the extraction product to obtain a supernatant; eluting the supernatant through a macroporous resin to obtain the pine mushroom extract. The preparation method involved in the present invention creatively uses pine mushroom superfine powder as an extraction raw material, adopts a low eutectic solvent and a complex enzyme co-extraction method, and cooperates with subsequent purification operations, so that the obtained pine mushroom extract product has excellent soothing and repairing effects, which is specifically reflected in the ability to significantly inhibit hyaluronidase activity and the aggregation of zebrafish embryonic neutrophils.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and relates to a preparation method of a pine mushroom extract with soothing and repairing effects, a product thereof and an application thereof, and specifically relates to a preparation method of a pine mushroom extract with soothing and repairing effects, a product thereof and an application thereof in preparing a product with soothing and repairing effects. Background Art

[0002] Skin sensitivity is becoming more and more common among the population. Harmful substances in the environment, such as dust, chemicals and heavy metals, constantly attack people's skin, causing damage to the skin barrier. At the same time, bad living habits such as staying up late, unhealthy diet and lack of exercise also aggravate the occurrence of skin sensitivity problems. Skin sensitivity symptoms are often more troublesome. Slight stimulation or touch may cause skin discomfort, such as tingling, redness, dryness and itching. These symptoms are not only related to people's appearance, but also have a significant impact on their daily life and mood. In order to cope with this problem, cosmetic raw materials for sensitive skin care are also being continuously researched and developed on the market. Medicinal and edible plants that are both safe and soothing and repairing are therefore widely used in sensitive skin care products, aiming to help sensitive skin return to a healthy state and relieve discomfort.

[0003] One of the main causes of skin sensitivity is the massive production of inflammatory factors. Matsutake is one of the natural medicinal mushrooms. Due to its high nutritional value and medicinal value, it is known as the "king of edible fungi". It has been used as medicine since ancient times. "Shennong's Herbal Classic" records that "Matsutake is sweet, flat and non-toxic. It mainly treats urinary problems, replenishes the five internal organs, replenishes qi and blood, consolidates and produces body fluids, calms the mind and soothes the nerves. Long-term use can make the body lighter and increase qi and strength. It is also called fairy mushroom". According to modern scientific research, Matsutake extract can inhibit the expression of inflammatory factors and inflammation-related proteins by blocking the upstream signaling pathway of mitogen-activated protein kinase (MAPK), further inhibiting the activation of nuclear factor κB (NF-κB) pathway. However, the current research on Matsutake extract in the existing technology mainly focuses on whitening effects, and there are few reports on soothing and repairing skin, especially on the preparation process of Matsutake extract. Therefore, it is a direction worth exploring to develop a preparation process of Matsutake extract with efficient soothing and repairing effects, and apply the resulting product to the soothing and repairing effects of the skin, which will help to bring better solutions for the improvement of sensitive skin. Summary of the invention

[0004] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a method for preparing a pine mushroom extract with soothing and repairing effects, its products and applications, and specifically to provide a method for preparing a pine mushroom extract with soothing and repairing effects, its products and applications in the preparation of products with soothing and repairing effects.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a method for preparing a Tricholoma matsutake extract having soothing and repairing effects, the preparation method comprising the following steps:

[0007] The ultrafine powder of Tricholoma matsutake is mixed with a low eutectic solvent and a complex enzyme for extraction to obtain an extraction product; the extraction product is centrifuged to obtain a supernatant; the supernatant is passed through a macroporous resin for elution to obtain the Tricholoma matsutake extract.

[0008] The preparation method involved in the present invention creatively uses superfine matsutake powder as the extraction raw material, adopts the method of co-extraction with a low eutectic solvent and a complex enzyme, and cooperates with subsequent purification operations, so that the obtained matsutake extract product has excellent soothing and repairing effects, which is specifically reflected in the ability to significantly inhibit hyaluronidase activity and the aggregation of zebrafish embryonic neutrophils.

[0009] Preferably, the raw materials for preparing the deep eutectic solvent are thymol and lauryl alcohol.

[0010] The low eutectic solvent used in the preparation method of the present invention is more preferably a low eutectic solvent obtained by reacting thymol and lauryl alcohol as preparation raw materials. Compared with other types of low eutectic solvents, the product extracted therefrom has better soothing and repairing effects.

[0011] Preferably, the low eutectic solvent is prepared by reacting thymol and lauryl alcohol as raw materials at 80-120°C (e.g., 80°C, 85°C, 90°C, 95°C, 100°C, 105°C, 110°C, 115°C, 120°C, etc.) for 1-5h (e.g., 1h, 1.5h, 2h, 2.5h, 3h, 3.5h, 4h, 5h, etc.).

[0012] Preferably, the molar ratio of thymol to lauryl alcohol is 1:(1-3), for example 1:1, 2:3, 1:2, 2:5, 1:3, etc.

[0013] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0014] Preferably, the complex enzyme is a combination of cellulase, pectinase and papain.

[0015] The composite enzyme used in the preparation method of the present invention is more preferably a combination of cellulase, pectinase and papain. Compared with other types of single enzymes or enzyme combinations, the product extracted by the composite enzyme has a better effect in soothing and repairing. Cellulase, pectinase and papain have a potential synergistic promotion relationship in improving the soothing effect of the product.

[0016] Preferably, the total enzyme activity of the complex enzyme is 30000-50000 U / g, for example, 30000 U / g, 35000 U / g, 40000 U / g, 45000 U / g, 48000 U / g, 50000 U / g, etc.

[0017] Preferably, the enzymatic activity ratio of the cellulase, pectinase and papain is (1-3):(1-3):(1-3), wherein the specific point values ​​in (1-3) can be selected from 1, 1.5, 2, 2.5, 3, etc.

[0018] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0019] Preferably, the ratio of the superfine matsutake powder to the low eutectic solvent is 1g:(100-200)mL, for example, 1g:100mL, 1g:120mL, 1g:150mL, 1g:160mL, 1g:180mL, 1g:200mL, etc.

[0020] Preferably, the concentration of the complex enzyme in the extraction system is 0.2-0.8 mg / mL, for example, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, etc.

[0021] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0022] Preferably, the preparation method of the superfine Matsutake powder comprises: slicing and drying the Matsutake fruiting body, freezing the slices, and then crushing them into particle sizes of 10-30 μm (e.g., 10 μm, 15 μm, 18 μm, 20 μm, 22 μm, 25 μm, 30 μm, etc.).

[0023] Preferably, the leaching is carried out at 35-55°C (e.g., 35°C, 38°C, 40°C, 42°C, 45°C, 48°C, 50°C, 55°C, etc.) for 0.5-3h (e.g., 0.5h, 1h, 1.5h, 2h, 2.5h, 3h, etc.).

[0024] Preferably, ultrasound-assisted extraction is also performed during the leaching process.

[0025] The use of ultrasound-assisted extraction during the extraction process can further enhance the soothing and repairing effects of the product.

[0026] Preferably, the power of the ultrasound-assisted extraction is 100-300W, for example, 100W, 150W, 200W, 250W, 300W, etc.

[0027] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0028] Preferably, the centrifugation is performed at 5000-10000 r / min (e.g. 5000 r / min, 6000 r / min, 7000 r / min, 8000 r / min, 9000 r / min, 10000 r / min, etc.) for 10-20 min (e.g. 10 min, 12 min, 15 min, 18 min, 20 min, etc.).

[0029] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0030] Preferably, the eluent used for elution is an ethanol aqueous solution with a gradient concentration, and the elution method is: first use 10-20% (for example, 10%, 12%, 15%, 18%, 20%, etc.) ethanol aqueous solution to elute 3-10BV (for example, 3BV, 5BV, 7BV, 8BV, 10BV, etc.), then use 30-40% (for example, 30%, 32%, 35%, 38%, 40%, etc.) ethanol aqueous solution to elute 3-10BV (for example, 3BV, 5BV, 7BV, 8BV, 10BV, etc.), and finally use 60-70% (for example, 60%, 62%, 65%, 68%, 70%, etc.) ethanol aqueous solution to elute, and collect the eluate.

[0031] The elution method used in the column chromatography in the preparation method of the present invention adopts the above-mentioned specific procedure, and the purified product has more excellent soothing and repairing effects.

[0032] Preferably, the eluate obtained by elution is concentrated and dried.

[0033] Preferably, the macroporous resin is selected from any one of D101 type, HP-20 type, X-5 type, AB-8 type or NKA-9 type.

[0034] Other specific point values ​​within the above numerical ranges can be selected and will not be described in detail here.

[0035] In a second aspect, the present invention provides a Tricholoma matsutake extract having soothing and repairing effects, which is prepared according to the preparation method described in the first aspect.

[0036] In a third aspect, the present invention provides use of the Tricholoma matsutake extract according to the second aspect in preparing a product with soothing and repairing effects.

[0037] Preferably, the product comprises a cosmetic product.

[0038] In a fourth aspect, the present invention provides use of the Tricholoma matsutake extract according to the second aspect in the preparation of a hyaluronidase inhibitor.

[0039] According to the research results of the present invention, the Matsutake extract involved in the present invention can inhibit the enzymatic activity of hyaluronidase at the in vitro level, that is, the Matsutake extract involved in the present invention can also be made into a simple experimental preparation. The above-mentioned hyaluronidase inhibitor is not used for skin soothing and repairing, but is used solely for the inhibition of hyaluronidase.

[0040] In a fifth aspect, the present invention provides use of the Tricholoma matsutake extract according to the second aspect in the preparation of a zebrafish embryo neutrophil aggregation inhibitor.

[0041] According to the research results of the present invention, the Matsutake extract involved in the present invention can inhibit the aggregation of zebrafish embryo neutrophils, that is, the Matsutake extract involved in the present invention can also be made into a simple experimental preparation. The above-mentioned inhibitor is not used for skin soothing and repairing, but is used solely for the study of the behavior of zebrafish embryo neutrophils.

[0042] Compared with the prior art, the present invention has the following beneficial effects:

[0043] The preparation method involved in the present invention creatively uses superfine matsutake powder as the extraction raw material, adopts the method of co-extraction with a low eutectic solvent and a complex enzyme, and cooperates with subsequent purification operations, so that the obtained matsutake extract product has excellent soothing and repairing effects, which is specifically reflected in the ability to significantly inhibit hyaluronidase activity and the aggregation of zebrafish embryonic neutrophils. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0045] Example 1

[0046] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof is as follows:

[0047] (1) Take Tricholoma matsutake fruiting bodies, clean them, slice them and dry them, add liquid nitrogen to freeze them for 5 min, and then grind them into a particle size of 20 μm by an ultrafine grinder to obtain ultrafine powder of Tricholoma matsutake for later use;

[0048] (2) thymol and lauryl alcohol were mixed in a molar ratio of 1:2 in a round-bottom flask, and stirred in an oil bath at 100° C. for 3 h to form a uniform and stable transparent liquid, i.e., a low eutectic solvent, which was set aside;

[0049] (3) Mixing the superfine powder of Tricholoma matsutake and the low eutectic solvent in a mass volume ratio of 1 g:150 mL, and adding 0.5 mg / mL of a composite enzyme, wherein the total enzyme activity of the composite enzyme is 40,000 U / g, and the composite enzyme is composed of cellulase, pectinase, and papain in an enzyme activity ratio of 1:1:1; placing the container containing the mixture in an ultrasonic instrument for hot extraction, and simultaneously performing ultrasonic assisted extraction at a power of 200 W at 45° C. for 1 h;

[0050] (4) The extracted product was centrifuged at 8000 r / min for 15 min, and the precipitate was discarded to obtain a supernatant;

[0051] (5) The supernatant was chromatographed on a macroporous resin AB-8 column using a water-alcohol system, first eluting with 20% ethanol aqueous solution for 5 BV, then eluting with 40% ethanol aqueous solution for 8 BV, and finally eluting with 60% ethanol aqueous solution, and the eluate was collected; the eluate was concentrated and freeze-dried to obtain a matsutake extract.

[0052] Example 2

[0053] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof is as follows:

[0054] (1) Take Tricholoma matsutake fruiting bodies, clean them, slice them and dry them, add liquid nitrogen to freeze them for 5 min, and then grind them into a particle size of 10 μm by an ultrafine grinder to obtain ultrafine powder of Tricholoma matsutake for later use;

[0055] (2) thymol and lauryl alcohol were mixed in a round-bottom flask at a molar ratio of 1:2, and stirred in an oil bath at 80° C. for 4 h until a uniform and stable transparent liquid, i.e., a deep eutectic solvent, was formed for later use;

[0056] (3) Mixing the superfine powder of Tricholoma matsutake and the low eutectic solvent in a mass volume ratio of 1 g:100 mL, and adding 0.3 mg / mL of a composite enzyme, wherein the total enzyme activity of the composite enzyme is 45000 U / g, and the composite enzyme is composed of cellulase, pectinase, and papain in an enzyme activity ratio of 1:2:2; placing the container containing the mixture in an ultrasonic instrument for hot extraction, and simultaneously performing ultrasonic assisted extraction at a power of 300 W at 50° C. for 2 h;

[0057] (4) The extracted product was centrifuged at 5000 r / min for 20 min, and the precipitate was discarded to obtain a supernatant;

[0058] (5) The supernatant was chromatographed on a macroporous resin AB-8 column using a water-alcohol system, first eluting 5 BV with a 10% ethanol aqueous solution, then eluting 8 BV with a 30% ethanol aqueous solution, and finally eluting with a 70% ethanol aqueous solution, and the eluate was collected; the eluate was concentrated and freeze-dried to obtain a matsutake extract.

[0059] Example 3

[0060] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof is as follows:

[0061] (1) Take Tricholoma matsutake fruiting bodies, clean them, slice them and dry them, add liquid nitrogen to freeze them for 5 min, and then grind them into a particle size of 30 μm by an ultrafine grinder to obtain ultrafine powder of Tricholoma matsutake for later use;

[0062] (2) thymol and lauryl alcohol were mixed in a round-bottom flask at a molar ratio of 1:2, and stirred in an oil bath at 120° C. for 2 h to form a uniform and stable transparent liquid, i.e., a low eutectic solvent, which was set aside;

[0063] (3) Mixing the superfine powder of Tricholoma matsutake and the low eutectic solvent in a mass volume ratio of 1 g:170 mL, and adding 0.7 mg / mL of a composite enzyme, wherein the total enzyme activity of the composite enzyme is 35000 U / g, and the composite enzyme is composed of cellulase, pectinase, and papain in an enzyme activity ratio of 2:1:1; placing the container containing the mixture in an ultrasonic instrument for hot extraction, and simultaneously performing ultrasonic assisted extraction at a power of 100 W at 40° C. for 3 h;

[0064] (4) The extracted product was centrifuged at 9000 r / min for 10 min, and the precipitate was discarded to obtain a supernatant;

[0065] (5) The supernatant was chromatographed on a macroporous resin D101 column using a water-alcohol system, first eluting with 15% ethanol aqueous solution for 5 BV, then eluting with 35% ethanol aqueous solution for 8 BV, and finally eluting with 65% ethanol aqueous solution, and the eluate was collected; the eluate was concentrated and freeze-dried to obtain a matsutake extract.

[0066] Example 4

[0067] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof is different from that of Embodiment 1 only in that the preparation method of the low eutectic solvent is different, specifically, in step (2), glycerol and betaine are mixed in a molar ratio of 6:1 and placed in a round-bottom flask, and stirred at a constant temperature of 80° C. in an oil bath for 3 h to form a uniform and stable transparent liquid, i.e., the low eutectic solvent, which is set aside. The operations of the other steps remain unchanged.

[0068] Example 5

[0069] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof is different from that of Embodiment 1 only in that the preparation method of the low eutectic solvent is different, specifically, step (2) is to mix glycolic acid and acetamide in a molar ratio of 4:1 in a round-bottom flask, and stir in an oil bath at 80°C for 3 hours to form a uniform and stable transparent liquid, i.e., a low eutectic solvent, and set aside. The operations of other steps remain unchanged.

[0070] Example 6

[0071] This embodiment provides a Matsutake extract, and the preparation method thereof is different from that of Example 1 only in that the formula of the composite enzyme in step (3) is different. Specifically, the composite enzyme is composed of pectinase and papain with an enzyme activity ratio of 1:1, the usage amount of the composite enzyme and the total enzyme activity remain unchanged, and the operations of other steps remain unchanged.

[0072] Example 7

[0073] This embodiment provides a Matsutake extract, and the preparation method thereof is different from that of Example 1 only in that the formula of the composite enzyme in step (3) is different, specifically, the composite enzyme is composed of cellulase and papain with an enzyme activity ratio of 1:1, the usage amount of the composite enzyme and the total enzyme activity remain unchanged, and the operations of other steps remain unchanged.

[0074] Example 8

[0075] This embodiment provides a Matsutake extract, and the preparation method thereof is different from that of Embodiment 1 only in that the formula of the composite enzyme in step (3) is different, specifically, the composite enzyme is composed of cellulase and pectinase with an enzyme activity ratio of 1:1, the usage amount of the composite enzyme and the total enzyme activity remain unchanged, and the operations of other steps remain unchanged.

[0076] Example 9

[0077] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof differs from that of embodiment 1 only in that the elution procedure of step (5) is different, specifically, eluting 5 BV with 20% ethanol aqueous solution, and then eluting with 40% ethanol aqueous solution, collecting the eluate; concentrating and freeze-drying the eluate to obtain the Tricholoma matsutake extract. The operations of the other steps remain unchanged.

[0078] Example 10

[0079] This embodiment provides a Tricholoma matsutake extract, and the preparation method thereof differs from that of Embodiment 1 only in that the elution procedure of step (5) is different, specifically, eluting 5 BV with 20% ethanol aqueous solution, eluting 8 BV with 40% ethanol aqueous solution, eluting 8 BV with 60% ethanol aqueous solution, and finally eluting with 80% ethanol aqueous solution, collecting the eluate; concentrating and freeze-drying the eluate to obtain the Tricholoma matsutake extract. The operations of the other steps remain unchanged.

[0080] Comparative Example 1

[0081] This comparative example provides a pine mushroom extract, and the preparation method thereof is different from that of Example 1 only in that no complex enzyme is added in step (3), specifically, the pine mushroom ultrafine powder and the low eutectic solvent are mixed in a mass volume ratio of 1 g:150 mL, and the container containing the mixture is placed in an ultrasonic instrument for hot extraction, and ultrasonic assisted extraction is performed at a power of 200 W at 45° C. for 1 hour. The operations of the other steps remain unchanged.

[0082] Comparative Example 2

[0083] This comparative example provides a Tricholoma matsutake extract, the preparation method of which is different from that of Example 1 only in that the low eutectic solvent extraction method is replaced by an alcohol extraction method, specifically, in step (3), the low eutectic solvent is replaced by a 60% ethanol aqueous solution in equal amounts. The operations of other steps remain unchanged.

[0084] Test Example 1

[0085] Hyaluronidase activity inhibition test:

[0086] Hyaluronidase is a specific cleavage enzyme of hyaluronic acid. Inhibiting its activity can ensure the normal content and function of hyaluronic acid. Therefore, the hyaluronidase inhibition rate can be used as an indicator to evaluate the anti-allergic activity of a substance, indicating that it has a soothing effect.

[0087] First, the pine mushroom extracts prepared in Examples 1-10 and Comparative Examples 1-2 were prepared into samples to be tested at different concentrations for later use. In a 96-well plate, the samples to be tested at different concentrations were mixed with a hyaluronidase solution (concentration of 5 U / mL) and shaken and mixed for incubation at 37°C for 10 min, and then a sodium hyaluronate solution (concentration of 0.03% (w / v)) was added and incubated at 37°C for 45 min, followed by a BSA solution (concentration of 0.1% (w / v)) and continued incubation for 10 min, and the OD600nm value was detected to obtain the inhibitory effect of the sample to be tested on hyaluronidase.

[0088] The calculation formula of hyaluronidase activity inhibition rate is:

[0089]

[0090] Among them, A is the OD600nm value of the control solution (using acetate buffer solution instead of sample solution); B is the OD600nm value of the control blank solution (using acetate buffer solution instead of sample solution and enzyme solution); C is the OD600nm value of the sample solution to be tested; D is the OD600nm value of the sample blank solution (using acetate buffer solution instead of enzyme solution).

[0091] The results are shown in Table 1.

[0092] Table 1

[0093]

[0094]

[0095] From the data results in Table 1, it can be seen that the Tricholoma matsutake extract prepared by the preparation method of the present invention has excellent hyaluronidase activity inhibition effect, reflecting its potential in soothing effect. And the type of low eutectic solvent and complex enzyme will also affect the performance of the above effect.

[0096] Test Example 2

[0097] Zebrafish embryo neutrophil aggregation inhibition assay:

[0098] Healthy zebrafish embryos 3 days after fertilization were selected and divided into a model control group, a positive control group and a test group, wherein 24 fish embryos were randomly selected from the model control group and placed in a 3 cm culture dish, and mL of fish embryo culture solution containing 0.16 mg / L copper sulfate pentahydrate was added; wherein 24 fish embryos were randomly selected from the positive control group and placed in a 3 cm culture dish, and mL of fish embryo culture solution containing 0.1 mg / L copper sulfate pentahydrate and 0.0036 mg / L indomethacin was added; wherein 24 fish embryos were randomly selected from the test group and placed in a 3 cm culture dish, and 5 mL of fish embryo culture solution containing 0.16 mg / L copper sulfate pentahydrate and 5 mg / mL of each group of test substances (Matsutake extracts prepared in Examples 1-10 and Comparative Examples 1-2) was added.

[0099] Each group was placed in a 28°C incubator for 45 minutes, and then each group of fish embryos was fixed in paraformaldehyde for 1 hour, treated with PBST 3 times, 5 minutes each time, and then treated with 50% ethanol for 3 minutes. After the fish embryos were stained with Sudan black staining solution for 1 hour, they were washed with 70% ethanol 4 times, 5 minutes each time, and then treated with PBST 2 times, 5 minutes each time. The fish embryos were treated with bleaching solution for 10 minutes, then treated with 70% ethanol solution for 5 minutes, PBST for 1 minute, treated with clear solution 1 for 15 minutes, clear solution 2 for 10 minutes, and then treated with PBST for 3 minutes.

[0100] The fish embryos were placed sideways, and then placed under a stereo microscope to observe the tail of the fish embryos. The number of neutrophils in the three-quarters tail region of each fish embryo from the anus was counted, and the neutrophil aggregation inhibition rate was calculated. The inhibition rate (%) = [(MS) / M] × 100%, wherein S represents the average number of neutrophils in the fish embryos of the test substance treatment group, and the unit is granules per tail (granules / tail); M represents the average number of neutrophils in the fish embryos of the model control group, and the unit is granules per tail (granules / tail); the average value of each test group was calculated and expressed, and the results are shown in Table 2.

[0101] Table 2

[0102]

[0103]

[0104] From the data results in Table 2, it can be seen that the Matsutake extract prepared by the preparation method adopted in the present invention has excellent zebrafish embryo neutrophil aggregation effect, reflecting its potential in soothing and repairing effect. And the type of low eutectic solvent and complex enzyme will also affect the performance of the above effect.

[0105] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0106] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0107] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A method for preparing a pine mushroom extract having soothing and repairing effects, characterized in that: The preparation method comprises the following steps: Mixing the superfine powder of Tricholoma matsutake with a low eutectic solvent and a complex enzyme for extraction to obtain an extraction product; centrifuging the extraction product to obtain a supernatant; and eluting the supernatant through a macroporous resin to obtain the Tricholoma matsutake extract; The raw materials for preparing the deep eutectic solvent are thymol and lauryl alcohol; the molar ratio of thymol to lauryl alcohol is 1:(1-3); The composite enzyme is a combination of cellulase, pectinase and papain; the enzyme activity ratio of the cellulase, pectinase and papain is (1-3):(1-3):(1-3); the total enzyme activity of the composite enzyme is 30000-50000 U / g; the concentration of the composite enzyme in the extraction system is 0.2-0.8 mg / mL; The ratio of the superfine matsutake powder to the low eutectic solvent is 1g: (100-200)mL; the extraction is carried out at 35-55°C for 0.5-3h; ultrasonic-assisted extraction is also carried out during the extraction process; the eluent used for elution is a gradient concentration ethanol aqueous solution, and the elution method is: first elute with 10-20% ethanol aqueous solution for 3-10 BV, then elute with 30-40% ethanol aqueous solution for 3-10 BV, and finally elute with 60-70% ethanol aqueous solution, and collect the eluent.

2. The method for preparing the Tricholoma matsutake extract with soothing and repairing effects according to claim 1, characterized in that: The low eutectic solvent is prepared by reacting thymol and lauryl alcohol as raw materials at 80-120° C. for 1-5 h.

3. The method for preparing the Tricholoma matsutake extract with soothing and repairing effects according to claim 1, characterized in that: The preparation method of the superfine powder of Tricholoma matsutake comprises: cutting and drying the fruiting body of Tricholoma matsutake, freezing the slices, and then crushing the slices to a particle size of 10-30 μm.

4. The method for preparing the Tricholoma matsutake extract with soothing and repairing effects according to claim 1, characterized in that: The power of the ultrasound-assisted extraction is 100-300 W.

5. The method for preparing the Tricholoma matsutake extract with soothing and repairing effects according to claim 1, characterized in that: The centrifugation is carried out at 5000-10000 r / min for 10-20 min.

6. The method for preparing the Tricholoma matsutake extract with soothing and repairing effects according to claim 1, characterized in that: The eluate obtained by the elution is concentrated and dried.

7. A Tricholoma matsutake extract with soothing and repairing effects obtained by the preparation method according to any one of claims 1 to 6.

8. Use of the Tricholoma matsutake extract according to claim 7 in preparing a product with soothing and repairing effects.

9. The use of the Tricholoma matsutake extract according to claim 8 in preparing a product with soothing and repairing effects, characterized in that: The products include cosmetics.

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