A supramolecular eutectic compound of ganoderma lucidum Tibetan cypress extract and preparation method thereof

Abstract

The invention discloses a supramolecular eutectic compound of a ganoderma lucidum and Tibetan cypress extract and a preparation method thereof, the preparation method comprising the steps of: adding a mixed ganoderma lucidum powder and Tibetan cypress powder to a solvent for soaking, adding acid cellulase and betaine to the solvent for hydrolysis; inactivating the cellulase, centrifuging and filtering to obtain a crude extract; taking the crude extract and blowing nitrogen to remove the solvent to obtain a ganoderma lucidum and Tibetan cypress extract paste; adding betaine to the paste to form a mixture, heating the molten mixture under an inert atmosphere, and stirring the reaction to obtain a supramolecular eutectic compound of a ganoderma lucidum and Tibetan cypress extract. The invention effectively improves the extraction efficiency of active ingredients in ganoderma lucidum and Tibetan cypress by using ultrasound and betaine-assisted enzymolysis technology; applying the supramolecular eutectic compound of a ganoderma lucidum and Tibetan cypress extract to the preparation of cosmetics or skin ointments can significantly improve the skin's anti-oxidation, anti-aging, anti-inflammatory, and moisturizing properties.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to a supramolecular eutectic compound of a ganoderma lucidum Tibetan cypress extract and a preparation method thereof. Background Art

[0002] Ganoderma lucidum extract has a variety of functions and effects in skin care and treatment, especially the ingredient ganoderic acid, which is favored for its significant antioxidant, moisturizing, whitening, and anti-inflammatory effects. Among them, in terms of antioxidant and anti-aging, it is manifested as follows: (1) Scavenging free radicals: active ingredients such as polysaccharides and triterpenoids (including ganoderic acid) in Ganoderma lucidum extract can effectively scavenge free radicals and reduce oxidative damage to skin cells; (2) Improving antioxidant capacity: ingredients in Ganoderma lucidum extract such as ganoderic acid C2 have significant antioxidant properties and can be used as moisturizers and anti-aging ingredients in cosmetics to help the skin stay young. In terms of moisturizing and nourishing, it is manifested as follows: natural moisturizing factor: ingredients such as amino acids in Ganoderma lucidum have good moisturizing properties, which can effectively improve the dry state of the skin, enhance the skin's moisturizing ability, and make the skin softer and more moisturized. In terms of whitening and spot removal, it has the following effects: (1) Inhibit melanin production: triterpenoids in Ganoderma lucidum can inhibit the activity of tyrosinase and reduce the formation of melanin, thereby reducing skin pigmentation and making the skin tone more even; (2) Brighten skin tone: Long-term use of cosmetics containing Ganoderma lucidum extract can make the skin brighter and more lustrous. In terms of anti-inflammatory and anti-allergic effects, it has the following effects: (1) Inhibit inflammatory response: active ingredients in Ganoderma lucidum can inhibit inflammatory response and reduce skin inflammation symptoms, and have a certain effect on sensitive skin and acne skin; (2) Enhance skin barrier: certain ingredients in Ganoderma lucidum extract can enhance skin barrier function, improve skin resistance, and reduce damage to the skin caused by external stimuli. In terms of endocrine regulation and detoxification, it has the following effects: (1) Regulate endocrine: various extracts of Ganoderma lucidum can regulate the endocrine system, help excrete toxins in the body, and improve skin condition; (2) Promote metabolism: active ingredients in Ganoderma lucidum can promote skin metabolism, increase collagen synthesis and elastic fiber regeneration, thereby improving skin sagging and wrinkle problems.

[0003] The application of Tibetan cypress in cosmetics is mainly reflected in the various active ingredients in its extracts, which give cosmetics multiple functions. Its extract ingredients mainly include: Tibetan cypress essential oil (containing cedrene, sesquiterpenes, cedryl alcohol and other active ingredients), cedryl alcohol (with significant aroma and antibacterial effects, which can calm nerves and relax muscles) and other ingredients (such as bisabolol, which is also present in Tibetan cypress extracts and has anti-inflammatory and antibacterial effects). It has multiple functions, such as: (1) Antibacterial and anti-inflammatory: The active ingredients in Tibetan cypress extracts can inhibit the growth of various pathogenic bacteria, which is helpful for preventing and treating skin inflammation caused by bacteria. (2) Antioxidant: Its antioxidant properties help protect skin cells from oxidative damage, thereby delaying the aging process of the skin. (3) Moisturizing: Certain ingredients in Tibetan cypress extracts can form a protective film on the skin surface, locking in moisture and preventing moisture loss, thereby maintaining the moisture of the skin. (4) Calming and soothing: For sensitive skin and skin discomfort caused by external stimuli, Tibetan cypress extract can play a calming and soothing role, reducing the skin's inflammatory response.

[0004] As mentioned above, Ganoderma lucidum and Tibetan cypress, as traditional Chinese medicinal materials, have a variety of bioactive ingredients, such as ganoderic acid, polysaccharides, flavonoids, etc., which have significant benefits to human skin. However, these active ingredients are poorly water-soluble during the extraction process, and are often difficult to effectively retain and stabilize, limiting their application in cosmetics, skin ointments and other fields. Supramolecular eutectic technology is an emerging molecular assembly method that can form a stable eutectic structure through intermolecular interactions, thereby effectively protecting and utilizing these active ingredients.

[0005] Therefore, how to provide a supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract that can be used as a cosmetic raw material and improve the penetration effect is a technical problem that needs to be urgently solved by those skilled in the art. Summary of the invention

[0006] To this end, the present invention provides a supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract and a preparation method thereof to solve the related technical problems existing in the prior art.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] According to a first aspect of the present invention, there is provided a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0009] Weighing ganoderma lucidum powder and Tibetan cypress powder respectively and mixing them according to a proportion, wherein the particle size of the ganoderma lucidum powder and Tibetan cypress powder is 300 mesh;

[0010] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, the pH of the solvent is adjusted to 4.5-5.5, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis;

[0011] Centrifuge and filter to remove the unhydrolyzed Ganoderma lucidum powder and Tibetan cypress powder, and take the filtrate for later use;

[0012] Taking the filtrate, adjusting the pH of the filtrate to 7.0-8.0, heating at 100° C. for 5-10 minutes to inactivate the cellulase, and centrifuging and filtering to obtain a crude extract;

[0013] The crude extract is nitrogen-blown to remove the solvent, thereby obtaining a Ganoderma Lucidum Tibetan Cypress extract paste;

[0014] A second amount of betaine is added to the paste to form a mixture, and the mixture is heated and melted under an inert atmosphere, and stirred for reaction to obtain a crude product of a supramolecular eutectic compound of a Ganoderma lucidum and Tibetan cypress extract.

[0015] Furthermore, the weight ratio of the Ganoderma lucidum powder to the Tibetan cypress powder is 1: (1-3).

[0016] Furthermore, the solvent is an ethanol solution with a mass fraction of 2%.

[0017] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time, and the ultrasonic treatment parameters are: ultrasonic power of 200-500W, ultrasonic frequency of 20-50kHz, pausing for 10s after every 5s of ultrasonic treatment, and the ultrasonic treatment time is 15-30h.

[0018] Furthermore, the amount of the acid cellulase added is 0.5-2wt%, the hydrolysis temperature is 25-36°C, and the hydrolysis reaction time is 15-30h; and the amount of the first mass of betaine added is 10-15wt%.

[0019] Furthermore, the crude extract is subjected to nitrogen blowing to remove the solvent, and the nitrogen blowing treatment parameters are: nitrogen flow rate is 0.5-3.0 L / min, nitrogen pressure is 0.1-0.3 MPa, heating temperature is 40-60°C, and stirring is at 200-400 rpm, and the nitrogen blowing treatment time is 2-5 h.

[0020] Furthermore, the mass ratio of the second mass of betaine to the paste is 1:(2-3).

[0021] Further, wherein, under an inert atmosphere, the processing parameters for heating and melting the mixture are:

[0022] The heating temperature is 80-110°C and the reaction time is 3-8h.

[0023] According to a second aspect of the present invention, there is provided a supramolecular eutectic compound of a Ganoderma lucidum and Tibetan cypress extract, which is prepared by the above method for preparing a supramolecular eutectic compound of a Ganoderma lucidum and Tibetan cypress extract.

[0024] According to a third aspect of the present invention, there is provided a use of the supramolecular eutectic compound of the above-mentioned Ganoderma lucidum Tibetan cypress extract in the preparation of cosmetics or skin ointments.

[0025] The present invention has the following advantages:

[0026] The present invention effectively improves the extraction efficiency of active ingredients in Ganoderma lucidum and Tibetan cypress by using ultrasound and betaine-assisted cellulose enzymolysis technology, and simultaneously hydrolyzes and co-extracts Ganoderma lucidum and Tibetan cypress, thereby reducing the extraction steps. Betaine forms a eutectic supramolecular structure with Ganoderma lucidum extract and Tibetan cypress extract, thereby promoting the dissolution of insoluble matter, and promoting the extraction and skin absorption of ganoderic acid, Tibetan cypress extract, etc. The supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract of the present invention is applied to the preparation of cosmetics or skin ointments, which can significantly improve the skin's anti-oxidation, anti-aging, anti-inflammatory, and moisturizing properties. DETAILED DESCRIPTION

[0027] The following is a description of the implementation of the present invention by specific embodiments. People familiar with the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] In order to solve the problems existing in the prior art of low efficiency in extracting Ganoderma lucidum and Tibetan cypress separately, and poor solubility of effective ingredients such as ganoderic acid and Tibetan cypress extract, the present application promotes the dissolution of insoluble matter from the aspects of co-extraction of Ganoderma lucidum and Tibetan cypress and formation of compounds by betaine, Ganoderma lucidum extract and Tibetan cypress extract, thereby improving the existing extraction efficiency and enhancing the use effect.

[0029] The present invention provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0030] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0031] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to a proportion, wherein the weight ratio of Ganoderma lucidum powder to Tibetan cypress powder was 1:(1-3), and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 meshes.

[0032] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the pH of the solvent is adjusted to 4.5-5.5, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. The amount of acid cellulase added is 0.5-2wt%, the hydrolysis temperature is 25-36°C, and the hydrolysis reaction time is 15-30h; the amount of the first mass of betaine added is 10-15wt%.

[0033] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time, and the ultrasonic treatment parameters are: ultrasonic power of 200-500W, ultrasonic frequency of 20-50kHz, pausing for 10s after every 5s of ultrasonic treatment, and the ultrasonic treatment time is 15-30h.

[0034] The unhydrolyzed Ganoderma lucidum powder and Tibetan cypress powder were removed by centrifugation and filtration, and the filtrate was set aside.

[0035] Take the filtrate, adjust the pH of the filtrate to 7.0-8.0, heat at 100° C. for 5-10 minutes to inactivate the cellulase, and centrifuge and filter to obtain a crude extract.

[0036] The crude extract is nitrogen-blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract is nitrogen-blown to remove the solvent, and the nitrogen blowing treatment parameters are: nitrogen flow rate of 0.5-3.0L / min, nitrogen pressure of 0.1-0.3MPa, heating temperature of 40-60°C, and stirring at 200-400rpm, and the nitrogen blowing treatment time is 2-5h.

[0037] Add the second mass of betaine to the paste to form a mixture, wherein the mass ratio of the second mass of betaine to the paste is 1:(2-3). Heat the molten mixture under an inert atmosphere, wherein the processing parameters of heating the molten mixture under the inert atmosphere are: heating temperature of 80-110° C., reaction time of 3-8 hours. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0038] According to the above technical solution, the present application provides the following embodiments based on actual experiments to illustrate the technical effects of the present application.

[0039] Example 1

[0040] This embodiment 1 provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0041] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0042] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to proportion, wherein the weight of Ganoderma lucidum powder was 100 g, the weight of Tibetan cypress powder was 100 g, and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 mesh.

[0043] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the amount of the solvent is 1000 mL, the pH of the solvent is adjusted to 5.0 by a hydrochloric acid solution, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. Among them, the amount of acid cellulase added is 0.5 g, the acid cellulase used is 50,000 U / g, the hydrolysis temperature is 30°C, and the hydrolysis reaction time is 20 hours; the amount of the first mass of betaine added is 10 g.

[0044] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: ultrasonic power of 300 W, ultrasonic frequency of 40 kHz, pausing for 10 seconds after each ultrasonic treatment of 5 seconds, and ultrasonic treatment time of 20 hours.

[0045] The unhydrolyzed ganoderma powder and Tibetan cypress powder are removed by centrifugation and filtration, and the filtrate is used for later use. In this embodiment, the centrifugal speed is 500 rpm, the centrifugal time is 30 min, the centrifugal temperature is 5-10° C., and the filtration is performed by suction filtration. Of course, in order to better remove solid impurities, multiple filtrations can be performed.

[0046] The filtrate is taken, the pH of the filtrate is adjusted to 7.0-8.0 by sodium hydroxide solution, the cellulase is inactivated by heating at 100° C. for 5-10 minutes, and the crude extract is obtained by centrifugation and filtration.

[0047] The crude extract was nitrogen blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract was nitrogen blown to remove the solvent, and the nitrogen blowing treatment parameters were: nitrogen flow rate of 2L / min, nitrogen pressure of 0.2MPa, heating temperature of 50°C, and stirring at 300rpm, and the nitrogen blowing treatment time was 3h.

[0048] Add the second mass of betaine to the paste to form a mixture, the mass ratio of the second mass of betaine to the paste is 1: (2-3), in this embodiment, weigh 5g of betaine and 10g of the paste, and mix them. Heat the molten mixture under an inert atmosphere, wherein the inert gas is nitrogen, and the processing parameters for heating the molten mixture are: heating temperature is 100°C, and reaction time is 5h. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0049] The crude product of the supramolecular eutectic compound is recrystallized. Specifically, the crude product of the supramolecular eutectic compound is redissolved in ethanol, the weight volume ratio of the crude product to ethanol is 1g:10mL, the insoluble impurities in the crude product are removed by centrifugal filtration, the filtrate is heated and concentrated to a supersaturated state, the temperature is reduced to about 5°C and allowed to stand for 3 hours until the crystals are completely precipitated. The above crystals are washed and recrystallized with ethanol for multiple times, and heated and dried for standby use to obtain the supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract. The above processing steps belong to the prior art, and the parameters such as the number of crystal washing times and the heating and drying temperature are not repeated.

[0050] Example 2

[0051] This embodiment 2 provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0052] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0053] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to a proportion, wherein the weight of Ganoderma lucidum powder was 100 g, the weight of Tibetan cypress powder was 150 g, and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 mesh.

[0054] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the amount of the solvent is 1000 mL, the pH of the solvent is adjusted to 4.5 by a hydrochloric acid solution, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. Among them, the amount of acid cellulase added is 1.0 g, the acid cellulase used is 50,000 U / g, the hydrolysis temperature is 28°C, and the hydrolysis reaction time is 20 hours; the amount of the first mass of betaine added is 12 g.

[0055] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: ultrasonic power of 400 W, ultrasonic frequency of 40 kHz, pausing for 10 seconds after each ultrasonic treatment of 5 seconds, and ultrasonic treatment time of 20 hours.

[0056] The unhydrolyzed ganoderma powder and Tibetan cypress powder are removed by centrifugation and filtration, and the filtrate is used for later use. In this embodiment, the centrifugal speed is 500 rpm, the centrifugal time is 30 min, the centrifugal temperature is 5-10° C., and the filtration is performed by suction filtration. Of course, in order to better remove solid impurities, multiple filtrations can be performed.

[0057] The filtrate is taken, the pH of the filtrate is adjusted to 7.0-8.0 by sodium hydroxide solution, the cellulase is inactivated by heating at 100° C. for 5-10 minutes, and the crude extract is obtained by centrifugation and filtration.

[0058] The crude extract was nitrogen blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract was nitrogen blown to remove the solvent, and the nitrogen blowing treatment parameters were: nitrogen flow rate of 1.5L / min, nitrogen pressure of 0.15MPa, heating temperature of 45°C, and stirring at 400rpm, and the nitrogen blowing treatment time was 3h.

[0059] Add the second mass of betaine to the paste to form a mixture, the mass ratio of the second mass of betaine to the paste is 1: (2-3), in this embodiment, weigh 5g of betaine and 11g of the paste, and mix them. Heat the molten mixture under an inert atmosphere, wherein the inert gas is nitrogen, and the processing parameters for heating the molten mixture are: heating temperature is 85°C, and reaction time is 6h. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0060] The crude product of the supramolecular eutectic compound is recrystallized. Specifically, the crude product of the supramolecular eutectic compound is redissolved in ethanol, the weight volume ratio of the crude product to ethanol is 1g:10mL, the insoluble impurities in the crude product are removed by centrifugal filtration, the filtrate is heated and concentrated to a supersaturated state, the temperature is reduced to about 5°C and allowed to stand for 3 hours until the crystals are completely precipitated. The above crystals are washed and recrystallized with ethanol for multiple times, and heated and dried for standby use to obtain the supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract. The above processing steps belong to the prior art, and the parameters such as the number of crystal washing times and the heating and drying temperature are not repeated.

[0061] Example 3

[0062] This embodiment 3 provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0063] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0064] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to a proportion, wherein the weight of Ganoderma lucidum powder was 100 g, the weight of Tibetan cypress powder was 200 g, and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 meshes.

[0065] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the amount of the solvent is 1000 mL, the pH of the solvent is adjusted to 5.5 by a hydrochloric acid solution, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. Among them, the amount of acid cellulase added is 1.5 g, the acid cellulase used is 50,000 U / g, the hydrolysis temperature is 30°C, and the hydrolysis reaction time is 25 hours; the amount of the first mass of betaine added is 12 g.

[0066] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: ultrasonic power of 450W, ultrasonic frequency of 35kHz, pause of 10s after each ultrasonic treatment of 5s, and ultrasonic treatment time of 25h.

[0067] The unhydrolyzed ganoderma powder and Tibetan cypress powder are removed by centrifugation and filtration, and the filtrate is used for later use. In this embodiment, the centrifugal speed is 500 rpm, the centrifugal time is 30 min, the centrifugal temperature is 5-10° C., and the filtration is performed by suction filtration. Of course, in order to better remove solid impurities, multiple filtrations can be performed.

[0068] The filtrate is taken, the pH of the filtrate is adjusted to 7.0-8.0 by sodium hydroxide solution, the cellulase is inactivated by heating at 100° C. for 5-10 minutes, and the crude extract is obtained by centrifugation and filtration.

[0069] The crude extract was nitrogen blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract was nitrogen blown to remove the solvent, and the nitrogen blowing treatment parameters were: nitrogen flow rate of 1.5L / min, nitrogen pressure of 0.3MPa, heating temperature of 50°C, and stirring at 250rpm, and the nitrogen blowing treatment time was 2.5h.

[0070] Add the second mass of betaine to the paste to form a mixture, the mass ratio of the second mass of betaine to the paste is 1: (2-3), in this embodiment, weigh 5g of betaine and 12g of the paste, and mix them. Heat the molten mixture under an inert atmosphere, wherein the inert gas is nitrogen, and the processing parameters of the heated molten mixture are: heating temperature is 95°C, and reaction time is 6h. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0071] The crude product of the supramolecular eutectic compound is recrystallized. Specifically, the crude product of the supramolecular eutectic compound is redissolved in ethanol, the weight volume ratio of the crude product to ethanol is 1g:10mL, the insoluble impurities in the crude product are removed by centrifugal filtration, the filtrate is heated and concentrated to a supersaturated state, the temperature is reduced to about 5°C and allowed to stand for 3 hours until the crystals are completely precipitated. The above crystals are washed and recrystallized with ethanol for multiple times, and heated and dried for standby use to obtain the supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract. The above processing steps belong to the prior art, and the parameters such as the number of crystal washing times and the heating and drying temperature are not repeated.

[0072] Example 4

[0073] This embodiment 4 provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0074] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0075] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to a proportion, wherein the weight of Ganoderma lucidum powder was 100 g, the weight of Tibetan cypress powder was 250 g, and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 meshes.

[0076] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the amount of the solvent is 1000 mL, the pH of the solvent is adjusted to 5.3 by a hydrochloric acid solution, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. Among them, the amount of acid cellulase added is 1.8 g, the acid cellulase used is 50,000 U / g, the hydrolysis temperature is 35°C, and the hydrolysis reaction time is 20 hours; the amount of the first mass of betaine added is 14 g.

[0077] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: ultrasonic power of 400 W, ultrasonic frequency of 40 kHz, pausing for 10 seconds after each ultrasonic treatment of 5 seconds, and ultrasonic treatment time of 20 hours.

[0078] The unhydrolyzed ganoderma powder and Tibetan cypress powder are removed by centrifugation and filtration, and the filtrate is used for later use. In this embodiment, the centrifugal speed is 500 rpm, the centrifugal time is 30 min, the centrifugal temperature is 5-10° C., and the filtration is performed by suction filtration. Of course, in order to better remove solid impurities, multiple filtrations can be performed.

[0079] The filtrate is taken, the pH of the filtrate is adjusted to 7.0-8.0 by sodium hydroxide solution, the cellulase is inactivated by heating at 100° C. for 5-10 minutes, and the crude extract is obtained by centrifugation and filtration.

[0080] The crude extract was nitrogen blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract was nitrogen blown to remove the solvent, and the nitrogen blowing treatment parameters were: nitrogen flow rate of 2.5L / min, nitrogen pressure of 0.2MPa, heating temperature of 55°C, and stirring at 300rpm, and the nitrogen blowing treatment time was 3.5h.

[0081] Add the second mass of betaine to the paste to form a mixture, the mass ratio of the second mass of betaine to the paste is 1: (2-3), in this embodiment, weigh 5g of betaine and 13g of the paste, and mix them. Heat the molten mixture under an inert atmosphere, wherein the inert gas is nitrogen, and the processing parameters of the heated molten mixture are: heating temperature is 110°C, and reaction time is 5h. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0082] The crude product of the supramolecular eutectic compound is recrystallized. Specifically, the crude product of the supramolecular eutectic compound is redissolved in ethanol, the weight volume ratio of the crude product to ethanol is 1g:10mL, the insoluble impurities in the crude product are removed by centrifugal filtration, the filtrate is heated and concentrated to a supersaturated state, the temperature is reduced to about 5°C and allowed to stand for 3 hours until the crystals are completely precipitated. The above crystals are washed and recrystallized with ethanol for multiple times, and heated and dried for standby use to obtain the supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract. The above processing steps belong to the prior art, and the parameters such as the number of crystal washing times and the heating and drying temperature are not repeated.

[0083] Example 5

[0084] This embodiment 5 provides a method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, comprising the following steps:

[0085] Select high-quality Ganoderma lucidum, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Ganoderma lucidum powder for later use; similarly, select Tibetan cypress, wash and dry it, then grind it through a 300-mesh sieve, and collect the sieved Tibetan cypress powder for later use.

[0086] Ganoderma lucidum powder and Tibetan cypress powder were weighed separately and mixed according to a proportion, wherein the weight of Ganoderma lucidum powder was 100 g, the weight of Tibetan cypress powder was 300 g, and the particle size of Ganoderma lucidum powder and Tibetan cypress powder was 300 meshes.

[0087] The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the amount of the solvent is 1000 mL, the pH of the solvent is adjusted to 4.8 by a hydrochloric acid solution, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis. The amount of acid cellulase added is 2 g, the acid cellulase used is 50,000 U / g, the hydrolysis temperature is 25°C, and the hydrolysis reaction time is 25 hours.

[0088] Furthermore, the mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for immersion and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: ultrasonic power of 500 W, ultrasonic frequency of 45 kHz, pausing for 10 seconds after each ultrasonic treatment of 5 seconds, and ultrasonic treatment time of 25 hours.

[0089] The unhydrolyzed ganoderma powder and Tibetan cypress powder are removed by centrifugation and filtration, and the filtrate is used for later use. In this embodiment, the centrifugal speed is 500 rpm, the centrifugal time is 30 min, the centrifugal temperature is 5-10° C., and the filtration is performed by suction filtration. Of course, in order to better remove solid impurities, multiple filtrations can be performed.

[0090] The filtrate is taken, the pH of the filtrate is adjusted to 7.0-8.0 by sodium hydroxide solution, the cellulase is inactivated by heating at 100° C. for 5-10 minutes, and the crude extract is obtained by centrifugation and filtration.

[0091] The crude extract was nitrogen blown to remove the solvent to obtain a Ganoderma lucidum Tibetan cypress extract paste; wherein, the crude extract was nitrogen blown to remove the solvent, and the nitrogen blowing treatment parameters were: nitrogen flow rate of 3L / min, nitrogen pressure of 0.2MPa, heating temperature of 55°C, and stirring at 400rpm, and the nitrogen blowing treatment time was 3h.

[0092] Add the second mass of betaine to the paste to form a mixture, the mass ratio of the second mass of betaine to the paste is 1: (2-3), in this embodiment, weigh 5g of betaine and 15g of the paste, and mix them. Heat the molten mixture under an inert atmosphere, wherein the inert gas is nitrogen, and the processing parameters for heating the molten mixture are: heating temperature is 100°C, and reaction time is 7h. Stir the reaction to obtain a crude product of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract.

[0093] The crude product of the supramolecular eutectic compound is recrystallized. Specifically, the crude product of the supramolecular eutectic compound is redissolved in ethanol, the weight volume ratio of the crude product to ethanol is 1g:10mL, the insoluble impurities in the crude product are removed by centrifugal filtration, the filtrate is heated and concentrated to a supersaturated state, the temperature is reduced to about 5°C and allowed to stand for 3 hours until the crystals are completely precipitated. The above crystals are washed and recrystallized with ethanol for multiple times, and heated and dried for standby use to obtain the supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract. The above processing steps belong to the prior art, and the parameters such as the number of crystal washing times and the heating and drying temperature are not repeated.

[0094] Comparative Example 1

[0095] In this comparative example 1, no Ganoderma lucidum powder was added, and other processing conditions were the same as those in Example 1.

[0096] Comparative Example 2

[0097] In this comparative example 2, no Tibetan cypress powder was added, and other processing conditions were the same as those in Example 1.

[0098] Comparative Example 3

[0099] In this comparative example 3, no betaine was added, and other processing conditions were the same as those in Example 1.

[0100] Test Example 1

[0101] Transdermal test experiment:

[0102] The same weight of the co-crystal compounds or products of Examples 1-5 and Comparative Examples 1-3 were taken to conduct a transdermal test to examine the penetration effects of the above products, as follows:

[0103] Select 20-day-old common piglets weighing 12 kg, take the skin on the back of the ear, peel off the subcutaneous fat layer and connective tissue, rinse it with saline several times, put it in saline, and store it in a refrigerator at 4°C for later use. Take it out before the experiment and soak it in saline for 0.5 h to room temperature. The Franz cell method was used for the transdermal experiment. The exposed piglet skin area in the Franz diffusion cell was 2.1 cm 2, the volume of the receiving tank is 15mL. Take 0.5g of the product weight in Examples 1-5 and Comparative Examples 1-3, dissolve it in 100mL of water, take 1mL of the solution to be tested and apply it to the skin surface, add 15mL of buffered saline solution (PBS buffer solution is selected in this experimental example, pH is about 6.5) to the receiving tank, place it in a constant temperature water bath, and stir continuously. Among them, the stirring speed is 300rpm, and the temperature is controlled at 30°C. Then, take 2mL of receiving liquid at different time points, and add the same volume of receiving liquid to the receiving tank, and the receiving liquid is subjected to liquid phase detection to measure the concentration of the receiving liquid. The test time is 24h, and samples are taken and tested every 6h, and the data is recorded.

[0104] The results are shown in Table 1:

[0105]

[0106] Table 1 shows the permeation per unit area of ​​each embodiment and comparative example, and the permeation permeation increases with time, but the increase in the comparative example is obviously lower than that of embodiments 1-5. At the same time, at the end of the 24h experiment, the permeation effect of embodiment 3 is significantly better than that of other embodiments and comparative examples. The supramolecular Ganoderma lucidum Tibetan cypress extract has a very good permeation effect, and is expected to achieve better use effect in terms of human skin absorption.

[0107] Test Example 2

[0108] Antioxidant and anti-aging test:

[0109] On the basis of Test Example 1, Example 3 and Comparative Example 1 were selected as the experimental groups of this test example, and ROS determination and DPPH free radical elimination rate were used as evaluation indicators for the antioxidant and anti-aging effects. Specifically, they are as follows:

[0110] Test preparation: Keratinocytes were selected and placed in multiple 6-well plates. The culture temperature was about 37°C and the culture was carried out in a humid environment incubator with 5% CO2 for 24 hours. The cultured cells were uniformly administered at the same dose in each well. Among them, Example 3 and Comparative Example 1 were used as the experimental groups of this test example, 100 μg / mL of vitamin C was selected as the positive control group, and pure water was selected as the negative control group. Three parallel experiments were performed for each group. The experimental results are as follows:

[0111] Table 2 shows the results of ROS determination.

[0112]

[0113] The above experimental results show that the compound of Example 3 of the present application has a better inhibitory effect on ROS than the comparative example 1 and the positive control group, which indicates that the Ganoderma lucidum Tibetan cypress supramolecular eutectic compound prepared by the preparation method of the present application can achieve a good anti-oxidation and anti-aging effect. At the same time, the comparative example 1 also reflects its anti-oxidation and anti-aging effect to a certain extent, which is not much different from the positive control group.

[0114] Table 3 shows the elimination rate of DPPH free radicals

[0115]

[0116] It can be seen from the above experimental results that the compound of Example 3 of the present application has the highest elimination rate of DPPH free radicals, which is better than that of Comparative Example 1 and the positive control group. Therefore, in terms of antioxidant and anti-aging performance, it can achieve better use effect.

[0117] Test Example 3

[0118] Anti-inflammatory test:

[0119] The cultured keratinocytes in Test Example 2 were selected and placed in multiple 6-well plates, and irradiated with ultraviolet light at an irradiation condition of 250 mJ / cm 2 . Each well was uniformly administered with the same dose, and the dosage was configured and diluted 10 times according to Test Example 1, wherein Example 3 and Comparative Example 1 were used as the experimental groups of this test example, 0.1 mg / mL of dexamethasone was used as the positive control group, and pure water was used as the negative control group. After irradiation, the culture was continued at a temperature of about 37°C and in a humid environment incubator with 5% CO2 for 24 hours. After the culture was completed, the inflammatory mediator (PGE2) of the cultured keratinocytes was detected using an enzyme-linked immunosorbent assay (ELISA), and the specific results are as follows:

[0120] Table 4 shows the anti-inflammatory test results

[0121]

[0122] It can be seen from the above experimental results that, compared with the negative control group, the application of the compounds of Example 3 of the present application and Comparative Example 1 achieved a better anti-inflammatory effect, and the effect of Example 3 of the present application was also better than that of Comparative Example 1, thus indicating that the application of the compound of Example 3 of the present application has a significant anti-inflammatory effect.

[0123] Test Example 4

[0124] Moisturizing effect test:

[0125] Prepare basic skin care agent: 20 mL of olive oil, 10 mL of glycerin, 1.0 g of simple emulsifier, 0.5 g of sorbitan ester, 70 mL of pure water, and 0.5 mL of cosmetic antibacterial agent. Mix well to form a basic skin care agent.

[0126] 0.05 g of the compound prepared in Example 3 was added to the above basic skin care agent to prepare an experimental example. The same weight of clean water was added to the above basic skin care agent to prepare a comparative example.

[0127] 30 testers who met the criteria were selected and randomly divided into two groups, namely the experimental group (using the basic skin care agent improved in the above experimental example) and the control group (using the basic skin care agent improved in the above comparative example). 2 The area of ​​size is the test area. The skin moisture content of the subjects was measured before and after use using a skin moisture tester. The average TEWL change rate of each group was calculated according to the formula. in, is the moisture content of the skin after applying the experimental skin care product, Skin moisture content after 3 hours of use, The average water content change rate of the skin of 15 people in the group. The specific experimental results are as follows:

[0128] Table 5 shows the test results of moisturizing effect

[0129]

[0130] It can be seen from the above experimental results that the basic skin care agent to which the compound of Example 3 of the present application is added greatly improves the moisturizing performance of the skin care agent.

[0131] Although the present invention has been described in detail above by general description and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims

1. A method for preparing a supramolecular eutectic compound of a Ganoderma lucidum Tibetan cypress extract, characterized in that: The steps include: Weighing ganoderma lucidum powder and Tibetan cypress powder respectively and mixing them according to a ratio, wherein the weight ratio of the ganoderma lucidum powder to the Tibetan cypress powder is 1:(1-3), and the particle size of the ganoderma lucidum powder and the Tibetan cypress powder is 300 mesh; The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking, wherein the solvent is an ethanol solution with a mass fraction of 2%, the pH of the solvent is adjusted to 4.5-5.5, and acid cellulase and the first mass of betaine are added to the solvent in proportion for hydrolysis, wherein the addition amount of the first mass of betaine is 10-15wt%, the addition amount of the acid cellulase is 0.5-2wt%, the hydrolysis temperature is 25-36°C, and the hydrolysis reaction time is 15-30h; Centrifuge and filter to remove the unhydrolyzed Ganoderma lucidum powder and Tibetan cypress powder, and take the filtrate for later use; Taking the filtrate, adjusting the pH of the filtrate to 7.0-8.0, heating at 100° C. for 5-10 minutes to inactivate the cellulase, and centrifuging and filtering to obtain a crude extract; The crude extract is nitrogen-blown to remove the solvent, thereby obtaining a Ganoderma Lucidum Tibetan Cypress extract paste; A second mass of betaine is added to the paste to form a mixture, wherein the mass ratio of the second mass of betaine to the paste is 1:(2-3); the mixture is heated and melted under an inert atmosphere, the heating temperature is 80-110° C., the reaction time is 3-8 hours, and the mixture is stirred to react to obtain a crude product of a supramolecular eutectic compound of a Ganoderma lucidum and Tibetan cypress extract; the crude product of the supramolecular eutectic compound is recrystallized to obtain a supramolecular eutectic compound of a Ganoderma lucidum and Tibetan cypress extract.

2. The method for preparing the supramolecular eutectic compound of the Ganoderma Lucidum Tibetan cypress extract according to claim 1, characterized in that: in, The mixed Ganoderma lucidum powder and Tibetan cypress powder are added to a solvent for soaking and ultrasonic treatment is performed at the same time. The ultrasonic treatment parameters are: The ultrasonic power is 200-500W, the ultrasonic frequency is 20-50kHz, and the ultrasonic treatment time is 15-30h after 5s of pausing for 10s.

3. The method for preparing the supramolecular eutectic compound of the Ganoderma Lucidum Tibetan cypress extract according to claim 1, characterized in that: in, The crude extract is subjected to nitrogen blowing to remove the solvent, and the nitrogen blowing treatment parameters are: The nitrogen flow rate is 0.5-3.0 L / min, the nitrogen pressure is 0.1-0.3 MPa, the heating temperature is 40-60°C, and the stirring is at 200-400 rpm. The nitrogen blowing treatment time is 2-5 h.

4. A supramolecular eutectic compound of Ganoderma lucidum Tibetan cypress extract, characterized in that: The composition is prepared by the method for preparing the supramolecular eutectic compound of Ganoderma lucidum and Tibetan cypress extract as described in any one of claims 1 to 3.

5. Use of the supramolecular eutectic compound of the Ganoderma lucidum Tibetan cypress extract as claimed in claim 4 in the preparation of cosmetics or skin ointments.

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