Preparation method and application of cordyceps sinensis cordycepin extract

Abstract

The invention relates to the field of biological pharmacy, and discloses a preparation method and application of cordyceps sinensis cordycepin extract.The preparation method comprises the following steps that S1, cordyceps sinensis sporocarp is dried to constant weight and subjected to low-temperature freezing treatment, the cooled cordyceps sinensis sporocarp is smashed into fine powder through a low-temperature smashing machine, and the smashed cordyceps sinensis fine powder is sieved; according to the method, microwave-assisted extraction is combined with ultrasonic treatment, the microwave-assisted extraction can rapidly heat, moisture in cells can be rapidly gasified, pressure is generated, and cell walls are broken, and the ultrasonic treatment further destroys cell structures and increases substance exchange inside and outside the cells through a cavitation effect and mechanical vibration, so that the cell walls are broken. The extraction time is remarkably shortened, meanwhile, the dissolution rate of the cordycepin is increased, close combination of the cordycepin and other components can be destroyed through the synergistic effect of microwaves and ultrasonic waves, and the extraction purity is further improved.

Description

Technical Field

[0001] The invention relates to the field of biopharmaceuticals, and in particular to a preparation method and application of a cordyceps sinensis cordycepin extract. Background Art

[0002] Cordyceps sinensis is a precious medicinal fungus, mainly distributed in high-altitude areas such as Tibet, Qinghai, and Sichuan in China. Cordyceps sinensis has a variety of pharmacological activities, such as enhancing immunity, anti-tumor, anti-oxidation, and anti-fatigue. One of its main active ingredients is cordycepin. Cordycepin is a natural nucleoside analogue with significant pharmacological effects, such as antiviral, anti-tumor, and anti-inflammatory. Therefore, the efficient extraction of cordycepin from Cordyceps sinensis has important research and application value.

[0003] Traditional methods for extracting cordycepin from Cordyceps sinensis mainly include water extraction, organic solvent extraction, and reflux extraction. The water extraction method is to crush the dried Cordyceps sinensis into powder, add an appropriate amount of water, soak it at room temperature or under heating conditions for several hours to several days, filter to remove solid residues, collect the filtrate, and concentrate the filtrate by reduced pressure evaporation or freeze drying to obtain a cordycepin extract. The water extraction method uses water as a solvent to dissolve cordycepin in water by heating or long-term soaking. However, water, as a polar solvent, has limited solubility for non-polar cordycepin, resulting in a low extraction rate; The organic solvent extraction method is to grind the dried Cordyceps sinensis into powder, add an appropriate amount of organic solvent, such as ethanol, methanol, acetone, soak it at room temperature or under heating conditions for several hours to several days, filter to remove solid residues, collect the filtrate, and concentrate the filtrate by reduced pressure evaporation or rotary evaporation to obtain a cordycepin extract. Although organic solvents have a strong ability to dissolve cordycepin, the cordycepin in Cordyceps sinensis will be tightly combined with other components, such as polysaccharides and proteins, resulting in part of the cordycepin being unable to be completely dissolved, thereby affecting the extraction rate; The reflux extraction method is to grind the dried Cordyceps sinensis into powder, add an appropriate amount of water or organic solvent, heat and extract for several hours in a reflux device, filter to remove solid residues, collect the filtrate, and concentrate the filtrate by reduced pressure evaporation or rotary evaporation to obtain a cordycepin extract. The reflux extraction method uses heating reflux to make water or organic solvents pass through the raw materials repeatedly. Although the extraction efficiency is higher than that of water extraction and organic solvent extraction, the reflux extraction method will extract a variety of components, including polysaccharides, proteins, lipids, etc., resulting in more impurities in the extract, which in turn affects the extraction purity; In order to solve the above problems, a preparation method and application of Cordyceps sinensis cordycepin extract are proposed.

[0004] The above information disclosed in this background technology is only used to increase the understanding of the background technology of the present invention and therefore, it may include information that does not constitute the prior art known to ordinary technicians in this field. Summary of the invention

[0005] In order to solve the problem of biopharmaceuticals, the present invention provides a preparation method and application of a cordyceps sinensis cordycepin extract.

[0006] The preparation method and application of a Cordyceps sinensis cordycepin extract provided by the present invention adopt the following technical scheme: A method for preparing a cordycepin extract of Cordyceps sinensis comprises the following steps: S1, drying the Cordyceps sinensis fruiting bodies to a constant weight, freezing them at low temperature, crushing the cooled Cordyceps sinensis fruiting bodies into fine powder using a low temperature pulverizer, and sieving the crushed Cordyceps sinensis fine powder; S2, select 430-550 parts by mass of 50% ethanol solution, add 0.1-0.5% by mass of citric acid to the 50% ethanol solution, stir for 10-15 minutes, select 15-30 parts by mass of Cordyceps sinensis powder and add it to the 50% ethanol solution, use microwave-assisted extraction, the extraction temperature is 70-80°C, the extraction time is 60-80 minutes, the microwave power is 150-200W, stir every 10-15 minutes, and obtain a crude extract of cordycepin; S3, select 10-20 parts by mass of D101 resin, soak the D101 resin in 95% ethanol for 12-24 hours, then wash it with deionized water until it is neutral, add the washed D101 resin to the crude cordycepin extract obtained in S2, stir for 12-24 hours, filter and collect the resin mixture; S4, select the resin mixture collected in S3 and wash it with deionized water, and then wash it with 5-10 times the amount of 30-50% ethanol solution for 2-4 times, and the obtained eluate is the cordycepin extract; S5, select the cordycepin extract in S4, add 0.1-0.5% by mass of activated carbon to the cordycepin extract, stir for 5-10 minutes, let stand and then separate the phases, remove the phase, add 2-4 times the amount of 95% ethanol to precipitate the cordycepin in the cordycepin extract and collect the cordycepin precipitate; S6. Dispersing the cordycepin precipitate in S5 in the cordycepin extract by ultrasonication for 10-20 min, and vacuum drying to obtain a cordycepin extract.

[0007] Preferably, in S2, 0.1-0.5% by mass of citric acid is added to the 50% ethanol solution to adjust the pH value of the 50% ethanol solution to 5.0-6.0.

[0008] Preferably, in S2, during microwave-assisted extraction, ultrasonic treatment is added, the ultrasonic frequency is 25-50 kHz, and the ultrasonic power is 100-200 W.

[0009] Preferably, in S6, the cordycepin precipitate is further centrifuged after ultrasonic dispersion, with a centrifugal speed of 3000-5000 r / min and a centrifugal time of 10-20 min.

[0010] A use of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising a cordyceps sinensis cordycepin extract eye cream, the cordyceps sinensis cordycepin extract eye cream comprising the following components in parts by weight: 1-5 parts of cordycepin extract, 5-10 parts of moisturizer, 0.5-1.0 parts of antioxidant, 2-3 parts of surfactant, and 0.5-1.0 parts of pH regulator, wherein the cordycepin extract is prepared by the preparation method of the cordyceps sinensis cordycepin extract according to any one of claims 1 to 4.

[0011] Preferably, the moisturizing agent comprises the following components in parts by weight: 5-10 parts of glycerin, 3-5 parts of butylene glycol, 0.5-1.0 parts of hyaluronic acid, 3-5 parts of sweet almond oil, and 2-4 parts of jojoba oil.

[0012] Preferably, the antioxidant is 0.5-1.0 parts of vitamin E.

[0013] Preferably, the surfactant is 2-3 parts of coconut glucoside.

[0014] Preferably, the pH regulator comprises the following components in parts by weight: 0.5-1.0 parts of triethanolamine and 0.1-0.2 parts of citric acid.

[0015] Preferably, the cordyceps sinensis cordycepin extract eye cream further comprises 0.1-0.5 parts by weight of retinol and 1-3 parts by weight of hexapeptide.

[0016] In summary, the present invention includes the following beneficial technical effects: By freezing the dried Cordyceps sinensis fruiting bodies at -20°C for 12-24 hours, compared with related technologies, low-temperature freezing treatment helps to rupture the cell walls of Cordyceps sinensis, which can further increase the release rate of cordycepin, thereby improving the extraction efficiency. At the same time, low-temperature freezing treatment can avoid the destruction of the active ingredients of cordycepin by high-temperature treatment, which is conducive to ensuring the biological activity of cordycepin extracts; By using microwave-assisted extraction combined with ultrasonic treatment, microwave-assisted extraction and ultrasonic treatment act on cells through thermal effect and mechanical effect respectively. Microwave-assisted extraction can quickly increase the temperature, so that the water in the cells is quickly vaporized, generating pressure, causing the cell wall to rupture. Ultrasonic treatment further destroys the cell structure through cavitation effect and mechanical vibration, and increases the material exchange inside and outside the cells. Compared with related technologies, it significantly shortens the extraction time and improves the dissolution rate of cordycepin. The synergistic effect of microwave and ultrasonic waves helps to destroy the close combination of cordycepin and other components, further improving the extraction purity. Compared with related technologies, the adsorption and elution of D101 resin effectively removed impurities such as polysaccharides and proteins, further purified the cordycepin extract, and ensured the high purity of the product; By adding 0.1-0.5% by mass of citric acid to a 50% ethanol solution, the pH value of the cordycepin crude extract is adjusted to 5.0-6.0. Compared with the related art, the appropriate pH value can improve the solubility of cordycepin in the solvent, optimize the dissolution environment of cordycepin, reduce the loss of cordycepin during the extraction process, thereby improving the extraction rate. The appropriate pH value helps to protect the activity of cordycepin and prevent its degradation during the extraction process; By precisely controlling the extraction temperature, time and microwave power, compared with related technologies, the damage to the structure of cordycepin caused by long-term high-temperature extraction is avoided, thus protecting the biological activity of cordycepin. By adding 0.1-0.5% by mass of activated carbon to the cordycepin extract before the cordycepin extract is allowed to stand for phase separation, compared with the related art, the activated carbon can effectively remove suspended impurities in the cordycepin extract, further improve the purity of the cordycepin extract, and the purified cordycepin extract is more stable during storage and transportation and is not easy to deteriorate; The cordycepin extract prepared by the present invention is successfully applied to cosmetics, especially eye cream products, which provides a new way for the high-value utilization of cordycepin. In the eye cream formula, the cordycepin extract is reasonably matched with retinol and hexapeptide. Compared with the related art, cordycepin and retinol both have significant antioxidant capacity, can synergistically remove free radicals, reduce the damage of oxidative stress to skin cells, and delay skin aging. Hexapeptide works together with cordycepin and retinol by inhibiting muscle contraction and promoting collagen synthesis, and can more effectively reduce the formation of fine lines and wrinkles. At the same time, cordycepin can promote the regeneration and repair of skin cells and accelerate the recovery of damaged skin. Retinol and hexapeptide can promote the renewal of epidermal cells and the synthesis of collagen, and further enhance the skin's repair ability. The reasonable combination of cordycepin extract, retinol and hexapeptide plays a synergistic role. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1It is a flowchart of a method for preparing a cordyceps sinensis cordycepin extract of the present invention; DETAILED DESCRIPTION

[0018] The following is combined with Figure 1 The present invention is described in further detail.

[0019] The embodiment of the invention discloses a preparation method and application of a cordyceps sinensis cordycepin extract.

[0020] Embodiment 1, a method for preparing a cordycepin extract of Cordyceps sinensis, comprising the following steps: S1, drying the Cordyceps sinensis fruiting bodies at 65° C. to a constant weight, freezing them at low temperature, crushing the cooled Cordyceps sinensis fruiting bodies into fine powder using a low temperature crusher, and passing the crushed Cordyceps sinensis fine powder through a 100 mesh sieve; It should be noted that the standard for drying to constant weight is that the difference in mass between two consecutive weighings does not exceed 0.1%. The specific steps are as follows: A. Spread the Cordyceps sinensis fruiting bodies evenly on the drying tray; B. Place the drying tray in a drying oven preheated to 65°C; C. Take out and weigh every hour and record the mass; D. When the difference in mass between two consecutive weighings does not exceed 0.1%, it is considered to have been dried to constant weight; It should be further explained that the dried Cordyceps sinensis fruiting bodies are subjected to low-temperature freezing treatment by freezing at -20°C for 12 hours to destroy the cell walls of the Cordyceps sinensis fruiting bodies and improve the extraction rate of cordycepin.

[0021] S2, select 430 parts by mass of 50% ethanol solution, add 0.1% by mass of citric acid to the 50% ethanol solution, stir for 10 minutes, select 15 parts by mass of Cordyceps sinensis powder and add it to the 50% ethanol solution, use microwave-assisted extraction, the extraction temperature is 70°C, the extraction time is 60 minutes, the microwave power is 150W, add ultrasonic treatment, the ultrasonic frequency is 25kHz, the ultrasonic power is 100W, stir every 10 minutes, and obtain a crude extract of cordycepin; It should be noted that microwave-assisted extraction and ultrasonic treatment act on cells through thermal effect and mechanical effect respectively. Microwave-assisted extraction can quickly increase the temperature, so that the water in the cells is quickly vaporized, generating pressure and causing the cell wall to rupture. Ultrasonic treatment further destroys the cell structure through cavitation effect and mechanical vibration, and increases the exchange of substances inside and outside the cells. The combination of microwave-assisted extraction and ultrasonic treatment can more thoroughly destroy the cell structure and promote the release of cordycepin, thereby significantly improving the extraction efficiency and extraction rate, playing a synergistic role. S3, select 10 parts by mass of D101 resin, soak the D101 resin in 95% ethanol for 12 hours, then wash it with deionized water until it is neutral, add the washed D101 resin to the crude cordycepin extract obtained in S2, stir for 12 hours, filter and collect the resin mixture; It should be noted that soaking D101 resin in 95% ethanol can remove impurities and residual production aids in D101 resin, activating D101 resin, and washing it with deionized water to neutrality can remove ethanol and other impurities in D101 resin, making D101 resin reach a neutral state, so as to better adsorb the target substance. The specific steps are as follows: put D101 resin into a container, add 95% ethanol until the resin is completely immersed, and soak for 12 hours; rinse the soaked D101 resin with deionized water until the effluent is clear and has no ethanol smell; use pH test paper or pH meter to detect the pH value of the effluent to ensure that it is close to the neutral pH 7; if the pH value is not within the neutral range, continue to rinse with deionized water until the pH value stabilizes at around 7 with an error value of 0.5.

[0022] It should be further explained that D101 resin can effectively adsorb cordycepin, remove impurities in the crude cordycepin extract, and improve the purity of the cordycepin extract.

[0023] S4. Select the resin mixture collected in S3 and wash it with deionized water. After washing, wash it twice with 5 times the amount of 30% ethanol solution. Collect the eluate after each washing. The obtained eluate is the cordycepin extract.

[0024] S5, select the cordycepin extract in S4, add 0.1% by mass of activated carbon to the cordycepin extract, stir for 5 minutes, let stand for 1 hour and then separate the phases, remove the phase, add 2 times the amount of 95% ethanol to precipitate the cordycepin in the cordycepin extract and collect the cordycepin precipitate; It should be noted that activated carbon has a strong adsorption capacity due to its porous structure, and can effectively adsorb pigments, impurities and other unnecessary organic or inorganic substances in the cordycepin extract, which can not only improve the color of the cordycepin extract and make it clearer and more transparent, but also improve the purity of cordycepin. After adding activated carbon, through stirring and standing, the activated carbon can be fully in contact with the cordycepin extract and exert its adsorption effect. In the subsequent phase separation and precipitation steps, the activated carbon and the adsorbed impurities can be more easily removed, providing a purer cordycepin extract for the subsequent step, which is beneficial to the precipitation and collection of cordycepin.

[0025] S6, dispersing the cordycepin precipitate in S5 in the cordycepin extract by ultrasonication for 10 min, centrifuging at a speed of 3000 r / min for 10 min, and vacuum drying to obtain a cordycepin extract; It should be noted that centrifugal treatment can effectively remove the residual solution and other impurities in the cordycepin precipitate, further improving the purity of cordycepin.

[0026] Embodiment 2, a method for preparing a cordycepin extract of Cordyceps sinensis, comprising the following steps: S1, same as embodiment 1; S2, select 500 parts by mass of 50% ethanol solution, add 0.3% by mass of citric acid to the 50% ethanol solution, stir for 13 minutes, select 25 parts by mass of Cordyceps sinensis powder and add it to the 50% ethanol solution, use microwave-assisted extraction, the extraction temperature is 75 ° C, the extraction time is 70 minutes, the microwave power is 150 W, add ultrasonic treatment, the ultrasonic frequency is 35 kHz, the ultrasonic power is 180 W, stir every 13 minutes, and obtain a crude extract of cordycepin; S3, select 15 parts by mass of D101 resin, soak the D101 resin in 95% ethanol for 20 hours, then wash it with deionized water until it is neutral, add the washed D101 resin to the crude cordycepin extract obtained in S2, stir for 20 hours, filter and collect the resin mixture; S4, selecting the resin mixture collected in S3 and washing it with deionized water, and then washing it with 8 times the amount of 40% ethanol solution for 3 times, collecting the eluate after each washing, and the obtained eluate is the cordycepin extract; S5, select the cordycepin extract in S4, add 0.3% by mass of activated carbon to the cordycepin extract, stir for 8 minutes, let stand for 1 hour and then separate the phases, remove the phase, add 3 times the amount of 95% ethanol to precipitate the cordycepin in the cordycepin extract and collect the cordycepin precipitate; S6. Dispersing the cordycepin precipitate in S5 in the cordycepin extract by ultrasonication for 15 min. After ultrasonic dispersion, centrifuging at a speed of 4000 r / min for 15 min. After centrifugation, vacuum drying is performed to obtain a cordycepin extract.

[0027] Embodiment 3, a method for preparing a cordycepin extract of Cordyceps sinensis, comprising the following steps: S1, same as embodiment 1; S2, select 550 parts by mass of 50% ethanol solution, add 0.5% by mass of citric acid to the 50% ethanol solution, stir for 15 minutes, select 30 parts by mass of Cordyceps sinensis powder and add it to the 50% ethanol solution, use microwave-assisted extraction, the extraction temperature is 80°C, the extraction time is 80 minutes, the microwave power is 200W, add ultrasonic treatment, the ultrasonic frequency is 50kHz, the ultrasonic power is 200W, stir every 15 minutes, and obtain a crude extract of cordycepin; It should be further explained that citric acid is a weak organic acid that can effectively adjust the pH value of 50% ethanol solution. The addition of 0.5% by mass of citric acid can adjust the pH value of 50% ethanol solution to 6.0. For the extraction of cordycepin, the appropriate pH value can increase the solubility of cordycepin, thereby improving the extraction efficiency.

[0028] S3, select 20 parts by mass of D101 resin, soak the D101 resin in 95% ethanol for 24 hours, then wash it with deionized water until it is neutral, add the washed D101 resin to the crude cordycepin extract obtained in S2, stir for 24 hours, filter and collect the resin mixture; S4, selecting the resin mixture collected in S3 and washing it with deionized water, and then washing it with 10 times the amount of 50% ethanol solution for 4 times, collecting the eluate after each washing, and the obtained eluate is the cordycepin extract; S5, select the cordycepin extract in S4, add 0.5% by mass of activated carbon to the cordycepin extract, stir for 10 minutes, let stand for 1 hour and then separate the phases, remove the phase, add 4 times the amount of 95% ethanol to precipitate the cordycepin in the cordycepin extract and collect the cordycepin precipitate; S6. Dispersing the cordycepin precipitate in S5 in the cordycepin extract by ultrasonication for 20 min. After ultrasonic dispersion, centrifuging at a speed of 5000 r / min for 20 min. After centrifugation, vacuum drying is performed to obtain a cordycepin extract.

[0029] Comparative Example 1: extracting cordycepin extract from Cordyceps sinensis by water extraction, comprising the following steps: S1, same as Example 1; S2, taking 20 parts by weight of Cordyceps sinensis powder and adding it to deionized water, heating it to boiling, keeping boiling for 6 hours, and filtering to obtain a crude extract; S3, adding 0.5% by mass of activated carbon to the crude extract, stirring for 10 minutes, standing for 1 hour and then separating the phases, removing the phase, adding 3 times the amount of 95% ethanol to precipitate cordycepin and collecting the cordycepin precipitate; S4. Vacuum-drying the cordycepin precipitate to obtain a cordycepin extract.

[0030] Comparative Example 2, using an organic solvent extraction method to extract cordycepin extract from Cordyceps sinensis, comprising the following steps: S1, adding Cordyceps sinensis powder to deionized water, heating to boiling, keeping boiling for 6 hours, and filtering to obtain a crude extract; S2, taking 20 parts by weight of Cordyceps sinensis powder and adding it to a 50% ethanol solution, stirring for 24 hours, and filtering to obtain a crude extract; S3, adding 0.5% by mass of activated carbon to the crude extract, stirring for 10 minutes, standing for 1 hour to separate the phases, removing the phase, adding 3 times the amount of 95% ethanol to precipitate cordycepin and collecting the cordycepin precipitate; S4. Vacuum-drying the cordycepin precipitate to obtain a cordycepin extract.

[0031] Comparative Example 3, using a reflux extraction method to extract cordycepin extract from Cordyceps sinensis, comprising the following steps: S1, drying the Cordyceps sinensis fruiting body at 65° C. to constant weight, freezing it at low temperature, crushing it into fine powder using a low-temperature pulverizer, and passing it through a 100-mesh sieve; S2, taking 20 parts by mass of Cordyceps sinensis powder and adding it to 450 g of 50% ethanol solution, reflux extraction for 5 hours, and filtering to obtain a crude extract; S3, adding 0.5% by mass of activated carbon to the crude extract, stirring for 10 minutes, standing for 1 hour to separate the phases, removing the phase, adding 3 times the amount of 95% ethanol to precipitate cordycepin and collecting the cordycepin precipitate; S4. Vacuum-drying the cordycepin precipitate to obtain a cordycepin extract.

[0032] Experimental Group 1: 20 g of cordycepin extract was extracted using the extraction methods in Examples 1 to 3 and Comparative Examples 1 to 3, respectively. The optimal extraction method was determined by proposing the indicators of cost, extraction rate, and extraction purity. The experimental data are as follows: method Cost (yuan / kg) Extraction rate (%) purity(%) Embodiment 1 500 85 92 Embodiment 2 550 90 94 Embodiment 3 600 95 96 Comparative Example 1 300 60 70 Comparative Example 2 400 75 80 Comparative Example 3 350 80 75 Analysis of the experimental results: the extraction rates of Examples 1 to 3 are significantly higher than those of Comparative Examples 1 to 3, especially Example 3, which reaches an extraction rate of 95%; the purity of Examples 1 to 3 is also significantly better than that of Comparative Examples 1 to 3, and the purity of Example 3 reaches 96%; although the cost of Examples 1 to 3 is slightly higher than that of Comparative Examples 1 to 3, the extraction rate and purity are proportional to the cost, and the extraction rate and purity are higher when the cost is relatively high, and the overall economic benefit is better. Comprehensively judging that Example 3 is the optimal extraction method.

[0033] Example 4: Use of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising a cordyceps sinensis cordycepin extract eye cream, The cordycepin extract eye cream includes the following components in parts by weight: 5 parts of cordycepin extract, 10 parts of glycerin, 5 parts of butylene glycol, 1.0 parts of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 parts of vitamin E, 3 parts of coco-glucoside, 1.0 parts of triethanolamine, 0.2 parts of citric acid, 0.5 parts of retinol, and 3 parts of hexapeptide, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0034] Example 5: Use of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising a cordyceps sinensis cordycepin extract eye cream, The cordycepin extract eye cream includes the following components in parts by weight: 3 parts of cordycepin extract, 7 parts of glycerin, 4 parts of butylene glycol, 0.7 parts of hyaluronic acid, 4 parts of sweet almond oil, 3 parts of jojoba oil, 0.7 parts of vitamin E, 2.5 parts of coco-glucoside, 0.7 parts of triethanolamine, 0.15 parts of citric acid, 0.3 parts of retinol, and 2 parts of hexapeptide, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0035] Example 6: Use of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising a cordyceps sinensis cordycepin extract eye cream, The cordycepin extract eye cream includes the following components in parts by weight: 1 part of cordycepin extract, 5 parts of glycerin, 3 parts of butylene glycol, 0.5 parts of hyaluronic acid, 3 parts of sweet almond oil, 2 parts of jojoba oil, 0.5 parts of vitamin E, 2 parts of coco-glucoside, 0.5 parts of triethanolamine, 0.1 parts of citric acid, 0.1 parts of retinol and 1 part of hexapeptide, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0036] Comparative Example 4, application of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising an eye cream containing the cordyceps sinensis cordycepin extract, The cordycepin extract eye cream includes the following components in parts by weight: 5 parts of cordycepin extract, 10 parts of glycerin, 5 parts of butylene glycol, 1.0 parts of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 parts of vitamin E, 3 parts of coco-glucoside, 1.0 parts of triethanolamine, 0.2 parts of citric acid, and 3 parts of hexapeptide, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0037] Comparative Example 5, application of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising an eye cream containing the cordyceps sinensis cordycepin extract, The cordycepin extract eye cream includes the following components in parts by weight: 5 parts of cordycepin extract, 10 parts of glycerin, 5 parts of butylene glycol, 1.0 parts of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 parts of vitamin E, 3 parts of cocoyl glucoside, 1.0 parts of triethanolamine, 0.2 parts of citric acid and 0.5 parts of retinol, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0038] Comparative Example 6, application of a cordyceps sinensis cordycepin extract in cosmetics, the cosmetics comprising an eye cream containing the cordyceps sinensis cordycepin extract, The cordycepin extract eye cream includes the following components in parts by weight: 5 parts of cordycepin extract, 10 parts of glycerin, 5 parts of butylene glycol, 1.0 part of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 part of vitamin E, 3 parts of cocoyl glucoside, 1.0 part of triethanolamine, and 0.2 part of citric acid, wherein the cordycepin extract is prepared by the above-mentioned preparation method of the cordycepin extract.

[0039] Comparative Example 7, an eye cream, the eye cream comprises the following components in parts by weight: 10 parts of glycerin, 5 parts of butylene glycol, 1.0 part of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 part of vitamin E, 3 parts of coconut glucoside, 1.0 part of triethanolamine, 0.2 parts of citric acid, 0.5 parts of retinol, and 3 parts of hexapeptide.

[0040] Comparative Example 8, an eye cream, the eye cream comprises the following components in parts by weight: 10 parts of glycerin, 5 parts of butylene glycol, 1.0 part of hyaluronic acid, 5 parts of sweet almond oil, 4 parts of jojoba oil, 1.0 part of vitamin E, 3 parts of cocoyl glucoside, 1.0 part of triethanolamine, and 0.2 part of citric acid.

[0041] Experimental group 2, according to the formulas in Examples 4, 5, 6 and Comparative Examples 4, 5, 6, 7 and 8, eye cream samples were prepared respectively, and the preparation process was as follows: I. Weigh each component according to the above formula, add them into the mixing tank in sequence, and stir evenly. II. Heat to 70-80℃, stir until completely dissolved. III. The components in Examples 4, 5 and 6 are cooled to 40-50° C., retinol and hexapeptide are added, and stirring is continued until completely dissolved.

[0042] IV. Cool to room temperature to obtain an eye cream sample.

[0043] Experimental group 2, subjects: 500 female subjects aged 30-35 years old from the same region were selected and divided into 10 groups, numbered 1-10; Usage: The subjects used the eye cream samples of Examples 4-6 and Comparative Examples 4-8 respectively, once in the morning and once in the evening after cleansing, taking a mung bean-sized amount each time and applying it on the key parts around the eyes. The subjects only used it around the left eye and did not use it around the right eye; Observation indicators: Record the eye condition of the subjects after 30 days of use, including the degree of reduction of fine lines, skin elasticity, moisturizing effect, and skin tone uniformity; The experimental data are as follows: Experimental results analysis: Example 4 has the highest reduction in fine lines, reaching 81%, while Comparative Example 8 has the lowest reduction in fine lines, only 35%. This indicates that the higher the content of cordycepin extract, the better the improvement effect on fine lines; The skin elasticity improvement effect of Example 4 was the best, reaching 85%, while the skin elasticity improvement effect of Comparative Example 8 was the worst, only 38%. This also shows that cordycepin extract has a significant effect on improving skin elasticity; The moisturizing effect of Example 4 was the best, reaching 95%, while the moisturizing effect of Comparative Example 8 was the worst, only 51%. This indicates that the synergistic effect of cordycepin extract and other moisturizing ingredients has a significant effect on skin moisturizing; The skin color uniformity improvement effect of Example 4 was the best, reaching 85%, while the skin color uniformity improvement effect of Comparative Example 8 was the worst, only 43%. This shows that the cordycepin extract has a significant effect on improving the uniformity of skin color; In summary, the content of cordycepin extract has a significant effect on the effect of eye cream. The higher the content, the more obvious the eye cream is in reducing fine lines, improving skin elasticity, moisturizing effect and improving skin tone uniformity. Ingredients such as retinol and hexapeptide also contribute significantly to the effect of eye cream. Comparative Examples 7 and 8 lacking these ingredients have significantly poorer effects. The formula of Example 4 has the best effect. The preparation method of the cordycepin extract of cordyceps sinensis proposed in the present invention not only improves the extraction rate and purity of cordycepin, but also has high economic benefits. The cordycepin extract extracted by the preparation method of the cordycepin extract of cordyceps sinensis proposed in the present invention is used in the cordyceps sinensis cordycepin extract eye cream, which can significantly improve the eye condition and can also be well applied.

[0044] Finally, a few points should be explained: First, in the description of the present invention, it should be noted that, unless otherwise specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, which may refer to mechanical connection or electrical connection, or internal communication between two components, or direct connection. "upper", "lower", "left", "right", etc. are only used to indicate relative positional relationships. When the absolute position of the object being described changes, the relative positional relationship may change; Secondly: In the drawings of the embodiments disclosed in the present invention, only the structures related to the embodiments disclosed in the present invention are involved, and other structures can refer to the general design. In the absence of conflict, the same embodiment and different embodiments of the present invention can be combined with each other; Finally: The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

[0045] The above are all preferred embodiments of the present invention, and are not intended to limit the protection scope of the present invention. Therefore, any equivalent changes made based on the structure, shape, and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a cordycepin extract of Cordyceps sinensis, characterized in that: The following steps are involved: S1. Dry the Cordyceps sinensis fruiting bodies to constant weight, freeze them at low temperature, crush the cooled Cordyceps sinensis fruiting bodies into fine powder using a low-temperature crusher, and sieve the crushed Cordyceps sinensis fine powder; S2, select 430-550 parts by mass of 50% ethanol solution, add 0.1-0.5% by mass of citric acid to the 50% ethanol solution, stir for 10-15 minutes, select 15-30 parts by mass of Cordyceps sinensis powder and add it to the 50% ethanol solution, use microwave-assisted extraction, the extraction temperature is 70-80°C, the extraction time is 60-80 minutes, the microwave power is 150-200W, stir every 10-15 minutes, and obtain a crude extract of cordycepin; S3, select 10-20 parts by mass of D101 resin, soak the D101 resin in 95% ethanol for 12-24 hours, then wash it with deionized water until it is neutral, add the washed D101 resin to the crude cordycepin extract obtained in S2, stir for 12-24 hours, filter and collect the resin mixture; S4, select the resin mixture collected in S3 and wash it with deionized water, and then wash it with 5-10 times the amount of 30-50% ethanol solution for 2-4 times, and the obtained eluate is the cordycepin extract; S5, select the cordycepin extract in S4, add 0.1-0.5% by mass of activated carbon to the cordycepin extract, stir for 5-10 minutes, let stand and then separate the phases, remove the phase, add 2-4 times the amount of 95% ethanol to precipitate the cordycepin in the cordycepin extract and collect the cordycepin precipitate; S6. Dispersing the cordycepin precipitate in S5 in the cordycepin extract by ultrasonication for 10-20 min, and vacuum drying to obtain a cordycepin extract.

2. The method for preparing a Cordyceps sinensis cordycepin extract according to claim 1, characterized in that: In S2, 0.1-0.5% by mass of citric acid is added to the 50% ethanol solution to adjust the pH value of the 50% ethanol solution to 5.0-6.

0.

3. The method for preparing a Cordyceps sinensis cordycepin extract according to claim 1, characterized in that: In S2, ultrasonic treatment was added during microwave-assisted extraction, with an ultrasonic frequency of 25-50 kHz and an ultrasonic power of 100-200 W.

4. The method for preparing a Cordyceps sinensis cordycepin extract according to claim 1, characterized in that: In S6, after the cordycepin precipitate is ultrasonically dispersed, it is further centrifuged at a speed of 3000-5000 r / min for a time of 10-20 min.

5. A use of a cordyceps sinensis cordycepin extract in cosmetics, characterized in that: The cosmetics include cordyceps sinensis cordycepin extract eye cream, which includes the following components in parts by weight: 1-5 parts of cordycepin extract, 5-10 parts of moisturizer, 0.5-1.0 parts of antioxidant, 2-3 parts of surfactant, and 0.5-1.0 parts of pH regulator, wherein the cordycepin extract is prepared by the preparation method of the cordyceps sinensis cordycepin extract according to any one of claims 1 to 4.

6. The use of the Cordyceps sinensis cordycepin extract in cosmetics according to claim 5, characterized in that: The moisturizing agent comprises the following components in parts by weight: 5-10 parts of glycerin, 3-5 parts of butylene glycol, 0.5-1.0 parts of hyaluronic acid, 3-5 parts of sweet almond oil, and 2-4 parts of jojoba oil.

7. The use of the Cordyceps sinensis cordycepin extract in cosmetics according to claim 5, characterized in that: The antioxidant is 0.5-1.0 parts of vitamin E.

8. The use of the Cordyceps sinensis cordycepin extract in cosmetics according to claim 5, characterized in that: The surfactant is 2-3 parts of coconut glucoside.

9. The use of the Cordyceps sinensis cordycepin extract in cosmetics according to claim 5, characterized in that: The pH regulator comprises the following components in parts by weight: 0.5-1.0 parts of triethanolamine and 0.1-0.2 parts of citric acid.

10. The use of the Cordyceps sinensis cordycepin extract in cosmetics according to claim 5, characterized in that: The cordyceps sinensis cordycepin extract eye cream also includes 0.1-0.5 parts by weight of retinol and 1-3 parts by weight of hexapeptide.

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