Anti-aging cosmetic, wine meal oil and preparation method of wine meal oil

Abstract

The invention relates to the field of cosmetics, in particular to an anti-aging cosmetic, wine meal oil and a preparation method thereof. The anti-aging cosmetic comprises wine meal oil; the grape wine meal oil is prepared from the following characteristic components: oleanolic acid, alpha-guaiacol, beta-tocotrienol, beta-sitosterol, campesterol, soyasaponol E, ganoderic acid F, ganoderma lucidum alcohol I and D-erythro-dihydrosphingosine; the total phenol content of the wine meal oil is greater than or equal to 1mg / g, the total sterol content of the wine meal oil is greater than or equal to 26mg / g, and the total flavone content of the wine meal oil is greater than or equal to 1mg / g; the wine meal oil is extracted from wine meal; the grape wine meal is mainly composed of grape skin, grape stems and grape seeds. The compounds have excellent anti-oxidation characteristics, and can improve the anti-aging effect of anti-aging cosmetics. The skin can be protected from oxidative damage, and a positive effect is achieved in cosmetics. The grape wine meal oil maximally retains the active ingredients of each part of grape wine meal.

Description

Technical Field

[0001] The present application relates to the field of cosmetics, and in particular, to an anti-aging cosmetic, grape wine dregs oil and a preparation method thereof. Background Art

[0002] With the development of the grape brewing industry, the output of wine dregs is increasing day by day, which mainly consists of grape skins, grape stems and grape seeds. At present, the secondary utilization of wine dregs is mainly to sort out the grape seeds and then produce grape seed oil, while the remaining grape skins and grape stems are often treated in an inefficient and extensive way, such as making low-value fertilizers, feed or directly landfilling them, which leads to problems such as waste of resources and environmental pollution. Summary of the invention

[0003] The purpose of the embodiments of the present application is to provide an anti-aging cosmetic, grape wine dregs oil and a preparation method thereof.

[0004] In a first aspect, the present application provides an anti-aging cosmetic, comprising:

[0005] Grape dregs oil; the characteristic components of grape dregs oil include: oleanolic acid, α-guaiacene, β-tocotrienol, β-sitosterol, campesterol, soybean saponin E, ganoderic acid F, ganoderic alcohol I and D-erythro-dihydrosphingosine; the total phenol content of grape dregs oil is ≥1mg / g, the total sterol content of grape dregs oil is ≥26mg / g, and the total flavonoid content of grape dregs oil is ≥1mg / g;

[0006] Grape dregs oil is extracted from grape dregs; grape dregs are mainly composed of grape skins, grape stems and grape seeds.

[0007] In the above technical scheme, the characteristic components of grape dregs oil are: oleanolic acid, α-guaiacene, β-tocotrienol, β-sitosterol, campesterol, soybean saponin E, ganoderic acid F, ganoderic alcohol I and D-erythro-dihydrosphingosine; these compounds have excellent antioxidant properties and can be used as anti-aging active ingredients of anti-aging cosmetics, thereby greatly enhancing the anti-aging effect of anti-aging cosmetics. In addition, the grape dregs oil showed an anti-aging effect comparable to astaxanthin in the nematode model, indicating that it helps protect the skin from oxidative damage and plays a positive role in cosmetics. The grape dregs oil retains the active ingredients of each part of the grape dregs to the greatest extent.

[0008] In other embodiments of the present application, the peroxide value of the above-mentioned grape wine dregs oil is ≤7mmol / Kg.

[0009] In other embodiments of the present application, the acid value of grape wine dregs oil is ≤4 mg / g.

[0010] In a second aspect, the present application provides a method for preparing grape wine dregs oil, the preparation method comprising:

[0011] The wine dregs powder is hot-soaked in an acid solution to obtain a first liquid;

[0012] The first liquid is subjected to continuous phase change extraction at 0.2MPa-0.8MPa and 40°C-50°C.

[0013] The above technical solution performs continuous phase change extraction on the first liquid at 0.2MPa-0.8MPa and 40℃-50℃, which enables the solvent to quickly circulate in liquid-gas-liquid cycle, and the circulating and fresh solvent can ensure sufficient and effective extraction of the material. In addition, the solvent is recovered under vacuum, which will not cause solvent residue and environmental pollution, thereby realizing low-cost green extraction.

[0014] In other embodiments of the present application, continuous phase change extraction comprises:

[0015] n-Butane is used as the extractant to carry out liquid phase, gas phase and liquid phase continuous cycle phase change extraction.

[0016] In other embodiments of the present application, the continuous phase change extraction includes multiple extraction cycles, and one extraction cycle includes:

[0017] The extractant gas is compressed into an extractant liquid in a vacuum, and the extractant liquid is passed through the first feed liquid for extraction; the wine dregs oil and the acid solution obtained by extraction flow out with the extractant liquid to obtain a mixed liquid; the extractant liquid is converted into an extractant gas and separated from the mixed liquid; the separated extractant gas repeats the extraction cycle;

[0018] Optionally, after separating the extractant gas, the acid solution is removed from the mixed solution to obtain a crude extract;

[0019] Optionally, the crude extract is subjected to a second extraction;

[0020] Optionally, the crude extract is subjected to a second extraction using n-hexane.

[0021] In other embodiments of the present application, the acid solution is an alcohol solution of an acid;

[0022] Optionally, the acid solution is a citric acid-ethanol solution;

[0023] Optionally, the citric acid-ethanol solution comprises, by mass percentage, 1% to 5% citric acid and 95% to 99% ethanol.

[0024] In other embodiments of the present application, the wine dregs powder is hot-soaked with an acid solution, comprising:

[0025] Based on the mass ratio, the grape dregs powder and the acid solution are hot-soaked in a solid-liquid ratio of (1:1) to (1:3).

[0026] In other embodiments of the present application, the wine dregs powder is hot-soaked with an acid solution, comprising:

[0027] Soak the grape dregs powder and the acid solution at 40℃~50℃;

[0028] Optionally, the soaking time is 1 h to 3 h.

[0029] In a third aspect, the present application provides a grape wine dregs oil obtained by using the preparation method of grape wine dregs oil provided in the second aspect. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0031] Figure 1 The total ion current spectrum of the wine dregs oil prepared in Example 1 analyzed by gas chromatography-mass spectrometry;

[0032] Figure 2 The total ion current spectrum of the grape wine meal oil prepared in Example 1 analyzed by liquid chromatography-mass spectrometry;

[0033] Figure 3 This is a graph showing the effect of the grape wine dregs oil prepared in Example 1 on the survival curve of nematodes under H2O2 oxidative stress conditions;

[0034] Figure 4 This is a graph showing the effect of the wine dregs oil prepared in Example 1 on the survival curve of nematodes under PQ oxidative stress conditions;

[0035] Figure 5 This is a graph showing the effect of the wine dregs oil prepared in Example 1 on the survival curve of nematodes under heat stress conditions;

[0036] Figure 6 This is a graph showing the effect of the wine dregs oil prepared in Example 1 on the survival curve of nematodes under light stress conditions. DETAILED DESCRIPTION

[0037] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments.

[0038] Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present application.

[0039] After research, it was found that during the production of grape seed oil, only a small amount of free phenol can be dissolved in grape seed oil, and most of the active compounds remain in the grains. Further research found that grape skins and grape stems are also rich in a variety of anti-cancer, antibacterial, and antioxidant active ingredients, such as polyphenols, flavonoids, and terpenes, which contain broad application prospects and great development value. Therefore, there is room for research on how to further increase the content of active ingredients in grape seed oil. How to utilize the remaining grape skins and grape seeds in wine grains in a high-value and resource-based manner and to extract wine grains in a green and efficient one-time manner are issues worth studying.

[0040] Traditional methods such as Soxhlet extraction, maceration and reflux extraction require the use of large amounts of potentially toxic solvents, and the required extraction temperature may lead to the destruction and loss of active substances. Ultrasound-assisted extraction, microwave-assisted extraction and supercritical extraction methods effectively improve the yield and retention rate, but they consume a lot of energy and are costly. Aqueous enzyme method, as an environmentally friendly and efficient oil extraction method, has attracted attention because it does not produce solvent residues, environmental pollution and safety issues during the extraction process. However, the technology has the problem of high cost of commercial enzyme preparations in practical applications, which limits the industrialization process.

[0041] The present application provides an anti-aging cosmetic, including:

[0042] Grape dregs oil; the characteristic components of grape dregs oil include: oleanolic acid, α-guaiacene, β-tocotrienol, β-sitosterol, campesterol, soybean saponin E, ganoderic acid F, ganoderic alcohol I and D-erythro-dihydrosphingosine; the total phenol content of grape dregs oil is ≥1mg / g, the total sterol content of grape dregs oil is ≥26mg / g, and the total flavonoid content of grape dregs oil is ≥1mg / g;

[0043] Grape dregs oil is extracted from grape dregs; grape dregs are mainly composed of grape skins, grape stems and grape seeds.

[0044] In the above technical scheme, the characteristic components of grape wine dregs oil are: oleanolic acid, α-guaiacene, β-tocotrienol, β-sitosterol, campesterol, soybean saponin E, ganoderic acid F, ganoderic alcohol I and D-erythro-dihydrosphingosine; these compounds have excellent antioxidant properties and can be used as anti-aging active ingredients of anti-aging cosmetics, thereby greatly enhancing the anti-aging effect of anti-aging cosmetics. In addition, the grape wine dregs oil showed an anti-aging effect equivalent to 60 μM astaxanthin in the nematode model, indicating that it helps to protect the skin from oxidative damage and plays a positive role in cosmetics. The grape wine dregs oil retains the active ingredients of each part of the grape wine dregs to the greatest extent.

[0045] Illustratively, in some embodiments of the present application, the total phenol content of wine dregs is 1 mg / g, 1.1 mg / g, 1.2 mg / g, 1.3 mg / g, 1.4 mg / g, 1.5 mg / g, 1.6 mg / g, 1.7 mg / g, 1.8 mg / g, 1.9 mg / g, 2.0 mg / g, 2.5 mg / g, 3.0 mg / g, 3.5 mg / g, 4.0 mg / g, 5.0 mg / g or a range between any two of the foregoing values.

[0046] Illustratively, in some embodiments of the present application, the total sterol content of wine dregs is 26 mg / g, 27 mg / g, 28 mg / g, 29 mg / g, 30 mg / g, 31 mg / g, 32 mg / g, 33 mg / g, 34 mg / g, 35 mg / g, 36 mg / g, 37 mg / g, 38 mg / g, 39 mg / g, 40 mg / g or a range between any two of the foregoing values.

[0047] Illustratively, in some embodiments of the present application, the total flavonoid content of wine dregs oil is 1 mg / g, 1.2 mg / g, 1.5 mg / g, 2 mg / g, 2.5 mg / g, 3 mg / g, 3.5 mg / g, 4 mg / g, 4.5 mg / g, 5 mg / g or a range between any two of the foregoing values.

[0048] In the above technical solution, the definition of "characteristic components" of wine dregs oil is: the relative content measured by gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry is greater than the potential anti-aging active ingredients.

[0049] The “potential” mentioned above is because some ingredients have been reported in the literature to have antioxidant effects (aging is caused by continuous oxidative damage), but have not been directly verified by anti-aging experiments.

[0050] For example, it has been reported that oleanolic acid can inhibit liver cell damage caused by oxidative stress, inhibit the generation of reactive oxygen species (ROS) and scavenge free radicals by increasing the activity of antioxidant enzymes (such as superoxide dismutase, catalase, glutathione peroxidase, etc.) and the level of reduced glutathione. These effects of oleanolic acid may be beneficial to anti-aging, that is, oleanolic acid is a "potential" anti-aging active ingredient.

[0051] It should be noted that the above-mentioned grape wine dregs also contain some other active ingredients, but these active ingredients have not been reported in the literature to have antioxidant and anti-aging activities, and therefore are not used as the "characteristic components" of the above-mentioned grape wine dregs. For example, in some embodiments of the present application, the above-mentioned grape wine dregs are analyzed by gas chromatography-mass spectrometry to obtain oleanolic acid (11.6108%), α-guaiacene (4.5109%), β-tocotrienol (2.8166%), β-sitosterol (7.0947%), and campesterol (1.3126%) as the "characteristic components" of grape wine dregs; and soybean saponin E (7.5149%), ganoderic acid F (3.2038%), ganoderic alcohol I (1.7824%), and D-erythro-dihydrosphingosine (20.1058%) are measured by liquid chromatography-mass spectrometry analysis as the "characteristic components" of grape wine dregs.

[0052] Furthermore, in some embodiments of the present application, the peroxide value of grape wine meal oil is ≤7mmol / Kg.

[0053] Peroxide value is an important indicator to measure the degree of oil oxidation. A lower peroxide value indicates that the oil has better oxidation stability. It is usually required to be less than or equal to 10mmol / Kg. In the above technical solution, the peroxide value of grape dregs oil is ≤7mmol / Kg, which has very excellent oxidation stability. Therefore, its application in anti-aging cosmetics can effectively improve the anti-aging effect of cosmetics.

[0054] Illustratively, in some embodiments of the present application, the peroxide value of wine dregs oil is 7mmol / Kg, 6mmol / Kg, 5mmol / Kg, 4mmol / Kg, 3mmol / Kg, 2mmol / Kg, 1mmol / Kg, 0.5mmol / Kg or a range between any two of the foregoing values.

[0055] In some embodiments of the present application, the method for determining the peroxide value is as follows:

[0056] Weigh 2-3g of the prepared sample, place it in a 250mL iodine volumetric flask, add 30mL of chloroform-glacial acetic acid solution, and gently shake the sample until it is completely dissolved. Accurately add 1.00mL of saturated potassium iodide solution, plug the bottle cap tightly, and gently shake for 0.5min, and place it in a dark place for 3min. Take it out and add 100mL of water. After shaking well, immediately titrate the precipitated iodine with a standard sodium thiosulfate titration solution. When it is titrated to a light yellow color, add 1mL of starch indicator, continue titrating and shake vigorously until the blue color of the solution disappears as the end point. At the same time, perform a blank test. The volume of the standard sodium thiosulfate titration solution consumed in the blank test is V0. The peroxide value calculation formula is as follows:

[0057]

[0058] Furthermore, in some embodiments of the present application, the acid value of the wine dregs oil is ≤4 mg / g.

[0059] Acid value is an indicator for evaluating the free fatty acid content in oils and fats. A lower acid value means that the oils and fats are less rancid and of higher quality; it is usually required to be less than or equal to 5mmol / Kg. In the above technical solution, the acid value of grape dregs oil is ≤4mg / g, the oils and fats are less rancid and of higher quality, so applying it to anti-aging cosmetics can effectively improve the anti-aging effect of cosmetics.

[0060] Illustratively, in some embodiments of the present application, the acid value of wine dregs is 4 mg / g, 3.5 mg / g, 3 mg / g, 2.5 mg / g, 2 mg / g, 1.5 mg / g, 1.2 mg / g, 1.1 mg / g, 1 mg / g or a range between any two of the foregoing values.

[0061] In some embodiments of the present application, the method for determining the acid value is as follows:

[0062] Weigh the oil sample into a 250mL conical flask, add 50-100mL of ether-isopropanol mixture and 3-4 drops of phenolphthalein indicator, shake thoroughly to dissolve the sample, and then titrate with standard titration solution. The endpoint of the titration is when the sample solution begins to turn slightly red and there is no obvious fading within 15 seconds. Record the milliliters of standard titration solution consumed in this titration, which is V. Take another clean 250mL conical flask and accurately add the same volume and type of organic solvent mixture and indicator as when measuring the sample, shake to mix, and the milliliters of standard titration solution consumed by titration are V0. The acid value calculation formula is as follows:

[0063]

[0064] Some embodiments of the present application provide a method for preparing grape wine dregs oil, the preparation method comprising:

[0065] The wine dregs powder is hot-soaked in an acid solution to obtain a first liquid;

[0066] The first liquid is subjected to continuous phase change extraction at 0.2MPa-0.8MPa and 40°C-50°C.

[0067] The above technical solution performs continuous phase change extraction on the first liquid at 0.2MPa-0.8MPa and 40℃-50℃, which enables the solvent to quickly circulate in liquid-gas-liquid cycle, and the circulating and fresh solvent can ensure sufficient and effective extraction of the material. In addition, the solvent is recovered under vacuum, which will not cause solvent residue and environmental pollution, thereby realizing low-cost green extraction. The above technical solution solves the current problems of insufficient and incomplete extraction of active ingredients in wine dregs, and part of the wine dregs being wasted or used at a low value. The wine dregs oil extracted by the above method retains the active ingredients of each part of the wine dregs to the greatest extent. At the same time, the method must be green, efficient, low-cost, environmentally friendly and low-energy.

[0068] Illustratively, in some embodiments of the present application, the first liquid is subjected to continuous phase change extraction at a pressure of 0.2MPa, 0.22MPa, 0.25MPa, 0.28MPa, 0.3MPa, 0.32MPa, 0.35MPa, 0.38MPa, 0.4MPa, 0.42MPa, 0.45MPa, 0.5MPa, 0.6MPa, 0.7MPa, 0.8MPa or a pressure range between any two of the foregoing values; and a temperature of 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C or a temperature range between any two of the foregoing values.

[0069] In the above technical solution, the first liquid is subjected to continuous phase change extraction at 0.2MPa-0.8MPa and 40°C-50°C.

[0070] Further, in some embodiments of the present application, continuous phase change extraction comprises:

[0071] n-Butane is used as the extractant to carry out liquid phase, gas phase and liquid phase continuous cycle phase change extraction.

[0072] n-Butane is an organic compound with the chemical formula C4H 10 , is a common alkane, a colorless, easily liquefied gas at room temperature and pressure. It can realize liquid-gas-liquid circulation (i.e. continuous phase change extraction) during continuous phase change extraction, and the circulating and fresh solvent can ensure the full and effective extraction of the material; thereby increasing the content of active ingredients in the extracted wine dregs.

[0073] Further, in some embodiments of the present application, the continuous phase change extraction includes multiple extraction cycles, and one extraction cycle includes:

[0074] The extractant gas is compressed into an extractant liquid in a vacuum, and the extractant liquid is made to flow through the first feed liquid for extraction; the wine dregs oil and the acid solution obtained by extraction flow out with the extractant liquid to obtain a mixed liquid; the extractant liquid is converted into an extractant gas and separated from the mixed liquid; the separated extractant gas repeats the extraction cycle.

[0075] Furthermore, in some embodiments of the present application, after separating the extractant gas, the acid solution is removed from the mixed liquid to obtain a crude extract.

[0076] Furthermore, in some embodiments of the present application, the crude extract is subjected to a second extraction.

[0077] Furthermore, in some embodiments of the present application, the crude extract is subjected to a second extraction using n-hexane.

[0078] Furthermore, in some embodiments of the present application, the acid solution is an alcohol solution of an acid.

[0079] The above technical solution uses an acid solution to soak the wine dregs powder, which can destroy the tight structure of the cell wall of the wine dregs powder by deacidification, and then use an organic solvent to extract the oil in the cells. At the same time, phenolic compounds are more easily dissolved and retained under acidic conditions, which promotes their further extraction into the oil phase.

[0080] Furthermore, in some embodiments of the present application, the acid solution is a citric acid-ethanol solution.

[0081] Furthermore, in some embodiments of the present application, the citric acid-ethanol solution includes, by mass percentage, 1% to 5% citric acid and 95% to 99% ethanol.

[0082] Illustratively, in some embodiments of the present application, the citric acid-ethanol solution comprises, by mass percentage, 1%, 1.2%, 1.5%, 2%, 2.5%, 2.8%, 3%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.8%, 5% or a range between any two of the foregoing citric acid; 95%, 96%, 97%, 98%, 99% or a range between any two of the foregoing ethanol.

[0083] Further, in some embodiments of the present application, the wine dregs powder is hot-soaked with an acid solution, comprising:

[0084] Based on the mass ratio, the grape dregs powder and the acid solution are hot-soaked in a solid-liquid ratio of (1:1) to (1:3).

[0085] Illustratively, in some embodiments of the present application, the wine dregs powder and the acid solution are soaked in a solid-liquid ratio of 1:1, 1:1.1, 1:1.2, 1:1.5, 1:1.8, 1:2, 1:2.1, 1:2.2, 1:2.5, 1:2.8, 1:3 or a range between any two of the foregoing values, based on mass ratio.

[0086] Further, in some embodiments of the present application, the wine dregs powder is hot-soaked with an acid solution, comprising:

[0087] Soak the grape lees powder and the acid solution at 40℃~50℃.

[0088] For example, in some embodiments of the present application, the wine dregs powder is hot-soaked with an acid solution, comprising:

[0089] Soak the wine lees powder and the acid solution at 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, 50°C or in a range between any two of the aforementioned values.

[0090] Furthermore, in some embodiments of the present application, the soaking time is 1 hour to 3 hours.

[0091] Illustratively, in some embodiments of the present application, the immersion time is 1 h, 1.1 h, 1.2 h, 1.3 h, 1.4 h, 1.5 h, 1.6 h, 1.7 h, 1.8 h, 1.9 h, 2.0 h, 2.1 h, 2.2 h, 2.3 h, 2.4 h, 2.5 h, 2.6 h, 2.7 h, 2.8 h, 2.9 h, 3 h or a range between any two of the foregoing values.

[0092] Some embodiments of the present application provide a grape wine dregs oil, which is obtained by using the preparation method of grape wine dregs oil provided in any of the aforementioned embodiments.

[0093] The features and performance of the present application are further described in detail below in conjunction with the embodiments:

[0094] Example 1

[0095] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0096] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0097] S2. Weigh 1000g of the grape dregs powder obtained in step S1 and fill it into a 3L extraction axe of a continuous phase change extraction device, add 2L of a citric acid-ethanol solution with a mass percentage of 5%, and soak it at 50°C for 120min. Then use n-butane as the extraction solvent, and extract under the conditions of an extraction pressure of 0.8MPa, an extraction temperature of 50°C, a decomposition temperature of 60°C, and a flow rate of 60L / h. The n-butane is compressed into a liquid in a vacuum and flows through the extraction axe to extract the grape dregs. The grape dregs oil and the citric acid-ethanol solution flow into the decomposition kettle along with the n-butane liquid, and the n-butane liquid is converted into a gas by heating and decompression, separated from the grape dregs oil and recovered to the storage tank. The above process is continuously circulated, and the extraction is 120min in total, and finally a crude extract is obtained in the decomposition axe.

[0098] S3, subjecting the crude extract obtained in step S2 to rotary evaporation to remove the ethanol solution, adding the crude extract to a separating funnel to extract with n-hexane, and subjecting the extract to rotary evaporation to obtain grape dregs oil.

[0099] Example 2

[0100] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0101] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0102] S2, weigh 1000g of the wine dregs powder obtained in step S1 and fill it into a 3L extraction axe of a continuous phase change extraction device, add 3L of citric acid-ethanol solution with a mass percentage of 1%, and soak at 45°C for 90min; then use n-butane as the extraction solvent, extract under the conditions of extraction pressure of 0.5MPa, extraction temperature of 45°C, decomposition temperature of 55°C, and flow rate of 60L / h. n-Butane is compressed into liquid in a vacuum and flows through the extraction axe to extract the wine dregs. The wine dregs oil and citric acid-ethanol solution flow into the decomposition kettle along with the n-butane liquid, and the n-butane liquid is converted into gas by heating and decompression, separated from the wine dregs oil and recovered to the storage tank. The above process is continuously circulated, and the extraction is 60min in total; finally, the crude extract is obtained in the decomposition axe.

[0103] S3, subjecting the crude extract obtained in step S2 to rotary evaporation to remove the ethanol solution, adding the crude extract to a separating funnel to extract with n-hexane, and subjecting the extract to rotary evaporation to obtain grape dregs oil.

[0104] Example 3

[0105] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0106] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0107] S2. Weigh 1000g of the grape dregs powder obtained in step S1 and fill it into a 3L extraction axe of a continuous phase change extraction device, add 1L of a citric acid-ethanol solution with a mass percentage of 3%, and soak at 40°C for 60min. Then use n-butane as the extraction solvent, and extract under the conditions of an extraction pressure of 0.2MPa, an extraction temperature of 40°C, a decomposition temperature of 50°C, and a flow rate of 60L / h. The n-butane is compressed into a liquid in a vacuum and flows through the extraction axe to extract the grape dregs. The grape dregs oil and the citric acid-ethanol solution flow into the decomposition kettle along with the n-butane liquid, and the n-butane liquid is converted into a gas by heating and decompression, separated from the grape dregs oil and recovered to the storage tank. The above process is continuously circulated, and the extraction is performed for a total of 90min; finally, a crude extract is obtained in the decomposition axe.

[0108] S3, subjecting the crude extract obtained in step S2 to rotary evaporation to remove the ethanol solution, adding the crude extract to a separating funnel to extract with n-hexane, and subjecting the extract to rotary evaporation to obtain grape dregs oil.

[0109] Comparative Example 1

[0110] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0111] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0112] S2. Take 30 g of crushed grape wine dregs powder, add 300 mL of n-hexane, and stir and extract at 50°C and normal pressure for 3 h, filter, and evaporate to remove n-hexane to obtain grape wine dregs oil.

[0113] Comparative Example 2

[0114] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0115] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0116] S2. Take 30g of wine dregs powder crushed to 40 mesh, add 60mL of 5% citric acid-ethanol solution with a mass percentage of , and soak at 50℃ for 2h. Then add 300mL of n-hexane and stir and extract at 50℃ for 3h under normal pressure, and filter to obtain a crude extract.

[0117] S3, subjecting the crude extract obtained in step S2 to rotary evaporation to remove the ethanol solution, adding the crude extract to a separating funnel to extract with n-hexane, and subjecting the extract to rotary evaporation to obtain grape dregs oil.

[0118] Comparative Example 3

[0119] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0120] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0121] S2, weigh 1000g of the wine dregs powder obtained in step S1 and fill it into the 3L extraction axe of the continuous phase change extraction device, use n-butane as the extraction solvent, and extract for 120min under the conditions of extraction pressure of 0.8MPa, extraction temperature of 50℃, decomposition temperature of 60℃, and flow rate of 60L / h. n-Butane is compressed into liquid in a vacuum and flows through the extraction axe to extract the wine dregs. The wine dregs oil and citric acid-ethanol solution flow into the decomposition kettle along with the n-butane liquid, and the n-butane liquid is converted into gas by heating and decompression, separated from the wine dregs oil and recovered to the storage tank. The above process is continuously circulated, and the extraction is 120min in total; finally, the wine dregs oil is obtained in the decomposition axe.

[0122] Comparative Example 4

[0123] Provided is a grape wine dregs oil, which is prepared according to the following steps:

[0124] S1. Grind and sieve the dried grape dregs to obtain grape dregs powder with a particle size of 40 meshes. The grape dregs consist of grape skins, grape stems and grape seeds.

[0125] S2. Weigh 1000g of the grape dregs powder obtained in step S1 and fill it into a 3L extraction axe of a continuous phase change extraction device, add 2L of a citric acid-ethanol solution with a mass percentage of 5%, and soak it at room temperature for 120min. Then use n-butane as the extraction solvent, and extract under the conditions of an extraction pressure of 0.8MPa, an extraction temperature of 50°C, a decomposition temperature of 60°C, and a flow rate of 60L / h. The n-butane is compressed into a liquid in a vacuum and flows through the extraction axe to extract the grape dregs. The grape dregs oil and the citric acid-ethanol solution flow into the decomposition kettle along with the n-butane liquid, and the n-butane liquid is converted into a gas by heating and decompression, separated from the grape dregs oil and recovered to a storage tank. The above process is circulated continuously, and the extraction is 120min in total; finally, the grape dregs oil is obtained in the decomposition axe.

[0126] S3, subjecting the crude extract obtained in step S2 to rotary evaporation to remove the ethanol solution, adding the crude extract to a separating funnel to extract with n-hexane, and subjecting the extract to rotary evaporation to obtain grape dregs oil.

[0127] Experimental Example 1 Extraction rate, active ingredient content and in vitro antioxidant activity of grape wine dregs oil

[0128] The technical effects of the embodiments and comparative examples of the present invention were compared, and the wine dregs oil extraction rate and component content were calculated.

[0129] (1) Calculation of yield: Grape dregs oil extraction rate (%) = Grape dregs oil mass (g) / Grape dregs crude fat mass (g) × 100%

[0130] (2) Determination of total phenol content:

[0131] Weigh 0.5g of oil and add 2ml of methanol-water solution (methanol: water = 90:10), then vortex for 5min, centrifuge at 3000rpm / min for 5 minutes, aspirate the supernatant, and repeat the above steps. Perform the extraction procedure for each oil three times. Mix all the extracts and concentrate to dryness, dissolve the dry matter in 1ml of methanol-water solution (methanol: water = 10:90) to obtain polyphenol extract. Accurately aspirate 0.5mL of the extract, add distilled water to 5ml, add 1mL of Folin-Ciocateu reagent and 4mL of 10% Na2CO3 solution respectively, shake well and place at room temperature for color development for 60min, use distilled water as a blank test, and measure the absorbance at 765nm respectively. Calculate the polyphenol content according to the regression equation of the gallic acid standard curve, and repeat the measurement 3 times for each sample.

[0132] (3) Determination of sterol content:

[0133] Weigh 0.5g of mercuric oxide, accurately add 2mL of concentrated sulfuric acid and 10mL of distilled water, ultrasonically dissolve, measure 1mL of the solution and place it in a 100mL volumetric flask, dilute to the mark with a mixed solution of glacial acetic acid and concentrated sulfuric acid (35:70), and obtain sulfoacetic acid mercuric reagent. Accurately weigh 0.1g of oil, dilute to the mark with methanol in a 10mL volumetric flask, take 2mL of the obtained oil-methanol solution and place it in a 10mL volumetric flask, dilute to the mark with sulfoacetic acid mercuric reagent, and measure its absorbance at 410nm using methanol as a blank. Calculate the sterol content according to the regression equation of the sitosterol standard curve, and repeat the measurement 3 times for each sample.

[0134] (4) Determination method of flavonoid content:

[0135] Weigh 0.5g of oil and add 2ml of methanol-water solution (methanol: water = 90:10), then vortex for 5 minutes, centrifuge at 3000rpm / min for 5min, absorb the supernatant, and repeat the above steps. The extraction procedure for each oil is performed three times. All extracts are mixed and concentrated to dryness, and the dry matter is dissolved in 5ml of methanol-water solution (methanol: water = 10:90) to obtain flavonoid extract. Pipette 1.0mL of extract into a 25mL graduated test tube, add 1mL of 5% sodium nitrite solution and shake well, wait for 6min, add 1mL of 10% Al(NO3)3 solution, add 10mL of 4% NaOH solution after 6min, and then add 60% ethanol solution to the scale. After standing for 15min, measure the absorbance at 510nm. Use distilled water as a blank control and rutin standard as a standard curve. Repeat the measurement for each sample 3 times.

[0136] (5) Determination of DPPH free radical scavenging rate

[0137] Weigh an appropriate amount of oil and dissolve it in anhydrous ethanol, then dilute it to obtain sample solutions of different mass concentrations for antioxidant activity test. 样品 :Pipette samples with different mass concentration gradients and mix them with DPPH working solution; A 空白 : Samples to be tested with different mass concentration gradients are taken and mixed with anhydrous ethanol; A 对照 : Mix the DPPH working solution with anhydrous ethanol. Mix well and react for 30 minutes at room temperature in the dark. After the reaction, use an ultraviolet spectrophotometer to measure the absorbance at 517nm. Repeat the measurement 3 times for each sample. The calculation formula for DPPH free radical scavenging rate is as follows:

[0138]

[0139] The effects of different extraction methods on the extraction rate, total phenols, flavonoids, sterols and in vitro antioxidant activity of wine dregs oil were determined. The results are shown in Table 1.

[0140] Table 1 Extraction rate, active ingredient content and DPPH free radical scavenging rate of wine dregs oil

[0141]

[0142]

[0143] Note: In the same column, different letters indicate significant difference between the two (p<0.05), and the same letters indicate no significant difference (p>0.05).

[0144] As can be seen from Table 1, the oil extraction rate of grape wine meal in the embodiment of the present invention is significantly higher than that in the control example. Specifically, the oil extraction rate of Example 1 reached 96.70±1.27%, which is the highest among all experimental groups. At the same time, the polyphenol, sterol and flavonoid contents of the grape wine meal oil extracted by the embodiment are relatively high, especially the best effect of Example 1, which are 1.13±0.18, 26.29±2.01, and 1.40±0.42 mg / g, respectively, which are significantly higher than the control examples. At the same concentration, the DPPH free radical scavenging rate of Example 1 is 98.22±1.54%, which is significantly higher than the control example, showing excellent antioxidant properties. These results show that the method of the present invention can effectively retain the active ingredients in the oil while extracting oil, and has a high in vitro antioxidant activity. The acid-thermal pretreatment combined with the use of low-temperature continuous phase change extraction fully extracts the active ingredients in each part of the grape wine meal and enriches them in the obtained grape wine meal oil.

[0145] Compared with Comparative Examples 1 and 2, Examples 1-3 are superior in DPPH free radical scavenging rate, oil extraction rate, polyphenol, flavonoid and sterol contents.

[0146] Compared with Examples 1 to 3, the extraction rate and DPPH free radical scavenging rate of Comparative Example 3 are significantly lower. Compared with Examples 1 to 3, the oil extraction rate, polyphenol, flavonoid and sterol content of Comparative Example 4 are all lower.

[0147] Experimental Example 2 Analysis of Physical and Chemical Indexes of Grape Seed Oil

[0148] (1) Determination method of peroxide value:

[0149] Weigh 2-3g of the prepared sample, place it in a 250mL iodine volumetric flask, add 30mL of chloroform-glacial acetic acid solution, and gently shake the sample until it is completely dissolved. Accurately add 1.00mL of saturated potassium iodide solution, plug the bottle cap tightly, and gently shake for 0.5min, and place it in a dark place for 3min. Take it out and add 100mL of water. After shaking well, immediately titrate the precipitated iodine with a standard sodium thiosulfate titration solution. When it is titrated to a light yellow color, add 1mL of starch indicator, continue titrating and shake vigorously until the blue color of the solution disappears as the end point. At the same time, perform a blank test. The volume of the standard sodium thiosulfate titration solution consumed in the blank test is V0. The peroxide value calculation formula is as follows:

[0150]

[0151] (2) Determination method of acid value:

[0152] Weigh the oil sample into a 250mL conical flask, add 50-100mL of ether-isopropanol mixture and 3-4 drops of phenolphthalein indicator, shake thoroughly to dissolve the sample, and then titrate with standard titration solution. The endpoint of the titration is when the sample solution begins to turn slightly red and there is no obvious fading within 15 seconds. Record the milliliters of standard titration solution consumed in this titration, which is V. Take another clean 250mL conical flask and accurately add the same volume and type of organic solvent mixture and indicator as when measuring the sample, shake to mix, and the milliliters of standard titration solution consumed by titration are V0. The acid value calculation formula is as follows:

[0153]

[0154] (3) Determination method of citric acid content:

[0155] Prepare 7 10mL centrifuge tubes and add 0.5mL of 0.4mol / L nitric acid solution, 2mL of 5.0×10 -4 mol / L phenol red solution, 0.1mL of 10mg / mL iron (III) solution, 0.1mL of 5% hydrogen peroxide solution, add 1mL of citric acid standard working solution (0.05mg / mL, 0.10mg / mL, 0.20mg / mL, 0.40mg / mL, 0.50mg / mL, 1.00mg / mL) to 6 of them, dilute to 8mL with distilled water, shake well, heat in 80℃ water bath for 8min, take out, quickly cool with 4℃ water for 3min, and compare with the one without citric acid, measure the absorbance A and A0 of the inhibitory system and catalytic system at 440nm wavelength, calculate △A=A-A0, and obtain the regression equation of the standard curve to calculate the sterol content. After the oil sample is extracted with water, the extract obtained is used to calculate the citric acid content in the sample according to the above experimental operation.

[0156] The effects of different extraction methods on the physicochemical indexes of wine dregs oil were determined. The results are shown in Table 2.

[0157] Table 2 Peroxide value, acid value and citric acid residue of wine dregs

[0158] Group Peroxide value (mmol / kg) Acid value (mg / g) Citric acid residue (%) Example 1 5.73±0.76 3.42±0.49 0.29±0.03 Example 2 6.96±0.65 3.43±0.18 0.22±0.01 Example 3 6.17±1.29 3.32±0.58 0.18±0.01 Comparative Example 1 17.86±0.84 3.67±0.18 Not detected Comparative Example 2 10.45±1.80 3.69±0.43 0.32±0.02 Comparative Example 3 8.85±1.94 3.33±0.09 Not detected Comparative Example 4 6.92±0.73 367±0.04 0.16±0.01

[0159] Peroxide value is an important indicator for measuring the degree of oxidation of oils and fats. A lower peroxide value indicates that the oxidation stability of oils and fats is better; acid value is an indicator for evaluating the content of free fatty acids in oils and fats. A lower acid value means that the degree of rancidity of oils and fats is lower and the quality is higher. The peroxide value and acid value of the grape wine dregs obtained in the embodiment of the present application are both low, far less than the requirements of the GB / T29990-2013 moisturizing oil standard (peroxide value is less than or equal to 10mmol / kg; acid value is less than or equal to 5mmol / kg). And there is a significant decrease compared with the comparative example. The peroxide value and acid value in comparative example 1 are both high. It can be seen that the solvent extraction method will promote the oxidation of oils and fats to a certain extent, and the quality of the obtained oils and fats is poor. At the same time, the citric acid residue of the grape wine dregs obtained in the embodiment of the present application is also within the standard range, and there is literature to prove that its presence in a small amount in oils and fats can play the role of an antioxidant. Therefore, the oil extraction method of the present application effectively controls the degree of oxidation and rancidity of oils and fats while ensuring a high extraction rate and active ingredient retention rate, thereby providing a high-quality and high-stability oil extraction solution.

[0160] The determination of the above-mentioned physical and chemical indicators shows that the extraction method of the present application is highly feasible and can achieve the goal of not sacrificing the quality of the oil while improving the content of active ingredients and in vitro antioxidant activity. The extracted oil meets the standard of GB / T 29990-2013 moisturizing oil.

[0161] The small amount of citric acid residue in the method of the present application indicates that although citric acid is used in the method of the present application, the extracted oil meets the standard of citric acid content in emollient oils of GB / T 29990-2013.

[0162] Experimental Example 3 Analysis of characteristic components of grape wine meal oil

[0163] The grape wine dregs oil obtained in Example 1 was analyzed for characteristic components.

[0164] (1) Gas chromatography-mass spectrometry analysis

[0165] S1. Take 100 μL of sample and add 300 μL of extraction solution (methanol:acetonitrile=2:1 (containing 0.05 mg / mL internal standard ribitol)); vortex for 30 seconds to mix, and then extract in an ice-water bath for 30 minutes; let the sample stand at -20°C for 30 minutes, and then centrifuge at high speed for 15 minutes (4°C, 13000 rcf); take the supernatant of the centrifuge, put it into a glass derivatization vial, and blow dry with nitrogen; add 80 μL of methoxyamine hydrochloride pyridine solution (15 mg / mL) to the glass derivatization vial, vortex for 2 minutes, and then carry out oximation reaction in a 37°C shaking incubator for 90 minutes; then add 80 μL of BSTFA (containing 1% TMCS) derivatization reagent, vortex for 2 minutes, and react at 70°C for 60 minutes; after taking out the sample, place it at room temperature for 30 minutes, and perform GC-MS metabolomics analysis.

[0166] S2. Chromatographic conditions: After derivatization, the sample was injected into the GC-MS system for analysis in split mode, with an injection volume of 1 μL and a split ratio of 10:1. The sample was separated by a TG-5SILMS capillary column (30m×0.25mm×0.25μm, Thermo26096-1420) and then entered the mass spectrometer detection. The injection port temperature was 300°C, the carrier gas was high-purity helium, the carrier gas flow rate was 1.0mL / min, and the septum purge flow was 3mL / min. Heating program: initial temperature 80°C, equilibrium 0min, then increased to 310°C at a rate of 20°C / min, and maintained for 8min, the total running time was 20min, and the solvent delay was 2min.

[0167] S3. Mass spectrometry conditions: electron impact (EI) source, electron energy 70 eV, scanning mode Full Scan, scanning mass range 35-500 m / z, ion source temperature 250°C.

[0168] Test results see Figure 1 And Table 3. Figure 1 This is the total ion current spectrum of wine meal oil analyzed by gas chromatography-mass spectrometry.

[0169] Table 3 Chemical components and relative contents of wine meal oil analyzed by GC-MS

[0170]

[0171]

[0172]

[0173]

[0174]

[0175] The results of gas chromatography-mass spectrometry analysis showed that a total of 103 compounds were identified in grape wine dregs, involving 12 categories of substances. Among them, 31 belonged to fatty acids and lipids, 14 belonged to amino acids, 8 belonged to sugars and their derivatives, 11 belonged to carboxylic acids, 3 belonged to aldehydes, 7 belonged to alcohols, 3 belonged to phenols and their derivatives, 3 belonged to flavonoids, 6 belonged to terpenes and terpenes-related compounds, 5 belonged to sterols, 7 vitamins and 5 other categories. It can be seen that grape wine dregs are very rich in active ingredients.

[0176] It is worth noting that relatively high contents include conjugated (9E, 11E)-linoleic acid (20.3205%) and α-linolenic acid (6.6875%). These polyunsaturated fatty acids can protect cells from oxidative damage by neutralizing free radicals, and can improve the skin's ability to retain moisture, thereby delaying skin aging. Another lipid substance with a high content is monopalmitoylglycerol (5.0188%). In addition to its anti-aging activity, it is also commonly used as an emulsifier and emollient in cosmetics, and has moisturizing and nourishing effects on the skin. Among terpenes and terpene-related compounds, oleanolic acid (11.6108%) has a high content and has significant anti-aging and antioxidant effects, and can improve and enhance dermal collagen; and α-guaiacene (4.5109%) is next. Although there are no reports on its anti-aging activity, its antibacterial and anti-inflammatory activities have been widely recognized. In terms of phytosterols, grape dregs are rich in β-sitosterol (7.0947%) and campesterol (1.3126%), which are representatives of high-value active ingredients in grape seeds, giving grape dregs excellent anti-aging effects. Another representative of high-value active ingredients is vitamin E, the main configuration of which in the grape dregs prepared by the present invention is β-tocotrienol (2.8166%), and it also contains tocopherols of different configurations, laying a foundation for grape dregs to be used as an effective antioxidant.

[0177] At the same time, the results also found that the grape wine dregs contained a variety of polyphenols and flavonoids, including protocatechuic acid, salicylic acid, caffeic acid, catechins, kaempferol and gallic acid, etc., with a total relative content of 0.5074%. In addition, other high-value anti-aging active ingredients were also discovered, such as squalene (0.0079%), inositol (0.0449%), phytol (0.0740%), L-erythro-sphingosine (0.5239%), lupeol (0.2087%), etc. The above substances are all high-quality antioxidants that can play an anti-aging role in the body or in the skin. At the same time, some substances also have anti-inflammatory activity, so they are used as raw materials for high-end cosmetics. Therefore, the grape wine dregs prepared by the present invention has a variety of potential health benefits and has important application value in the fields of health foods, cosmetics, etc.

[0178] (2) Liquid chromatography-mass spectrometry analysis

[0179] S1. Accurately transfer 10 μL of sample to a 1.5 mL centrifuge tube; add 400 μL of extract (methanol: water = 4:1 (v:v)) containing four internal standards (L-2-chlorophenylalanine (0.02 mg / mL) etc.); vortex mix for 30 seconds, perform low-temperature ultrasonic extraction for 10 minutes (5°C, 40 kHz); centrifuge for 15 minutes (13000 g, 4°C), transfer the supernatant to an injection vial with an inner tube and analyze on the analyzer.

[0180] S2. Chromatographic conditions: the chromatographic column was ACQUITY UPLC HSS T3 (100 mm × 2.1 mm id, 1.8 μm; Waters, Milford, USA); the mobile phase A was 95% water + 5% acetonitrile (containing 0.1% formic acid), the mobile phase B was 47.5% acetonitrile + 47.5% isopropanol + 5% water (containing 0.1% formic acid), the injection volume was 3 μL, and the column temperature was 40°C.

[0181] S3. Mass spectrometry conditions: The sample was electrospray ionized, and the mass spectrometry signals were collected in positive and negative ion scanning modes, respectively. The scanning range was 70-1050 m / z, the spray voltage (positive mode) was 3500 V, the spray voltage (negative mode) was -3500 V, and the capillary temperature was 325°C.

[0182] Test results see Figure 2 And Table 4. Figure 2 This is the total ion current spectrum of wine meal oil analyzed by liquid chromatography-mass spectrometry.

[0183] Table 4 Chemical components and relative contents of wine dregs analyzed by HPLC-MS

[0184]

[0185]

[0186]

[0187]

[0188]

[0189] The results of liquid chromatography-mass spectrometry analysis showed that a total of 123 compounds were identified in wine dregs, involving 15 categories of substances. Among them, 20 belonged to phenols and their derivatives, 12 belonged to amino acids and their derivatives, 2 belonged to sugars, 20 belonged to terpenes and terpenes-related compounds, 16 belonged to organic acids, 12 belonged to flavonoids, 6 belonged to alcohols, 19 belonged to fatty acids and lipids, 3 belonged to glycosides, as well as 1 amide, 1 alkaloid, 2 nitrogen-containing heterocyclic compounds, 2 vitamins, 3 aldehyde compounds and 4 ketone compounds.

[0190] Among them, soybean saponin E has the highest relative content, reaching 29.7707%, and has multiple biological activities such as lowering blood lipids, immunomodulation, anti-oxidation, and anti-tumor, making it have broad application prospects in the fields of medicine, health foods, and cosmetics. Secondly, the second most abundant compound is D-erythro-dihydrosphingosine (20.1058%), which is an isomer of sphingosine and is a key component of cell membrane phospholipids. It is essential for maintaining the skin barrier and nerve cell function. The third most abundant compound is soybean cerebroside I (7.5149%), a glucocerebroside that has not only been shown to have multiple pharmacological effects such as anti-tumor and anti-inflammatory, but also shows a strong inhibitory effect on tyrosinase. Interestingly, two active ingredients that appear in Ganoderma lucidum were also found in grape dregs, and the content is relatively considerable, namely, ganoderic acid F (3.2038%) and ganoderic alcohol I (1.7824%), which are reported to have anti-tumor, anti-inflammatory and immunomodulatory effects. In addition, 9 kinds of ceramides were found to exist in the grape dregs of the present invention, with Cer (8:0_2O / 10:0) having the highest content (5.7270%), wherein "8:0" indicates that the length of the fatty acid chain is 8 carbon atoms, and all of them are saturated carbon atoms (no double bonds), "2O" indicates that two hydroxyl groups (OH) are connected to the sphingosine base, and "10:0" indicates that the length of another fatty acid chain is 10 carbon atoms, and all of them are saturated carbon atoms. Another ceramide HexCer refers to hexosylceramide, which contains two fatty acid chains and one sugar group, with HexCer (14:3_20 / 2 8:1_20) having the highest content (1.2074%). Ceramide has a variety of physiological and pharmacological functions, such as regulating cellular immunity, delaying aging, anti-tumor, etc., and in the cosmetics industry, ceramide may help promote the degradation of the stratum corneum of the skin, thin the stratum corneum, promote the division and proliferation of epidermal cells, and thus delay the aging process of epidermal cells of the skin.

[0191] Also worth noting are the polyphenols and flavonoids in wine dregs, with total relative contents of 0.9710% and 2.4048% respectively. Among the polyphenols, gentisic acid (0.1112%), 2-hydroxycinnamic acid (0.1340%), gallic acid (0.1242%) and other compounds are relatively high, while protocatechuic aldehyde, ferulic acid, vanillin, dihydrocaffeic acid and other compounds are also detected, which are all common and effective antioxidants in plants, and some substances, such as dihydrocaffeic acid, can even prevent UV-induced skin damage, implying its positive role in resisting photoaging. Among the flavonoids, isorhamnetin (0.8964%), kaempferol (0.4201%), 6"-O-p-coumaryl trifoliol (0.1950%), quercetin (0.1752%) and other substances are present in high levels. Myricetin, which also has anti-photoaging effects, and dihydroquercetin, which has anti-tyrosinase activity, were also found. Therefore, the above results further demonstrate the health benefits of grape dregs oil, especially its antioxidant and anti-aging effects, and it has great application value in health foods and cosmetics.

[0192] Effect example: Anti-aging effect of grape dregs oil

[0193] The anti-aging activity of the wine dregs oil obtained in Example 1 was evaluated using the Caenorhabditis elegans model, and its effect on extending the lifespan of nematodes in the presence of oxidative stress damage was tested.

[0194] Caenorhabditis elegans has shown significant advantages in anti-aging research and has been widely accepted as a research model due to its easy manipulation, simple observation, simple physiological structure, rapid life cycle, clear genetic information, high similarity to human genes, and complete genome sequencing.

[0195] Oxidative stress is caused by excessive reactive oxygen species (ROS) in the body and is considered to be the main factor driving aging in nematodes. The antioxidant capacity of nematodes is closely related to their lifespan, and the enhancement of antioxidant capacity is often accompanied by an extension of lifespan. Hydrogen peroxide (H2O2), as an effective mimetic, can simulate the process of cell apoptosis caused by free radicals, and therefore plays a key role in the preparation of cell oxidative damage models. Paraquat (PQ) induces oxidative stress reactions in cells, produces excessive ROS, and causes mitochondrial function damage, which makes it widely used in the study of mitochondrial oxidative damage. At the same time, ultraviolet (UVB) exposure is the main exogenous factor causing photoaging. UVB radiation can directly penetrate the atmosphere and interact with cells in the skin, producing a large number of free radicals and reactive oxygen molecules, causing intracellular oxidative stress and accelerating the skin aging process. There is a positive intrinsic connection between enhancing the ability to withstand stress and extending lifespan. Active substances can improve the ability of nematodes to resist heat stress in a 35°C environment by regulating heat stress proteins and extend the lifespan of nematodes.

[0196] Therefore, by treating nematodes with samples, subsequently placing them under corresponding stress conditions, and recording the survival status of the nematodes, the potential effect of wine meal oil in anti-aging can be evaluated.

[0197] (1) H2O2-induced oxidative stress injury experiment

[0198] After synchronization of nematodes, L4 larvae were selected and transferred to the culture medium of the blank group and the sample group. After the sample group was fed with wine lees at a concentration of 1 mg / mL for 3 days, 30 to 50 nematodes were selected from each culture medium and exposed to NGM culture medium containing H2O2 for oxidative stress induction intervention. Three parallel plates were set up in the experiment, and the number of nematode survival, escape, and death was recorded every 30 minutes until all nematodes died. The survival curve of nematodes under H2O2 oxidative stress environment was obtained, and 60μM astaxanthin was used as a positive control.

[0199] The results are as follows Figure 3 As shown in the figure, compared with the blank group, the survival curve of the wine dregs group was significantly shifted to the right (P < 0.0001), among which the average life span of the blank group was 1.53h, and the average life span of the wine dregs group and the astaxanthin group were 2.03h and 2.00h, respectively, which were increased by 33.09% and 30.86% compared with the blank group, indicating that wine dregs significantly improved the resistance of nematodes to H2O2 oxidative stress damage and could effectively prolong the life span of nematodes. At the same time, the anti-aging activity of 1mg / mL wine dregs was not significantly different from that of 60μM astaxanthin (P > 0.05), indicating that its anti-aging activity was good.

[0200] (2) PQ-induced oxidative stress injury experiment

[0201] After synchronization of nematodes, L4 larvae were selected and transferred to the culture medium of the blank group and the sample group. After 3 days of drug action, 50 to 60 nematodes were selected from each culture medium and exposed to NGM culture medium containing PQ for oxidative stress induction intervention. Three parallel plates were set up in the experiment. Every 24 hours or so, the number of nematode survival, escape, and death was recorded until all nematodes died, and the survival curve of nematodes under PQ oxidative stress environment was obtained.

[0202] The results are as follows Figure 4 As shown in the figure, similar to the results of the H2O2 oxidative stress experiment, the survival curve of the drug group was significantly shifted to the right compared with the blank group (P < 0.001), indicating that wine dregs oil can prolong the survival time of nematodes under PQ oxidative damage conditions. Among them, the average lifespan of the blank group was 2.39 days, while the average lifespans of the wine dregs oil group and the astaxanthin group were 2.88 days and 3.43 days, respectively, and the average survival time was extended by 20.37% and 43.30%, respectively, further indicating that wine dregs oil can effectively enhance the resistance of nematodes to oxidative stress and thus prolong the lifespan of nematodes.

[0203] (3) Heat-induced stress injury experiment

[0204] Wild-type Caenorhabditis elegans in the reproductive period (3d to 5d old) were collected for synchronization. After synchronization, the nematodes were transferred to NGM with or without drugs for 5 days and cultured at 20°C. During this period, the nematodes were transferred to new plates every day. On the 6th day of the adult stage, the nematodes were placed in a 35°C incubator for 12 hours, and then placed in a 20°C incubator for 12 hours. The survival rate of the nematodes was observed and recorded. The experiment was set up in 3 parallels, and the number of nematodes in each treatment group was not less than 120.

[0205] Depend on Figure 5 It can be seen that the nematode survival rate in the blank group was significantly lower than that in the other two groups, only 18.89%, which indicates that heat stress has a negative impact on the survival of nematodes without treatment. The nematode survival rate in the wine dregs oil treatment group was significantly higher than that in the blank group by 38.89 percentage points (P < 0.05), indicating that wine dregs oil can improve the survival rate of nematodes under heat stress conditions. At the same time, the nematode survival rate in the positive control group was slightly lower than that in the wine dregs oil treatment group, but the difference between the two groups was not statistically significant. This shows the effectiveness of wine dregs oil in improving the survival rate of nematodes under heat stress and its potential advantages in resisting heat aging.

[0206] (4) Light-induced stress damage experiment

[0207] Wild-type Caenorhabditis elegans in the reproductive period (3-5 days old) were collected for synchronization. After synchronization, the nematodes were transferred to NGM with or without drugs for 3 days. The nematodes in each group were collected, washed 3 times with M9 buffer, and transferred to NGM without OP50. The NGM plate with nematodes was placed directly under the UVB lamp of the irradiator, one dish at a time, and the irradiation dose was set to 250mJ / cm 2 After UV irradiation, transfer each group of nematodes to OP50 NGM containing FUDR and continue to place them in a 20°C incubator. Pick out dead nematodes every 12 hours, and record the number of dead and alive nematodes to obtain a survival curve of nematodes, as shown in Figure 2. Figure 6 The experiment was set up in triplicate, with no less than 120 nematodes in each treatment group.

[0208] Depend on Figure 6 It can be seen that the survival rate of nematodes in the blank group decreased rapidly under light stress conditions, and the survival rate was 0% on the 4th day. In contrast, the nematodes in the wine dregs oil group and the astaxanthin group showed a significantly improved survival rate under light stress conditions, and the survival curves were significantly shifted to the right (P < 0.001). The average lifespan of the blank group was 2.63 days, while the average lifespans of the wine dregs oil group and the astaxanthin group were 2.87 days and 3.05 days, respectively, and the average survival time was extended by 9.19% and 16.11%, respectively. Therefore, wine dregs oil has a good effect on improving the survival ability of nematodes in UVB irradiation environment, and its anti-photoaging activity has been confirmed.

[0209] Conclusion: The results show that the wine dregs oil prepared by the present invention has high-quality anti-aging effect.

[0210] In summary, the present invention adopts the acid-heat pretreatment method, combined with the low-temperature continuous phase change extraction equipment, to extract and prepare grape wine dregs oil with high active ingredient content, good in vitro antioxidant activity, rich in anti-aging active ingredients, and anti-aging efficacy. The comprehensive utilization of oil in grape wine dregs is achieved, which is green, safe, efficient, and suitable for industrial production and application.

[0211] The contents of the three active ingredients (total phenols, total flavonoids, and total sterols) and the in vitro antioxidant activity measured by the method of this application are excellent, indicating the effectiveness and feasibility of the innovative method of this application. This application provides a new extraction method, which can improve the anti-aging activity of the extract without sacrificing the quality of the oil. The product of this innovative extraction method can be fully understood by analyzing the characteristic components of the extract, and its application in cosmetics can effectively enhance the anti-aging effect of cosmetics.

[0212] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. An anti-aging cosmetic, characterized in that: include: Grape dregs; The characteristic components of the grape wine dregs oil include: oleanolic acid, α-guaiacene, β-tocotrienol, β-sitosterol, campesterol, soybean saponin E, ganoderic acid F, ganoderic alcohol I and D-erythro-dihydrosphingosine; the total phenol content of the grape wine dregs oil is ≥1 mg / g, the total sterol content of the grape wine dregs oil is ≥26 mg / g, and the total flavonoid content of the grape wine dregs oil is ≥1 mg / g; The grape wine meal oil is obtained by extracting grape wine meal; the grape wine meal consists of grape skins, grape stems and grape seeds.

2. The anti-aging cosmetic according to claim 1, characterized in that: The peroxide value of the grape wine meal oil is ≤7mmol / Kg.

3. The anti-aging cosmetic according to claim 1, characterized in that: The acid value of the grape wine meal oil is ≤4 mg / g.

4. A method for preparing grape wine dregs oil, characterized in that: The preparation method comprises: The wine dregs powder is hot-soaked in an acid solution to obtain a first liquid; Subjecting the first liquid to continuous phase change extraction at 0.2 MPa to 0.8 MPa and 40° C. to 50° C.; The wine dregs powder consists of grape skins, grape stems and grape seeds.

5. The method for preparing grape wine dregs oil according to claim 4, characterized in that: The continuous phase change extraction comprises: n-Butane is used as the extractant to carry out liquid phase, gas phase and liquid phase continuous cycle phase change extraction.

6. The method for preparing grape wine dregs oil according to claim 4 or 5, characterized in that: The continuous phase change extraction comprises a plurality of extraction cycles, one of the extraction cycles comprising: compressing the extractant gas into an extractant liquid in a vacuum, and allowing the extractant liquid to flow through the first feed liquid for extraction; the wine dregs oil and the acid solution obtained by extraction flow out with the extractant liquid to obtain a mixed liquid; converting the extractant liquid into an extractant gas, and separating it from the mixed liquid; and repeating the extraction cycle with the separated extractant gas; Optionally, after separating the extractant gas, the acid solution is removed from the mixed solution to obtain a crude extract; Optionally, subjecting the crude extract to a second extraction; Optionally, the crude extract is subjected to a second extraction using n-hexane.

7. The method for preparing grape wine dregs oil according to claim 4, characterized in that: The acid solution is an alcohol solution of an acid; Optionally, the acid solution is a citric acid-ethanol solution; Optionally, the citric acid-ethanol solution comprises, by mass percentage, 1% to 5% citric acid and 95% to 99% ethanol.

8. The method for preparing grape wine dregs oil according to claim 4, characterized in that: The method of hot soaking the wine dregs powder with an acid solution comprises: Based on the mass ratio, the grape dregs powder and the acid solution are hot-soaked in a solid-liquid ratio of (1:1) to (1:3).

9. The method for preparing grape wine dregs oil according to claim 4, characterized in that: The method of hot soaking the wine dregs powder with an acid solution comprises: Soaking the grape wine dregs powder and the acid solution at 40° C. to 50° C.; Optionally, the soaking time is 1 hour to 3 hours.

10. A grape wine dregs oil, characterized in that: The wine dregs oil is obtained by the preparation method of any one of claims 4 to 9.

Images