Plant composition with skin anti-aging activity and preparation thereof

Abstract

The invention discloses a plant composition with skin anti-aging activity and a preparation thereof. The plant composition is prepared from the following active ingredients in percentage by weight: 84.0 to 89.0 percent of haematococcus pluvialis extract, 8.0 to 12.0 percent of lithospermum oil, 0.5 to 0.8 percent of melaleuca alternifolia leaf oil, 0.5 to 1.0 percent of hydrolyzed olive leaf extract, 0.1 to 0.5 percent of rhodiola rosea root extract and 1.2 to 2.5 percent of bitter orange leaf oil. The plant composition with the skin anti-aging activity is used as an effective component, other auxiliaries are added, and emulsion, cream or essence oil is prepared. According to the plant composition and the preparation, the fermented camelina seed oil, the fermented safflower seed oil, the fermented meadowfoam seed oil and the like are used as solvents for extracting active components in haematococcus pluvialis and are also used as functional components and matrixes of the preparation, and compared with application of original plant oil, the plant composition and the preparation have better skin feeling and biological activity; the extraction rate of active ingredients in haematococcus pluvialis is higher, and transdermal absorption of the active ingredients is facilitated.

Description

1. Technical Field:

[0001] The present invention belongs to the technical field of cosmetics, and specifically relates to a plant composition with skin anti-aging activity, which has plant-based ingredients such as Haematococcus pluvialis extract and lithospermum oil as main functional ingredients, and a preparation thereof. 2. Background technology:

[0002] The skin is the largest organ in the human body and a crucial barrier to entry. It blocks the invasion of foreign particles and pathogens, prevents fluid loss, and regulates body temperature, secretion, and excretion. With rising living standards, people are paying particular attention to the care of their facial skin. Facial skin not only reflects a person's health and mental outlook, but also influences their psychological and social well-being. Healthy facial skin not only enhances one's appearance but also boosts social confidence. Facial skin aging is an inevitable physiological consequence of aging. However, with increasing social pressures, increasing dependence on electronic devices, environmental pollution, and increased exposure to various chemicals, facial skin aging is occurring at a younger age. As cosmetics become increasingly essential, demand for their efficacy has evolved from simple moisturizing and hydrating to multiple benefits such as whitening, spot reduction, anti-aging, and wrinkle reduction. Common chemical additives in cosmetics, such as preservatives, artificial fragrances and pigments, and penetration enhancers, are believed to be key factors in disrupting the skin barrier and inducing skin allergies. Plant-derived ingredients are generally considered mild and low-irritation. Therefore, as consumers pay more attention to the green and safety of products, plant-based cosmetics have gained attention and their market share has increased. The development and utilization of cosmetics with plant-derived functional raw materials has become an important development direction for the cosmetics industry in the future.

[0003] Astaxanthin is a popular cosmetic ingredient, also known as a "super antioxidant." It's one of the most potent natural antioxidants discovered to date. Its potent antioxidant properties can protect against the damage of reactive oxygen species (ROS) to collagen and elastin in the dermis, thereby exerting anti-wrinkle effects. Furthermore, its structure results in strong UV absorption close to UVA wavelengths, which can protect against photoaging. (UVA is currently the primary cause of photoaging. Due to its longer wavelength, it can penetrate the skin's surface and penetrate deep into the dermis, damaging collagen and elastin, leading to sagging, wrinkling, and hyperpigmentation.) Therefore, astaxanthin can exert its anti-aging effects through multiple mechanisms. Astaxanthin is widely found in organisms, including shrimp, crab, and fish. However, the highest natural source of astaxanthin discovered so far is Haematococcus pluvialis. Because Haematococcus pluvialis has thick cell walls and its active ingredients, such as astaxanthin, are easily oxidized and destroyed, efficient utilization of astaxanthin and other active ingredients from Haematococcus pluvialis requires a comprehensive extraction process.

[0004] Existing extraction processes for astaxanthin and other active ingredients from Haematococcus pluvialis mostly involve three steps: cell wall disruption, extraction, and drying / purification. This entire process is time-consuming and can lead to oxidative destruction of the active ingredients in the algae. Haematococcus pluvialis cell wall disruption primarily involves physical and chemical cell wall disruption. Patents CN 101381337A, CN104513185A, CN103787941A, and CN10762897A, respectively, propose methods for cell wall disruption using airflow pulverization, supersonic airflow pulverization, high-pressure homogenization, and steam explosion. Patents CN103232375A, CN107011225A, and CN103044303A, respectively, propose cell wall disruption using an acidic aqueous solution, a concentrated sulfuric acid-alcohol system, and enzyme treatment. The extraction methods of astaxanthin mainly include organic solvent extraction and oil solvent extraction. Using a mixed low-toxic organic solvent and common vegetable oil as the extraction solvent faces the problem of solvent separation. Excessively high temperature and complex extraction process will inevitably affect the stability of active ingredients such as astaxanthin in Haematococcus pluvialis. For example, patent CN 202410037536.4 discloses a method for extracting and applying astaxanthin, using shrimp shell enzymatic hydrolysate as Haematococcus pluvialis culture medium, and extracting astaxanthin by organic solvent extraction-rotary evaporation drying. Patent CN202211238411.5 discloses a method for preparing an antioxidant extract, which uses a polar solvent and a non-polar solvent mixed with the Haematococcus pluvialis after breaking the wall as the extraction solvent. The polar solvent is ethanol, and the non-polar solvent is one of ethyl acetate or hexane. However, the extract obtained by this method still needs to be evaporated to remove the solvent before it can be used as a cosmetic raw material. Therefore, how to optimize the extraction process of Haematococcus pluvialis, on the one hand, to achieve simultaneous completion of cell wall breaking and active ingredient extraction, while minimizing or eliminating subsequent enrichment and purification processes, will show great advantages in the application of Haematococcus pluvialis in the cosmetics field.

[0005] The human epidermis is a "sand brick structure." "Bricks" mainly refer to epidermal cells, and "sand" mainly refers to the lipid components between epidermal cells. Skin lipids mainly include sebaceous gland lipids (lipid components secreted to the skin surface by sebaceous glands) and intercellular lipids, which together constitute the lipid barrier of human skin and play an important role in resisting environmental pollutants, participating in inflammatory responses, delaying aging, and regulating skin color. Triglycerides and free fatty acids are important components of skin lipids. Clinical studies have observed that the triglyceride content in human sebum gradually decreases after adulthood, resulting in a decrease in the skin's hydration capacity, causing aging phenomena such as dull skin, desquamation, and wrinkles. The skin of people with oily skin ages significantly slower than that of people with dry skin. The effect of triglycerides on the ultraviolet-induced photoaging model of human immortalized keratinocytes showed that triglycerides can significantly reduce the expression of extracellular matrix metalloproteinase-1, playing an important role in the repair mechanism of skin damage after photoaging and acute ultraviolet exposure. Free fatty acids are important lipids that maintain the skin barrier. They ensure the skin's weakly acidic environment, prevent the colonization of external pathogens, and thus maintain the balance of the skin's microecology.

[0006] Plant oils are important raw materials in the cosmetics industry, their primary component being triglycerides. Diffusion through the stratum corneum is the primary route of transdermal absorption of cosmetics, including through the intercellular space, through the cellular space, and through the skin's appendages. The intercellular space accounts for approximately 30% of the total volume of the stratum corneum, but because it has less resistance than corneocytes, it plays a major role in transdermal penetration. The intercellular spaces of the epidermis are primarily composed of lipids, making it easier for fat-soluble, non-polar substances to penetrate the lipid layer of the intercellular space and be absorbed transdermally, reaching deeper layers of the epidermis and even the dermis, thereby exerting their skin care benefits. Using plant oils as both a base oil and a functional active ingredient in cosmetics, and simultaneously as an extraction solvent for astaxanthin and other active ingredients in Haematococcus pluvialis, plant oils can be directly used as a compounding ingredient, promoting the penetration and absorption of functional ingredients from Haematococcus pluvialis, achieving multiple goals simultaneously. This not only streamlines the process but also enhances the overall functional activity of the cosmetic. Different plant oils differ in their fatty acid composition, the types and amounts of concomitants, and other factors, leading to varying effects in skin care. Limnanthes alba seed oil and camelina seed oil are two cosmetic ingredients with compositions that differ significantly from other vegetable oils. Limnanthes alba seed oil is one of the most stable vegetable oils known, with a high content of very-long-chain (≥20C) unsaturated fatty acids. Camelina seed oil is also one of the few vegetable oils discovered to contain a high content of very-long-chain unsaturated fatty acids, containing up to 14% C20:1 and 0.3-1.2% nervonic acid (C24:1), which plays a vital role in maintaining the skin's lipid barrier. Furthermore, nervonic acid has unique advantages in wound healing. Safflower seed oil is a unique vegetable oil ingredient in cosmetics, known as the "king of linoleic acid," with the highest linoleic acid content among vegetable oils. Linoleic acid participates in the synthesis of phospholipids in the human body, is a component of cell membranes and mitochondria, and is also a component of epidermal lipid ceramides. Therefore, linoleic acid is crucial for human skin metabolism and the repair of damaged skin tissue. Safflower seed oil also contains tocopherols, flavonoids, phytosterols, and other substances, and has excellent stability.

[0007] Biofermentation technology uses microbial metabolism to convert complex ingredients in raw materials into small molecules that are easily absorbed by the skin, improving the absorption rate and efficacy of active ingredients. It is also a key area for the development of new cosmetic raw materials. During the fermentation process of cosmetic vegetable oils by oleaginous yeast, some triglycerides are hydrolyzed to produce free fatty acids, while a series of endogenous aromatic compounds are also metabolized. Cosmetics are an ideal application for the advantages of fermented vegetable oils. Compared to the original vegetable oils, fermented vegetable oils exhibit significant changes in sensory properties, including increased polarity and decreased surface tension. These oils can serve not only as cosmetic bases but also as a primary active ingredient, enhancing the functional activity of cosmetics. To date, there are no published studies on cosmetic formulations using Haematococcus pluvialis extracts extracted using fermented vegetable oils as a solvent as an active ingredient, nor are there any references to the combination of fermented camelina seed oil, fermented meadowfoam seed oil, and fermented safflower seed oil. 3. Summary of the invention:

[0008] The technical problem to be solved by the present invention is: in response to the current market demand for green and safe plant-based anti-aging cosmetics, the present invention provides a plant composition and preparation thereof with anti-aging activity, comprising a Haematococcus pluvialis extract as the primary active ingredient. Specifically, the present invention provides a plant composition and preparation thereof with skin anti-aging activity. The plant composition of the present invention uses fermented camelina seed oil, fermented safflower seed oil, and fermented meadowfoam seed oil as solvents for extracting the active ingredients from Haematococcus pluvialis, and also as both the active ingredient and matrix of the preparation. Compared with the use of crude plant oils, the plant composition exhibits improved skin feel and biological activity, a higher extraction rate of the active ingredients from Haematococcus pluvialis, and enhanced transdermal absorption of the active ingredients.

[0009] In order to solve the above problems, the technical solution adopted by the present invention is:

[0010] The invention provides a plant composition with skin anti-aging activity. The plant composition comprises active ingredients of Haematococcus pluvialis extract, lithospermum officinale oil, Melaleuca alternifolia leaf oil, hydrolyzed olive leaf extract, Rhodiola rosea root extract and petitgrain oil. Expressed in percentage by mass, the proportions of the active ingredients are as follows: 84.0-89.0% of Haematococcus pluvialis extract, 8.0-12.0% of lithospermum officinale oil, 0.5-0.8% of Melaleuca alternifolia leaf oil, 0.5-1.0% of hydrolyzed olive leaf extract, 0.1-0.5% of Rhodiola rosea root extract and 1.2-2.5% of petitgrain oil.

[0011] According to the above-mentioned plant composition with anti-aging activity for the skin, the Haematococcus pluvialis extract is obtained by subjecting Haematococcus pluvialis powder to high-pressure homogenization and wall-breaking extraction using fermented vegetable oil as an extraction solvent; the fermented vegetable oil is at least one of fermented camelina seed oil, fermented meadowfoam seed oil and fermented safflower seed oil.

[0012] According to the above-mentioned plant composition with skin anti-aging activity, the specific extraction method of the Haematococcus pluvialis extract is:

[0013] a. Using Yarrowia lipolytica as the fermentation strain, the fermentation strain was inoculated into a liquid culture medium for aerobic fermentation on a shaker at a temperature of 28 to 35° C. for 16 to 36 hours to obtain a seed solution;

[0014] b. adding the obtained seed liquid to the sterilized liquid culture medium at a volume ratio of 3-5% for primary fermentation at a fermentation temperature of 28-35° C. for 24-48 h to obtain a primary fermentation liquid;

[0015] c. Sterilize each vegetable oil separately, then add each sterilized vegetable oil into the obtained primary fermentation liquid, and continue fermenting for 24 to 72 hours at a fermentation temperature of 28 to 35°C;

[0016] The volume ratio between the primary fermentation liquid and the sterilized vegetable oil is 1:1-2;

[0017] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is removed and the bacterial cells are separated;

[0018] e. Wash the obtained cells with phosphate buffer, centrifuge at 4000-6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0019] The obtained clean yeast cells are added with dried Haematococcus pluvialis powder and mixed to obtain Haematococcus pluvialis powder containing yeast cells; the Haematococcus pluvialis powder containing yeast cells is then added to the obtained fermented vegetable oil and mixed. After the mixture is evenly mixed, a high-pressure homogenizer is used to homogenize the mixture 1 to 2 times at a pressure of 1000 to 1500 bar (to break the yeast cells and the Haematococcus pluvialis powder and to extract the fat-soluble active ingredients in the yeast cells and the Haematococcus pluvialis powder); after homogenization, the mixture is centrifuged at 8000 rpm for 10 minutes, and the supernatant is collected to obtain the Haematococcus pluvialis extract.

[0020] According to the above-mentioned plant composition with skin anti-aging activity, the vegetable oil in step c is camelina seed oil, meadowfoam seed oil or safflower seed oil; and in step e, the ratio of Haematococcus pluvialis powder to fermented vegetable oil is 1 g:15-20 mL.

[0021] According to the above-mentioned plant composition with skin anti-aging activity, the rotation speed of the shaker is controlled to be 160-220 rpm during the fermentation.

[0022] In addition, a preparation of a plant composition having skin anti-aging activity is provided, wherein the preparation is prepared from the plant composition having skin anti-aging activity into an emulsion, a cream or an essential oil.

[0023] According to the above-mentioned preparation of the plant composition with skin anti-aging activity, the plant composition preparation emulsion with skin anti-aging activity is composed of the following components in percentage by mass: 6.7-13.2% of the above-mentioned plant composition with skin anti-aging activity, 1.0-2.0% of glyceryl stearate, 1.0-1.5% of cetearyl alcohol, 2.0-5.0% of isohexadecane, 62.9-81.2% of water / birch sap, 5.0-10.0% of glycerin, 0.1-0.2% of sodium polyacrylate, 0.2-0.4% of allantoin, 0.2-0.4% of carbomer 940, 1.0-1.5% of tocopherol, 0.4-0.6% of p-hydroxyacetophenone and 1.0-2.5% of 1,2-hexanediol.

[0024] According to the above-mentioned preparation of the plant composition with anti-aging activity for skin, the plant composition preparation cream with anti-aging activity for skin is prepared from the following components in percentage by weight: 9.0-18.0% of the above-mentioned plant composition with anti-aging activity for skin, 1.5-2.5% of glyceryl stearate, 3.0-9.0% of cetearyl alcohol, 4.0-6.0% of isohexadecane, 55.0-74.5% of water / birch sap, 4.0-10.0% of glycerol, 0.1-0.2% of sodium polyacrylate, 0.2-0.4% of allantoin, 0.1-0.2% of xanthan gum, 1.0-1.5% of tocopherol, 0.4-0.6% of p-hydroxyacetophenone and 1.0-2.0% of 1,2-hexanediol.

[0025] According to the above-mentioned preparation of the plant composition with anti-aging activity for skin, the essence oil of the plant composition preparation with anti-aging activity for skin is composed of the following components in percentage by weight: 14.0-26.0% of the above-mentioned plant composition with anti-aging activity for skin, 72.0-85.0% of base oil and 0.5-3.5% of tocopherol.

[0026] According to the above-mentioned preparation of the plant composition with skin anti-aging activity, the base oil is at least one of camelina seed oil, linseed oil, rapeseed oil, jojoba seed oil, perilla seed oil, peony seed oil, meadowfoam seed oil, safflower seed oil, camellia seed oil and sweet almond oil.

[0027] In the technical solution of the present invention, the preparation method of the liquid culture medium is as follows: 2 g of glucose, 0.3 g of yeast extract, 2 g of peptone and 0.3 g of malt extract are added to every 100 mL of distilled water.

[0028] In the technical solution of the present invention, the hydrolyzed olive leaf extract is obtained by subjecting the crushed olive leaves to ethanol ultrasonic extraction, concentration, spray drying, β-glucosidase hydrolysis, enzyme inactivation treatment, centrifugation and freeze drying in sequence.

[0029] In the technical solution of the present invention, the Rhodiola rosea root extract is obtained by subjecting the crushed Rhodiola rosea root to 70-80% ethanol extraction, followed by concentration and spray drying.

[0030] The plant composition preparation with skin anti-aging activity of the present invention does not contain preservatives, artificial fragrances and other ingredients that may cause skin discomfort, is relatively mild, and is suitable for use on various skin types.

[0031] The positive beneficial effects of the present invention are:

[0032] 1. The present invention utilizes fermented meadowfoam seed oil, fermented camelina seed oil, and fermented safflower seed oil in combination, leveraging their unique fatty acid composition to provide a unique functional ingredient for the skin barrier. The fermented plant oil matrix reduces triglyceride levels while increasing free fatty acid levels, further facilitating transdermal absorption of active ingredients in cosmetics. This helps maintain a mildly acidic environment in the skin, preventing the colonization of pathogenic bacteria and thus maintaining a balanced skin microbiome.

[0033] 2. The plant composition provided by the present invention uses fermented plant oil as the active ingredient and matrix. Compared with the original plant oil, it has lower surface tension, higher affinity with water, is easier to emulsify, and is easier to use in various cosmetic matrices. The prepared cosmetics are non-greasy and have better spreadability.

[0034] 3. The present invention uses fermented vegetable oil as an extraction solvent and compound raw material for astaxanthin and other active ingredients in Haematococcus pluvialis. Compared with the original vegetable oil, the extraction rate of astaxanthin in Haematococcus pluvialis is higher. At the same time, the process route is simple, avoiding the subsequent step of removing the solvent, so as to fully ensure the stability of the astaxanthin components.

[0035] 4. The plant compositions provided by the present invention are all extracted from pure nature, which can increase the area ratio of collagen fibers in the dermis of the skin and have strong anti-aging activity. In addition, no artificial flavors and preservatives are added to the preparations, making them safer to use. 4. Description of the accompanying drawings:

[0036] Figure 1 GC-MS total ion current chromatograms of free fatty acid components before and after fermentation of single vegetable oil;

[0037] Figure 1 A-comparison of camelina seed oil before and after fermentation; B-comparison of safflower seed oil before and after fermentation; C-comparison of meadowfoam seed oil before and after fermentation; (a)-fermented vegetable oil; (b)-original vegetable oil.

[0038] Figure 2 Full wavelength scanning spectrum of astaxanthin standard solution (340-800nm);

[0039] Figure 2 In the figure, red is the fermented vegetable oil configuration of Example 1, and black is the control solvent configuration of Example 1.

[0040] Figure 3 HE and Masson staining results of skin tissues of rats in each group;

[0041] Figure 3 In the figure, I-HE staining results; II-Masson staining results; (a) - control group; (b) - model group; (c) - Example 9 group; (d) - Example 10 group. V. Specific implementation methods:

[0042] The present invention is further described below with reference to the following examples, but the scope of protection of the technical solution of the present invention is not limited thereto.

[0043] Example 1:

[0044] The plant composition with skin anti-aging activity comprises 87.7 g of Haematococcus pluvialis extract, 8.0 g of lithospermum officinale oil, 0.7 g of Melaleuca alternifolia leaf oil, 0.6 g of hydrolyzed olive leaf extract, 0.5 g of Rhodiola rosea root extract and 2.5 g of petitgrain leaf oil.

[0045] The extraction method of the Haematococcus pluvialis extract is:

[0046] a. Take 1 strain of Yarrowia lipolytica and add it to 150 mL of sterilized liquid culture medium (containing 3 g of glucose, 0.45 g of yeast extract, 3 g of peptone, and 0.45 g of malt extract). Ferment it aerobically on a shaking table at 30°C and 160 rpm for 16 h to obtain a seed solution.

[0047] b. Add 75 mL of the obtained seed solution to the liquid culture medium after high-pressure sterilization for primary fermentation at a fermentation temperature of 30° C., a rotation speed of 200 r / min, and a fermentation time of 24 h to obtain a primary fermentation liquid;

[0048] The liquid culture medium was prepared by adding 50 g of glucose, 7.5 g of yeast extract, 50 g of peptone, and 7.5 g of malt extract to 2.5 L of distilled water;

[0049] c. The obtained primary fermentation broth was divided into 1.8 L, 0.3 L and 0.4 L, and then introduced into sterilized camelina seed oil, safflower seed oil and white meadowfoam seed oil in a volume ratio of 1:1, respectively, and continued to ferment for 48 hours at 30°C and 200 r / min;

[0050] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is discarded and the bacterial cells are separated;

[0051] e. Wash the obtained cells with phosphate buffer, centrifuge at 6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0052] The clean yeast cells were taken and 167 g of dry Haematococcus pluvialis powder was added and mixed to obtain Haematococcus pluvialis powder containing yeast cells. The Haematococcus pluvialis powder containing yeast cells was then added to the fermented vegetable oil and mixed. After uniform mixing, the mixture was homogenized twice using a high-pressure homogenizer at a pressure of 1000 bar (the homogenization not only broke the yeast cells and Haematococcus pluvialis powder, but also extracted fat-soluble components and active ingredients from the yeast cells and Haematococcus pluvialis powder). After homogenization, the mixture was centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected to obtain a Haematococcus pluvialis powder / fermented vegetable oil extract, i.e., a Haematococcus pluvialis extract.

[0053] Example 2:

[0054] The plant composition with skin anti-aging activity comprises 84 g of Haematococcus pluvialis extract, 12 g of lithospermum oil, 0.5 g of Melaleuca alternifolia leaf oil, 0.5 g of hydrolyzed olive leaf extract, 0.5 g of Rhodiola rosea root extract and 2.5 g of petitgrain leaf oil.

[0055] The extraction method of the Haematococcus pluvialis extract is:

[0056] a. Take 1 strain of Yarrowia lipolytica and add it to 150 mL of sterilized liquid culture medium (containing 3 g of glucose, 0.45 g of yeast extract, 3 g of peptone, and 0.45 g of malt extract). Ferment it aerobically on a shaking table at 30°C and 160 rpm for 16 h to obtain a seed solution.

[0057] b. Add 75 mL of the obtained seed solution to the liquid culture medium after high-pressure sterilization for primary fermentation at a fermentation temperature of 32° C., a rotation speed of 200 r / min, and a fermentation time of 36 h to obtain a primary fermentation liquid;

[0058] The liquid culture medium was prepared by adding 30 g of glucose, 4.5 g of yeast extract, 30 g of peptone, and 4.5 g of malt extract to 1.5 L of distilled water;

[0059] c. The obtained primary fermentation broth was divided into 1.0 L, 0.3 L and 0.2 L portions, and then introduced into sterilized camelina seed oil, safflower seed oil and meadowfoam seed oil in a volume ratio of 1:1, respectively, and fermented for 36 h at 32 ° C and 200 r / min;

[0060] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is discarded and the bacterial cells are separated;

[0061] e. Wash the obtained cells with phosphate buffer, centrifuge at 6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0062] The clean yeast cells obtained were taken, and 75 g of dry Haematococcus pluvialis powder was added and mixed to obtain Haematococcus pluvialis powder containing yeast cells. The Haematococcus pluvialis powder containing yeast cells was then added to the fermented vegetable oil obtained, mixed, and after uniform mixing, homogenized once using a high-pressure homogenizer at a pressure of 1300 bar (the homogenization not only breaks the yeast cells and the Haematococcus pluvialis powder, but also extracts the fat-soluble components and active ingredients in the yeast cells and the Haematococcus pluvialis powder). After homogenization, the mixture was centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected to obtain the Haematococcus pluvialis powder / fermented vegetable oil extract, i.e., the Haematococcus pluvialis extract.

[0063] Example 3:

[0064] The plant composition with skin anti-aging activity comprises 89 g of Haematococcus pluvialis extract, 8.1 g of lithospermum officinale oil, 0.5 g of Melaleuca alternifolia leaf oil, 1.0 g of hydrolyzed olive leaf extract, 0.2 g of Rhodiola rosea root extract and 1.2 g of petitgrain leaf oil.

[0065] The extraction method of the Haematococcus pluvialis extract is:

[0066] a. Take 1 strain of Yarrowia lipolytica and add it to 150 mL of sterilized liquid culture medium (containing 3 g of glucose, 0.45 g of yeast extract, 3 g of peptone, and 0.45 g of malt extract). Ferment it aerobically on a shaking table at 28°C and 200 r / min for 36 h to obtain a seed solution.

[0067] b. Add 66 mL of the obtained seed solution to the liquid culture medium after high-pressure sterilization for primary fermentation at a fermentation temperature of 28° C., a rotation speed of 200 r / min, and a fermentation time of 24 h to obtain a primary fermentation liquid;

[0068] The liquid culture medium was prepared by adding 44 g of glucose, 6.6 g of yeast extract, 44 g of peptone, and 6.6 g of malt extract to 2.2 L of distilled water;

[0069] c. The obtained primary fermentation broth was divided into 1.6 L, 0.3 L and 0.3 L, and then introduced into sterilized camelina seed oil, safflower seed oil and white meadowfoam seed oil according to a volume ratio of 1:2, respectively, and continued to ferment for 72 h at 28 ° C and 200 r / min;

[0070] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is discarded and the bacterial cells are separated;

[0071] e. Wash the obtained cells with phosphate buffer, centrifuge at 6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0072] The clean yeast cells were taken, 220 g of dry Haematococcus pluvialis powder was added and mixed to obtain Haematococcus pluvialis powder containing yeast cells; the Haematococcus pluvialis powder containing yeast cells was then added to the fermented vegetable oil, mixed, and after uniform mixing, homogenized twice using a high-pressure homogenizer at a pressure of 1000 bar (the homogenization not only breaks the yeast cells and Haematococcus pluvialis powder, but also extracts fat-soluble components and active ingredients from the yeast cells and Haematococcus pluvialis powder); after homogenization, the mixture was centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected to obtain the Haematococcus pluvialis powder / fermented vegetable oil extract, i.e., the Haematococcus pluvialis extract.

[0073] Example 4:

[0074] The plant composition with skin anti-aging activity comprises 86 g of Haematococcus pluvialis extract, 10 g of lithospermum oil, 0.8 g of Melaleuca alternifolia leaf oil, 0.8 g of hydrolyzed olive leaf extract, 0.3 g of Rhodiola rosea root extract and 2.1 g of petitgrain leaf oil.

[0075] The extraction method of the Haematococcus pluvialis extract is:

[0076] a. Take 1 strain of Yarrowia lipolytica and add it to 150 mL of sterilized liquid culture medium (containing 3 g of glucose, 0.45 g of yeast extract, 3 g of peptone, and 0.45 g of malt extract). Ferment it aerobically on a shaking table at 28°C and 200 r / min for 36 h to obtain a seed solution.

[0077] b. Add 60 mL of the obtained seed solution to the liquid culture medium after high-pressure sterilization for primary fermentation at a fermentation temperature of 30° C., a rotation speed of 220 r / min, and a fermentation time of 36 h to obtain a primary fermentation liquid;

[0078] The liquid culture medium was prepared by adding 24 g of glucose, 3.6 g of yeast extract, 24 g of peptone, and 3.6 g of malt extract to 1.2 L of distilled water;

[0079] c. The obtained primary fermentation broth was divided into 0.72 L, 0.25 L and 0.23 L, and then introduced into sterilized camelina seed oil, safflower seed oil and white meadowfoam seed oil according to a volume ratio of 1:1, respectively, and continued to ferment for 72 h at 35 ° C and 200 r / min;

[0080] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is discarded and the bacterial cells are separated;

[0081] e. Wash the obtained cells with phosphate buffer, centrifuge at 6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0082] The clean yeast cells obtained were added with 60 g of dry Haematococcus pluvialis powder and mixed to obtain Haematococcus pluvialis powder containing yeast cells. The Haematococcus pluvialis powder containing yeast cells was then added to the fermented vegetable oil obtained and mixed. After uniform mixing, the mixture was homogenized once using a high-pressure homogenizer at a pressure of 1500 bar (the homogenization not only broke the yeast cells and Haematococcus pluvialis powder, but also extracted fat-soluble components and active ingredients from the yeast cells and Haematococcus pluvialis powder). After homogenization, the mixture was centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected to obtain a Haematococcus pluvialis powder / fermented vegetable oil extract, i.e., a Haematococcus pluvialis extract.

[0083] Example 5:

[0084] The plant composition with skin anti-aging activity comprises 84.4 g of Haematococcus pluvialis extract, 12 g of lithospermum oil, 0.6 g of Melaleuca alternifolia leaf oil, 0.5 g of hydrolyzed olive leaf extract, 0.5 g of Rhodiola rosea root extract and 2.0 g of petitgrain leaf oil.

[0085] The extraction method of the Haematococcus pluvialis extract is:

[0086] a. Take 1 strain of Yarrowia lipolytica and add it to 150 mL of sterilized liquid culture medium (containing 3 g of glucose, 0.45 g of yeast extract, 3 g of peptone, and 0.45 g of malt extract). Ferment it aerobically on a shaking table at 30°C and 200 r / min for 36 h to obtain a seed solution.

[0087] b. Add 75 mL of the obtained seed solution to the liquid culture medium after high-pressure sterilization for primary fermentation at a fermentation temperature of 30° C., a rotation speed of 200 r / min, and a fermentation time of 24 h to obtain a primary fermentation liquid;

[0088] The liquid culture medium was prepared by adding 30 g of glucose, 4.5 g of yeast extract, 30 g of peptone, and 4.5 g of malt extract to 1.5 L of distilled water;

[0089] c. The obtained primary fermentation broth was divided into 0.7 L, 0.4 L and 0.4 L, and then introduced into sterilized camelina seed oil, safflower seed oil and white meadowfoam seed oil according to a volume ratio of 1:2, respectively, and continued to ferment for 48 hours at 30°C and 200 r / min;

[0090] d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is discarded and the bacterial cells are separated;

[0091] e. Wash the obtained cells with phosphate buffer, centrifuge at 6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells;

[0092] The obtained clean yeast cells were added with 200 g of dry Haematococcus pluvialis powder to obtain Haematococcus pluvialis powder containing yeast cells. The Haematococcus pluvialis powder containing yeast cells was then added to the obtained fermented vegetable oil and mixed. After uniform mixing, the mixture was homogenized twice using a high-pressure homogenizer at a pressure of 1200 bar (the homogenization not only broke the yeast cells and Haematococcus pluvialis powder, but also extracted fat-soluble components and active ingredients from the yeast cells and Haematococcus pluvialis powder). After homogenization, the mixture was centrifuged at 8000 rpm for 10 minutes, and the supernatant was collected to obtain the Haematococcus pluvialis powder / fermented vegetable oil extract, i.e., the Haematococcus pluvialis extract.

[0093] Example 6:

[0094] An anti-aging facial cream was prepared using the plant composition having skin anti-aging activity described in Example 1. The weight percentages of the components in the anti-aging facial cream were as follows: 14% of the plant composition described in Example 1, 2% of glyceryl stearate, 3.7% of cetearyl alcohol, 4% of isohexadecane, 67% of birch sap, 6% of glycerol, 0.1% of sodium polyacrylate, 0.2% of allantoin, 0.1% of xanthan gum, 1.5% of tocopherol, 0.4% of p-hydroxyacetophenone, and 1% of 1,2-hexanediol.

[0095] Example 7:

[0096] An anti-aging emulsion was prepared using the plant composition having skin anti-aging activity described in Example 2. The weight percentages of the components in the anti-aging emulsion were as follows: 7.5% of the plant composition described in Example 2, 2% of glyceryl stearate, 1.5% of cetearyl alcohol, 5% of isohexadecane, 73.6% of water, 6% of glycerin, 0.2% of sodium polyacrylate, 0.3% of allantoin, 0.4% of carbomer 940, 1% of tocopherol, 0.5% of p-hydroxyacetophenone, and 2% of 1,2-hexanediol.

[0097] Example 8:

[0098] An anti-aging lotion was prepared using the plant composition having skin anti-aging activity described in Example 3. The weight percentages of the components in the anti-aging lotion were as follows: 9.5% of the plant composition described in Example 3, 1% of glyceryl stearate, 1% of cetearyl alcohol, 5% of isohexadecane, 71.5% of birch sap, 8% of glycerin, 0.1% of sodium polyacrylate, 0.3% of allantoin, 0.2% of carbomer 940, 1% of tocopherol, 0.4% of p-hydroxyacetophenone, and 2% of 1,2-hexanediol.

[0099] Example 9:

[0100] An anti-aging facial cream was prepared using the plant composition having skin anti-aging activity described in Example 4. The weight percentages of the components in the anti-aging facial cream were as follows: 18% of the plant composition described in Example 4, 2.5% of glyceryl stearate, 6% of cetearyl alcohol, 5% of isohexadecane, 59.5% of water, 5% of glycerol, 0.1% of sodium polyacrylate, 0.4% of allantoin, 0.1% of xanthan gum, 1.0% of tocopherol, 0.4% of p-hydroxyacetophenone, and 2.0% of 1,2-hexanediol.

[0101] Example 10:

[0102] An anti-aging essential oil was prepared using the plant composition with skin anti-aging activity described in Example 5. The weight percentages of the components in the anti-aging essential oil were as follows: 20% of the plant composition described in Example 5, 20% of canola oil, 29% of sweet almond oil, 30% of jojoba seed oil, and 1% of tocopherol.

[0103] Example 11:

[0104] An anti-aging essential oil was prepared using the plant composition with skin anti-aging activity described in Example 1. The weight percentages of the components in the anti-aging essential oil were: 15% of the plant composition described in Example 1, 8% of linseed oil, 10% of camellia seed oil, 30% of meadowfoam seed oil, 34.5% of jojoba seed oil, and 2.5% of tocopherol.

[0105] The relevant experiments and experimental results of the relevant raw materials and products used in the technical solution of the present invention are as follows:

[0106] 1. Determination of free fatty acids in vegetable oils before and after fermentation: GC-MS was used to determine the free fatty acids in each vegetable oil before and after fermentation and their relative content.

[0107] 1. Sample pretreatment method: Accurately pipette 300 μL of the fermented vegetable oil of the single vegetable oil obtained by the extraction method in Example 1 and 300 μL of the single unfermented vegetable oil, respectively, add 300 μL of methanol and 50 μL of glacial acetic acid solution, vortex extraction for 3 minutes, centrifuge at 4°C and 10,000 rpm for 5 minutes, pipette 150 μL of the supernatant after centrifugation, add 8 mL of 2% sodium hydroxide methanol solution, heat in an 85°C water bath until the oil droplets disappear, then add 7 mL of 15% boron trifluoride methanol solution, continue heating for 2 minutes, cool to room temperature, add 10 mL of n-hexane, shake for 2 minutes, add 5 mL of saturated sodium chloride aqueous solution, let stand for stratification, draw the upper n-hexane solution, add an appropriate amount of anhydrous sodium sulfate for dehydration, let stand for 30 minutes, take the supernatant, filter and test.

[0108] 2. Determination method:

[0109] Gas chromatography conditions: Shimadzu GC-MS, using an Rtx-5MS capillary column (30 m × 0.25 mm, 0.25 μm), split mode with a split ratio of 20:1, an inlet temperature of 330°C, helium carrier gas at a total flow rate of 62.1 mL / min. The temperature program was as follows: initial temperature at 180°C, held for 5 min, then increased to 230°C at a rate of 2°C / min, held for 1 min, and then increased to 330°C at a rate of 10°C / min, held for 7 min.

[0110] Mass spectrometry conditions: full scan mode, EI ionization source, ion source temperature 250°C, interface temperature 230°C, solvent delay 2 min, scan range 40-1000 m / z.

[0111] 3. Measurement results:

[0112] The results of free fatty acid composition determination of single vegetable oil before and after fermentation show that (see Appendix Figure 1Before fermentation, free fatty acid levels in camelina, safflower, and meadowfoam seed oils were extremely low. However, after fermentation, the total ion current chromatograms showed significant changes compared to the original vegetable oils, revealing the presence of a variety of polar free fatty acid components.

[0113] The main polar components of free fatty acids in each vegetable oil after fermentation and their peak area percentages are detailed in Table 1. As shown in Table 1, the free polar components in fermented camelina seed oil, fermented safflower seed oil, and fermented meadowfoam seed oil differed significantly. Fermented camelina seed oil was dominated by linolenic acid and linoleic acid, accounting for 76% of the total polar components. Fermented safflower seed oil and fermented meadowfoam seed oil were both dominated by linoleic acid, accounting for 76.23% and 60.86% of the total polar components, respectively.

[0114] Table 1 Composition and percentage of main free fatty acids in various fermented vegetable oils

[0115]

[0116] 2. Determination of acid value of fermented vegetable oil:

[0117] The fermented vegetable oils obtained in step d of the extraction methods described in Examples 1, 2, and 4 were combined. For Example 1, a direct mixture of camelina seed oil, safflower seed oil, and meadowfoam seed oil was used in a ratio of 18:3:4. For Example 2, a direct mixture of camelina seed oil, safflower seed oil, and meadowfoam seed oil was used in a ratio of 10:3:2. For Example 4, a direct mixture of camelina seed oil, safflower seed oil, and meadowfoam seed oil was used in a ratio of 72:25:23. The acid value of each sample was determined using the first method cold solvent indicator titration method in accordance with national standard GB 5009.229. The results are shown in Table 2.

[0118] Table 2 Acid value determination results of each sample

[0119] sample Acid value (mg KOH / 100g) Example 1 Mixed fermentation of vegetable oil 24.42±1.17 Example 2 Mixed fermentation of vegetable oil 18.33±0.38 Example 4 Mixed fermentation of vegetable oil 36.85±0.63 Example 1 comparison 0.99±0.07 Example 2 comparison 0.87±0.04 Example 4 comparison 0.80±0.05

[0120] As shown in the acid value determination results in Table 2, the acid values of the samples obtained from Examples 1, 2, and 4, all of which were physically mixed without any other treatment, were relatively low. However, after fermentation with Yarrowia lipolytica, the acid value of the vegetable oil increased significantly. This is because the lipase secreted by the yeast during fermentation partially degrades triglycerides into free fatty acids.

[0121] 3. Determination of astaxanthin content in Haematococcus pluvialis extract:

[0122] 1. Sample preparation: The test sample was the Haematococcus pluvialis extract prepared in Example 1.

[0123] Preparation of a control sample: Weigh an appropriate amount of Haematococcus pluvialis powder and add the control solvent of Example 1 (camelina seed oil, safflower seed oil, and meadowfoam seed oil mixed in a ratio of 18:3:4) at a ratio of 1:15 (g:mL). Perform high-pressure homogenization with the same parameters as Example 1 to prepare a Haematococcus pluvialis extract to obtain the control sample of Example 1.

[0124] The test samples and control samples were diluted 100-fold before measurement.

[0125] 2. Standard curve drawing: The astaxanthin content was determined by UV-visible spectrophotometry. An appropriate amount of astaxanthin standard was prepared into a 240 mg / L mother liquor using the control solvent of Example 1. An appropriate amount of the mother liquor was pipetted and diluted to 4, 6, 8, 10, and 12 mg / L using the combined fermented vegetable oil obtained in step d of the extraction method for the Haematococcus pluvialis extract of Example 1 as a solvent. A full wavelength scan was performed at a concentration of 10 mg / L, and the optimal measurement wavelength was selected to draw an astaxanthin standard curve for determining the astaxanthin content in the Haematococcus pluvialis extract of Example 1. An appropriate amount of the mother liquor was pipetted and diluted to 4, 6, 8, 10, and 12 mg / L using the control solvent of Example 1 as a solvent. A full wavelength scan was performed at a concentration of 10 mg / L, and the optimal measurement wavelength was selected to draw an astaxanthin standard curve for determining the astaxanthin content in the control sample.

[0126] 3. Measurement results: The astaxanthin standard solution prepared by combining the control solvent of Example 1 and the fermented vegetable oil obtained in step d of Example 1 was subjected to UV-visible spectral scanning in the range of 340-800 nm. The maximum absorption peak was 486 nm (see the attached figure for details). Figure 2 ), and the absorbance value of the astaxanthin standard solution at the same concentration (10 mg / L) is 0.612. However, it can also be seen that there are certain differences in the full wavelength absorption of the astaxanthin standard obtained by scanning with different solvents. The standard curve for astaxanthin content determination drawn with the absorbance value at a wavelength of 486 nm is Y=0.0687X-0.0886(R 2 =0.9983). Calculation showed that the astaxanthin content in the control sample was (729.02±13.12) mg / L, while the astaxanthin content in the Haematococcus pluvialis extract of Example 1 was (892.48±4.73) mg / L, which was approximately 22% higher than that in the control sample.

[0127] Astaxanthin in Haematococcus pluvialis exists in both esterified and free forms, with the esterified form being predominant and stored in triglyceride-rich cytoplasmic lipid bodies. Therefore, using vegetable oil as a solvent is more beneficial for extracting astaxanthin from Haematococcus pluvialis than conventional extraction reagents such as anhydrous ethanol and acetone. Because fermented vegetable oil is slightly more polar than crude vegetable oil, it offers better extraction efficiency for both forms of astaxanthin.

[0128] 4. Effects of the Examples on the Skin of Aging Model Rats:

[0129] 1. Animals: Twenty-four male SD rats, weighing 180 ± 20 g, were purchased from Henan Sikebes Biotechnology Co., Ltd. Rats were housed under appropriate conditions, with a temperature range of 20–25°C, relative humidity maintained at 40–70%, and a 12 h light / dark cycle.

[0130] 2. Experimental methods:

[0131] 2.1 Animal Grouping and Modeling: After a one-week acclimation period, rats were randomly divided into four groups: a control group, a model group, an Example 9 group, and an Example 10 group. Each rat was housed individually and depilated using an electric shaver, with the depilated area measuring 4 cm × 4 cm. Depilation was performed at irregular intervals depending on the rat's hair growth. The depilation procedure was performed without damaging the skin.

[0132] A rat aging model was established by subcutaneous injection of D-galactose into the neck of rats. All rats, except the control group, received a subcutaneous injection of D-galactose (200 mg / kg / day) once daily for 42 consecutive days. The control group received normal saline. The injection volume was 5 mL / kg.

[0133] 2.2 Animal Administration: Rats were administered transdermally via application to the depilated area. During the experimental period, the control and model groups were treated with 0.1 mL of distilled water daily, while the Example 9 and Example 10 groups were treated with 0.2 g per rat daily for 42 consecutive days. Skin changes in each group were observed.

[0134] 2.3 Sampling: After the experiment, rats were sacrificed by cervical dislocation. The hairless skin on the back was quickly removed and excess subcutaneous fat was carefully removed. Part of the skin was fixed in 10% formaldehyde solution for histomorphological analysis. Part of the skin tissue was used for the analysis of antioxidant markers in the skin.

[0135] 2.4 Determination of antioxidant index: Take part of the skin tissue and add pre-cooled saline to prepare a sample homogenate at a ratio of 1:9 (mg:μL). Centrifuge at 4°C and 5000 rpm for 10 minutes and collect the supernatant. Detect the SOD and MDA levels in each sample according to the kit instructions.

[0136] 2.5 Skin Histomorphological Examination: Skin tissues were fixed with paraformaldehyde, routinely dehydrated, paraffin-impregnated, embedded, and sectioned. Hematoxylin and eosin staining and Masson's trichrome staining were performed to observe changes in the epidermis and dermis of the skin tissues of the rats in each group, as well as changes in the collagen fiber structure in the dermis. For Masson's trichrome sections, the collagen fiber area ratio was measured using Image-Pro Plus 6.0 in four fields of view from the same section at the same magnification.

[0137] 2.6 Statistical analysis: Experimental data are expressed as mean ± standard deviation. Statistical analysis was performed using SPSS (V.25.0). When data were normally distributed and showed homogeneous variance, one-way analysis of variance was performed, followed by a post hoc LSD test for three-group comparisons. For non-normally distributed data, the Tamhane's t2 test was used, with a P value < 0.05 considered significant.

[0138] 3. Experimental results:

[0139] 3.1 D-galactose-induced skin aging manifestations: The D-galactose skin injection aging model is a commonly used experimental skin aging model. Its principle is that a large amount of reactive oxygen species is produced through D-galactose metabolism, leading to cellular oxidative damage, metabolic disorders and functional impairment. At the same time, the continuous accumulation of D-galactose metabolites leads to increased cellular osmotic pressure, further inducing oxidative stress, and ultimately leading to body aging.

[0140] Compared to the control group, the skin of the model group rats showed more pronounced wrinkles and dryness. During the experiment, the skin of the model group rats was noticeably loose and thinner. The skin of the rats in the Example 9 and Example 10 treatment groups was relatively firm, with fewer wrinkles compared to the model group.

[0141] 3.2 Effects of different preparations on antioxidant indicators of the skin of aging rats (see Table 3 for details): SOD and MDA are one of the two most commonly used indicators of the body's antioxidant capacity. They are also important biomarkers for assessing the oxidative stress state of the skin and are closely related to skin aging. SOD is responsible for scavenging superoxide free radicals and converting them into hydrogen peroxide (H2O2) and oxygen (O2), thereby protecting cells from oxidative damage. If the activity of SOD decreases and free radicals cannot be cleared in time, it can lead to the accumulation of free radicals, degradation of skin cell collagen, and accelerated skin aging. MDA is the end product of lipid peroxidation, which can cross-link with collagen, destroy its structure, and lead to decreased skin elasticity and wrinkle formation.

[0142] The results of this experiment show that after continuous percutaneous injection of D-galactose, the SOD activity in the skin tissue of the model group rats was significantly decreased compared with the control group rats (P < 0.01), while the MDA level was significantly increased (P < 0.01). Different preparations were intervened at the same time as modeling, which can significantly inhibit the skin aging induced by D-galactose, as shown by the significant increase in the SOD activity of the skin tissue of the rats in the 9th group and the 10th group of Examples compared with the model group (P < 0.01; P < 0.01), while the MDA level was significantly decreased compared with the model group (P < 0.01; P < 0.01). The results show that the Example 9 and Example 10 groups can effectively scavenge free radicals in the body, reduce oxidative stress in the body, enhance antioxidant defense, reduce oxidative damage, and rebuild the oxidation-antioxidation balance, thereby protecting the structure and function of the skin.

[0143] Table 3 Effects of different preparations on the antioxidant indexes of rats

[0144]

[0145] (Note: Compared with the control group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01).

[0146] 3.3 Effects of different preparations on the skin tissue structure of aging rats: The results of HE and Masson staining of skin tissue of rats in different groups are detailed in the attached Figure 3 .Depend on Figure 3 It can be seen that the skin tissue structure of the control group rats is normal, the epidermis is uniform in thickness and clearly demarcated from the dermis, and structures such as hair follicles, sebaceous glands and hair roots can be seen in the dermis, and collagen fibers are neatly arranged. The skin structure of the model group rats showed obvious pathological changes, with thinning and irregular epidermis, loose collagen fibers and collagen fibers missing or broken in some areas. The collagen fiber ratio was reduced by about 36.09% compared with the normal group, with a significant difference (P < 0.01). Compared with the model group, the skin tissue structure of the rats in Example 9 and Example 10 groups was relatively complete, the epidermis was relatively thickened, and the collagen fibers in the dermis were arranged more tightly and neatly. The collagen fiber ratio increased by 30.07% and 22.69% respectively compared with the model group. Existing studies have found that collagen fibers in the dermis are an important factor in forming the skin reticular scaffold, maintaining the thickness and firmness of the skin. The decrease in skin collagen fiber density and cross-linking can lead to insufficient support of the dermis, thinning, sagging, and loss of elasticity of the skin, and the epidermis is prone to folding and forming wrinkles. In this experiment, the Example group can significantly increase the collagen fiber area ratio (see Table 4 for detailed results), indicating that it has a good anti-aging effect on the skin.

[0147] Table 4 Collagen fiber area ratio of rat dermal tissue

[0148]

[0149] (Note: Compared with the control group, * P<0.05, ** P<0.01; compared with the model group, # P<0.05, ## P<0.01).

[0150] 5. Consumer Testing

[0151] Consumer testing of the product of the present invention was conducted using a questionnaire survey. Questions related to product efficacy included, "How moisturized is your skin after use?", "How effective do you think this product is at anti-aging your skin?", and "Do you experience any allergic or discomfort symptoms after using the product?"

[0152] 1. Sample information: The test sample is Example 6 (50 g / bottle).

[0153] 2. Test population information:

[0154] Target population: mainly women aged 30 to 60, with anti-aging needs.

[0155] Exclusion criteria: Those with severely sensitive skin and those who have received cosmetic treatments within the past 6 months.

[0156] Tested consumers: 33 people in total, 26 women and 7 men. Ages ranged from 30 to 45 years old, with an average age of 35.8±3.8 years for the entire population, 36.4±5.4 years for men, and 35.6±3.3 years for women.

[0157] 3. Requirements during the test period: During the test period, no oral or external use of other preparations claiming to have anti-wrinkle, skin firming, or similar claims is permitted; cosmetic surgery that may affect the anti-wrinkle efficacy test is prohibited; testers should primarily engage in indoor activities and avoid prolonged exposure to outdoor light.

[0158] 4. Test conditions and steps:

[0159] Instructions for Sample Use: Use in place of your daily face cream. Apply an appropriate amount evenly to your face after cleansing morning and evening. Samples should be used for four weeks. Please provide feedback and reissue the sample upon completion of the testing period. Complete and submit the questionnaire after four weeks.

[0160] 5. Consumer Test Results: After the test period, questionnaires were collected and analyzed. Regarding the question "How moisturized is my skin after use?", 28 consumers (84.8%) chose "Quite moisturized, my skin feels quite comfortable," and 5 consumers (15.2%) chose "Very moisturized, my skin feels very hydrated." Regarding the question "How effective do you think this product is on your skin in anti-aging?", 21 consumers (63.6%) chose "Quite good, with some effect," 3 consumers (9.1%) chose "Very good, with significant effect," and 9 consumers (27.3%) chose "Average, with no significant effect." Regarding the question "Did you experience any allergic or discomfort symptoms after using the product?", 33 consumers (100%) chose "None, very satisfied."

[0161] The above test results show that the fermented vegetable oil prepared using the method described in the present invention exhibits significant changes in physical and chemical properties compared to the original vegetable oil. The present invention utilizes a mixture of fermented camelina seed oil, fermented safflower seed oil, and fermented meadowfoam seed oil as one of its key functional ingredients, along with a solvent for extracting astaxanthin and other active ingredients from Haematococcus pluvialis. This yields high astaxanthin extraction efficiency. Furthermore, the oil is compounded with other plant-based functional raw materials, and both animal and human testing have demonstrated excellent anti-aging activity.

Claims

1. A plant composition having skin anti-aging activity, characterized in that: The active ingredients in the plant composition are Haematococcus pluvialis extract, lithospermum officinale oil, Melaleuca alternifolia leaf oil, hydrolyzed olive leaf extract, Rhodiola rosea root extract and petitgrain oil. Expressed in percentage by mass, the proportions of the active ingredients are: 84.0-89.0% of Haematococcus pluvialis extract, 8.0-12.0% of lithospermum officinale oil, 0.5-0.8% of Melaleuca alternifolia leaf oil, 0.5-1.0% of hydrolyzed olive leaf extract, 0.1-0.5% of Rhodiola rosea root extract and 1.2-2.5% of petitgrain oil.

2. The plant composition with skin anti-aging activity according to claim 1, characterized in that: The Haematococcus pluvialis extract is obtained by subjecting Haematococcus pluvialis powder to high-pressure homogenization and wall-breaking extraction using fermented vegetable oil as an extraction solvent; the fermented vegetable oil is at least one of fermented camelina seed oil, fermented meadowfoam seed oil, and fermented safflower seed oil.

3. The plant composition having skin anti-aging activity according to claim 2, characterized in that: The specific extraction method of the Haematococcus pluvialis extract is: a. Using Yarrowia lipolytica as the fermentation strain, the fermentation strain was inoculated into a liquid culture medium for aerobic fermentation on a shaker at a temperature of 28 to 35° C. for 16 to 36 hours to obtain a seed solution; b. adding the obtained seed liquid to the sterilized liquid culture medium at a volume ratio of 3-5% for primary fermentation at a fermentation temperature of 28-35° C. for 24-48 h to obtain a primary fermentation liquid; c. Sterilize each vegetable oil separately, then add each sterilized vegetable oil into the obtained primary fermentation liquid, and continue fermenting for 24 to 72 hours at a fermentation temperature of 28 to 35°C; The volume ratio between the primary fermentation liquid and the sterilized vegetable oil is 1:1-2; d. After the fermentation in step c is completed, each fermentation product is taken out and centrifuged at 6000 rpm for 10 minutes. After centrifugation, the supernatants are mixed to obtain fermented vegetable oil; at the same time, the lower aqueous phase is removed and the bacterial cells are separated; e. Wash the obtained cells with phosphate buffer, centrifuge at 4000-6000 rpm for 10 min, discard the supernatant after centrifugation, and repeat washing twice to obtain clean cells; The obtained clean bacterial cells are added with dried Haematococcus pluvialis powder and mixed to obtain Haematococcus pluvialis powder containing bacterial cells; the Haematococcus pluvialis powder containing bacterial cells is then added to the obtained fermented vegetable oil and mixed. After the mixture is evenly mixed, a high-pressure homogenizer is used to homogenize the mixture 1 to 2 times at a pressure of 1000 to 1500 bar; after homogenization, the mixture is centrifuged at 8000 rpm for 10 minutes, and the supernatant is collected to obtain the Haematococcus pluvialis extract.

4. The plant composition having skin anti-aging activity according to claim 3, characterized in that: The vegetable oil in step c is camelina seed oil, meadowfoam seed oil or safflower seed oil; and the ratio of Haematococcus pluvialis powder to fermented vegetable oil in step e is 1 g:15-20 mL.

5. The plant composition with skin anti-aging activity according to claim 3, characterized in that: During the fermentation, the rotation speed of the shaking table is controlled to be 160-220 rpm.

6. A preparation of a plant composition having skin anti-aging activity, characterized in that: The preparation is prepared from the plant composition with skin anti-aging activity according to claim 1 into an emulsion, a cream or an essential oil.

7. The preparation of the plant composition having skin anti-aging activity according to claim 6, characterized in that: The plant composition preparation emulsion with skin anti-aging activity is composed of the following components in percentage by mass: 6.7-13.2% of the plant composition with skin anti-aging activity according to claim 1, 1.0-2.0% of glyceryl stearate, 1.0-1.5% of cetearyl alcohol, 2.0-5.0% of isohexadecane, 62.9-81.2% of water / birch sap, 5.0-10.0% of glycerol, 0.1-0.2% of sodium polyacrylate, 0.2-0.4% of allantoin, 0.2-0.4% of carbomer 940, 1.0-1.5% of tocopherol, 0.4-0.6% of p-hydroxyacetophenone and 1.0-2.5% of 1,2-hexanediol.

8. The preparation of the plant composition having skin anti-aging activity according to claim 6, characterized in that: The plant composition preparation cream with skin anti-aging activity is prepared from the following components in percentage by mass: 9.0-18.0% of the plant composition with skin anti-aging activity according to claim 1, 1.5-2.5% of glyceryl stearate, 3.0-9.0% of cetearyl alcohol, 4.0-6.0% of isohexadecane, 55.0-74.5% of water / birch sap, 4.0-10.0% of glycerol, 0.1-0.2% of sodium polyacrylate, 0.2-0.4% of allantoin, 0.1-0.2% of xanthan gum, 1.0-1.5% of tocopherol, 0.4-0.6% of p-hydroxyacetophenone and 1.0-2.0% of 1,2-hexanediol.

9. The preparation of the plant composition having skin anti-aging activity according to claim 6, characterized in that: The essential oil of the plant composition preparation with skin anti-aging activity is composed of the following components in percentage by mass: 14.0-26.0% of the plant composition with skin anti-aging activity according to claim 1, 72.0-85.0% of base oil and 0.5-3.5% of tocopherol.

10. The preparation of the plant composition having skin anti-aging activity according to claim 9, characterized in that: The base oil is at least one of camelina seed oil, linseed oil, rapeseed oil, jojoba seed oil, perilla seed oil, peony seed oil, meadowfoam seed oil, safflower seed oil, camellia seed oil and sweet almond oil.

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