A preparation method of camellia seed extract

Abstract

The present invention belongs to the technical field of cosmetic raw materials, and in particular to a method for preparing a camellia seed extract. The preparation method comprises the following steps: (1) crushing camellia seeds, adding cellulase, pectinase and α-amylase, enzymolysis, and then using composite bacteria, fermentation, vacuum drying, and obtaining fermented camellia seed powder; (2) subcritical extraction of the fermented camellia seed powder using an extractant to obtain a first extract; (3) the extract obtained in step (2) is dried and crushed to 20-60 mesh to obtain a second powder, and the second powder is subjected to a supercritical carbon dioxide extraction kettle, and a second extract is obtained after solvent recovery and drying; (4) the first extract and the second extract are mixed uniformly to obtain a camellia seed extract. The preparation method of the present invention combines various effective extraction methods, can effectively improve the yield and the content of active components, thereby obtaining a product of better quality; and is applied to cosmetics, so that the cosmetics have a better effect.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetic raw materials, and particularly relates to a preparation method of a camellia seed extract. Background Art

[0002] Camellia oleifera is the main woody oil tree in my country, known as the "Oriental Tree". It is an evergreen small tree belonging to the genus Camellia of the Theaceae family and is a unique woody plant oil resource in my country. Camellia seed is another name for camellia seed, which is the fruit of the oil tea tree. Camellia seed extract usually refers to the natural ingredients extracted from camellia seeds, which are rich in a variety of beneficial substances, such as unsaturated fatty acids such as oleic acid and linoleic acid, and antioxidant ingredients such as vitamin E. In cosmetics and personal care products, camellia seed extract is widely used because of its good moisturizing and hydrating properties. It can help improve dry skin, enhance skin barrier function, and has a certain anti-aging effect.

[0003] There are some technical problems encountered in the preparation process of camellia seed extract. These problems mainly involve raw material processing, extraction efficiency, product quality and other aspects, which will affect the quality of the product: (1) Raw material processing: Oil content: Camellia seeds contain a high proportion of oil, which will affect the purity of the extract and the concentration of active ingredients. Removal of impurities: Camellia seeds may contain impurities or inactive ingredients, which need to be removed before or during the extraction process. (2) Extraction efficiency: Selection of extraction method: Different extraction methods have different application scopes and limitations. Choosing a suitable extraction method is crucial to improving extraction efficiency. Parameter optimization: During the extraction process, factors such as temperature, pressure, and solvent type need to be precisely controlled to achieve the best extraction effect. (3) Product quality: Active ingredient protection: Some active ingredients are sensitive to heat, so measures need to be taken during the extraction process to protect these ingredients from destruction. Finished product stability: The extract needs to remain stable for a long time, which may require the addition of stabilizers or specific processing techniques. (4) Sensitivity issues: Some plant proteins in camellia seeds may also become allergens, especially for those who are already sensitive to certain plant proteins.

[0004] In the prior art, the preparation process of camellia seed extract generally adopts the distillation extraction method. The distillation extraction method is simple to operate and has low equipment cost, so it is widely used. However, the distillation extraction method is to transport water vapor to the distillation through a boiler, and evaporate the components to be extracted at high temperature. High-temperature distillation will destroy the active ingredients in the camellia seeds themselves, reduce the stability and activity of the camellia seed extract, and is not conducive to later use. Summary of the invention

[0005] The purpose of the present invention is to provide a method for preparing a camellia seed extract.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] A method for preparing a camellia seed extract, comprising the following steps:

[0008] (1) crushing camellia seeds to less than 100-200 meshes to obtain a first powder, adding water to the first powder to obtain a mixture, adding cellulase, pectinase and α-amylase, performing enzymolysis at 30-35° C. for 2-3 days, inactivating the enzymes, continuously adding composite bacteria, performing anaerobic fermentation at 30-35° C. for 4-5 days, and vacuum drying to a moisture content of 2-5% to obtain fermented camellia seed powder;

[0009] (2) subcritically extracting the fermented camellia seed powder using an extractant at an extraction temperature of 40-45° C., an extraction pressure of 2.2-2.5 MPa, and an extraction time of 50-70 min to obtain an extract and an extraction product, and subjecting the extract to alcohol precipitation to remove impurities to obtain a supernatant, and to obtain a first extract after removing alcohol;

[0010] (3) drying the extract obtained in step (2) and crushing it into 20-60 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 20-25 MPa, the extraction temperature to 50-55° C., the separation pressure to 3-7 MPa, the separation temperature to 25-30° C., performing supercritical carbon dioxide extraction, extracting for 100-120 min, adding ethanol aqueous solution, continuing extraction for 3-5 hours, and obtaining a second extract after recovering the solvent and drying;

[0011] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0012] Furthermore, in step (1), the weight of water is 2-4 times the weight of the first powder.

[0013] Furthermore, in step (1), the concentration of cellulase in the mixture is 60U / mL-80U / mL.

[0014] Furthermore, in step (1), the concentration of pectinase in the mixture is 90 U / mL-110 U / mL.

[0015] Furthermore, in step (1), the concentration of α-amylase in the mixture is 130 U / mL-150 U / mL.

[0016] Furthermore, the composite bacteria in step (1) include Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei.

[0017] Furthermore, in step (1), the concentration of plant lactobacillus in the mixture is 10 8 -109 CFU / mL.

[0018] Furthermore, in the step (1), the concentration of Lactobacillus helveticus in the mixture is 10 6 -10 7 CFU / mL.

[0019] Furthermore, in the step (1), the concentration of Lactobacillus casei in the mixture is 10 10 -10 11 CFU / mL.

[0020] Furthermore, in the step (2), a fluid composed of butane and dimethyl ether in a weight ratio of 1:(2-4) is used as the extractant.

[0021] Furthermore, in step (3), a 60%-70% ethanol aqueous solution is added in an amount of 5-7 times the weight of the second powder.

[0022] The present invention also provides application of the camellia seed extract prepared by the preparation method in preparing cosmetics.

[0023] Compared with the prior art, the advantages and beneficial effects of the present invention are:

[0024] 1. The preparation method of the present invention combines various effective extraction methods, can effectively improve the yield and the content of active components, will not destroy the effective components, thereby obtaining extremely high-quality products. The camellia seed extract of the present invention can be used to prepare cosmetics, is highly safe, and is not prone to allergies.

[0025] 2. Camellia seed extract contains tea saponin, tea polysaccharide and tea protein. Adding them to skin care products, especially shampoo products, has many benefits. Tea saponin has good emulsification, foaming, dispersion, penetration and lubrication activities, which can help clean the skin and hair, while providing moisturizing effects. It also has anti-inflammatory effects and can reduce skin inflammation and redness. Tea polysaccharides have good moisturizing properties and can form a protective film on the skin surface to lock in moisture, prevent moisture loss, keep the skin hydrated, help resist free radical damage to the skin, and slow down the aging of the skin. The amino acids and peptides in tea protein help promote the repair and regeneration of skin cells and improve skin texture. Tea saponin, tea polysaccharide and tea protein can provide a variety of benefits in skin care products, including but not limited to moisturizing, anti-oxidation, antibacterial, anti-inflammatory and promoting skin repair. The application of these natural ingredients enables skin care products to care for the skin more safely and gently, and also meets the needs of modern consumers for natural and healthy skin care. At present, when extracting camellia seeds, they are usually directly extracted after being crushed. However, the percentage of the above three components in the obtained product is low, and the effect of using it in skin care products is not ideal.

[0026] The present invention uses a composite enzyme composed of cellulase, pectinase and α-amylase to enzymolyze the crushed camellia seeds, which can increase the content of tea polysaccharides and tea proteins in the camellia seed extract. The enzymolysis process decomposes proteins, polysaccharides, etc. into smaller molecules, thereby increasing the solubility of these components and making them easier to extract. However, the extraction effect of tea saponin is not ideal. Analysis shows that the composite enzyme may not have a particularly high specificity for the glycosidic bonds of tea saponin. On the basis of enzymolysis, the present invention uses composite bacteria for fermentation to increase the extraction amount of tea saponin. It is also found that when a specific amount of three bacteria are used for fermentation, the storage stability of the camellia seed extract is also improved.

[0027] 3. After obtaining the fermented camellia powder, the present invention first uses a conventional alcohol extraction method to extract the fermented camellia powder, and the obtained camellia seed extract contains more impurities. The present invention first uses subcritical extraction on the fermented camellia powder, and then performs supercritical carbon dioxide extraction, which can reduce the impurities in the camellia seed extract. At the same time, the percentage of moldy grains is also reduced. The camellia seed extract obtained by the above extraction method is brown and accompanied by the fragrance of camellia seeds, and is a high-quality product.

[0028] 4. The camellia seed extract prepared by the present invention can be applied to cosmetics, which have a variety of beneficial skin care ingredients, such as antioxidants, unsaturated fatty acids, tea polyphenols, camellia cheese, tea saponin, etc. Camellia seed extract is rich in antioxidant ingredients, which can help resist the damage of free radicals and delay the process of skin aging; it is rich in unsaturated fatty acids, which can provide deep nourishment and moisturizing effects for the skin and help maintain the moisture balance of the skin. It is particularly suitable for skin care needs such as whitening, anti-wrinkle and freckle removal, and can improve problems such as uneven skin color and fine lines. The camellia seed extract of the present invention is a safe ingredient suitable for all skin types, including sensitive skin. Camellia seed extract can also be used to make sunscreen products, products for repairing damaged skin, etc. DETAILED DESCRIPTION

[0029] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] The enzymes and bacteria used in the present invention are all commercially available products, as follows:

[0031] Cellulase, ficin, pectinase, bromelain, α-amylase, and alkaline protease were purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0032] Lactobacillus plantarum, accession number: CCTCCAB2010210, was purchased from China Center for Type Culture Collection.

[0033] Lactobacillus acidophilus, bacterial activity 20 billion CFU / g. Collection number: CCTCCAB2010208. Purchased from China Center for Type Culture Collection.

[0034] Bifidobacterium lactis, accession number: SHBCCD24675, was purchased from Shanghai Microbiological Collection Center.

[0035] Lactobacillus casei, accession number: SHBCCD24737, was purchased from Shanghai Microbiological Collection Center.

[0036] Lactobacillus bulgaricus, accession number: CCTCCCB20082295; purchased from China Center for Type Culture Collection.

[0037] Lactobacillus helveticus, accession number: CCTCCAB2010205; purchased from China Center for Type Culture Collection.

[0038] Example 1

[0039] This embodiment provides a method for preparing a camellia seed extract, the preparation method comprising the following steps:

[0040] (1) Crush camellia seeds to less than 100 meshes to obtain a first powder, add water to the first powder, the weight of the water is 3 times the weight of the first powder to obtain a mixture, add cellulase, pectinase and α-amylase, the concentration of cellulase in the mixture is 70 U / mL, the concentration of pectinase in the mixture is 100 U / mL, the concentration of α-amylase in the mixture is 140 U / mL, enzymolysis is carried out at 32° C. for 2.5 days, after the enzymes are inactivated, continue to add composite bacteria, the composite bacteria include Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei, the concentration of Lactobacillus plantarum in the mixture is 10 8 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 7 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 10 CFU / mL, anaerobic fermentation at 32°C for 4.5 days, vacuum drying to a moisture content of 3% to obtain fermented camellia seed powder;

[0041] (2) subcritically extracting the fermented camellia seed powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:3 as the extractant, the extraction temperature is 42° C., the extraction pressure is 2.4 MPa, and the extraction time is 60 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities to obtain a supernatant, and after removing alcohol, a first extract is obtained;

[0042] (3) drying the extract obtained in step (2) and then crushing it into 50 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 22 MPa, the extraction temperature to 52° C., the separation pressure to 5 MPa, and the separation temperature to 27° C., performing supercritical carbon dioxide extraction for 110 min, adding a 65% ethanol aqueous solution in an amount of 6 times the weight of the second powder, continuing the extraction for 4 hours, and obtaining a second extract after recovering the solvent and drying;

[0043] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0044] Example 2

[0045] This embodiment provides a method for preparing a camellia seed extract, the preparation method comprising the following steps:

[0046] (1) Crush camellia seeds to less than 100 meshes to obtain a first powder, add water to the first powder, the weight of the water is twice the weight of the first powder to obtain a mixture, add cellulase, pectinase and α-amylase, the concentration of cellulase in the mixture is 60 U / mL, the concentration of pectinase in the mixture is 90 U / mL, and the concentration of α-amylase in the mixture is 130 U / mL, perform enzymolysis at 30° C. for 2 days, inactivate the enzymes, and continue to add composite bacteria, the composite bacteria including Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei, the concentration of Lactobacillus plantarum in the mixture is 10 8 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 6 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 10 CFU / mL, anaerobic fermentation at 30°C for 4 days, vacuum drying to a moisture content of 2% to obtain fermented camellia seed powder;

[0047] (2) subcritically extracting the fermented camellia seed powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:2 as the extractant, the extraction temperature is 40° C., the extraction pressure is 2.2 MPa, and the extraction time is 50 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities to obtain a supernatant, and after removing alcohol, a first extract is obtained;

[0048] (3) drying the extract obtained in step (2) and crushing it into 20 meshes to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 20 MPa, the extraction temperature to 50° C., the separation pressure to 3 MPa, and the separation temperature to 25° C., performing supercritical carbon dioxide extraction for 100 min, adding a 60% ethanol aqueous solution in an amount of 5 times the weight of the second powder, and continuing the extraction for 3 hours, and obtaining a second extract after recovering the solvent and drying;

[0049] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0050] Example 3

[0051] This embodiment provides a method for preparing a camellia seed extract, the preparation method comprising the following steps:

[0052] (1) Crush camellia seeds to less than 200 meshes to obtain a first powder, add water to the first powder, the weight of the water is 4 times the weight of the first powder to obtain a mixture, add cellulase, pectinase and α-amylase, the concentration of cellulase in the mixture is 80 U / mL, the concentration of pectinase in the mixture is 110 U / mL, the concentration of α-amylase in the mixture is 150 U / mL, perform enzymolysis at 35° C. for 3 days, inactivate the enzymes, and continue to add composite bacteria, the composite bacteria including Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei, the concentration of Lactobacillus plantarum in the mixture is 10 9 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 7 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 11 CFU / mL, anaerobic fermentation at 35°C for 5 days, vacuum drying to a moisture content of 5% to obtain fermented camellia seed powder;

[0053] (2) subcritically extracting the fermented camellia seed powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:4 as the extractant, the extraction temperature is 45° C., the extraction pressure is 2.5 MPa, and the extraction time is 70 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities, obtaining a supernatant, and after removing alcohol, obtaining a first extract;

[0054] (3) drying the extract obtained in step (2) and then crushing it into 60 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 25 MPa, the extraction temperature to 55° C., the separation pressure to 7 MPa, and the separation temperature to 30° C., performing supercritical carbon dioxide extraction for 120 min, adding a 70% ethanol aqueous solution in an amount of 7 times the weight of the second powder, and continuing the extraction for 5 hours, and obtaining a second extract after recovering the solvent and drying;

[0055] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0056] Comparative Example 1

[0057] The difference between this comparative example and Example 1 is that no enzymatic hydrolysis and fermentation are performed.

[0058] A method for preparing a camellia seed extract, comprising the following steps:

[0059] (1) crushing camellia seeds to a size less than 100 meshes to obtain a first powder;

[0060] (2) subcritically extracting the first powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:3 as the extractant, the extraction temperature is 42° C., the extraction pressure is 2.4 MPa, and the extraction time is 60 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities, thereby obtaining a supernatant, and after removing alcohol, a first extract is obtained;

[0061] (3) drying the extract obtained in step (2) and then crushing it into 50 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 22 MPa, the extraction temperature to 52° C., the separation pressure to 5 MPa, and the separation temperature to 27° C., performing supercritical carbon dioxide extraction for 110 min, adding a 65% ethanol aqueous solution in an amount of 6 times the weight of the second powder, continuing the extraction for 4 hours, and obtaining a second extract after recovering the solvent and drying;

[0062] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0063] Comparative Example 2

[0064] The difference between this comparative example and Example 1 is that no fermentation is performed.

[0065] A method for preparing a camellia seed extract, comprising the following steps:

[0066] (1) grinding camellia seeds to less than 100 mesh to obtain a first powder, adding water to the first powder, the weight of the water being 3 times the weight of the first powder to obtain a mixture, adding cellulase, pectinase and α-amylase, the concentration of cellulase in the mixture being 70 U / mL, the concentration of pectinase in the mixture being 100 U / mL, and the concentration of α-amylase in the mixture being 140 U / mL, performing enzymolysis at 32° C. for 7 days, inactivating the enzymes, and vacuum drying to a moisture content of 3% to obtain fermented camellia seed powder;

[0067] (2) subcritically extracting the fermented camellia seed powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:3 as the extractant, the extraction temperature is 42° C., the extraction pressure is 2.4 MPa, and the extraction time is 60 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities to obtain a supernatant, and after removing alcohol, a first extract is obtained;

[0068] (3) drying the extract obtained in step (2) and then crushing it into 50 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 22 MPa, the extraction temperature to 52° C., the separation pressure to 5 MPa, and the separation temperature to 27° C., performing supercritical carbon dioxide extraction for 110 min, adding a 65% ethanol aqueous solution in an amount of 6 times the weight of the second powder, continuing the extraction for 4 hours, and obtaining a second extract after recovering the solvent and drying;

[0069] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0070] Comparative Example 3

[0071] The difference between this comparative example and Example 1 is that no enzymatic hydrolysis is performed.

[0072] A method for preparing a camellia seed extract, comprising the following steps:

[0073] (1) crushing camellia seeds to less than 100 meshes to obtain a first powder, adding water to the first powder, the weight of the water being 3 times the weight of the first powder, to obtain a mixture, adding composite bacteria, the composite bacteria comprising Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei, the concentration of Lactobacillus plantarum in the mixture being 10 8 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 7 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 10 CFU / mL, anaerobic fermentation at 32°C for 7 days, vacuum drying to a moisture content of 3% to obtain fermented camellia seed powder;

[0074] (2) subcritically extracting the fermented camellia seed powder using an extractant, using a fluid composed of butane and dimethyl ether in a weight ratio of 1:3 as the extractant, the extraction temperature is 42° C., the extraction pressure is 2.4 MPa, and the extraction time is 60 min to obtain an extract and an extraction product, and the crude extract obtained by removing the solvent from the extract is subjected to alcohol precipitation to remove impurities to obtain a supernatant, and after removing alcohol, a first extract is obtained;

[0075] (3) drying the extract obtained in step (2) and then crushing it into 50 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 22 MPa, the extraction temperature to 52° C., the separation pressure to 5 MPa, and the separation temperature to 27° C., performing supercritical carbon dioxide extraction for 110 min, adding a 65% ethanol aqueous solution in an amount of 6 times the weight of the second powder, continuing the extraction for 4 hours, and obtaining a second extract after recovering the solvent and drying;

[0076] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0077] Comparative Example 4

[0078] The difference between this comparative example and Example 1 is that the concentration of cellulase in the mixture is 90 U / mL, the concentration of pectinase in the mixture is 130 U / mL, and the concentration of α-amylase in the mixture is 60 U / mL.

[0079] Comparative Example 5

[0080] The difference between this comparative example and Example 1 is that cellulase is replaced by ficin; pectinase is replaced by bromelain; α-amylase is replaced by alkaline protease.

[0081] Comparative Example 6

[0082] The difference between this comparative example and Example 1 is that the concentration of Lactobacillus plantarum in the mixture is 10 7 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 10 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 8 CFU / mL.

[0083] Comparative Example 7

[0084] The difference between this comparative example and Example 1 is that Lactobacillus plantarum is replaced by Lactobacillus acidophilus; Lactobacillus helveticus is replaced by Lactobacillus bulgaricus; and Lactobacillus casei is replaced by Bifidobacterium lactis.

[0085] Comparative Example 8

[0086] The difference between this comparative example and Example 1 is that the supercritical carbon dioxide extraction method is different.

[0087] A method for preparing a camellia seed extract, comprising the following steps:

[0088] (1) Crush camellia seeds to less than 100 meshes to obtain a first powder, add water to the first powder, the weight of the water is 3 times the weight of the first powder to obtain a mixture, add cellulase, pectinase and α-amylase, the concentration of cellulase in the mixture is 70 U / mL, the concentration of pectinase in the mixture is 100 U / mL, the concentration of α-amylase in the mixture is 140 U / mL, enzymolysis is carried out at 32° C. for 2.5 days, after the enzymes are inactivated, continue to add composite bacteria, the composite bacteria include Lactobacillus plantarum, Lactobacillus helveticus and Lactobacillus casei, the concentration of Lactobacillus plantarum in the mixture is 10 8 CFU / mL, the concentration of Lactobacillus helveticus in the mixture is 10 7 CFU / mL, the concentration of Lactobacillus casei in the mixture is 10 10 CFU / mL, anaerobic fermentation at 32°C for 4.5 days, vacuum drying to a moisture content of 3% to obtain fermented camellia seed powder;

[0089] (2) subcritically extracting the fermented camellia seed powder using an extractant, using anhydrous ethanol as the extractant, at an extraction temperature of 60° C., an extraction pressure of 3.5 MPa, and an extraction time of 60 min to obtain an extract and an extraction product, and performing alcohol precipitation to remove impurities from the crude extract obtained by removing the solvent from the extract to obtain a supernatant, and after removing alcohol, obtaining a first extract;

[0090] (3) drying the extract obtained in step (2) and then crushing it into 50 mesh to obtain a second powder, placing the second powder into a supercritical carbon dioxide extraction kettle, adjusting the extraction pressure to 22 MPa, the extraction temperature to 52° C., the separation pressure to 5 MPa, and the separation temperature to 27° C., performing supercritical carbon dioxide extraction for 110 min, adding a 65% ethanol aqueous solution in an amount of 6 times the weight of the second powder, continuing the extraction for 4 hours, and obtaining a second extract after recovering the solvent and drying;

[0091] (4) The first extract and the second extract are mixed evenly to obtain a camellia seed extract.

[0092] Performance Testing

[0093] The camellia seed extracts prepared in Examples 1-3 and Comparative Examples 1-8 were subjected to performance tests.

[0094] 1. Referring to GB / T11765-2018, the dry basis weight percentage of tea saponin, tea polysaccharide and tea protein in camellia seed extract was determined, and the calculation formula = dry basis weight of tea saponin (dry basis weight of tea polysaccharide or dry basis weight of tea protein) ÷ total dry basis weight of camellia seed extract * 100%. The method of the present invention adopts the vanillyl alcohol-concentrated sulfuric acid colorimetric method to detect tea saponin, the phenol-sulfuric acid method to detect tea polysaccharide, and the BAC reagent method to detect tea protein.

[0095] Yield = mass of camellia seed extract ÷ mass of camellia seeds * 100%.

[0096] The results are shown in Table 1.

[0097] Table 1 Performance test results

[0098]

[0099]

[0100] 2. The camellia seed extracts of Examples 1-3 and Comparative Examples 1-8 were sealed and stored, and placed in the following four environments for observation to test the storage stability. The results are shown in Table 2.

[0101] Normal temperature: The sample was left at 25°C, 60% RH (relative humidity) for 90 days.

[0102] Cold resistance: -10℃ for 10 days.

[0103] Hot and cold cycle: -10℃, 25%RH; 25℃, 60%RH; 38℃, 75%RH, 24h at each temperature, 10 cycles in total.

[0104] High temperature acceleration: The test conditions are 50℃±1℃, 25%RH±5%RH for 60 days.

[0105] If there is no obvious change in the material, it is recorded as passed. If there is one or more changes such as precipitation, odor change or darkening of color, it is recorded as failed.

[0106] Table 2 Stability test results

[0107]

[0108]

[0109] From the above performance test results, it can be seen that the camellia seed extracts of Examples 1-3 have high quality and high yield, especially the comprehensive performance of Example 1 is the most outstanding.

[0110] 3. Test the whitening effect of camellia seed extract

[0111] Thirty male subjects aged 30-40 with dull skin and acne scars were recruited and randomly divided into three groups. Camellia seed extract was applied to the face every night. No other cosmetics were used on the test area during the test. The test was continued for 7 days. The skin melanin and hemoglobin tester Mexameter MX18 of German CK Company was used to test the average MI value of the cheek area on the 0th day and the 7th day (MI value represents the signal of melanin, and the measurement range of the instrument is 0-999. The higher the value, the higher the content of melanin in the skin). The results are shown in Table 3.

[0112] Table 3 Whitening effect

[0113]

[0114] However, the comparative examples were significantly inferior to the embodiments in the corresponding performance tests because they did not adopt the necessary technical solutions. In comparative example 1, no enzymolysis and fermentation were performed, and in comparative examples 2-3, only enzymolysis or fermentation solutions were used. It can be seen that the quality and yield effects decreased. The conditions of enzymolysis and fermentation in comparative examples 4-7 were different, which affected the extraction amount of tea saponin, tea polysaccharides and tea protein. Comparative example 8 used different extraction solutions, which would affect the product yield. The above experimental results further prove the importance of the technical solutions defined in the present invention for its technical effects.

[0115] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing camellia seed extract, characterized in that: The preparation method comprises the following steps: (1) Crush camellia seeds to less than 100-200 meshes to obtain a first powder, add water to the first powder to obtain a mixture, add cellulase, pectinase and α-amylase, perform enzymolysis at 30-35° C. for 2-3 days, inactivate the enzymes, continue to add composite bacteria, perform anaerobic fermentation at 30-35° C. for 4-5 days, sterilize, and vacuum dry to a moisture content of 2-5% to obtain fermented camellia seed powder; (2) subcritically extracting the fermented camellia seed powder using an extractant at an extraction temperature of 40-45° C., an extraction pressure of 2.2-2.5 MPa, and an extraction time of 50-70 min to obtain an extract and an extraction product, and subjecting the extract to alcohol precipitation to remove impurities to obtain a supernatant, and to obtain a first extract after removing alcohol; (3) The extraction product obtained in step (2) is dried and crushed into 20-60 mesh to obtain a second powder, and the second powder is placed in a supercritical carbon dioxide extraction kettle, and the extraction pressure is adjusted to 20-25 MPa, the extraction temperature is 50-55°C, the separation pressure is 3-7 MPa, and the separation temperature is 25-30°C, and supercritical carbon dioxide extraction is performed for 100-120 min. An ethanol aqueous solution is added and the extraction is continued for 3-5 hours. After the solvent is recovered and dried, a second extract is obtained; (4) mixing the first extract and the second extract evenly to obtain a camellia seed extract; In the step (1), the concentration of cellulase in the mixture is 60U / mL-80U / mL; the concentration of pectinase in the mixture is 90U / mL-110U / mL; and the concentration of α-amylase in the mixture is 130U / mL-150U / mL; The composite bacteria in step (1) include Lactobacillus plantarum CCTCCAB2010210, Lactobacillus helveticus CCTCCAB2010205 and Lactobacillus casei SHBCCD24737; the concentration of Lactobacillus plantarum CCTCCAB2010210 in the mixture is 10 8 -10 9 CFU / mL; the concentration of Lactobacillus helveticus CCTCCAB2010205 in the mixture is 10 6 -10 7 CFU / mL; the concentration of Lactobacillus casei SHBCCD24737 in the mixture is 10 10 -10 11 CFU / mL; In the step (2), a fluid composed of butane and dimethyl ether in a weight ratio of 1:(2-4) is used as the extractant.

2. The method for preparing the camellia seed extract according to claim 1, characterized in that: In the step (3), a 60%-70% ethanol aqueous solution is added in an amount of 5-7 times the weight of the second powder.

3. Use of the camellia seed extract prepared according to the preparation method according to any one of claims 1 to 2 in the preparation of cosmetics.

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