Camellia polypeptide composition with anti-wrinkle, repairing and brightening effects as well as preparation method and application thereof

Abstract

The invention provides a preparation method and application of a camellia polypeptide composition with anti-wrinkle, repairing and brightening effects. The camellia polypeptide composition comprises a component A and a component B, wherein the component A comprises a wrapping solubilizer and polyhydric alcohol; component B: a plurality of polypeptides; the component C comprises carnosine, camellia polypeptide and water. The camellia polypeptide disclosed by the invention is prepared by the following steps: performing supercritical deoiling on camellia seeds, performing alkali extraction, acid precipitation and enzymolysis to prepare polypeptide, performing membrane filtration, taking micromolecule polypeptide as a main component, promoting synthesis of type I, type III and type V collagen, promoting cell migration and scavenging free radicals, and has the effects of resisting wrinkles, repairing and resisting oxidation; according to the invention, the problem of water solubility of palmitoyl peptide and acetyl peptide is solved, so that palmitoyl peptide and acetyl peptide can form a stable composition together with water-soluble polypeptide. The anti-wrinkle, repairing and whitening effects are achieved in multiple ways and multiple directions, the effects of improving wrinkles, repairing the skin, brightening the skin color and delaying senescence can be better achieved, and the anti-wrinkle, repairing and whitening composition can be widely applied to various types of cosmetics.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and specifically relates to a camellia polypeptide composition with anti-wrinkle, repair and brightening effects, a preparation method thereof, and an application thereof. Background Art

[0002] Skin aging is a physiological or pathological phenomenon of degenerative changes in skin morphology and function, and its main characteristics are increased and deepened skin wrinkles, dryness, roughness, sagging, loss of elasticity, pigmentation, dull skin, etc. Skin aging is a complex biological process, affected by various internal and external factors such as genetics, environment and lifestyle. External factors, such as light, environmental pollution, etc., will cause the skin to produce a large number of free radicals, inflammatory factors, etc., accelerate the production of matrix metalloproteinases, tyrosinases, etc., cause the degradation of various proteins in the skin, melanin production. At the same time, it accelerates the non-enzymatic condensation reaction of the carbonyl group of reducing sugar and protein, and the advanced glycation end products (AGEs) produced cause browning, fluorescence and cross-linking damage of collagen, and finally lead to dull, sagging and loss of elasticity of the skin. With the increase of age, the degradation of the extracellular matrix ECM of skin cells, especially the continuous loss of collagen in the dermis layer, leads to the appearance of wrinkles, loss of elasticity and sagging of the skin. Therefore, free radicals, inflammatory factors, glycation reactions, collagen degradation, etc. are the main factors leading to skin aging.

[0003] Bioactive polypeptides, also known as functional peptides, such as palmitoyl peptides, acetyl peptides, carnosine, etc. are recognized as highly effective and safe active ingredients for preventing and resisting skin aging, and are widely used in the field of beauty skin care, such as anti-skin aging, skin repair, skin brightening, etc. However, the application of palmitoyl peptides and acetyl peptides is limited by their solubility.

[0004] Since each active peptide has a unique structure and function, a relatively clear mechanism of action and target on the skin, and significant efficacy, it is necessary to combine peptides with multiple different effects to achieve multi-faceted and three-dimensional anti-skin aging. CN 116370331 A discloses a polypeptide anti-wrinkle composition and its application, including carnosine, acetyl hexapeptide-8, palmitoyl pentapeptide-4, and dipalmitoyl hydroxyproline, which has the effect of skin aging, but does not consider the solubility of different peptides, resulting in poor permeability. CN201910957516.8 discloses a polypeptide anti-aging composition, in which acetyl hexapeptide-8 is 10-20%, palmitoyl tripeptide-1 is 5-7%, palmitoyl tetrapeptide-7 is 5-8%, acetyl tetrapeptide-2 is 1-5%, palmitoyl tripeptide-5 is 3-6%, pepper seed extract is 0.01-0.1%, butanediol is 10-20%, propylene glycol is 5-10%, inositol is 1-5%, Koelreuteria paniculata fruit extract is 0.05-0.1%, and the balance is deionized water. The addition amount of the functional peptide in this patent is very high. Due to the high price of polypeptides, the cost is very high. At the same time, the polyol system is not good enough for the solubility of peptides, and the peptide bioavailability is not considered either. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a camellia polypeptide composition with anti-wrinkle, repair, and brightening effects, its preparation method, and application. First of all, the present invention introduces low-cost camellia polypeptides and organically combines them with multiple functional peptides to achieve skin anti-aging effects under the condition of very low addition amount of functional peptides in the whole composition. Secondly, palmitoyl tripeptide-5 and palmitoyl pentapeptide-4 in the present invention can promote the synthesis of type I and type III collagen; acetyl hexapeptide-8 can quickly anti-wrinkle by regulating nerve conduction; palmitoyl tetrapeptide-7 can anti-skin aging by scavenging inflammatory factors; carnosine can anti-skin aging by scavenging free radicals; nonapeptide-1 can inhibit melanin production and brighten the skin tone; camellia polypeptides can promote the synthesis of type I, type III, and type V collagen, especially can significantly increase the synthesis of type V collagen, and can also scavenge free radicals. Therefore, this combination can achieve multi-faceted and three-dimensional anti-skin aging; furthermore, the present invention adopts the encapsulation solubilization technology to solve the water solubility problems of palmitoyl peptides, acetyl peptides, nonapeptide-1, etc., and at the same time obtains a water-soluble liquid with clear and transparent appearance, small particle size, and uniform particle size, increasing its use convenience.

[0006] The purpose of the present invention is achieved by the following technical solutions:

[0007] The present invention provides a camellia polypeptide composition with anti-wrinkle, repair, and brightening effects, which includes the following components in mass percentage:

[0008]

[0009] The camellia polypeptide composition of the present invention comprises three components, namely component A, component B and component C. Among them, component A includes a wrapping solubilizer and a polyol; component B includes palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, nonapeptide-1; component C includes carnosine, camellia polypeptide and water.

[0010] The highlight of the present invention lies in the design of the polypeptide combination. According to the internal and external causes and intuitive manifestations of aging, the polypeptide combination is designed, and through a variety of polypeptide combinations, the problem of skin aging is comprehensively solved; for different causes and external surfaces of skin aging, different polypeptides are selected for solution, so as to achieve comprehensive three-dimensional anti-aging.

[0011] As an embodiment of the present invention, the wrapping solubilizer includes a penetration wrapping solubilizer and a particle size wrapping solubilizer; the penetration wrapping solubilizer is acetylcholine, and the particle size wrapping solubilizer is one or a combination of two of PPG-13-decyltetradeceth-24 and polyglyceryl-4 oleate. Among the wrapping solubilizers, the weight content of the penetration solubilizer is not less than 50%, preferably 50%-70%.

[0012] The present invention forms an inclusion body with small particle size (particle size less than 100nm), uniform distribution and good permeability through the combined use of a penetration wrapping solubilizer and a particle size wrapping solubilizer. The penetration wrapping solubilizer (acetylcholine) is a biological surfactant and a component of the cell membrane, with good permeability, but the inclusion body formed by it has a relatively large particle size and poor stability; while the particle size wrapping solubilizer (PPG-13-decyltetradeceth-24, polyglyceryl-4 oleate) can form very small inclusion particle sizes, but its permeability is not as good as that of the penetration wrapping solubilizer (acetylcholine). Therefore, the combined use makes the inclusion body not only have a small particle size but also good biological permeability.

[0013] As an embodiment of the present invention, the polyol includes one or several of 1,3-propanediol, 1,3-butanediol, 1,2-pentanediol, 1,2-hexanediol.

[0014] As an embodiment of the present invention, the camellia polypeptide is prepared by the following method:

[0015] A1. The camellia seed powder is subjected to a supercritical CO2 extraction process to remove the oil therein, and a defatted camellia seed cake is obtained;

[0016] A2. The defatted camellia seed cake is added to deionized water, the pH is adjusted to 8.50-10.00, and then heated and stirred for extraction. After filtration, a camellia seed protein extract is obtained;

[0017] A3. The pH of the camellia seed protein extract is adjusted to 3.0-4.0, and then filtered to obtain camellia seed protein 1;

[0018] A4. Wash the camellia seed protein 1 with an ethanol solution to remove the camellia saponin in the protein, and then dry it to remove the ethanol, obtaining camellia seed protein 2;

[0019] A5. Add the camellia seed protein 2 into deionized water, and then add papain, alkaline protease, neutral protease and trypsin respectively to obtain a mixed solution. Heat and stir for enzymatic hydrolysis, heat up again for heat preservation and stir, and filter to obtain the camellia seed protein filtrate;

[0020] A6. Add activated carbon to the camellia seed protein filtrate, heat and stir, and filter to obtain the said camellia polypeptide.

[0021] As an embodiment of the present invention, in step A1, the supercritical CO2 extraction process is as follows: pressure: 25 Mpa; temperature: 50 °C; time: 3 h. The present invention uses the supercritical technology to remove the camellia seed oil during the extraction process, avoiding the low extraction rate caused by oil emulsification during the process of water extracting protein. After the supercritical treatment, the protein extraction rate is higher.

[0022] As an embodiment of the present invention, in step A2, the dosage of deionized water is 30 times the mass of the defatted camellia seed cake. The pH is adjusted using a sodium hydroxide solution, and the mass fraction of the sodium hydroxide solution is 3 - 5%. The temperature for heating and stirring extraction is 55 - 65 °C, and the time is 1 - 3 h. After heating and stirring extraction, cool it to room temperature and then filter to obtain the camellia seed protein extract.

[0023] As an embodiment of the present invention, in step A3, the pH is adjusted using a citric acid solution. The mass fraction of the citric acid solution is 5 - 8%.

[0024] As an embodiment of the present invention, in step A4, the mass fraction of the ethanol solution is above 80%. The dosage of the ethanol solution is 10 - 20 times the mass of the camellia seed protein. After washing, dry it to remove the ethanol. The removed camellia saponin is toxic to the protease and will cause its inactivation. Adding trypsin to the protease combination can improve the enzymatic hydrolysis efficiency.

[0025] As an embodiment of the present invention, in step A5, the dosage of deionized water is 2 - 4 times the mass of the defatted camellia seed cake in step A1. The dosages of papain, alkaline protease, neutral protease and trypsin are 0.5 - 2%, 0.1 - 1%, 0.1 - 1%, 0.3 - 2% of the mass of the defatted camellia seed cake in step A1 respectively.

[0026] As an embodiment of the present invention, in step A5, the temperature for heating and stirring enzymatic hydrolysis is 55 - 65 °C, and the time is 1 - 3 h. The temperature for heating up again for heat preservation is 80 - 100 °C, and the time is 5 - 20 min. Wait until it cools to room temperature and then filter.

[0027] When different enzymes are used in the enzymatic hydrolysis process, the polypeptide structures hydrolyzed by the enzymes are different. The polypeptides obtained from camellia seed powder in the present invention have a clear amino acid sequence, mainly tetrapeptide (Val-Val-Val-Lys), pentapeptide (Leu-Thr-Leu-Leu-Arg) and hexapeptide (Gly-Tyr-Leu-Arg-Leu-Lys), with a content of more than 80%, and the molecular weight is about 500, and the efficacy is better. In addition to the anti-wrinkle effect, camellia polypeptides also have antioxidant, anti-glycation, soothing and whitening effects, and at the same time have a synergistic effect with other peptides.

[0028] As an embodiment of the present invention, in step A6, the dosage of activated carbon is 0.1-0.3% of the mass of the camellia seed protein filtrate. The temperature of heating and stirring is 55-65 °C, and the time is 20-40 min. Filtration is carried out using an ultrafiltration membrane with a pore size of 500 Da. The solution is filtered after cooling to room temperature, and then passed through the ultrafiltration membrane to obtain camellia polypeptides.

[0029] Activated carbon can adsorb components such as flavonoids and polyphenols in the camellia polypeptide solution, thereby affecting the encapsulation efficiency. At the same time, the presence of flavonoids and polyphenols will also affect the particle size distribution after encapsulation.

[0030] The present invention also provides a preparation method of a camellia polypeptide composition with anti-wrinkle, repair and brightening effects, including the following steps:

[0031] S1. Preparation of component A: Stir and mix the encapsulating solubilizer and polyol evenly according to the mass ratio;

[0032] S2. Preparation of component B: Stir and mix palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 evenly according to the mass ratio;

[0033] S3. Preparation of component C: Stir and mix carnosine, camellia polypeptides and water evenly according to the mass ratio;

[0034] S4. Preparation of the composition: Slowly add component B to component A, stir while adding to ensure that component A and component B are mixed evenly; then heat component C to 50-70 °C, and slowly drop the mixture of component A and component B into component C, stir while dropping, and finally obtain the camellia polypeptide composition.

[0035] The present invention also provides the application of the camellia polypeptide composition with anti-wrinkle, repair and brightening effects in cosmetics.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] (1) Skin aging is mainly caused by factors such as free radicals, inflammation, glycosylation, and collagen loss. This invention designs and combines through four anti-aging mechanisms: scavenging free radicals (carnosine, camellia polypeptides), anti-glycation (camellia polypeptides), inhibiting inflammation (palmitoyl tetrapeptide-7), and promoting collagen regeneration (camellia polypeptides, palmitoyl tripeptide-5, palmitoyl pentapeptide-4), achieving scientific and reasonable design, as comprehensive, safe and effective as possible to achieve long-lasting anti-aging effects;

[0038] (2) The most obvious external features of skin aging are skin wrinkles and dullness. This invention aims at these two major skin problems, adding a neuropeptide (acetyl hexapeptide-8) to the composition to achieve instant anti-wrinkle effect; at the same time adding nonapeptide-1 and camellia polypeptides to inhibit melanin production and quickly brighten the skin tone; enabling the composition of this invention to achieve rapid anti-aging effects;

[0039] (3) This invention combines supercritical extraction technology, enzymatic hydrolysis technology and membrane separation to obtain the main structures (with a content of more than 80%) of tetrapeptide Leu-Thr-Leu-Leu-Arg (leucine-threonine-leucine-leucine-arginine), pentapeptide Val-Val-Val-Lys (valine-valine-valine-lysine), and hexapeptide Gly-Tyr-Leu-Arg-Leu-Lys (glycine-tyrosine-leucine-arginine-leucine-lysine), which have significant anti-aging effects.

[0040] (4) This invention solves the water solubility problems of palmitoyl peptides, acetyl peptides, nonapeptide-1, etc. through the combination of acetylcholine and PPG-13-decyltetradeceth-24, polyglyceryl-4 oleate combination wrapping solubilizer technology. At the same time, the particle size of the wrapped body is small (with a particle size below 100nm), uniform, and has good permeability, enabling high content of active peptides to be added to the composition, while maintaining a homogeneous clear and transparent state and good bioavailability.

[0041] (5) The addition of polyols increases the solubility of polypeptides and also solves the problem of system anti-corrosion.

[0042] (6) After supercritical treatment, camellia seeds can completely remove oil, greatly improving the protein extraction rate. The protein extraction rate is not only higher than that of camellia seeds without oil removal, but also higher than that of commercially available camellia seed cakes (camellia seeds after oil extraction); at the same time, saponins, flavonoids and polyphenols and other components are removed through ethanol washing and activated carbon adsorption, improving the activity of protease and the wrapping effect. Detailed implementation methods

[0043] The present invention will be described in detail below in conjunction with embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several adjustments and improvements can still be made. These all belong to the protection scope of the present invention.

[0044] The present invention provides a preparation method and application of a camellia polypeptide composition with anti-wrinkle, repair and brightening effects, including three components A, B, and C. Component A includes a wrapping solubilizer and polyol; Component B includes palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1; Component C includes carnosine, camellia polypeptide, and water. The camellia polypeptide in the present invention is prepared from camellia seeds after supercritical deoiling, through alkali extraction and acid precipitation, enzymatic hydrolysis to prepare polypeptides, and then membrane filtration to select small-molecule polypeptides with a molecular weight below 500. Its amino acid sequences are mainly Leu-Thr-Leu-Leu-Arg (leucine-threonine-leucine-leucine-arginine), Val-Val-Val-Lys (valine-valine-valine-lysine), Gly-Tyr-Leu-Arg-Leu-Lys (glycine-tyrosine-leucine-arginine-leucine-lysine), which can promote the synthesis of type I, III, and V collagen, promote cell migration, scavenge free radicals, and have anti-wrinkle, repair, and antioxidant effects; Palmitoyl tripeptide-5 and palmitoyl pentapeptide-4 can promote the synthesis of type I collagen through the TGF-β target and have anti-wrinkle effects; Palmitoyl tetrapeptide-7 can inhibit inflammatory factors, have a soothing effect, and can repair the skin; Nonapeptide-1 can inhibit the production of melanin and have a brightening effect; Acetyl hexapeptide-8 can regulate nerve conduction to achieve a rapid anti-wrinkle effect; Carnosine has a strong antioxidant ability, can neutralize free radicals, protect skin cells from oxidative damage, and thus delay skin aging. The present invention successfully solves the water solubility problem of palmitoyl peptides and acetyl peptides by various methods, enabling them to form a stable composition with water-soluble polypeptides. The composition of the present invention can achieve anti-wrinkle, repair, and whitening effects through multiple channels and aspects, and can better achieve the effects of improving wrinkles, repairing the skin, brightening the skin tone, and delaying aging, and can be widely applied to various types of cosmetics.

[0045] Examples 1-5

[0046] This example relates to a preparation method of a camellia polypeptide composition with anti-wrinkle, repair and brightening effects.

[0047] The components and dosages of the compositions described in Examples 1-5 are shown in Table 1 below:

[0048] Table 1

[0049]

[0050]

[0051] The preparation methods of the compositions described in Examples 1-5 are as follows:

[0052] 1. The encapsulating solubilizer in Component A is acetylcholine and PPG-13-decyltetradeceth-24, and their mass ratio is 1:1; the polyol is 1,3-propanediol.

[0053] 2. The preparation method of camellia polypeptide in Component C is as follows:

[0054] (1) Camellia seeds are crushed and sieved to obtain camellia seed powder; the oil is removed by supercritical CO2 extraction process (pressure: 25 Mpa; temperature: 50 °C; time: 3 h) to obtain defatted camellia seed cake;

[0055] (2) Weigh a certain amount of defatted camellia seed cake, add deionized water 30 times its mass, then adjust the pH of the system to 8.50 with 4% sodium hydroxide solution, and then heat to 60 °C and stir for extraction for 2 h;

[0056] (3) After the above solution is cooled to room temperature, it is filtered to obtain camellia seed protein extract, then the pH of the system is adjusted to 3.0 with 5% citric acid solution, and then filtered to obtain camellia seed protein;

[0057] (4) The obtained camellia seed protein is washed with 80% ethanol 10 times its mass to remove camellia saponins in the protein, and then dried to remove ethanol;

[0058] (5) Add deionized water 2 times the mass of the defatted camellia seed cake in step 2 to the camellia seed protein, and then add 0.5%, 0.1%, 0.1%, and 0.3% of papain, alkaline protease, neutral protease, and alkaline protease of the mass of the defatted camellia seed cake in step 2 respectively, then heat the system to 60 °C, stir for enzymatic hydrolysis for 2 h, then raise the temperature to 90 °C, keep warm and stir for 10 min, and filter after cooling to room temperature;

[0059] (6) Add 0.3% activated carbon of its mass to the above solution, heat to 60 °C, stir for 30 min, filter after cooling to room temperature, and then pass through an ultrafiltration membrane with a pore size of 500 Da to obtain camellia polypeptide.

[0060] 3. The preparation method of the composition is as follows:

[0061] (1) Mix acetylcholine, PPG-13-decyltetradeceth-24 and 1,3-propanediol evenly according to the ratio in Table 1;

[0062] (2) Mix palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 evenly according to the ratio in Table 1, and then slowly add Component B to Component A while stirring to ensure uniform mixing of Component A and Component B.

[0063] (3) Mix carnosine, camellia polypeptide, and water evenly according to the ratio in Table 1, then heat Component C to 50 °C, and slowly drop the mixture of Component A and Component B into Component C while stirring. Finally, a camellia polypeptide composition is obtained.

[0064] Then slowly drop the mixture of Component A and Component B into Component C while stirring. Finally, a camellia polypeptide composition is obtained.

[0065] Example 2

[0066] 1. The encapsulating solubilizer in Component A is acetylcholine and polyglyceryl-4 oleate, and their mass ratio is 2:1. The polyol is 1,3-butanediol

[0067] 2. The preparation method of camellia polypeptide in Component C is as follows:

[0068] (1) Crush camellia seeds and sieve them to obtain camellia seed powder; remove the oil therein by supercritical CO2 extraction process (pressure: 25 Mpa; temperature: 50 °C; time: 3 h) to obtain defatted camellia seed cake;

[0069] (2) Weigh a certain amount of defatted camellia seed cake, add deionized water 30 times its mass, then adjust the pH of the system to 10.00 with 4% sodium hydroxide solution, and then heat to 60 °C and stir for extraction for 2 h;

[0070] (3) After the above solution is cooled to room temperature, filter it to obtain camellia seed protein extract, then adjust the pH of the system to 4.0 with 8% citric acid solution, and then filter to obtain camellia seed protein;

[0071] (4) Wash the obtained camellia seed protein with absolute ethanol 20 times its mass to remove camellia saponin in the protein, and then dry it to remove ethanol;

[0072] (5) Add deionized water 4 times the mass of the defatted camellia seed cake in step 2 to the camellia seed protein, and then add papain, alkaline protease, neutral protease, and alkaline protease at 2%, 1%, 1%, and 2% of the mass of the defatted camellia seed cake in step 2 respectively. Then heat the system to 60 °C and stir for enzymatic hydrolysis for 2 h, then raise the temperature to 90 °C and keep stirring for 10 min. After cooling to room temperature, filter.

[0073] (6) Add activated carbon accounting for 0.3% of its mass to the above solution, heat to 60 °C, stir for 30 min, filter after cooling to room temperature, and then pass through an ultrafiltration membrane with a pore size of 500 Da to obtain camellia polypeptide.

[0074] 3. The preparation method of the composition is as follows:

[0075] (1) Mix acetylcholine, polyglyceryl-4 oleate, and 1,3-butanediol evenly according to the proportions in Table 1;

[0076] (2) Mix palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 evenly according to the proportions in Table 1, and then slowly add component B to component A while stirring to ensure uniform mixing of component A and component B;

[0077] (3) Mix carnosine, camellia polypeptide, and water evenly according to the proportions in Table 1, then heat component C to 70 °C, and slowly drop the mixture of component A and component B into component C while stirring to finally obtain the camellia polypeptide composition.

[0078] Example 3

[0079] 1. The encapsulating solubilizer in component A is acetylcholine, polyglyceryl-4 oleate, and PPG-13-decyltetradeceth-24, and their mass ratio is 2:1:1; the polyol is a mixture of 1,3-propanediol, 1,3-butanediol, and 1,2-pentanediol, and their mass ratio is 3:5:1

[0080] 2. The preparation method of camellia polypeptide in component C is as follows:

[0081] (1) Crush camellia seeds and sieve to obtain camellia seed powder; remove the oil therein by supercritical CO2 extraction process (pressure: 25 Mpa; temperature 50 °C; time: 3 h) to obtain defatted camellia seed cake;

[0082] (2) Weigh a certain amount of defatted camellia seed cake, add deionized water 30 times its mass, then adjust the pH of the system to 9.00 with 4% sodium hydroxide solution, and then heat to 60 °C and stir for 2 h;

[0083] (3) Filter after the above solution cools to room temperature to obtain camellia seed protein extract, then adjust the pH of the system to 3.5 with 8% citric acid solution, and then filter to obtain camellia seed protein;

[0084] (4) Wash the obtained camellia seed protein with 90% ethanol 15 times its mass to remove camellia saponin in the protein, and then dry to remove ethanol;

[0085] (5) Add deionized water that is 4 times the mass of the defatted camellia seed cake obtained in step 2 to the camellia seed protein, and then add papain, alkaline protease, neutral protease, and alkaline protease at 1.5%, 0.8%, 0.8%, and 1% of the mass of the defatted camellia seed cake obtained in step 2 respectively. Then heat the system to 60 °C and stir for enzymatic hydrolysis for 2 h. Subsequently, raise the temperature to 90 °C, keep warm and stir for 10 min, and filter after cooling to room temperature;

[0086] (6) Add activated carbon at 0.2% of its mass to the above solution, heat to 60 °C, stir for 30 min, filter after cooling to room temperature, and then pass through an ultrafiltration membrane with a pore size of 500 Da to obtain camellia polypeptide.

[0087] 3. The preparation method of the composition is as follows:

[0088] (1) Mix acetylcholine, polyglyceryl-4 oleate, PPG-13-decyltetradeceth-24, and polyol evenly according to the proportions in Table 1;

[0089] (2) Mix palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 evenly according to the proportions in Table 1, and then slowly add component B to component A while stirring to ensure that component A and component B are mixed evenly;

[0090] (3) Mix carnosine, camellia polypeptide, and water evenly according to the proportions in Table 1. Then heat component C to 60 °C, and slowly drip the mixture of component A and component B into component C while stirring. Finally, obtain the camellia polypeptide composition.

[0091] Example 4

[0092] 1. The encapsulating solubilizer in component A is a mixture of acetylcholine, polyglyceryl-4 oleate, and PPG-13-decyltetradeceth-24, and their mass ratio is 3:1:1; the polyol is a mixture of 1,3-propanediol, 1,3-butanediol, 1,2-pentanediol, and 1,2-hexanediol, and their mass ratio is 3:5:1:1

[0093] 2. The preparation method of camellia polypeptide in component C is as follows:

[0094] (1) Crush the camellia seeds and sieve to obtain camellia seed powder; remove the oil therein by supercritical CO2 extraction process (pressure: 25 Mpa; temperature: 50 °C; time: 3 h) to obtain defatted camellia seed cake;

[0095] (2) Weigh a certain amount of defatted camellia seed cake, add deionized water that is 30 times its mass, then adjust the pH of the system to 8.50 with 4% sodium hydroxide solution, and then heat to 60 °C and stir for extraction for 2 h;

[0096] (3) After the above solution is cooled to room temperature, it is filtered to obtain the camellia seed protein extract. Then, the pH of the system is adjusted to 4.0 with 6% citric acid solution, and then filtered to obtain camellia seed protein.

[0097] (4) The obtained camellia seed protein is washed with 90% ethanol 15 times its mass to remove camellia saponins in the protein, and then dried to remove ethanol.

[0098] (5) Deionized water 3.5 times the mass of the defatted camellia seed cake in step 2 is added to the camellia seed protein, and then papain, alkaline protease, neutral protease and alkaline protease 1.5%, 0.5%, 0.5% and 1.5% of the mass of the defatted camellia seed cake in step 2 are added respectively. Then, the system is heated to 60 °C, stirred and enzymatically hydrolyzed for 2 h. Subsequently, the temperature is raised to 90 °C, kept warm and stirred for 10 min, and filtered after cooling to room temperature.

[0099] (6) 0.15% activated carbon by its mass is added to the above solution, heated to 60 °C, stirred for 30 min, filtered after cooling to room temperature, and then passed through an ultrafiltration membrane with a pore size of 500 Da to obtain camellia polypeptide.

[0100] 3. The preparation method of the composition is as follows:

[0101] (1) Acetylcholine, polyglyceryl-4 oleate, PPG-13-decyltetradeceth-24 and polyol are mixed evenly according to the proportions in Table 1.

[0102] (2) Palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, nonapeptide-1 are mixed evenly according to the proportions in Table 1, and then component B is slowly added to component A while stirring to ensure that component A and component B are mixed evenly.

[0103] (3) Carnosine, camellia polypeptide and water are mixed evenly according to the proportions in Table 1. Then, component C is heated to 65 °C, and the mixture of component A and component B is slowly added dropwise to component C while stirring. Finally, the camellia polypeptide composition is obtained.

[0104] Example 5

[0105] 1. The encapsulating solubilizer in component A is acetylcholine and PPG-13-decyltetradeceth-24, and their mass ratio is 2:1; the polyol is 1,3-propanediol and 1,3-butanediol, and their mass ratio is 1:1.

[0106] 2. The preparation method of camellia polypeptide in component C is as follows:

[0107] (1) The camellia seeds are crushed and sieved to obtain camellia seed powder; the oil in them is removed by supercritical CO2 extraction process (pressure: 25 Mpa; temperature: 50 °C; time: 3 h) to obtain defatted camellia seed cake;

[0108] (2) Weigh a certain amount of defatted camellia seed cake, add deionized water 30 times its mass, then adjust the pH of the system to 8.50 with 4% sodium hydroxide solution, and then heat to 60 °C and stir for extraction for 2 h;

[0109] (3) After the above solution is cooled to room temperature, filter to obtain camellia seed protein extract, then adjust the pH of the system to 3.5 with 8% citric acid solution, and then filter to obtain camellia seed protein;

[0110] (4) Wash the obtained camellia seed protein with 90% ethanol 15 times its mass to remove camellia saponins in the protein, and then dry to remove ethanol;

[0111] (5) Add deionized water 3 times the mass of the defatted camellia seed cake in step 2 to the camellia seed protein, and then add 1.5%, 1%, 1%, and 2% of papain, alkaline protease, neutral protease, and trypsin based on the mass of the defatted camellia seed cake in step 2 respectively. Then heat the system to 60 °C and stir for enzymatic hydrolysis for 2 h, and then raise the temperature to 90 °C and keep stirring for 10 min. After cooling to room temperature, filter;

[0112] (6) Add 0.20% activated carbon based on the mass of the above solution, heat to 60 °C and stir for 30 min. After cooling to room temperature, filter, and then pass through an ultrafiltration membrane with a pore size of 500 Da to obtain camellia polypeptide.

[0113] 3. The preparation method of the composition is as follows:

[0114] (1) Mix acetylcholine, PPG-13-decyltetradeceth-24, and polyol evenly according to the ratio in Table 1;

[0115] (2) Mix palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 evenly according to the ratio in Table 1, and then slowly add component B to component A while stirring to ensure that component A and component B are mixed evenly;

[0116] (3) Mix carnosine, camellia polypeptide, and water evenly according to the ratio in Table 1, then heat component C to 60 °C, and slowly drop the mixture of component A and component B into component C while stirring, and finally obtain the camellia polypeptide composition.

[0117] Comparative Example 1

[0118] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, the supercritical defatting process in Step 2 of Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0119] Comparative Example 2

[0120] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, the 90% ethanol washing process in Step 2 of Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0121] Comparative Example 3

[0122] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, trypsin in the enzymatic hydrolysis process in Step 2 of Example 5 is replaced by three other enzymes (the ratio is their addition ratio), and other camellia polypeptide preparation processes remain unchanged.

[0123] Comparative Example 4

[0124] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, neutral protease in the enzymatic hydrolysis process in Step 2 of Example 5 is replaced by three other enzymes (the ratio is their addition ratio), and other camellia polypeptide preparation processes remain unchanged.

[0125] Comparative Example 5

[0126] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, papain in the enzymatic hydrolysis process in Step 2 of Example 5 is replaced by three other enzymes (the ratio is their addition ratio), and other camellia polypeptide preparation processes remain unchanged.

[0127] Comparative Example 6

[0128] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, alkaline protease in the enzymatic hydrolysis process in Step 2 of Example 5 is replaced by three other enzymes (the ratio is their addition ratio), and other camellia polypeptide preparation processes remain unchanged.

[0129] Comparative Example 7

[0130] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, the enzymatic hydrolysis process in Step 2 of Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0131] Comparative Example 8

[0132] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, the activated carbon adsorption process in Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0133] Comparative Example 9

[0134] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, the ultrafiltration membrane filtration process in Step 2 of Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0135] Comparative Example 10

[0136] This comparative example is a comparative example for the preparation of camellia polypeptide in Example 5. In this comparative example, camellia seed cake (camellia seeds after oil extraction) purchased from the market is used as the raw material, and at the same time, the supercritical deoiling process in Step 2 of Example 5 is cancelled, and other camellia polypeptide preparation processes remain unchanged.

[0137] Comparative Example 11

[0138] This comparative example is a comparative example for Example 5. In this comparative example, the activated carbon adsorption process in Step 2 of Example 5 is cancelled, and other processes remain unchanged; finally, under this process, the polypeptide composition cannot be completely encapsulated and cannot form a clear and transparent homogeneous system, not meeting the requirements.

[0139] Comparative Example 12

[0140] This comparative example is a comparative example for Example 5. In this comparative example, the content of the encapsulating solubilizer remains unchanged, and there is only acetylcholine, and its process remains unchanged.

[0141] Comparative Example 13

[0142] This comparative example is a comparative example for Example 5. In this comparative example, the content of the encapsulating solubilizer remains unchanged, and there is only PPG-13-decyltetradeceth-24, and its process remains unchanged.

[0143] Comparative Example 14

[0144] This comparative example is a comparative example for Example 5. In this comparative example, the content of the encapsulating solubilizer remains unchanged, and there is only polyglyceryl-4 oleate, and its process remains unchanged.

[0145] Comparative Example 15

[0146] This comparative example is a comparative example for Example 5. In this comparative example, the camellia polypeptide in Component C is cancelled and replaced with six other polypeptides of the same concentration, and other conditions remain unchanged.

[0147] Comparative Example 16

[0148] This comparative example is a comparative example for Example 5. In this comparative example, the carnosine in Component C is cancelled and replaced with six other polypeptides of the same concentration, and other conditions remain unchanged.

[0149] Comparative Example 17

[0150] This comparative example is a comparative example for Example 5. In this comparative example, the carnosine and camellia polypeptide in Component C are cancelled and replaced with five other polypeptides of the same concentration, and other conditions remain unchanged.

[0151] Comparative Example 18

[0152] This comparative example is a comparative example of Example 5. In this comparative example, acetyl hexapeptide-8 in component B is cancelled and replaced with six other polypeptides of the same concentration, and the others remain unchanged.

[0153] Comparative Example 19

[0154] This comparative example is a comparative example of Example 5. In this comparative example, nonapeptide-1 in component B is cancelled and replaced with six other polypeptides of the same concentration, and the others remain unchanged.

[0155] Comparative Example 20

[0156] This comparative example is a comparative example of Example 5. In this comparative example, palmitoyl tetrapeptide-7 in component B is cancelled and replaced with six other polypeptides of the same concentration, and the others remain unchanged.

[0157] Comparative Example 21

[0158] This comparative example is a comparative example of Example 5. In this comparative example, palmitoyl tripeptide-5 and palmitoyl pentapeptide-4 in component B are cancelled and replaced with five other polypeptides of the same concentration, and the others remain unchanged.

[0159] Comparative Example 22

[0160] This comparative example is a comparative example of Example 5. In this comparative example, all components in component B are cancelled and replaced with six other polypeptides of the same concentration, and the others remain unchanged.

[0161] Comparative Example 23

[0162] This comparative example is a comparative example of Example 5. In this comparative example, the encapsulating solubilizer in component A is cancelled and replaced with water, and the others remain unchanged.

[0163] Comparative Example 24

[0164] This comparative example is a comparative example of Example 5. In this comparative example, the polyol solvent in component A is cancelled and replaced with water, and the others remain unchanged.

[0165] Comparative Example 25

[0166] This comparative example is a comparative example of Example 5. In this comparative example, the encapsulating solubilizer and polyol in component A are cancelled and replaced with water, and the others remain unchanged.

[0167] Comparative Example 26

[0168] This comparative example is a comparative example of Example 5. In this comparative example, carnosine in component B and component C is cancelled and replaced with camellia polypeptide of the same concentration, and the others remain unchanged.

[0169] Test results of nitrogen content and main peptide sequence in camellia polypeptides of examples and comparative examples 1-3

[0170] Samples of the examples and Comparative Examples 1-3 were tested for their nitrogen content using the Kjeldahl method to calculate the total content of polypeptides therein. At the same time, the amino acid sequences of the prepared camellia polypeptides were identified by HPLC / MS / MS method. The specific detection method is as follows: The samples were separated and detected using EASY-nLC 1200; The chromatographic column was: Reprosil-Pur 120 C18-AQ (75μm×250mm×1.9μm); MS conditions: The full scan range was 100-1500 m / Z, the resolution of the first-stage mass spectrometry was set to 60000, AGC was Custom (AGC is the automatically gain control setting customized by Custom), Maximum IT (ion injection time, the time required to inject ions from the ion source into the mass analyzer) was 50 ms. The resolution of the second-stage mass spectrometry was set to 15000, AGC was Custom, Maximum IT was 22 ms, and the peptide fragmentation collision energy was set to 32 to generate the original mass spectrometry detection data. Finally, the polypeptide sequence was analyzed by the method of PEAKSDenovo.

[0171] Table 2 Test results of nitrogen content and polypeptide sequence 3 of camellia polypeptides in the examples and Comparative Examples 1-10

[0172]

[0173]

[0174]

[0175] Note: The relative content refers to the peak area of the polypeptide / the peak area of all polypeptides * 100%

[0176] Particle size test results of Examples 1-5

[0177] Samples of Examples 1-5 were tested for D10, D50, and D90 values of the sample particle size using a laser particle size analyzer (Bettersize 3000), and then the SPAN value of the sample particle size was calculated. The calculation formula of the SPAN value is as follows:

[0178] SPAN = (D90 - D10) / (D50 - D10)

[0179] The average particle size and particle size distribution width of the samples were evaluated by D50 and SPAN.

[0180] Table 3 Particle size test results of Examples 1-5 and Comparative Examples 12-14

[0181] Sample D50 (nm) SPAN Example 1 75 12.57 Comparative Example 2 65 1.95 Comparative Example 3 25 1.45 Comparative Example 4 35 1.55 Comparative Example 5 45 1.75 Comparative Example 12 110 3.75 Comparative Example 13 35 2.78 Comparative Example 14 48 2.36

[0182] Stability Investigation Results of Examples and Comparative Examples

[0183] Table 3 Stability Investigation Results of Compositions of Examples and Comparative Examples 15 - 26 for 3 Months

[0184]

[0185]

[0186] Antimicrobial Challenge Test of Examples and Comparative Examples: The samples of examples and comparative examples were detected according to the general rules of microbial test methods, total colony count, and test methods for molds and yeasts in Chapter 5 of "Technical Specifications for Cosmetics Safety" (2015 Edition). The test methods and judgment criteria for this antimicrobial challenge experiment referred to USP51 of the United States Pharmacopeia, EP7.0 of the European Pharmacopeia, and CTFA methods. The one - inoculation method was adopted, inoculating on the 0th day, and the results were detected on the 7th, 14th, 21st, and 28th days after inoculation. The test results are shown in Table 3.

[0187] Table 4 Microbial Investigation Results of Compositions of Examples and Comparative Examples 15 - 26

[0188]

[0189]

[0190] Note: The aseptic test standard is bacteria < 10 (cfu / ml); fungi < 10 (cfu / ml).

[0191] Efficacy Tests of Examples and Comparative Examples

[0192] Antioxidant Capacity Test

[0193] The compositions prepared from all the above - mentioned examples and comparative examples were subjected to an antioxidant test for the DPPH free - radical scavenging rate. The test method referred to the group standard T / SHRH 006 - 2018 issued by the Shanghai Daily Chemical Cosmetics Industry Association, "Cosmetics - Free Radical (DPPH) Scavenging Experiment Method". Shanghai Daily Chemical Cosmetics Industry Association, 2018. When the free - radical scavenging rate is 50%, the concentration of the free - radical scavenger is the IC50 value. The IC50 value is a commonly used index to evaluate the effect of free - radical scavengers. The smaller its value, the smaller the concentration dose of the free - radical scavenger used to achieve a 50% free - radical scavenging rate, and the better its free - radical scavenging effect.

[0194] Table 5 Antioxidant Test Results of Compositions of Examples and Comparative Examples 12 - 26

[0195]

[0196]

[0197] Note: " / " indicates no significant relevant efficacy.

[0198] Anti-glycation ability test

[0199] There are two sources of advanced glycation end products: 1. Glycation reactions accompanied by oxidation; 2. Reactions between lipid peroxidation products and proteins. There are many advanced glycation end products, most of which are fluorescent, with an absorption peak at 370 nm and an emission peak at 445 nm. Using this characteristic, advanced glycation end products can be tested by fluorescence method. The test method refers to the literature Anti-Glycation Activities of Phenolic Constituents from Silybum marianum (Milk Thistle) Flower in Vitro and on Human Explants; Isoferulic acid, a new anti-glycation agent, inhibits fructose- and glucose-mediated protein glycation in vitro (Meeprom, Sompong et al. 2013)

[0200] Table 6 Anti-glycation test results of the compositions of Examples and Comparative Examples 12 - 26

[0201]

[0202]

[0203] Repair efficacy test

[0204] Epidermal keratinocytes are important cells that make up the epidermal layer. When the skin surface is damaged, keratinocytes will be stimulated to migrate and repair the damaged part. When the cells grow to a monolayer state in vitro, an artificial blank area is created on the monolayer cells, and the cells at the scratch edge will gradually enter the blank area to heal the scratch, simulating the in vivo cell migration process to a certain extent. By measuring the cell migration rate after sample treatment, it is evaluated whether the test substance has a repair efficacy. The efficacy of the compositions described in all Examples and Comparative Examples is verified by testing the migration rate of the compositions on epidermal keratinocytes.

[0205] Table 7 Cell migration rate test results of the compositions of Examples and Comparative Examples 12 - 26

[0206]

[0207]

[0208] Brightening effect test

[0209] Melanin is a high-molecular biological pigment, which is essentially a kind of protein and usually exists in the form of polymer in animal skin or hair. Even plants and protists have this pigment, and its content and distribution will directly affect the colors of skin, hair and eyes. Melanin is mainly composed of two kinds of quinone polymers, namely eumelanin and pheomelanin. Its biosynthesis is a series of biochemical reaction processes initiated by the hydroxylation of tyrosine in the body catalyzed by tyrosinase. Tyrosinase is the rate-limiting enzyme for melanin production, and its quantity and activity determine the speed and output of melanin production. B16-F10 melanoma cell line is a typical cell line used in the study of cellular melanin production. In this experiment, the relative content of melanin production was measured to evaluate the melanin inhibitory ability of the test samples, that is, the brightening effect. The test method refers to Kim J H, Baek S H, Kim D H, et al. Downregulation of Melanin Synthesis by Haginin A and Its Application to In Vivo Lightening Model[J]. Journal of Investigative Dermatology, 2008, 128(5): 1227-35.

[0210] Table 8 Test results of the inhibitory rate of cellular melanin of the compositions in the examples and comparative examples 12-26

[0211]

[0212]

[0213] Anti-inflammatory effect test

[0214] The above-mentioned all compositions in the examples and comparative examples were used in the experiment of generally clearing TNF-α in macrophages to verify the anti-inflammatory effect of the compositions. The experimental method refers to T / SHRH034—2021, Test method for soothing effect of cosmetics - Determination of the content of TNF-α inflammatory factor in vitro, LPS-induced macrophage RAW264.7 test method. Shanghai Daily Cosmetics Industry Association, 2021.

[0215] Table 9 Test results of the inhibitory rate of TNF-α of the compositions in the examples and comparative examples 12-26

[0216]

[0217]

[0218] Anti-wrinkle effect test

[0219] The anti-wrinkle effects of all the above-mentioned examples and comparative example compositions were verified through an experiment on promoting the synthesis of type I, III, and V collagen in fibroblasts. The experimental method referred to T / SHRH 031—2020, Testing for firming and anti-wrinkle effects of cosmetics - Determination of type I collagen content in vitro fibroblasts. Shanghai Daily Cosmetics Industry Association, 2018; Cansn Güngrmü, Dürdane Kolankaya. Characterization of type I, III and V collagens in high-density cultured tenocytes by triple-immunofluorescence technique[J]. Cytotechnology, 2008, 58(3): 145-152.

[0220] Table 10 Anti-wrinkle test results of the compositions of examples and comparative examples 12 - 26

[0221]

[0222]

[0223] In-vivo immediate anti-wrinkle efficacy test

[0224] All the above-mentioned example and comparative example compositions were diluted with water to 1%, and applied to the crow's feet of the subjects. The volume of the crow's feet was measured with Antera 3D before and after application, and the change rate of the crow's feet volume before and after application was calculated. The test method was as follows: For each group of diluted samples of examples and comparative examples, 5 healthy women aged 30 ± 2 years old were selected. The sample was applied to the crow's feet. The volume of the crow's feet was measured with an instrument before applying the sample and 5 min, 30 min, and 2 h after using the sample. The test was conducted in parallel 3 times, and the average value was taken, and the change rate of the crow's feet volume was calculated.

[0225] Table 10 Immediate anti-wrinkle test results of the compositions of examples and comparative examples 12 - 26

[0226]

[0227]

[0228] Application Example 1

[0229] This application example provides the application and preparation method of the camellia polypeptide composition in a skin care cream formula. The specific formula is as follows:

[0230] Phase A

[0231]

[0232] Phase B

[0233]

[0234]

[0235] Phase C

[0236] SIMULGEL NS 0.8%;

[0237] Phase D

[0238] Camellia polypeptide composition 5.0%;

[0239] Fragrance 0.1%;

[0240] Its preparation method is as follows:

[0241] A. At room temperature, add item A to the water pot in sequence, stir well until evenly dispersed, then heat up to 90 ± 2 °C, stir evenly and pump into the emulsifying pot, homogenize for 5 minutes, and stir evenly;

[0242] B. At room temperature, add item B to the oil pot in sequence, then heat up to 85 ± 2 °C, after complete dissolution, pump into the emulsifying pot.

[0243] C. Homogenize for 10 minutes, add item C, keep warm for 15 minutes, and stir evenly;

[0244] D. At 50 ± 2 °C, add item D and stir evenly.

[0245] Application Example 2

[0246] This application example provides the application and preparation method of the camellia polypeptide composition in the emulsion formula. The specific formula is as follows:

[0247] Phase A

[0248]

[0249] Phase B

[0250] Caprylic / capric triglyceride 5%;

[0251] MONTANOV 68MB 0.5%;

[0252] Phase C

[0253] Tromethamine 0.04%

[0254] Phase D

[0255] SIMULGEL NS 0.3%;

[0256] Phase E

[0257] Polypeptide composition: 5.0%

[0258] Fragrance: 0.06%

[0259] The preparation method is as follows:

[0260] A. At room temperature, add item A to the water pot in sequence, stir well until evenly dispersed, then heat up to 90 ± 2 °C, stir evenly and pump it into the emulsifying pot, homogenize for 5 minutes, and stir evenly;

[0261] B. At room temperature, add item B to the oil pot in sequence, then heat up to 85 ± 2 °C, after complete dissolution, pump it into the emulsifying pot.

[0262] C. Homogenize for 10 minutes, add item C, keep warm for 15 minutes, and stir evenly;

[0263] D. At 50 ± 2 °C, add item D and stir evenly.

[0264] Application Example 3

[0265] This application example provides the application and preparation method of camellia polypeptide composition in the aqueous formulation. The specific formulation is as follows:

[0266] Phase A

[0267]

[0268] Phase B

[0269] Tromethamine: 0.01%

[0270] Phase C

[0271] Polypeptide composition: 5.0%

[0272] Phase D

[0273] Fragrance solubilizer: 0.05%

[0274] Fragrance: 0.01%

[0275] A. At room temperature, add item A to the water pot in sequence, stir well until evenly dispersed, then heat up to 90 ± 2 °C, stir evenly and pump it into the emulsifying pot, homogenize for 5 minutes, and stir evenly;

[0276] B. At room temperature, add item B to the oil pot in sequence, then heat up to 85 ± 2 °C, after complete dissolution, pump it into the emulsifying pot.

[0277] C. Homogenize for 10 minutes, add item C, keep warm for 15 minutes, and stir evenly;

[0278] D. At 50 ± 2 °C, add item D and stir evenly.

[0279] The specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the above specific embodiments, and those skilled in the art can make various changes or modifications within the scope of the claims, which does not affect the essence of the present invention. Without conflict, the embodiments of the present application and the features in the embodiments can be combined with each other arbitrarily.

Claims

1. A camellia polypeptide composition with anti-wrinkle, repair and brightening effects, characterized in that: Calculated by mass percentage, it includes the following components:

2. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 1, characterized in that: Encapsulation solubilizers include penetration encapsulation solubilizers and particle size encapsulation solubilizers; The osmotic encapsulation solubilizer is acetylcholine; The particle size encapsulating solubilizer is one of PPG-13-decyltetradecyl alcohol polyether-24 and polyglycerol-4 oleate or a combination of two thereof.

3. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 1, characterized in that: The polyol includes one or more of 1.3-propylene glycol, 1.3-butylene glycol, 1.2-pentanediol, and 1.2-hexanediol.

4. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 1, characterized in that: The camellia polypeptide is prepared by the following method: A1. Processing camellia seed powder with supercritical CO2 extraction to remove oil therein and obtain de-oiled camellia seed cake; A2, adding de-oiled camellia seed cake to deionized water, adjusting the pH to 8.50-10.00, then heating and stirring for extraction, and filtering to obtain camellia seed protein extract; A3, adjusting the pH of the camellia seed protein extract to 3.0-4.0, and then filtering to obtain camellia seed protein 1; A4, washing the camellia seed protein 1 with an ethanol solution to remove camellia saponin in the protein, and then drying to remove ethanol to obtain the camellia seed protein 2; A5, adding camellia seed protein 2 to deionized water, and then respectively adding papain, alkaline protease, neutral protease and trypsin to obtain a mixed solution, heating and stirring for enzymolysis, heating and keeping warm again with stirring, and filtering to obtain a camellia seed protein filtrate; A6. Add activated carbon to the camellia seed protein filtrate, heat and stir, and filter to obtain the camellia polypeptide.

5. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 4, characterized in that: In step A4, the mass fraction of the ethanol solution is above 80%; And / or, the amount of ethanol solution used is 10-20 times the mass of camellia seed protein, and after washing, the ethanol is removed by drying.

6. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 4, characterized in that: The dosages of papain, alkaline protease, neutral protease and trypsin are 0.5-2%, 0.1-1%, 0.1-1% and 0.3-2% of the mass of the de-oiled camellia seed cake in step A1, respectively.

7. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 4, characterized in that: In step A5, the temperature of the heated and stirred enzymatic hydrolysis is 55-65° C. and the time is 1-3 h; And / or, the temperature of heating and keeping again is 80-100° C. and the time is 5-20 min.

8. The camellia polypeptide composition with anti-wrinkle, repairing and brightening effects according to claim 4, characterized in that: In step A6, the amount of activated carbon used is 0.1-0.3% of the mass of the camellia seed protein filtrate; And / or, the heating and stirring temperature is 55-65° C. and the time is 20-40 min.

9. A method for preparing the camellia polypeptide composition according to claim 1, characterized in that: The steps include: S1. Preparation of component A: Stir and mix the encapsulating solubilizer and polyol in a mass ratio to make them uniform; S2, preparation of component B: palmitoyl tripeptide-5, palmitoyl pentapeptide-4, palmitoyl tetrapeptide-7, acetyl hexapeptide-8, and nonapeptide-1 were stirred and mixed evenly according to the mass ratio; S3, preparation of component C: stir and mix carnosine, camellia polypeptide and water in a mass ratio; S4. Preparation of the composition: slowly add component B to component A, stirring while adding to ensure that component A and component B are evenly mixed; Then, component C is heated to 50-70° C., and then the mixture of component A and component B is slowly dripped into component C while stirring, to finally obtain a camellia polypeptide composition.

10. Use of the camellia polypeptide composition according to claim 1 in preparing cosmetics.

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