A composition containing recombinant collagen and its application in freeze-dried facial mask

Abstract

The present invention relates to a composition containing recombinant collagen and its application in a freeze-dried facial mask, wherein the composition containing recombinant collagen comprises recombinant collagen, schizophyllan and plant extracts; the plant extracts comprise a combination of Haematococcus pluvialis extract, Portulaca oleracea extract and Lycium ruthenicum extract. The present invention creatively finds that the three plant extract components of Haematococcus pluvialis extract, Portulaca oleracea extract and Lycium ruthenicum extract are used in combination with recombinant collagen and schizophyllan, and can interact with each other, and have a synergistic promoting effect in moisturizing, firming and anti-oxidation effects that is difficult for technicians in this field to predict; and there is also a significant synergistic enhancing effect between the three plant extract components. The composition containing recombinant collagen provided by the present invention is very suitable for use in skin care products, is mild and non-irritating, and has good stability.

Description

Technical Field

[0001] The invention belongs to the field of cosmetics and medical dressings, and particularly relates to a composition containing recombinant collagen and application of the composition in a freeze-dried facial mask. Background Art

[0002] The skin is composed of the epidermis, dermis and subcutaneous tissue. The epidermis is the part that is in direct contact with the outside world. It consists of the stratum corneum, the transparent layer, the granular layer (containing transparent keratin particles), the spinous cell layer and the basal layer. The first three layers are the key to keeping the skin hydrated. The stratum corneum not only prevents the dissipation of water, but also absorbs water from the external environment; the transparent layer and the granular layer together form a waterproof barrier, which makes it difficult for water in the skin to penetrate from the outside, and also prevents the water below the stratum corneum from penetrating into the stratum corneum. There is a sebum membrane with a thickness of about 5nm on the surface of the stratum corneum, which is formed by the sebum secreted by the sebaceous glands, the water in the sweat and the skin metabolites. The multiple structures maintain the integrity of the skin barrier and prevent the loss of water through the epidermis. The root causes of many skin problems are closely related to insufficient skin moisture and poor moisturizing and water locking, such as sagging skin, sensitive redness, spots, dull skin color and clogged pores caused by water-oil imbalance. Therefore, it is of great significance to provide a gentle and effective skin care product to replenish skin moisture and relieve skin problems.

[0003] As an important category of skin care products, facial masks can be used to hydrate the skin, and also have multiple functions such as moisturizing, nourishing, nourishing and deep cleansing. The working principle of facial masks is to use the short time of covering the face to temporarily isolate the outside air and pollution, increase the skin temperature, expand the skin pores, promote sweat gland secretion and metabolism, increase the oxygen content of the skin, and help the skin to eliminate the products of epidermal cell metabolism and accumulated oil substances. The moisture in the mask penetrates into the stratum corneum and increases the skin's moisture content. However, the rich nutrients contained in the mask material are easy to deteriorate, so preservatives are generally needed to avoid the risk of shopping in Weihai City. For sensitive skin, the use of preservatives may cause skin allergies, tingling and other adverse effects.

[0004] Freeze-dried mask is a type of facial mask. It refers to the fusion of facial mask essence and mask cloth, using vacuum freeze-drying technology to remove the moisture in the essence and form a solid dry film. Freeze-drying technology is one of the best methods to damage tissues and components that are sensitive to biological characteristics. The biggest feature of freeze-dried mask is that it can maintain the activity of raw materials while preserving easily oxidized and volatile components. After being dissolved in water or other solvents, it can be fully and quickly released, and there is no need to add preservatives, flavors and other additives. It is more gentle and non-irritating. Therefore, how to develop a moisturizing and antioxidant skin care product that is mild and has good efficacy has become a problem that needs to be solved urgently. Summary of the invention

[0005] In view of the deficiencies of the prior art, the object of the present invention is to provide a composition containing recombinant collagen and its application in a freeze-dried facial mask.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a composition containing recombinant collagen, wherein the composition containing recombinant collagen comprises recombinant collagen, Schizophyllan polysaccharide and a plant extract;

[0008] The plant extract comprises a combination of a Haematococcus pluvialis extract, a Portulaca oleracea extract and a Lycium nigra extract.

[0009] In the present invention, recombinant collagen, Schizophyllan polysaccharide and plant extract components are used in combination, and it is found that the recombinant collagen, Schizophyllan polysaccharide and plant extract can cooperate with each other and have better effects in moisturizing and anti-oxidation.

[0010] Preferably, the Schizophyllan is octenylsuccinic acid-modified Schizophyllan.

[0011] Preferably, the method for preparing the octenylsuccinic acid modified Schizophyllan comprises: adding octenylsuccinic anhydride to a Schizophyllan solution for esterification reaction to obtain the modified Schizophyllan.

[0012] Collagen can maintain skin elasticity and promote cell growth. It can be used in skin care products to have moisturizing, repairing and anti-aging effects. Recombinant collagen is collagen prepared using DNA recombination technology. It can be directly absorbed by the human body and participate in the construction of collagen. It has the effect of promoting skin tissue regeneration and repairing damaged skin. Octenylsuccinic acid modified Schizophyllum polysaccharide is an amphiphilic polysaccharide that uses octenylsuccinic acid octenylsuccinic anhydride to undergo esterification reaction with Schizophyllum polysaccharide to introduce hydrophobic groups. The modified amphiphilic polysaccharide can self-assemble into micelles, particles or aggregates through intramolecular or intermolecular hydrophobic interactions, thereby improving its loading capacity and emulsification capacity for hydrophobic substances. The two are used in combination with natural plant extract components to protect and stabilize some highly bioactive components in the plant extract components, and exert excellent moisturizing and antioxidant effects.

[0013] Among the plant extract ingredients, Haematococcus pluvialis extract is a single-cell microalgae. The algae can accumulate astaxanthin in large quantities and appear red, so it is named Haematococcus pluvialis. The astaxanthin in Haematococcus pluvialis extract is pure L-astaxanthin, which is a fat-soluble component with a strong antioxidant effect and helps to delay skin aging; Portulaca oleracea extract is a mixture of active ingredients extracted from the whole plant of Portulaca oleracea, mainly including flavonoids, saponins, polysaccharides, etc., with excellent antioxidant, antibacterial, whitening, moisturizing and other effects; the anthocyanin component rich in black wolfberry has multiple effects such as antioxidant, anti-inflammatory, neuroprotective, anti-obesity and antibacterial.

[0014] In the present invention, the above three specific plant extracts are used in combination, and it is found that they have significant synergistic effects in moisturizing and anti-oxidation.

[0015] Preferably, the composition containing recombinant collagen comprises, by weight, 0.2-5 parts of recombinant collagen, 0.01-1 parts of Schizophyllan and 0.01-1.5 parts of plant extract;

[0016] The above “0.2-5 parts” can be, for example, 0.2 parts, 0.5 parts, 0.8 parts, 1 parts, 1.2 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, 5 parts, etc.;

[0017] The above-mentioned “0.01-1 part” can be, for example, 0.01 part, 0.05 part, 0.1 part, 0.15 part, 0.2 part, 0.3 part, 0.5 part, 0.7 part, 0.8 part, 0.9 part, 1 part, etc.;

[0018] The above-mentioned "0.01-1.5 parts" can be, for example, 0.01 parts, 0.05 parts, 0.08 parts, 0.1 parts, 0.15 parts, 0.2 parts, 0.25 parts, 0.3 parts, 0.4 parts, 0.5 parts, 0.6 parts, 0.7 parts, 0.8 parts, 1 parts, 1.2 parts, 1.5 parts, etc. Other specific point values ​​within the numerical range can be selected, which will not be elaborated here one by one.

[0019] In the present invention, when the dosage ratio of the recombinant collagen, Schizophyllan and the plant extract is within the above specific range, it has a better moisturizing and anti-oxidation effect.

[0020] Preferably, the molecular weight of the recombinant collagen is 5-20kDa, for example, it can be 5kDa, 6kDa, 7kDa, 8kDa, 9kDa, 10kDa, 12kDa, 14kDa, 15kDa, 16kDa, 18kDa, 20kDa, etc. Other specific point values ​​within this numerical range can be selected, which will not be described here one by one.

[0021] Preferably, the recombinant collagen includes any one of type I recombinant collagen, type II recombinant collagen and type III recombinant collagen.

[0022] Preferably, the amino acid sequence of the recombinant collagen includes amino acids 950-1075 of type I collagen and amino acids 594-729 and 1193-1199 of type III collagen.

[0023] Preferably, the molecular weight of the schizophyllan polysaccharide is 10-100kDa, for example, it can be 10kDa, 15kDa, 20kDa, 25kDa, 30kDa, 40kDa, 50kDa, 60kDa, 70kDa, 80kDa, 90kDa, 100kDa, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described one by one here.

[0024] Preferably, the mass ratio of the Haematococcus pluvialis extract, the Portulaca oleracea extract and the Lycium ruthenicum extract is (0.05-3):(0.1-5):(0.1-5);

[0025] The above “0.05-3” may be, for example, 0.05, 0.1, 0.15, 0.2, 0.5, 0.8, 1, 1.2, 1.5, 1.8, 2, 2.5, 3, etc.;

[0026] The above-mentioned "0.1-5" can be, for example, 0.1, 0.2, 0.5, 0.8, 1, 1.2, 1.5, 1.8, 2, 2.5, 3, 3.5, 4, 5, etc. Other specific point values ​​within this numerical range can be selected, and they will not be elaborated here one by one.

[0027] In the present invention, when the dosage ratio of each component in the plant extract is within the above specific range, it has a better moisturizing and antioxidant effect.

[0028] The Haematococcus pluvialis extract involved in the present invention is not limited to its specific preparation process and source, and conventional extraction and preparation processes and regular commercially available products can be used in the scheme of the present invention.

[0029] The purslane extract involved in the present invention is not limited to its specific preparation process. When it is prepared by the following process, its synergistic effect with the other two components in moisturizing and anti-oxidation is more significant.

[0030] Preferably, the purslane extract is prepared by a preparation method comprising the following steps:

[0031] Mixing purslane, complex enzyme and water, performing ultrasonication, filtering to obtain a filtrate, and concentrating and drying the filtrate to obtain the purslane extract;

[0032] The complex enzyme comprises a combination of cellulase, amylase and bromelain.

[0033] In the present invention, it is unexpectedly found that when a composite enzyme solution containing cellulase, amylase and bromelain is used to perform enzymatic hydrolysis and extraction of active substances in purslane, the combination of the three enzymes can synergistically enhance the effect and obtain a better extraction effect. The prepared purslane extract has higher activity and better effect. In addition, the enzymatic hydrolysis is carried out under ultrasonic conditions, which can further enhance the enzymatic hydrolysis effect and obtain a purslane extract with better effect.

[0034] Preferably, the mass ratio of the cellulase, amylase and bromelain is (1-6):(2-7):(0.1-2);

[0035] The above “1-6” may be, for example, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, etc.;

[0036] The above-mentioned “2-7” may be, for example, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, etc.;

[0037] The above-mentioned "0.1-2" can be, for example, 0.1, 0.2, 0.4, 0.5, 0.6, 0.8, 1, 1.2, 1.4, 1.5, 1.6, 1.8, 2, etc. Other specific point values ​​within this numerical range can be selected, and they will not be elaborated here one by one.

[0038] In the present invention, when the dosage ratio of the three enzymes in the composite enzyme is within the above specific range, the enzymatic extraction effect of Purslane can be further improved.

[0039] Preferably, the addition amount of the complex enzyme is 0.05‰-2‰ of the mass of Purslane, for example, it can be 0.05‰, 0.1‰, 0.15‰, 0.2‰, 0.25‰, 0.3‰, 0.5‰, 0.6‰, 0.7‰, 0.8‰, 0.9‰, 1‰, 1.2‰, 1.5‰, 1.8‰, 2‰, etc. Other specific point values ​​within this numerical range can be selected, which will not be described one by one here.

[0040] Preferably, the liquid ratio of purslane to water is 1g:(30-80)mL, for example, it can be 1g:30mL, 1g:35mL, 1g:40mL, 1g:45mL, 1g:50mL, 1g:55mL, 1g:60mL, 1g:65mL, 1g:70mL, 1g:72mL, 1g:75mL, 1g:80mL, etc. Other specific point values ​​within this numerical range can be selected, which will not be described here one by one.

[0041] Preferably, the ultrasonic power is 200-400 W, for example 200 W, 250 W, 300 W, 350 W, 400 W, etc., and the enzymatic hydrolysis time is 5-30 min, for example 5 min, 7 min, 10 min, 12 min, 15 min, 20 min, 22 min, 25 min, 30 min, etc. Other specific point values ​​within the numerical range can be selected, which will not be described here one by one.

[0042] The black wolfberry extract involved in the present invention is not limited to its specific preparation process. When it is prepared by the following process, its synergistic effect with the other two components in terms of moisturizing and anti-oxidation is more significant.

[0043] Preferably, the black wolfberry extract is prepared by a preparation method comprising the following steps:

[0044] The black wolfberry is subjected to supercritical CO2 extraction, gasified CO2 and anhydrous ethanol are filled into the extractor, and the obtained extract is concentrated and dried to obtain the black wolfberry extract.

[0045] When supercritical CO2 extraction is used to extract the black wolfberry raw material, the extraction effect can be further improved. Compared with the conventional alcohol extraction method, the nutrients and bioactive substances in the black wolfberry can be extracted more efficiently, and the obtained extract has a better effect.

[0046] Preferably, the temperature of the supercritical CO2 extraction is 10-40°C, for example, it can be 10°C, 15°C, 18°C, 20°C, 22°C, 25°C, 28°C, 30°C, 32°C, 35°C, 38°C, 40°C, etc.; the extraction pressure is 8-30MPa, for example, it can be 8MPa, 9MPa, 10MPa, 12MPa, 15MPa, 18MPa, 20MPa, 22MPa, 25MPa, 28MPa, 30MPa, etc.; the extraction time is 1-5h, for example, it can be 1h, 1.2h, 1.5h, 1.8h, 2h, 2.2h, 2.5h, 2.8h, 3h, 3.2h, 3.5h, 3.8h, 4h, 4.2h, 4.5h, 4.8h, 5h, etc., and other specific point values ​​within the numerical range can be selected, which will not be repeated here.

[0047] In a second aspect, the present invention provides a use of the composition containing recombinant collagen according to the first aspect in cosmetics;

[0048] The cosmetics include lotions, creams, skin care water, skin care essences or facial masks.

[0049] In a third aspect, the present invention provides a freeze-dried facial mask, wherein the raw materials for preparing the freeze-dried facial mask include, by mass percentage, 1%-10% of the composition containing recombinant collagen described in the first aspect, for example, it can be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, etc. Other specific point values ​​within the numerical range can be selected and will not be described one by one here.

[0050] Preferably, the raw materials for preparing the freeze-dried facial mask further include any one or a combination of at least two of a moisturizer, an antioxidant, a skin conditioner, a thickener, a stabilizer, a chelating agent or water.

[0051] Preferably, the moisturizing agent includes any one of sodium hyaluronate, trehalose, glycerin, butylene glycol, hydroxyethyl urea or betaine, or a combination of at least two thereof.

[0052] Preferably, the mass percentage of the moisturizing agent in the freeze-dried facial mask is 0.1%-15%, for example, it can be 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15%, etc. Other specific point values ​​within this numerical range can be selected, which will not be described here one by one.

[0053] Compared with the prior art, the present invention has the following beneficial effects:

[0054] In the present invention, recombinant collagen is combined with Schizophyllum polysaccharide to form a complex, which not only improves the stability of the application of recombinant collagen, but also can further improve the stability of the active substances in the plant extract components in skin care products; and further use three specific plant extract components of Haematococcus pluvialis extract, Portulaca oleracea extract, and Lycium ruthenicum extract, and it is found that the three components can interact with each other and positively influence each other, and have a synergistic promotion effect in terms of moisturizing and anti-oxidation effects that is difficult for technicians in this field to predict, which can further enhance the moisturizing and antioxidant effects of the product. DETAILED DESCRIPTION

[0055] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0056] The recombinant collagen involved in the following was purchased from Best (Guangzhou) Co., Ltd. under the model of HydroCol-y recombinant collagen; the schizophyllan polysaccharide was obtained from Best Pharmaceutical Company under the model of SPG is a schizophyllan polysaccharide product modified with octenylsuccinate; the Haematococcus pluvialis extract comes from Zhongran Company's product model Astabio AW1, and the key active ingredient is astaxanthin; cellulase, amylase and bromelain are all purchased from Shanghai MacLean Biochemical Technology Co., Ltd., with product models C758963, A800731, and B832358 respectively; the remaining raw materials can be used as long as they are purchased from regular dealers.

[0057] The weight percentages of the Haematococcus pluvialis extracts mentioned below are all calculated based on the actual content of the effective ingredients contained in the commercially available raw materials. The commercially available raw materials may also selectively contain any one or a combination of at least two of solvents, moisturizers, thickeners, fillers, diluents, stabilizers, pH adjusters, antibacterial agents, antioxidants or impurities within the permitted range.

[0058] Preparation Example 1

[0059] This preparation example provides a purslane extract, which is prepared by the following method:

[0060] (1) After freeze-drying and grinding purslane, the compound enzyme and water are mixed, the amount of compound enzyme added is 1‰ of the mass of purslane, the solid-liquid ratio of purslane to water is 1 g:50 mL, wherein the compound enzyme is composed of cellulase, amylase and bromelain in a mass ratio of 3:5:1, and ultrasonic enzymatic hydrolysis and extraction are performed at a constant temperature of 55° C., wherein the ultrasonic condition is 380 W ultrasonic for 10 min;

[0061] (2) After the ultrasonic extract is filtered, the filtrate is collected, the filter residue is mixed with water, the ultrasonic extraction process in step (1) and the filtration step in step (2) are repeated once, the combined filtrate is collected, and freeze-dried to obtain a purslane extract.

[0062] This preparation example also provides a black wolfberry extract, which is prepared by the following method:

[0063] The black wolfberry is freeze-dried and crushed, then added into a supercritical CO2 fluid extraction device, the extractor is filled with gasified CO2 and anhydrous ethanol, the extraction temperature is 25°C, the pressure is 18MPa, the time is 2.5h, and the extract is concentrated and dried to obtain the black wolfberry extract.

[0064] Preparation Example 2

[0065] This preparation example provides a purslane extract, which is prepared by the following method:

[0066] (1) After freeze-drying and grinding purslane, the compound enzyme and water are mixed, wherein the amount of the compound enzyme added is 0.2‰ of the mass of purslane, the solid-liquid ratio of purslane to water is 1 g:35 mL, wherein the compound enzyme is composed of cellulase, amylase and bromelain in a mass ratio of 1.5:6:0.2, and ultrasonic enzymatic hydrolysis and extraction are performed at a constant temperature of 60°C, wherein the ultrasonic condition is 300W ultrasonic for 20 min;

[0067] (2) After the ultrasonic extract is filtered, the filtrate is collected, the filter residue is mixed with water, the ultrasonic extraction process in step (1) and the filtration step in step (2) are repeated once, the combined filtrate is collected, and freeze-dried to obtain a purslane extract.

[0068] This preparation example also provides a black wolfberry extract, which is prepared by the following method:

[0069] The black wolfberry is freeze-dried and crushed, then added into a supercritical CO2 fluid extraction device, the extractor is filled with gasified CO2 and anhydrous ethanol, the extraction temperature is 15°C, the pressure is 10MPa, the time is 4.5h, and the extract is concentrated and dried to obtain the black wolfberry extract.

[0070] Preparation Example 3

[0071] This preparation example provides a purslane extract, which is prepared by the following method:

[0072] (1) After freeze-drying and grinding purslane, the compound enzyme and water are mixed, wherein the amount of the compound enzyme added is 1.5‰ of the mass of purslane, the solid-liquid ratio of purslane to water is 1 g:70 mL, wherein the compound enzyme is composed of cellulase, amylase and bromelain in a mass ratio of 5:2:2, and ultrasonic enzymatic hydrolysis and extraction are performed at a constant temperature of 45° C., wherein the ultrasonic condition is 240 W ultrasonic for 25 min;

[0073] (2) After the ultrasonic extract is filtered, the filtrate is collected, the filter residue is mixed with water, the ultrasonic extraction process in step (1) and the filtration step in step (2) are repeated once, the combined filtrate is collected, and freeze-dried to obtain a purslane extract.

[0074] This preparation example also provides a black wolfberry extract, which is prepared by the following method:

[0075] The black wolfberry is freeze-dried and crushed, then added into a supercritical CO2 fluid extraction device, the extractor is filled with gasified CO2 and anhydrous ethanol, the extraction temperature is 32°C, the pressure is 25MPa, the time is 1.5h, and the extract is concentrated and dried to obtain the black wolfberry extract.

[0076] Preparation Example 4

[0077] This preparation example provides a purslane extract, which uses the freeze-dried and ground purslane of Preparation Example 1. The preparation method thereof is different from that of Preparation Example 1 only in the enzyme composition in the complex enzyme, that is, cellulase is not contained, and the reduced amount thereof is proportionally distributed to amylase and bromelain. Other processing processes and related parameters are referred to Preparation Example 1.

[0078] Preparation Example 5

[0079] This preparation example provides a purslane extract, which uses the freeze-dried and ground purslane of Preparation Example 1. The preparation method thereof is different from that of Preparation Example 1 only in the enzyme composition in the complex enzyme, that is, amylase is not included, and the reduced amount thereof is proportionally distributed to cellulase and bromelain. Other processing processes and related parameters are referred to Preparation Example 1.

[0080] Preparation Example 6

[0081] This preparation example provides a purslane extract, which uses the freeze-dried and ground purslane of Preparation Example 1. The preparation method thereof is different from that of Preparation Example 1 only in the enzyme composition in the composite enzyme, that is, bromelain is not contained, and the reduced amount thereof is proportionally distributed to cellulase and amylase. Other processing processes and related parameters are referred to Preparation Example 1.

[0082] Preparation Example 7

[0083] This preparation example provides a black wolfberry extract, which is prepared by the following method:

[0084] The black wolfberry was freeze-dried and crushed, and then mixed with a 70% ethanol aqueous solution at a solid-liquid ratio of 1 g:20 mL, heated under reflux at 80° C. for 2 h, the extract was filtered through three layers of gauze, the filtrate was concentrated and freeze-dried to obtain the black wolfberry extract.

[0085] Example 1

[0086] The present invention provides a composition, which consists of 0.5 parts of recombinant collagen, 1 part of schizophyllan polysaccharide and 1.5 parts of plant extracts in parts by mass, wherein the plant extracts are Haematococcus pluvialis extract, Portulaca oleracea extract (Preparation Example 1) and Lycium ruthenicum extract (Preparation Example 1) in a mass ratio of 2:2.5:2.

[0087] Example 2

[0088] The present invention provides a composition, which consists of 4 parts of recombinant collagen, 0.1 parts of schizophyllan polysaccharide and 0.2 parts of plant extract in parts by mass, wherein the plant extract is a Haematococcus pluvialis extract, a Portulaca oleracea extract (Preparation Example 2) and a Lycium ruthenicum extract (Preparation Example 2) in a mass ratio of 0.5:1:5.

[0089] Example 3

[0090] The present invention provides a composition, which consists of 2 parts of recombinant collagen, 0.5 parts of schizophyllan polysaccharide and 0.8 parts of plant extract in parts by mass, wherein the plant extract is a Haematococcus pluvialis extract, a Portulaca oleracea extract (Preparation Example 3) and a Lycium ruthenicum extract (Preparation Example 3) in a mass ratio of 2.5:3:1.

[0091] Example 4

[0092] This example provides a composition, which is different from Example 1 only in that the Portulaca oleracea extract (Preparation Example 1) is replaced with an equal amount of Portulaca oleracea extract (Preparation Example 4).

[0093] Example 5

[0094] This example provides a composition, which is different from Example 1 only in that the Portulaca oleracea extract (Preparation Example 1) is replaced with an equal amount of Portulaca oleracea extract (Preparation Example 5).

[0095] Example 6

[0096] This example provides a composition, which is different from Example 1 only in that the Portulaca oleracea extract (Preparation Example 1) is replaced with an equal amount of Portulaca oleracea extract (Preparation Example 6).

[0097] Example 7

[0098] This example provides a composition, which differs from Example 1 only in that the Lycium ruthenicum extract (Preparation Example 1) is replaced with an equal amount of Lycium ruthenicum extract (Preparation Example 7).

[0099] Comparative Example 1

[0100] This comparative example provides a composition, which differs from Example 1 only in that it does not contain the Haematococcus pluvialis extract, and the reduced amount of the extract is proportionally distributed to the two components of the Portulaca oleracea extract and the Lycium ruthenicum extract.

[0101] Comparative Example 2

[0102] This comparative example provides a composition, which differs from Example 1 only in that it does not contain the purslane extract, and the reduced amount of the purslane extract is proportionally distributed to the two components of the Haematococcus pluvialis extract and the Lycium ruthenicum extract.

[0103] Comparative Example 3

[0104] This comparative example provides a composition, which differs from Example 1 only in that it does not contain the black fruit wolfberry extract, and the reduced amount of the extract is proportionally distributed to the two components of the Haematococcus pluvialis extract and the Portulaca oleracea extract.

[0105] Comparative Example 4

[0106] This comparative example provides a composition, which differs from Example 1 only in that it does not contain recombinant collagen, and its reduced amount is proportionally distributed to Schizophyllan and the three plant extract components.

[0107] Comparative Example 5

[0108] This comparative example provides a composition, which differs from Example 1 only in that it does not contain Schizophyllan and its reduced amount is proportionally distributed to the recombinant collagen and the three plant extract components.

[0109] Comparative Example 6

[0110] This comparative example provides a composition, which differs from Example 1 only in that it does not contain plant extracts, and the reduced amount thereof is proportionally distributed to the recombinant collagen and Schizophyllan.

[0111] Application Example 1

[0112] This application example provides a freeze-dried facial mask, the formula raw materials of which are (mass percentage): 0.7% of the composition (Example 1), 0.1% of sodium hyaluronate, 0.2% of trehalose and 99% of water.

[0113] The preparation method is as follows:

[0114] S1: Mix sodium hyaluronate, trehalose and part of water, stir thoroughly at 800°C for 40 min to obtain phase A;

[0115] S2: Mix Schizophyllum polysaccharide and part of the water, stir thoroughly at 80°C for 30 minutes, cool to 65°C, add Portulaca oleracea extract, Lycium ruthenicum extract, and Haematococcus pluvialis extract in sequence, and stir thoroughly for 15 minutes;

[0116] S3: After the stirred liquid in S2 is cooled to 45°C, recombinant collagen is added and stirred for 20 minutes to obtain phase B.

[0117] S4: Add phase B to phase A and stir thoroughly for 20 minutes to obtain the freeze-dried mask stock solution;

[0118] S5: Stir the freeze-dried mask stock solution at a rate of 15000 rpm for 20 min to generate small and uniform bubbles inside the mixed solution;

[0119] S6: Filling the stirred mixed solution into a facial mask mold and freeze-drying it to obtain a freeze-dried facial mask.

[0120] Application Example 2-7

[0121] This application example provides six freeze-dried facial masks, the formulas of which differ from those of the application example only in that the composition prepared in Example 1 is replaced with equal amounts of the compositions prepared in Examples 2-7, respectively, and other ingredients and contents remain unchanged. The preparation method refers to that in Application Example 1.

[0122] Comparative Application Examples 1-6

[0123] This comparative application example provides six freeze-dried facial masks, the formulas of which differ from those of the application examples only in that the composition prepared in Example 1 is replaced with equal amounts of the compositions prepared in Comparative Examples 1-6, respectively, and the other ingredients and contents remain unchanged. The preparation method refers to that in Application Example 1.

[0124] Test Example 1

[0125] In vitro antioxidant activity test

[0126] (1) DPPH-free radical scavenging

[0127] A 96-well plate was used, with three replicates in each group, and the system was 200 μL. Sample group: Take the composition samples prepared in Examples 1-7 and Comparative Examples 1-6, mix and dilute them to 5% with 100 μL of pure water, and add 100 μL of 0.1 mM DPPH solution to the reaction system; Control group: Take 100 μL of pure water, and then add 100 μL of 0.1 mM DPPH solution. After the reaction system is constructed, shake it in the dark for 10 minutes, and use an enzyme reader to test the absorbance at 517 nm.

[0128] The calculation method of clearance rate is as follows: Clearance rate (%) = [(A0-Ax) / A0] × 100%, A0 is the absorbance of the control group, and Ax is the absorbance of the sample group.

[0129] (2) ABTS-free radical scavenging

[0130] Preparation of ABTS working solution: Mix ABTS and potassium persulfate solution to prepare 7 mM ABTS stock solution, store in dark for 12 h, shake well and mix with 10 mM PBS (pH=7.4) so ​​that the absorbance of the solution at 734 nm is 0.8, to obtain ABTS working solution.

[0131] A 96-well plate was used, with three replicates in each group, and the system was 200 μL. Sample group: Take the extract samples prepared in Examples 1-7 and Comparative Examples 1-6, mix and dilute to 5% with 10 μL of pure water, and add 190 μL of ABTS working solution to the reaction system; Control group: Take 10 μL of pure water, and then add 190 μL of ABTS working solution. After the reaction system is constructed, shake well and incubate for 6 minutes, and test the absorbance at 734 nm with an enzyme marker.

[0132] The calculation method of clearance rate is as follows: Clearance rate (%) = [(A0-Ax) / A0] × 100%, A0 is the absorbance of the control group, and Ax is the absorbance of the sample group.

[0133] The test results are shown in Table 1:

[0134] Table 1

[0135] DPPH-free radical scavenging rate ABTS-free radical scavenging rate Example 1 99.7％ 99.3％ Example 2 97.3％ 98.7％ Example 3 97.6％ 98.4％ Example 4 85.3％ 80.9％ Example 5 83.7％ 80.5％ Example 6 86.2％ 81.0％ Example 7 80.6％ 79.4％ Comparative Example 1 60.9％ 55.7％ Comparative Example 2 75.4％ 70.9％ Comparative Example 3 65.0％ 60.2％ Comparative Example 4 78.8％ 79.9％ Comparative Example 5 70.6％ 66.7％ Comparative Example 6 12.6％ 10.3％

[0136] The ability of the product to scavenge DPPH and ABTS free radicals can reflect the antioxidant capacity of the substance to a certain extent. The higher the free radical scavenging rate, the stronger the antioxidant capacity. Therefore, the antioxidant effect of the composition can be judged by testing its ability to scavenge free radicals.

[0137] It can be seen from the data in Table 1 that the scavenging rates of DPPH-radicals and ABTS-radicals of Examples 1-3 are higher than those of Comparative Examples 4-6 as a whole, indicating that when the composition lacks any one of the recombinant collagen, Schizophyllum polysaccharide and plant extract, the scavenging effect of the DPPH-radicals and ABTS-radicals of the composition will decrease; the scavenging rates of DPPH-radicals and ABTS-radicals of Example 1 are higher than those of Comparative Examples 1-3, indicating that the three plant extract components of Haematococcus pluvialis extract, Portulaca oleracea extract and Lycium ruthenicum extract also have a certain synergistic effect in the scavenging effect of the two free radicals.

[0138] Test Example 2

[0139] Moisturizing and anti-oxidation comprehensive effect test

[0140] (1) A total of 130 healthy Chinese female subjects with dull, yellowish, and uneven facial skin were selected. Their ages ranged from 25 to 60 years old, with an average age of 46.3±5.6 years old.

[0141] The test day was Sunday, recorded as Day 0. After cleansing and using non-functional skin care water at night, the subjects applied the fully moistened and unfolded freeze-dried mask on their faces to make it fit the skin completely. Be careful not to leave bubbles or incompletely fitted areas on the mask. After 15 minutes, the mask was removed and gently massaged to help the mask essence absorb. The subjects then sat quietly for 30 minutes before the test. The test lasted for four weeks, with freeze-dried mask products used once every Wednesday and Sunday night. On Sunday night of the fourth week, after using the freeze-dried mask product and massaging it for absorption, the subjects sat quietly for 30 minutes before the test again. The cleansing and skin care water products used by the subjects in each group were the same products.

[0142] (2) Test contents: Corneometer CM 825 (Courage & Khazaka, Germany) was used to test the moisture content of the stratum corneum of the skin;

[0143] Skin elasticity tester Cutometer MPA580 (Courage & Khazaka, Germany) was used to test skin elasticity and firmness;

[0144] The test products are freeze-dried facial mask products prepared in Application Examples 1-7 and Comparative Application Examples 1-6.

[0145] The test environment is: temperature: 20±2℃, humidity 50±5%, which meets the requirements of the program design; the improvement after using the product (%) = (data after use - data before use) / data before use × 100%; the results are shown in Table 2:

[0146] Table 2

[0147] Change rate of stratum corneum moisture content Skin elasticity R2 change rate Change rate of firmness F4 Application Example 1 38.73％ 17.24％ 18.16％ Application Example 2 36.54％ 15.96％ 17.05％ Application Example 3 37.67％ 16.25％ 17.84％ Application Example 4 32.55％ 14.15％ 16.00％ Application Example 5 31.59％ 13.70％ 16.17％ Application Example 6 31.58％ 12.96％ 15.75％ Application Example 7 30.92％ 11.99％ 15.18％ Comparative application example 1 25.48％ 9.51％ 10.96％ Comparative Application Example 2 27.91％ 10.47％ 12.65％ Comparative Application Example 3 26.51％ 9.77％ 11.48％ Comparative Application Example 4 28.16％ 10.95％ 13.00％ Comparative Application Example 5 26.88％ 9.86％ 11.72％ Comparative Application Example 6 20.10％ 6.51％ 7.64％

[0148] From the data in the table we can see that:

[0149] (1) It can be seen from Application Examples 1 to 3 that the composition containing recombinant collagen provided by the present invention has significant moisturizing, firming and anti-oxidation effects, among which Application Example 1 has the best effect. Since skin oxidation can cause skin dryness and loss of elasticity, etc., the test results of skin moisture content and elasticity can also reflect the degree of skin oxidation to a certain extent.

[0150] (2) By comparing Application Example 1 with Application Examples 4-6, it can be seen that the purslane extract in the present invention is enzymatically extracted by a composite enzyme composed of cellulase, amylase and bromelain. The three enzymes can synergistically promote the enzymatic hydrolysis effect after being used in combination, further improving the extraction effect of the relevant active ingredients in purslane, and obtaining a purslane extract with better effect.

[0151] (3) By comparing Application Example 1 with Application Example 7, it can be seen that the black wolfberry extract prepared by supercritical CO2 extraction in the present invention has a better effect than the black wolfberry extract prepared by conventional alcohol extraction.

[0152] (4) By comparing Application Example 1 with Comparative Application Examples 1-3, it can be seen that when Comparative Application Examples 1-3 do not contain any of the three components of Haematococcus pluvialis extract, Portulaca oleracea extract and Lycium ruthenicum extract, the moisturizing and anti-oxidation effects of the prepared compositions are worse than those of Application Example 1. It can be seen that the three components of Haematococcus pluvialis extract, Portulaca oleracea extract and Lycium ruthenicum extract used in the present invention can cooperate with each other, synergistically enhance the effect, and improve the skin condition.

[0153] (5) By comparing Application Example 1 with Comparative Application Examples 4-6, it can be seen that the present invention uses recombinant collagen and Schizophyllum polysaccharide in combination to form a complex. While improving the stability of the application of recombinant collagen, it can also further improve the stability of the active substances in the plant extract components in skin care products. The combination of the three components synergistically improves the moisturizing and antioxidant effects of the product.

[0154] Test Example 3

[0155] Product safety testing

[0156] According to the inclusion criteria, 35 women and 35 men were selected to participate in the test, and randomly divided into 7 groups (5 men and 5 women in each group) to receive the trial. Take an appropriate amount of the freeze-dried mask stock solution sample in Application Example 1-7 and place it in the medicine chamber of the spot test tape. Apply the spot tester with the sample to the skin on the curved side of the forearm of the subject with medical tape for 24 hours. 30 minutes after removing the test spot tester, observe the skin reaction after the indentation disappears. For those with negative results, observe again 24 hours and 48 hours after the test, and record the skin reaction results. The reaction degree grading standard is as shown in the following table.

[0157]

[0158] The results show that the freeze-dried mask products prepared in Application Examples 1-7 showed negative reactions at 24h and 48h after the test, and there was no irritation or erythema. It can be seen that the mask products prepared by the present invention have no irritation to the skin and no positive reaction, which proves that the product is highly safe, less irritating, and milder.

[0159] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0160] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0161] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composition containing recombinant collagen, characterized in that: The composition containing recombinant collagen comprises recombinant collagen, Schizophyllan polysaccharide and plant extract; The plant extract comprises a combination of a Haematococcus pluvialis extract, a Portulaca oleracea extract and a Lycium nigra extract.

2. The composition containing recombinant collagen according to claim 1, characterized in that: Calculated by weight, the composition containing recombinant collagen includes 0.2-5 parts of recombinant collagen, 0.01-1 parts of schizophyllan and 0.01-1.5 parts of plant extract.

3. The composition containing recombinant collagen according to claim 1 or 2, characterized in that: The molecular weight of the recombinant collagen is 5-20 kDa; Preferably, the molecular weight of the Schizophyllan is 10-100 kDa.

4. The composition containing recombinant collagen according to claim 1, characterized in that: The mass ratio of the Haematococcus pluvialis extract, the Portulaca oleracea extract and the Lycium ruthenicum extract is (0.05-3):(0.1-5):(0.1-5).

5. The composition containing recombinant collagen according to claim 1 or 4, characterized in that: The purslane extract is prepared by a preparation method comprising the following steps: Mixing purslane, complex enzyme and water, performing ultrasonication, filtering to obtain a filtrate, and concentrating and drying the filtrate to obtain the purslane extract; The complex enzyme comprises a combination of cellulase, amylase and bromelain.

6. The composition containing recombinant collagen according to claim 5, characterized in that: The mass ratio of the cellulase, amylase and bromelain is (1-6):(2-7):(0.1-2); Preferably, the added amount of the complex enzyme is 0.05‰-2‰ of the mass of purslane; Preferably, the solid-liquid ratio of purslane to water is 1 g:(30-80) mL.

7. The composition containing recombinant collagen according to claim 1 or 4, characterized in that: The black wolfberry extract is prepared by a preparation method comprising the following steps: The black wolfberry is subjected to supercritical CO2 extraction, gasified CO2 and anhydrous ethanol are filled into the extractor, and the obtained extract is concentrated and dried to obtain the black wolfberry extract.

8. Use of the composition containing recombinant collagen according to any one of claims 1 to 7 in cosmetics; The cosmetics include lotions, creams, skin care water, skin care essences or facial masks.

9. A freeze-dried facial mask, characterized in that: The raw materials for preparing the freeze-dried facial mask include, by mass percentage, 1%-10% of the composition containing recombinant collagen according to any one of claims 1-6.

10. The freeze-dried facial mask according to claim 9, characterized in that: The raw materials for preparing the freeze-dried facial mask also include any one of moisturizers, antioxidants, skin conditioners, thickeners, stabilizers, chelating agents or water, or a combination of at least two of them.

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