CN112370369A

CN112370369A - Marine protein skin moisturizer for resisting skin aging

Info

Publication number: CN112370369A
Application number: CN202011171979.0A
Authority: CN
Inventors: 梁志勇; 孙倩; 丁洪雪; 宋江红
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-10-28
Filing date: 2020-10-28
Publication date: 2021-02-19

Abstract

The invention discloses a marine protein skin moisturizer for resisting skin aging, and belongs to the technical field of marine organisms. The invention takes sea cucumber viscera which are wastes of deep processing of sea cucumber as raw materials, integrates the modern bioseparation technology, the biological enzymolysis technology and the mixed bacteria microbial fermentation technology, develops and develops the marine protein skin care solution with high skin aging resistance, and the raw materials comprise 75.56 percent of marine protein extract, 0.04 percent of sodium hyaluronate, 10 percent of glycerol, 4003 percent of polyethylene glycol, 2 percent of emulsifier, 1.5 percent of thickener, 5 percent of squalane, 0.5 percent of tocopheryl acetate, 0.2 percent of triethanolamine, 2 percent of tranexamic acid and 0.2 percent of carbomer. The product can obviously increase the content of hyaluronic acid, hydroxyproline, total collagen and elastin in the skin, increase the water content of the skin and obviously improve the anti-aging effect of the skin, and is a novel skin care product based on the resource recycling of marine wastes.

Description

Marine protein skin moisturizer for resisting skin aging

Technical Field

The invention relates to a marine protein skin moisturizer for resisting skin aging, in particular to a method for preparing a marine protein skin moisturizer with high oxidation resistance and skin aging resistance by using sea cucumber viscera and Chinese herbal medicines as raw materials through synergistic fermentation of microorganisms, and belongs to the technical field of marine organisms.

Background

The sea cucumber is a precious sea food, has tender meat quality and rich nutrition, is a typical high-protein low-fat food, and has extremely high nutritional value and medicinal value. At present, the main edible part of the sea cucumber is the sea cucumber body wall, so the research on the sea cucumber is focused on the active substances on the sea cucumber body wall. The sea cucumber viscera which are byproducts in the sea cucumber processing are mostly discarded, the utilization rate is low, the research on the sea cucumber viscera at home and abroad is less, and the developed related products are very little. However, the viscera of the sea cucumber not only contain chemical components similar to the body wall, but also are rich in taurine, arginine, phospholipid, unsaturated fatty acid, various high-activity trace elements, pigments, active enzymes and functional bacteria, and the edible value and the medicinal value of the viscera of the sea cucumber are higher than those of the body wall of the sea cucumber within a certain range. At present, the lack and neglect of the knowledge of the active substances in the internal organs of the sea cucumbers cause serious waste and loss of resources. The high-value comprehensive utilization of the viscera of the sea cucumber becomes a technical problem to be solved urgently in the field.

Aging is a necessary rule of life process, and is a comprehensive process involving the influence of various factors inside and outside and the participation of multiple organs. The skin is the largest organ of the human body, and is responsible for protection, sensation, secretion, excretion, thermoregulation, immunoregulation, etc., and the skin gradually ages as other organs are aged. The main factors affecting skin aging are classified into intrinsic factors (such as age, gene mutation, cell metabolism, and hormonal environment) and extrinsic factors (such as chemicals, toxins, environmental pollution, ultraviolet rays, and ion rays), wherein the extrinsic factors cause skin aging due to solar radiation. Skin aging is manifested by increased wrinkles and pigmentation, loose and rough skin, and inelasticity. The exposure of the aging skin becomes the external expression of human aging, the aging skin is most easily perceived in the aging process of the human body, the face is most obvious, and the psychological burden of people is increased, so that the aging skin is delayed, the youth and the face are kept for a long time, the skin is a topic which is widely concerned by all walks of life in all ages, and the anti-aging product has a wide market. At present, various skin care products, skin operations such as wrinkle removal, laser treatment and the like are not lacked in the market, but most of common people often do not have time for skin care due to large working pressure and fast pace of life, and the existing anti-aging products have poor quality and are frequently lack of adverse reactions, so that Chinese herbal medicines with natural sources, small adverse reactions and long history become a new favorite in the field of beauty treatment and anti-aging. The preparation of the marine protein skin moisturizer for resisting skin aging by using the modern biotechnology has very important significance.

Disclosure of Invention

In order to overcome the defects of the prior art and meet the requirements of consumers on skin care lotion for resisting skin aging and beautifying the skin, the application provides the marine protein skin care lotion for resisting skin aging, and the marine protein skin care lotion with the obvious skin aging resisting effect is developed by integrating the modern bioseparation technology, the biological enzymolysis technology and the mixed bacteria microbial fermentation technology. The product can significantly increase the content of hyaluronic acid, hydroxyproline, total collagen and elastin in skin, increase the water content of skin, and increase the content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Malondialdehyde (MDA) and hydroxyproline (Hyp) in skin tissue.

The invention realizes the technical effects through the following technical scheme:

the marine protein skin lotion for resisting skin aging is characterized by comprising 75.56% of marine protein extracting solution, 0.04% of sodium hyaluronate, 10% of glycerol, 3% of polyethylene glycol-4003%, 2% of emulsifier, 1.5% of thickener, 5% of squalane, 0.5% of tocopherol acetate, 0.2% of triethanolamine, 2% of tranexamic acid and 0.2% of carbomer in a ratio by mass.

The emulsifier is one or more of glycerol polymethacrylate, propylene glycol and PVM/MA copolymer.

The thickener is one or more of laureth, C13-14 isoparaffin, and sodium polyacrylate.

The marine protein extracting solution is prepared by the following steps:

(1) cleaning: cleaning the processed fresh sea cucumber viscera with tap water, and then cleaning with purified water and draining for later use;

(2) homogenizing: adding the sea cucumber viscera obtained in the step (1) into a high-speed homogenizer, adding purified water, and performing high-speed homogenization until all the sea cucumber viscera can pass through a round sieve with the aperture of 2 mm;

(3) and (3) sterilization: sterilizing the homogenized internal organs of the sea cucumber by a high-pressure steamer, and cooling for later use;

(4) compound enzymolysis: adding a complex enzyme preparation with the mass ratio of 2% into the sterilized slurry, and stirring and performing enzymolysis for 30min at the temperature of 40-60 ℃;

(5) aerobic fermentation: adding a compound microbial agent with the mass ratio of 3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 8h at the temperature of 28-35 ℃;

(6) and (3) filtering: filtering to remove fermentation thallus;

(7) enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;

(8) centrifuging: centrifuging at 10000r/min to obtain supernatant as sea marine protein extractive solution.

Preferably, astaxanthin with the mass ratio of 2 percent of the viscera of the sea cucumber is added in the process of homogenizing the viscera of the sea cucumber.

The compound protease in the step (4) is a combination of trypsin, bromelain and flavourzyme, and the mass ratio of the compound protease to the flavourzyme is 2:2: 1.

The compound microbial agent in the step (5) is a mixed microbial agent of bacillus subtilis and bacillus licheniformis, and the mass ratio of the compound microbial agent to the bacillus licheniformis is 3: 1.

The ventilation rate is 0.2-0.3 m when the aerobic fermentation process in the step (5) is carried out in a fermentation tank³/m³.min。

The preparation method of the bacillus subtilis and bacillus licheniformis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus subtilis and bacillus licheniformis seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

The invention provides an anti-skin-aging marine protein skin moisturizer which is prepared by the following steps:

(1) the raw materials are matched: the raw materials are accurately mixed according to the proportion;

(2) mixing: mixing marine protein extract, sodium hyaluronate, glycerol, squalane, tocopheryl acetate, triethanolamine and tranexamic acid uniformly;

(3) thickening: adding a thickening agent and carbomer into the uniformly mixed raw materials;

(4) emulsification: adding polyethylene glycol-400 and an emulsifier, and carrying out emulsification by keeping the temperature for 30min at the temperature of 30-40 ℃;

(5) homogenizing: homogenizing for 10-20 min on a homogenizer;

(6) subpackaging: and subpackaging the homogenized skin moisturizer to obtain the marine protein skin moisturizer.

The invention provides a marine protein skin moisturizer for resisting skin aging, which can obviously increase the content of hyaluronic acid, hydroxyproline, total collagen and elastin in skin, increase the water content of skin, and improve the content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), Malondialdehyde (MDA), hydroxyproline (Hyp) and the like in skin tissues. The safety evaluation of the product of the invention is carried out on the skin acute toxicity of SD rats, the skin irritation of New Zealand rabbits and the skin allergy of guinea pigs. The experimental result shows that the product of the invention has no toxic reaction to the intact skin, no stimulation to the intact skin and no obvious effect on the allergic reaction of the guinea pig skin, thereby proving that the product is safe and has no toxicity.

Compared with the prior art, the marine protein skin moisturizer for resisting skin aging provided by the invention has the following remarkable advantages:

(1) according to the method, the sea cucumber visceral protein is taken as a main raw material, and a modern bioseparation technology, a biological enzymolysis technology and a mixed bacteria microbial fermentation technology are integrated, so that the skin aging resistance of the sea cucumber visceral protein is remarkably improved, the high-valued recycling of marine deep processing wastes is realized, and a new way is developed for the industrial development of skin care products;

(2) the application utilizes aerobic bacteria (bacillus subtilis and bacillus licheniformis) to jointly ferment the sea cucumber visceral protein to extract the sea cucumber protein, makes full use of the synergistic effect among microorganisms, and improves the anti-skin aging capacity of the sea cucumber protein (including sea cucumber polypeptide), wherein: the bacillus subtilis can kill pathogenic bacteria in fermentation liquor in the high-temperature fermentation process, has strong abilities of producing amylase, protease and the like, and can degrade macromolecular starch and protein substances into microorganisms for utilization, thereby improving the oxidation resistance and the nutritional and health-care effects of active peptide; the addition of the bacillus licheniformis can obviously improve the oxidation resistance of the sea cucumber protein, the product is improved through the fermentation effect of the bacillus licheniformis, the stability of the product is improved, and the experimental result shows that the shelf life of the product is not influenced by adding no preservative into the skin care lotion;

(3) the invention adopts the synergistic effect of composite protease (trypsin, bromelain and flavourzyme) enzymolysis and composite microorganism (bacillus subtilis and bacillus licheniformis) fermentation to convert the viscera of the sea cucumber into the marine protein extracting solution with high skin aging resistance, thereby obtaining the obvious technical effect which can not be expected by the technical personnel in the field;

(4) the skin care cream is scientifically matched with marine protein extracting solution (for skin care, anti-aging and moisturizing), sodium hyaluronate (for moisturizing), glycerol (for moisturizing), polyethylene glycol-400 (for emulsification), glycerol polymethacrylate/propylene glycol/PVM/MA copolymer (for emulsification), squalane (for skin care), tocopherol acetate (for antioxidation), triethanolamine (for regulating the pH value), tranexamic acid (for whitening), carbomer (for thickening) and other raw materials, and has the effects of skin tendering, skin care, face nourishing, whitening and the like through the synergistic effect of the raw materials.

Detailed Description

In the present application, the activation method of bacillus subtilis and bacillus licheniformis is as follows: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus subtilis and bacillus licheniformis seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

In the embodiment, the synzyme is a combination of trypsin, bromelain and flavourzyme, and the mass ratio of the synzyme is 2:2: 1; the composite microbial agent is a mixed microbial agent of bacillus subtilis and bacillus licheniformis, and the mass ratio of the composite microbial agent to the bacillus licheniformis is 3: 1.

Example 1

A marine protein skin care lotion for resisting skin aging comprises the following specific process steps:

(1) preparing a marine protein extracting solution: cleaning the processed fresh sea cucumber viscera with tap water, and then cleaning with purified water and draining for later use; adding the obtained sea cucumber viscera into a high-speed homogenizer, adding purified water and 2% of astaxanthin, and carrying out high-speed homogenization until all the sea cucumber viscera can pass through a circular sieve with the aperture of 2 mm; sterilizing the homogenized internal organs of the sea cucumber by a high-pressure steamer; adding a complex enzyme preparation with the mass ratio of 2% into the sterilized slurry, and stirring and performing enzymolysis for 30min at the temperature of 40-60 ℃; adding a compound microbial agent with the mass ratio of 3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 8h at the temperature of 28-35 ℃; after fermentation, filtering to remove fermentation thalli; inactivating enzyme at 100 deg.C for 10 min; centrifuging at 10000r/min to obtain supernatant as sea marine protein extractive solution;

(2) the raw materials are matched: accurately mixing raw materials according to the proportion of 75.56% of marine protein extract, 0.04% of sodium hyaluronate, 10% of glycerol, 4003% of polyethylene glycol, 2% of glycerol polymethacrylate, 1.5% of laureth, 5% of squalane, 0.5% of tocopherol acetate, 0.2% of triethanolamine, 2% of tranexamic acid and 0.2% of carbomer;

(3) mixing: mixing marine protein extract, sodium hyaluronate, glycerol, squalane, tocopheryl acetate, triethanolamine and tranexamic acid uniformly;

(4) thickening: adding laureth and carbomer into the uniformly mixed raw materials;

(5) emulsification: adding polyethylene glycol-400 and glycerol polymethacrylate, and carrying out emulsification by keeping the temperature for 30min at 30-40 ℃;

(6) homogenizing: homogenizing for 10-20 min on a homogenizer;

(7) subpackaging: subpackaging the homogenized skin moisturizer to obtain the marine protein skin moisturizer;

the influence of the skin care lotion containing the marine protein in example 1 on the skin aging of nude mice was determined, and the animal grouping and model making methods were as follows: the male nude mice are layered according to physical quality, and are randomly divided into a normal control group, a model group, an experimental group in example 1, a control 1 group (the marine protein preparation process adopts single fermentation of bacillus subtilis, the rest is the same as in example 1) and a control 2 group (the marine protein preparation process adopts single fermentation of bacillus licheniformis, the rest is the same as in example 1), and each group is 10. Model group and each sample group were injected with 10% D-galactose 1.0g.kg subcutaneously in the neck and back per day^-1Irradiating the mold for 40min at a vertical height of 40cm by using an ultraviolet lamp with the wavelength of 350-400 nm, and continuously molding for 40 d; the normal control group was injected with an equal volume of physiological saline and given normal light. Starting from the 11d of model building, smearing corresponding samples on the skin of the nude mice in each group, and smearing normal control group and model group with the same volume of physiological saline for 2 times a day for 30d continuously;

and (3) biochemical index determination: after the last administration, the animals are sacrificed to take 1cm multiplied by 1cm of back skin, after being cut into pieces, 9 times of normal saline is added to prepare 10 percent (m/v) of skin homogenate, and after being centrifuged at 900 multiplied by g for 10min, the supernatant is taken for standby. Determining the content of hyaluronic acid, hydroxyproline, total collagen, type III collagen and elastin in the skin according to the method of the kit instruction;

measurement of moisture content in skin: taking 1cm multiplied by 1cm of partial back skin at the end of the experiment, removing epidermis and subcutaneous tissues, and precisely weighing the skin as wet quality; drying in an incubator at 80 ℃ for 48h, and precisely weighing to obtain the dry mass of the skin. Calculating skin moisture content according to the following formula, wherein the skin moisture content (%) ═ (skin wet mass-skin dry mass)/skin wet mass x 100%;

the test results are as follows:

the experimental structure shows that the skin-resistant effect of the example 1 on the nude mice is not obviously different from that of a normal group and is extremely obvious from that of a model group. The preparation process of the marine protein extracting solution is changed, so that the anti-skin-aging effect of the product is obviously reduced.

Example 2

(1) preparing a marine protein extracting solution: cleaning the processed fresh sea cucumber viscera with tap water, and then cleaning with purified water and draining for later use; adding the obtained viscera of the sea cucumber into a high-speed homogenizer, adding purified water and 5% of astaxanthin, and carrying out high-speed homogenization until all the viscera can pass through a circular sieve with the aperture of 2 mm; sterilizing the homogenized internal organs of the sea cucumber by a high-pressure steamer; adding a complex enzyme preparation with the mass ratio of 2% into the sterilized slurry, and stirring and performing enzymolysis for 30min at the temperature of 40-60 ℃; adding a compound microbial agent with the mass ratio of 3% of the enzymolysis liquid into the enzymolysis liquid after enzymolysis, and carrying out aerobic fermentation for 8h at the temperature of 28-35 ℃; after fermentation, filtering to remove fermentation thalli; inactivating enzyme at 100 deg.C for 10 min; centrifuging at 10000r/min to obtain supernatant as sea marine protein extractive solution;

(6) homogenizing: homogenizing for 10-20 min on a homogenizer;

the influence of the skin care lotion containing the marine protein in example 2 on the skin aging of nude mice was determined, and the animal grouping and model making methods were as follows: the male nude mice were stratified according to their physical qualities and were randomly divided into test groups of example 2, control 3 groups (the marine protein preparation process employed Bacillus subtilis single fermentation, the remainder was the same as example 1), and control 4 groups (the marine protein preparation process employed Bacillus licheniformis single fermentation, the remainder was the same as example 1), each of which contained 10 mice. Each sample group was injected subcutaneously with 10% D-galactose 1.0 g/kg of D-galactose per day on the back and neck^-1And irradiating the mold for 40min at a vertical height of 40cm by using an ultraviolet lamp with the wavelength of 350-400 nm, and continuously molding for 40 d. Starting from the 11d of model building, smearing corresponding samples on the skin of the nude mice in each group, and smearing normal control group and model group with the same volume of physiological saline for 2 times a day for 30d continuously;

the biochemical index and the skin water content were measured in the same manner as in example 1;

the test results are as follows:

Example 3

(2) the raw materials are matched: accurately mixing the raw materials according to the proportion of 75.56% of marine protein extract, 0.04% of sodium hyaluronate, 10% of glycerol, 4003% of polyethylene glycol, 2% of propylene glycol, 1.5% of sodium polyacrylate, 5% of squalane, 0.5% of tocopheryl acetate, 0.2% of triethanolamine, 2% of tranexamic acid and 0.2% of carbomer;

(3) mixing: mixing marine protein extract, sodium hyaluronate, glycerol, squalane, tocopherol acetate, triethanolamine, tranexamic acid;

(4) thickening: adding sodium polyacrylate and carbomer into the uniformly mixed raw materials;

(5) emulsification: adding polyethylene glycol-400 and propylene glycol, and carrying out emulsification by keeping the temperature for 30min at the temperature of 30-40 ℃;

(6) homogenizing: homogenizing for 10-20 min on a homogenizer;

the influence of the skin care lotion containing the marine protein in example 3 on skin aging of nude mice is determined, and the experimental result is not significantly different from that in example 1.

The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; such modifications and substitutions do not depart from the spirit and scope of the present invention as set forth in the appended claims.

Claims

1. The marine protein skin lotion for resisting skin aging is characterized by comprising 75.56% of marine protein extracting solution, 0.04% of sodium hyaluronate, 10% of glycerol, 3% of polyethylene glycol-4003%, 2% of emulsifier, 1.5% of thickener, 5% of squalane, 0.5% of tocopherol acetate, 0.2% of triethanolamine, 2% of tranexamic acid and 0.2% of carbomer in a ratio by mass.

2. The marine protein skin lotion for resisting skin aging of claim 1, wherein the emulsifier is one or more of glycerol polymethacrylate, propylene glycol, and PVM/MA copolymer.

3. The marine protein skin lotion for resisting skin aging of claim 1, wherein the thickener is one or more of laureth, C13-14 isoparaffin, and sodium polyacrylate.

4. The marine protein skin moisturizer for resisting skin aging according to claim 1, wherein the marine protein extract is prepared by the following steps:

(6) and (3) filtering: filtering to remove fermentation thallus;

(7) enzyme deactivation: inactivating enzyme at 100 deg.C for 10 min;

5. The skin-aging resistant marine protein lotion according to claim 4, wherein the step (2) comprises adding 2% by mass of astaxanthin to the viscera of the sea cucumber during the homogenization of the viscera of the sea cucumber.

6. The marine protein skin moisturizer for resisting skin aging according to claim 4, wherein the compound protease in the step (4) is a combination of trypsin, bromelain and flavourzyme, and the mass ratio of the trypsin to the bromelain to the flavourzyme is 2:2: 1.

7. The marine protein skin moisturizer for resisting skin aging according to claim 4, wherein the compound microbial agent in the step (5) is a mixed microbial agent of bacillus subtilis and bacillus licheniformis, and the mass ratio of the compound microbial agent to the bacillus licheniformis is 3: 1.

8. The marine protein skin moisturizer for resisting skin aging of claim 1, wherein the ventilation rate of the aerobic fermentation process in the step (4) is 0.2 to 0.3m³/m³.min。

9. The marine protein skin moisturizer for resisting skin aging of claim 7, wherein the preparation method of the bacillus subtilis and bacillus licheniformis seed solution comprises the following steps: under the aseptic condition, LB culture medium is adopted to culture for 24 hours at the temperature of 28-32 ℃ to prepare liquid bacillus subtilis and bacillus licheniformis seed liquid, and the effective viable count of the seed liquid is required to be more than or equal to 10⁹cfu/mL。

10. The marine protein skin moisturizer for resisting skin aging according to claim 1, wherein the marine protein skin moisturizer is prepared by the following steps:

(5) homogenizing: homogenizing for 10-20 min on a homogenizer;