CN114634563A

CN114634563A - Active collagen extracting solution, preparation method and system thereof, skin care product and preparation method thereof

Info

Publication number: CN114634563A
Application number: CN202210142767.2A
Authority: CN
Inventors: 邓天发
Original assignee: Huizhou Foryou Medical Devices Co Ltd
Current assignee: Huizhou Foryou Medical Devices Co Ltd
Priority date: 2022-02-16
Filing date: 2022-02-16
Publication date: 2022-06-17

Abstract

When the active collagen extracting solution is prepared, the epidermis layer and the subcutaneous fat layer of a fresh leather are removed, and a first semi-finished product is obtained by adopting a first organic solvent and water for cleaning; crushing the first semi-finished product to obtain a second semi-finished product; sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product; and adopting HCl solution to pre-soak the third semi-finished product, adding softening solution to soften, wherein the temperature in the softening process is 20-40 ℃, the pH value is 2.5-4.5, enzymolysis liquid obtained through enzymolysis reaction is a fourth semi-finished product, and the solution obtained by filtering the fourth semi-finished product is the active collagen extracting solution, and the whole preparation process does not need low temperature conditions, so that the production difficulty of the active collagen extracting solution is reduced, and the active collagen extracting solution with low production difficulty, the preparation method and the system thereof, the skin care product and the preparation method thereof are realized.

Description

Active collagen extracting solution, preparation method and system thereof, skin care product and preparation method thereof

Technical Field

The application relates to the technical field of cosmetics, in particular to an active collagen extracting solution, a preparation method and a system thereof, a skin care product and a preparation method thereof.

Background

In the prior art, the extraction process conditions of the active collagen structure are harsh, and the process characteristics of low temperature are achieved, so that the problem of high production difficulty of the active collagen extracting solution is caused, and meanwhile, the problem of high production difficulty of skin care products comprising the active collagen extracting solution is further influenced, so that how to provide the active collagen extracting solution, the preparation method and system, the skin care products and the preparation method is to solve the technical problem of high production difficulty of the active collagen extracting solution and the skin care products comprising the active collagen extracting solution in the prior art.

Disclosure of Invention

The application provides an active collagen extracting solution, a preparation method and a system thereof, a skin care product and a preparation method thereof, and aims to solve the technical problem that the production difficulty of the active collagen extracting solution and the skin care product comprising the active collagen extracting solution is high in the prior art.

In order to solve the above problems, the present application provides an active collagen extract solution, which includes an active collagen main body and a matrix, wherein the active collagen main body includes triple-helix active collagen, collagen peptide and hydrolyzed gel, in the active collagen main body, the mass fraction of the triple-helix active collagen is greater than or equal to 90% and less than 100%, the sum of the mass fractions of the collagen peptide and the hydrolyzed gel is greater than 0 and less than or equal to 10%, and the matrix is water.

Wherein the pH value of the active collagen extracting solution is 3.5-5.5.

The present application also provides a method for preparing an active collagen extract solution for preparing the active collagen extract solution as described above, the method comprising:

providing fresh leather, removing a skin layer and a subcutaneous fat layer, and cleaning by adopting a first organic solvent and water to obtain a first semi-finished product;

the first semi-finished product is crushed into muddy flesh to obtain a second semi-finished product;

sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product; and

adopt HCl solution right the third semi-manufactured goods carry out presoaking, obtain the presoaking mixture, add the softening solution and soften, the temperature of softening process is 20 degrees centigrade-40 degrees centigrade, and pH value is 2.5-4.5, adds first group protease or the second group protease and carries out the enzymolysis reaction, and the enzymolysis liquid that obtains through the enzymolysis reaction is fourth semi-manufactured goods, right the solution that the fourth semi-manufactured goods filtered the acquisition does active collagen extract.

Wherein the first organic solvent is 75% ethanol.

Wherein the second organic solvent is a combination of at least two of acetone, ethanol and petroleum ether, and the mass ratio of the second organic solvent to the second semi-finished product is 0.5-3.5: 1.

wherein the second organic solvent is a mixture of acetone and ethanol, and the volume ratio of the acetone to the ethanol is 1: 0.5-3.5, or the second organic solvent is a mixture of petroleum ether and ethanol, and the volume ratio of the petroleum ether to the ethanol is 1: 0.5-3.5.

Wherein the salt solution is a NaCl solution with the mass fraction of 3% -15%, and the mass ratio of the salt solution to the second semi-finished product is 0.5-3.5: 1.

in the step of sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product, the mass ratio of the water to the second semi-finished product is (2.5-7): 1.

the concentration of the HCl solution is 0.001-0.02 mol/L, the softening solution is an acetic acid solution, a citric acid solution or a mixed solution of acetic acid and citric acid, the concentration of the softening solution is 0.01-0.3 mol/L, the first group of protease is pepsin, and the second group of protease comprises papain and/or bromelain.

Wherein the second group of proteases further comprises one or a combination of trypsin, ficin and cysteine protease.

Wherein the volume ratio of the softening solution to the pre-soak mixture is 20-60: 1, the mass ratio of the first group of protease to the third semi-finished product is 4-10 ‰: 1, the mass ratio of the second group of protease to the third semi-finished product is 2-6 ‰: 1.

the present application also provides a system for preparing an active collagen extracting solution, the system for preparing an active collagen extracting solution being used for preparing the active collagen extracting solution as described above, the system for preparing an active collagen extracting solution comprising:

the splitting device group is used for removing an epidermal layer and a subcutaneous fat layer from the fresh leather, and cleaning the fresh leather by adopting a first organic solvent and water to obtain a first semi-finished product;

a mincing device for mincing the first semi-finished product into muddy flesh to obtain a second semi-finished product;

the impurity removal equipment set is used for sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product; and

and the extraction equipment set is used for carrying out pre-soaking, softening and enzymolysis reaction on the third semi-finished product, the enzymolysis liquid obtained through the enzymolysis reaction is a fourth semi-finished product, and the solution obtained by filtering the fourth semi-finished product is the active collagen extracting solution.

The splitting device comprises a splitting machine, a splitting machine and a cleaning device, wherein the splitting machine is used for splitting fresh leather to obtain the leather strips, the splitting machine is used for removing the skin layers and subcutaneous fat layers of the leather strips to obtain first intermediate products, and the cleaning device is used for adopting the first organic solvent and water to clean the first intermediate products to obtain first semi-finished products.

The cleaning device comprises a cleaning groove, a leather strip feeding port, a first organic solvent feeding port, a water feeding port and a leather strip discharging port, wherein the leather strip feeding port, the first organic solvent feeding port, the water feeding port and the leather strip discharging port are communicated with the cleaning groove, and the leather strip feeding port, the first organic solvent feeding port, the water feeding port and the leather strip discharging port are the same opening or different openings.

Wherein, the impurity removing equipment group comprises a first cleaning cylinder, a first impurity removing and filtering equipment, a second cleaning cylinder, a second impurity removing and filtering equipment, a third cleaning cylinder and a third impurity removing and filtering equipment, the first cleaning cylinder is used for mixing the second organic solvent with the second semi-finished product, the first impurity removing and filtering equipment is used for filtering the mixture in the first cleaning cylinder to filter out liquid to obtain a second intermediate product, the second cleaning cylinder is used for mixing the salt solution with the second intermediate product, the second impurity removing and filtering equipment is used for filtering the mixture in the second cleaning cylinder to filter out liquid to obtain a third intermediate product, the third cleaning cylinder is used for mixing the water with the third intermediate product, the third impurity removing and filtering equipment is used for filtering the mixture in the third cleaning cylinder, and filtering off liquid to obtain a third semi-finished product, wherein the first cleaning cylinder, the second cleaning cylinder and the third cleaning cylinder are the same cleaning cylinder or different cleaning cylinders, and the first impurity removal filtering equipment, the second impurity removal filtering equipment and the third impurity removal filtering equipment are the same impurity removal filtering equipment or different impurity removal filtering equipment.

The impurity removing equipment group further comprises a meter, and the meter is used for calculating the mass or volume of the second semi-finished product added into the first cleaning cylinder, the second organic solvent added into the first cleaning cylinder, the salt solution added into the second cleaning cylinder and the water added into the third cleaning cylinder.

The impurity removing equipment group further comprises a processor, wherein the processor is used for controlling the mass ratio of the second organic solvent to the second semi-finished product, controlling the mass ratio of the saline solution to the second semi-finished product and controlling the mass ratio of the water added into the impurity removing equipment to the second semi-finished product.

Wherein, draw equipment group including drawing jar, agitator and extract filtration equipment, it is right to draw the jar to be used for adopting HCl solution the third semi-manufactured goods soaks in advance, obtains the presoaking mixture, adds the softening solution and softens, adds first group protease perhaps the enzymolysis reaction is carried out to second group protease, obtains through the enzymolysis reaction the enzymolysis liquid does the fourth semi-manufactured goods, the agitator is used for right the mixture that draws in the jar stirs, extract filtration equipment is used for right the fourth semi-manufactured goods are filtered, and the solution that obtains does active collagen extract.

Wherein the extraction apparatus set further comprises a meter for calculating the volume of the softening solution added to the extraction tank and the mass of the first or second set of proteases, a thermometer for detecting the temperature of the mixture within the extraction tank, and a pH meter for detecting the pH of the mixture within the extraction tank.

Wherein the extraction kit further comprises a processor for controlling the volume ratio of the softening solution to the pre-soak mixture, controlling the mass ratio of the first group of proteases to the third semi-finished product, controlling the mass ratio of the second group of proteases to the third semi-finished product, controlling the pH of the softening process, and controlling the temperature of the softening process.

Wherein the extraction equipment set further comprises a display component, and the processor is further configured to control the display component to display the mass calculated by the meter, the temperature detected by the thermometer, and the pH value detected by the pH meter.

The extracting solution filtering device comprises a first sub device and a second sub device, the mesh number of the filter bag of the first sub device is 30-50, the mesh number of the filter bag of the second sub device is 150-.

The application also provides a skin care product, the skin care product includes active collagen extract, sodium hyaluronate, balancing agent, emollient, formulation regulator, preservative and water, active collagen extract is as before active collagen extract, active collagen extract the sodium hyaluronate the balancing agent the emollient, the formulation regulator, preservative with the part by mass relation of water is 2 parts-10 parts active collagen extract, 2 parts-6 parts sodium hyaluronate, 1 part-4 parts the balancing agent, 100 parts-200 parts the emollient, 2 parts-20 parts the formulation regulator, 1 part-4 parts the preservative and 1760 parts-1900 parts water.

Wherein the sodium hyaluronate is one or a combination of more than one of sodium hyaluronate with the molecular weight of 120-150 ten thousand, 80-100 ten thousand, 20-40 ten thousand and 1-10 ten thousand daltons.

Wherein the balancing agent comprises one or more of dipotassium glycyrrhizinate, sodium dihydrogen phosphate and disodium hydrogen phosphate.

Wherein the emollient is one or more of glycerol, butanediol, propylene glycol, glyceryl caprylate, 1, 2-hexanediol, and glyceryl polyether-26.

Wherein the dosage form regulator is one or the combination of two of hydroxyethyl cellulose and xanthan gum.

Wherein, the preservative comprises one or the combination of two of caprylyl hydroximic acid and ethylhexyl glycerin.

Wherein the pH of the skin care product is 4.5-7.

The application also provides a preparation method of the skin care product, which comprises the following steps:

providing an active collagen extracting solution, sodium hyaluronate, a balancing agent, a softening agent, a formulation regulator, a preservative and water, wherein the active collagen extracting solution is the active collagen extracting solution, and the mass parts of the active collagen extracting solution, the sodium hyaluronate, the balancing agent, the softening agent, the formulation regulator, the preservative and the water are in a relation of 2-10 parts of the active collagen extracting solution, 2-6 parts of the sodium hyaluronate, 1-4 parts of the balancing agent, 100-200 parts of the softening agent, 2-20 parts of the formulation regulator, 1-4 parts of the preservative and 1760-1900 parts of water;

mixing and stirring a part of the water, the sodium hyaluronate, the balancing agent and the dosage form regulator at 40-60 ℃ according to the mass part relation to obtain a first mixture;

adding the softening agent and the preservative into the first mixture according to the mass part relationship, and mixing and stirring to obtain a second mixture; and

and when the second mixture is cooled to room temperature, adding the active collagen extracting solution into the second mixture according to the mass part relation, stirring, adding the other part of the water, mixing and stirring to obtain the skin care product.

The beneficial effects of the embodiment of the application are that: the application provides an active collagen extracting solution, a preparation method and a system thereof, and a skin care product and a preparation method thereof, wherein when the active collagen extracting solution is prepared, fresh leather is provided, a skin layer and a subcutaneous fat layer are removed, and a first semi-finished product is obtained by cleaning with a first organic solvent and water; the first semi-finished product is crushed into muddy flesh to obtain a second semi-finished product; sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product; and adopting HCl solution to carry out presoaking on the third semi-finished product to obtain a presoaking mixture, adding softening solution to soften, wherein the temperature of the softening process is 20-40 ℃, the pH value is 2.5-4.5, adding a first group of protease or a second group of protease to carry out enzymolysis reaction, the enzymolysis liquid obtained through the enzymolysis reaction is the fourth semi-finished product, and the solution obtained by filtering the fourth semi-finished product is the active collagen extracting solution, so that the whole preparation process does not need low temperature conditions, the production difficulty of the active collagen extracting solution is reduced, and the active collagen extracting solution with low production difficulty, the preparation method and the system thereof, and the skin care product and the preparation method thereof are realized.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, it is obvious that the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without inventive efforts, wherein:

FIG. 1 is a comparison graph of SDS-PAGE gel electrophoresis of an example A, SDS-PAGE protein high molecular weight standard protein and a sigma bovine skin type I collagen as a comparative example of the prepared active collagen extract according to the method for preparing the active collagen extract provided by the present application;

fig. 2 is a Circular Dichroism (CD) spectrum of example a of the active collagen extract prepared according to the method for preparing an active collagen extract provided by the present application.

FIG. 3 is a schematic structural view of a first embodiment of a system for producing an active collagen extract provided herein;

FIG. 4 is a schematic structural view of a second embodiment of the system for producing an active collagen extract according to the present application.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. It is to be understood that the specific embodiments described herein are merely illustrative of and not restrictive on the broad application. It should be further noted that, for the convenience of description, only some of the structures related to the present application are shown in the drawings, not all of the structures. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.

The present application provides an active collagen extract. The active collagen extracting solution comprises an active collagen main body and a substrate, wherein the active collagen main body comprises triple-helix active collagen, collagen peptide and hydrolyzed gel, in the active collagen main body, the mass fraction of the triple-helix active collagen is more than or equal to 90% and less than 100%, the sum of the mass fractions of the collagen peptide and the hydrolyzed gel is more than 0 and less than or equal to 10%, and the substrate is water.

The pH of the active collagen extract is 3.5-5.5, for example, 3.5, 3.65, 3.75, 3.8, 3.95, 4, 4.2, 4.35, 4.5, 4.68, 4.85, 4.9, 5, 5.1, 5.15, 5.25, 5.3 or 5.5.

The application also provides a preparation method of the active collagen extracting solution. The preparation method of the active collagen extracting solution is used for preparing the active collagen extracting solution.

The preparation method of the active collagen extracting solution comprises the following steps:

and B11, providing fresh leather, removing the epidermis layer and the subcutaneous fat layer, and cleaning by using a first organic solvent and water to obtain a first semi-finished product.

Specifically, the leather can be mammalian leather such as cow leather, pig leather, sheep leather, horse leather, donkey leather and the like. Cutting fresh fur into fur strips with width of 20-35 cm, removing epidermis layer and subcutaneous fat layer with a skin splitting machine to leave only dermis layer, washing with 75% ethanol for 1-2 times (each time for about 30 min-1 hr), and washing with purified water for 1-2 times (each time for about 15 min-30 min).

B12, mincing the first semi-finished product into muddy flesh to obtain a second semi-finished product.

Specifically, the first semi-finished product is placed in a vertical type chopper and then is crushed into muddy meat, and the size of the muddy meat is generally less than 0.5mm-3 mm.

And B13, sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product.

Specifically, the second semi-finished product and the second organic solvent are mixed and stirred, the second organic solvent is filtered out, the meat paste is reserved, then the meat paste and the salt solution are mixed and stirred, the salt solution is filtered out, the meat paste is reserved, and then the meat paste and the water are mixed and stirred, the water is filtered out, and the meat paste is collected.

In some embodiments, the second organic solvent is a combination of at least two of acetone, ethanol, and petroleum ether. Specifically, the second organic solvent may be a mixture of acetone and ethanol, a mixture of petroleum ether and ethanol, a mixture of acetone and petroleum ether, or a mixture of acetone, ethanol and petroleum ether.

When the second organic solvent is a mixture of acetone and ethanol, the volume ratio of acetone to ethanol may be 1: 0.5-3.5, e.g., 1: 0.5,1: 0.8,1: 1,1: 1.45,1: 1.5,1: 1.82,1: 1.9,1: 2,1: 2.15, 1: 2.3,1: 2.5,1: 2.75,1: 3,1: 3.2 or 1: 3.5, etc.

When the second organic solvent is a mixture of petroleum ether and ethanol, the volume ratio of petroleum ether to ethanol may be 1: 0.5-3.5, e.g., 1: 0.5,1: 0.75,1: 1,1: 1.1,1: 1.35,1: 1.5,1: 1.75,1: 1.9, 1: 2,1: 2.1,1: 2.35,1: 2.95,1: 3,1: 3.25,1: 3.3 or 1: 3.5, etc.

The mass ratio of the second organic solvent to the second semi-finished product is 0.5-3.5: 1, e.g., 0.5: 1,0.95: 1, 1: 1,1.4: 1,1.5: 1,1.65: 1,1.7: 1,1.9: 1,2: 1,2.15: 1,2.4: 1,2.75: 1,2.95: 1,3: 1 or 3.5: 1, etc.

The salt solution is 3-15% of NaCl solution in percentage by mass. The salt solution can be 3%, 3.5%, 4%, 5%, 5.5%, 6%, 7%, 7.2%, 8%, 9.5%, 10%, 11%, 12.2%, 13%, 14% or 15% by mass.

The mass ratio of the salt solution to the second semi-finished product is 0.5-3.5: 1, for example, 0.5: 1,0.8: 1,1: 1, 1.45: 1,1.5: 1,1.65: 1,1.75: 1,1.9: 1,2: 1,2.65: 1,2.8: 1,3: 1,3.15: 1, 3.2: 1 or 3.5: 1, etc.

In the step of sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product, the mass ratio of the water to the second semi-finished product is (2.5-7): 1, e.g., 2.5: 1,3: 1,3.3: 1, 3.4: 1,3.5: 1,3.65: 1,3.7: 1,3.92: 1,4: 1,4.25: 1,4.4: 1,4.5: 1,4.9: 1,5: 1,5.3: 1,5.73: 1,6: 1,6.5: 1,6.75: 1 or 7: 1, etc.

In one embodiment, the minced meat paste is placed in a cleaning cylinder, acetone/ethanol or petroleum ether/ethanol with the mass of 1-3 times of the meat paste is added for cleaning for 1-2 h, the organic solvent is filtered out, the meat paste is retained, 5% -10% NaCl solution with the mass of 1-3 times of the meat paste is added into the cleaning cylinder again for cleaning for 1-2 h, the solution is filtered out, the meat paste is retained, finally purified water with the mass of 3-6 times of the meat paste is added for cleaning for 2 times, and the meat paste is collected.

B14, adopt HCl solution right the third semi-manufactured goods carry out presoaking, obtain the presoaking mixture, add the softening solution and soften, the temperature of softening process is 20 degrees centigrade-40 degrees centigrade, pH is 2.5-4.5, add first group protease or second group protease and carry out the enzymolysis reaction, the enzymolysis liquid that obtains through the enzymolysis reaction is the fourth semi-manufactured goods, right the solution that the fourth semi-manufactured goods filtered and obtain does active collagen extract.

The concentration of the HCl solution is 0.001mol/L to 0.02mol/L, for example, 0.001mol/L, 0.008mol/L, 0.01mol/L, 0.013mol/L, 0.017mol/L, or 0.02 mol/L. The mass ratio of the HCl solution to the third semi-finished product is 0.01-1: 1, for example, 0.01: 1,0.1: 1,0.15: 1,0.3: 1,0.5: 1,0.65: 1,0.82: 1,0.9: 1,0.95: 1 or 1: 1, etc.

The softening solution is acetic acid solution, citric acid solution or mixed solution of acetic acid and citric acid. The concentration of the softening solution is 0.01mol/L to 0.3mol/L, for example, 0.01mol/L, 0.015mol/L, 0.1mol/L, 0.15mol/L, 0.2mol/L, 0.25mol/L, 0.28mol/L or 0.3mol/L, etc. The volume ratio of the softening solution to the pre-soaking mixture is 20-60: 1, e.g., 20: 1,25: 1,30: 1,48: 1,50: 1,55: 1 or 60: 1, etc.

The temperature of the softening process can be 20 degrees centigrade, 22 degrees centigrade, 25 degrees centigrade, 26 degrees centigrade, 27 degrees centigrade, 28 degrees centigrade, 30 degrees centigrade, 32 degrees centigrade, 35 degrees centigrade, 38 degrees centigrade or 40 degrees centigrade, etc.

The pH of the softening process may be 2.5, 2.6, 2.7, 2.85, 2.9, 3, 3.2, 3.35, 3.5, 3.7, 3.95, 4, 4.15, 4.35, 4.4, or 4.5, etc.

The first group of proteases are pepsin. The mass ratio of the first group of protease to the third semi-finished product is 4-10 ‰: 1, e.g., 4 ‰: 1, 4.5 ‰: 1, 5 ‰: 1, 7 per mill: 1, 8.5 permillage: 1, 9 per mill: 1 or 10 ‰: 1, etc.

The second group of proteases comprises papain and/or bromelain. The second group of proteases may also include one or a combination of trypsin, ficin, cysteine proteases. The mass ratio of the second group of protease to the third semi-finished product is 2-6 ‰: 1, e.g., 2 ‰: 1, 2.5 ‰: 1, 3 permillage: 1, 3.5 permillage: 1, 4 permillage: 1, 5 ‰: 1 or 6 per mill: 1, etc.

In the filtering process of step B14, a two-step filtering method may be adopted, that is, the enzymatic hydrolysate obtained by the enzymatic hydrolysis reaction is filtered twice. In the first filtration, filter bags with mesh number of 30-50 can be used. In some embodiments, the mesh number of the filter bags used in the first filtration can be 30, 35, 45, 48, 50, etc. In the second filtration, a filter bag with 150-300 meshes can be used. In some embodiments, the mesh number of the filter bags used in the second filtration may be 150, 180, 200, 275, 300, or the like.

In some embodiments, the meat paste is placed in an extraction tank, a small amount of 0.01-0.015 mol/L HCl solution is added for pre-soaking to obtain a pre-soaking mixture, 0.05-0.3 mol/L citric acid solution with the volume 30-60 times that of the pre-soaking mixture is added, the pre-soaking mixture is soaked and softened at 25-28 ℃ for 12-24 h, the pH is controlled at 3.0-4.0 in the soaking process, 3-5 thousandths of papain, bromelain and trypsin are added according to the mass of the meat paste, after stirring and reacting for 24-48 h, an enzymolysis reaction solution is sequentially filtered by a 48-mesh filter bag and a 200-mesh filter bag, and the solution is collected to obtain the active collagen extracting solution.

In some embodiments, the meat paste is placed in an extraction tank, a small amount of 0.005mol/L-0.015mol/L HCl solution is added for pre-soaking to obtain a pre-soaking mixture, 0.05mol/L-0.3mol/L acetic acid solution with the volume 30 times to 50 times of that of the pre-soaking mixture is added, the pre-soaking mixture is soaked and softened for 12 hours to 36 hours at the temperature of 25 ℃ to 28 ℃, the pH value is controlled to be 3.0 to 4.5 in the soaking process, pepsin with the mass of 5 per thousand to 8 per thousand of the meat paste is added, after stirring and reacting for 24 hours to 48 hours, an enzymolysis reaction solution is sequentially filtered by a 48-mesh filter bag and a 200-mesh filter bag, and the solution is collected to obtain the active collagen extracting solution.

Referring to table 1, table 1 shows quality indexes of the active collagen extract prepared by the method for preparing the active collagen extract provided by the present application:

TABLE 1

Example a of an active collagen extract prepared using the method for preparing an active collagen extract provided herein was analyzed as follows:

example A:

cutting 1Kg of fresh leather into leather strips with the width of 25cm, removing epidermis layers and subcutaneous fat layers by a leather splitting machine, only keeping the dermis layers, then cleaning for 30 minutes by 75% ethanol, cleaning for 2 times by purified water, each time for 15 minutes, and draining water for later use; putting the cleaned skin pieces into a vertical type chopper, and then stirring the cleaned skin pieces into muddy flesh, wherein the size of the muddy flesh is about 0.5-1 mm; placing the minced meat paste into a cleaning cylinder, adding 1.5L of mixed organic reagent (V acetone: V ethanol is 1: 1), cleaning for 60min, filtering out the organic solvent, adding 1.5L of NaCl solution with the mass fraction of 5% into the cleaning tank again, cleaning for 60min, filtering out the solution, finally adding 5L of purified water, cleaning for 2 times, and collecting the meat paste; placing the meat paste in an extraction tank, firstly adding a small amount of 0.01mol/L HCl solution for pre-soaking, adding 50L of 0.1mol/L acetic acid solution with the pH value of 3.0, soaking and softening for 12h at 25 ℃, then adding 5g of pepsin, stirring and reacting for 24h, filtering enzymolysis reaction liquid by adopting a 48-mesh filter bag and a 200-mesh filter bag in sequence, and collecting filtrate to obtain the active collagen extracting solution.

Referring to table 2, table 2 shows the quality test results of the active collagen extract of example a:

TABLE 2

Therefore, the active collagen extracting solution prepared by the preparation method of the active collagen extracting solution meets the quality standard.

Referring to fig. 1, fig. 1 is a graph showing a comparison of SDS-PAGE gels of an example A, SDS-PAGE protein high molecular weight standard protein of the active collagen extract prepared by the method for preparing the active collagen extract provided by the present application, and a comparative example of sigma bovine type I collagen.

As can be seen from FIG. 1, the active collagen extract obtained by the present application has a similar composition to sigma cow leather type I collagen, and has distinct alpha 1, alpha 2 and beta bands, the band near the relative molecular mass of 245kDa is a beta chain, the two bands near 100kDa are alpha 1 and alpha 2 chains, respectively, and the alpha 1, alpha 2 and beta chains of collagen and the triple helix structure formed by the two bands are relatively completely preserved during the treatment process.

Further, referring to fig. 2, fig. 2 is a Circular Dichroism (CD) spectrum of example a of the active collagen extracting solution prepared according to the method for preparing an active collagen extracting solution provided by the present application. The CD spectroscopy is used for the characterization of the triple-helical structure of the collagen, the CD spectrogram of the physiological collagen has a negative peak at a wavelength of about 195nm and a positive peak at a wavelength of about 220nm, the positive absorption peak is a typical characteristic of a circular dichroism spectrum with a levorotatory polyproline configuration, and the negative peak position is added, so that the completeness of the triple-helical structure of the collagen is indicated. If no positive peak at 222nm is detected, this indicates the absence of triple helix structure. The larger the ratio of the positive peak to the negative peak is, the more complete the triple helix structure is, and the higher the content is, so that the relative quantitative analysis of the triple helix content can be carried out by utilizing the ratio of the positive peak to the negative peak.

As can be seen from FIG. 2, the Circular Dichroism (CD) spectrogram of the active collagen extract obtained in the present application has a negative peak at 197nm and a positive peak at 222nm, and the ratio of the positive peak to the negative peak is large, which indicates that the triple helix structure is relatively complete.

In addition, in order to quantitatively determine the content of the active collagen containing triple-helix structure in the active collagen extract obtained in the present application, a trypsin sensitivity analysis method in technical consensus, characterization and quality evaluation of medical collagen products, is adopted to quantitatively detect a sample, and the trypsin sensitivity can detect a denatured collagen part. Trypsin digests the denatured parts of collagen (collagen peptides and hydrolyzed collagen), while triple helix collagen resists digestion by most proteases, and the denatured collagen content was analyzed by determining the hydroxyproline content of the digest, with the results shown in table 3:

sample numbering	Number	1	Number 2	No. 3	Mean value of
						Triple helix active collagenContent/%	92.5	91.7	90.9	91.7

TABLE 3

Further, the active collagen extract of example a was subjected to cell proliferation assay:

the cell proliferation assay for active collagen extraction of example A was performed as described in the literature (Chen R N, Ho H O, Sheu M T. Characterization of collagen materials crosslinked using a microbial transglutaminase [ J ]. Biomaterials,2005,26(20): 4229-4235).

Using the murine fibroblast cell line NIH/3T3 cell line, the active collagen extract was aseptically plated in 48-well plates approximately 2mm thick and lyophilized for use with a lyophilizer. Then, ultraviolet irradiation is carried out on the freeze-dried test sample of the 48-pore plate in an ultra-clean workbench for 30 minutes, the concentration of mouse fibroblasts which are in a good digestion state and in a logarithmic phase is adjusted to be 1 multiplied by 104/ml, the mouse fibroblasts are inoculated on the freeze-dried test sample, 400 mu L of the mouse fibroblasts is added to each pore, and the test sample is set as an experimental group;

cells of the same density were directly inoculated into a well plate (no collagen sample in the well plate), 400. mu.L per well, and set as a control group;

the 48-well plate containing the lyophilized sample was placed in an ultraclean bench and irradiated with ultraviolet light for 30 minutes, and 400. mu.L of the culture medium (containing no cells) was directly added to each well, thereby setting the plate as a zero-adjustment group.

5% CO at 37 deg.C₂And incubating and culturing under the saturated humidity condition, replacing culture solution at intervals of 24h, and regularly observing cell morphology by using an inverted microscope.

After culturing for 24h (1 day), 72h (3 days), and 120h (5 days), the cell viability was examined by MTT assay kit, and the cell proliferation promoting ability of the sample was evaluated based on the absorbance value (A570) at 570nm (630 nm as a reference wavelength).

Experimental group a570 ═ experimental group samples were measured for absorbance values-zero set absorbance values.

Remarking: the above test was averaged over 3 multiple wells.

Referring to table 4, table 4 shows the results of the cell proliferation assay.

Time (h)	Experimental group	Zero setting group	Control group
					0	0.072	0.048	0.041
24	0.204	0.063	0.175
				72	0.455	0.052	0.345
120	0.852	0.084	0.964

TABLE 4

From the above test data, the light absorption value of the experimental group increases with time, and the growth and proliferation conditions of the cells in the pore plate of the experimental group are better, which indicates that the active collagen extracting solution prepared by the embodiment has better capability of promoting cell proliferation.

Referring to fig. 3-4, the present application further provides a system for preparing an active collagen extract. The system 10 for preparing an active collagen extracting solution is used for preparing the active collagen extracting solution.

The preparation system 10 of the active collagen extracting solution comprises a splitting equipment group 11, a chopping equipment 12, an impurity removing equipment group 13 and an extracting equipment group 14.

The splitting equipment set 11 is used for removing the epidermis layer and the subcutaneous fat layer of the fresh leather, and cleaning the fresh leather by adopting a first organic solvent and water to obtain a first semi-finished product.

Specifically, the splitting equipment set 11 includes a cutter 111, a splitting machine 112, and a cleaning device 113.

The cutting machine 111 is used for cutting the fresh leather to obtain the leather strips. The cutting machine 111 is used for cutting the fresh leather into leather strips with the width of 20cm-35 cm.

The peeler 112 is configured to remove the epidermis layer and the subcutaneous fat layer of the skin strip to obtain a first intermediate product.

The cleaning device 113 is configured to clean the first intermediate product with the first organic solvent and the water to obtain the first semi-finished product.

The cutter 111 has a cutting feed port and a cutting discharge port. The splitting machine 112 has a splitting inlet and a splitting outlet. The cleaning device 113 comprises a cleaning tank, a leather strip feeding port, a first organic solvent feeding port, a water feeding port and a leather strip discharging port. The leather strip feeding port, the first organic solvent feeding port, the water feeding port and the leather strip discharging port are all communicated with the cleaning tank. The leather material strip feeding port, the first organic solvent feeding port, the water feeding port and the leather material strip discharging port are the same opening or different openings.

Feeding fresh leather into the cutting machine 111 from the cutting feed opening, cutting the cowhide by the cutting machine 111 to form the leather strips, and outputting the leather strips from the cutting feed opening; the leather strips output from the cutting discharge port enter the splitting machine 112 from the splitting feeding port, and are processed by the splitting machine 112 to obtain the first intermediate product, and the first intermediate product is output from the splitting discharge port; and the first intermediate product output from the sheet leather discharge port enters the cleaning tank from the leather material strip feeding port, the first organic solvent is added from the first organic solvent feeding port for cleaning, after the cleaning is finished, the first organic solvent is removed, water is added from the water feeding port for cleaning, after the cleaning is finished, the water is removed, the first semi-finished product is obtained, and the first semi-finished product is output from the leather material strip discharge port.

In some embodiments, the hide strip feed port, the first organic solvent feed port, the water feed port, and the hide strip discharge port are the same opening. In other embodiments, the leather strip feeding port, the first organic solvent feeding port, the water feeding port and the leather strip discharging port can be different openings.

The cleaning device 113 may further include a liquid outlet for discharging the first organic solvent and the water.

The mincing device 12 is used for mincing the first semi-finished product into muddy flesh to obtain a second semi-finished product. The meat paste is the second semi-finished product.

Specifically, the first semi-finished product is placed in a vertical type chopper and then is crushed into muddy flesh. The size of the meat paste is generally less than 0.5mm-3 mm.

And the impurity removing equipment group 13 is used for sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product.

The trash equipment group 13 includes a first cleaning cylinder 131a, a first trash filtering equipment 131b, a second cleaning cylinder 132a, a second trash filtering equipment 132b, a third cleaning cylinder 133a, and a third trash filtering equipment 133 b.

The first cleaning cylinder 131a is used for mixing the second organic solvent and the second semi-finished product. The first impurity removing and filtering device 131b is used for filtering the mixture in the first cleaning cylinder 131a, and filtering out liquid to obtain a second intermediate product.

The second washing cylinder 132a is used for mixing the salt solution and the second intermediate product. The second impurity removing and filtering device 132b is used for filtering the mixture in the second cleaning cylinder 132a, and filtering out liquid to obtain a third intermediate product.

The third washing cylinder 133a is used to mix the water and the third intermediate product. And the third impurity removing and filtering device 133b is used for filtering the mixture in the third cleaning cylinder 133a to remove liquid, so as to obtain the third semi-finished product.

The first cleaning cylinder 131a, the second cleaning cylinder 132a and the third cleaning cylinder 133a may be the same cleaning cylinder or different cleaning cylinders, and the first impurity removing and filtering device 131b, the second impurity removing and filtering device 132b and the third impurity removing and filtering device 133b may be the same impurity removing and filtering device or different impurity removing and filtering devices.

As shown in fig. 3, the first cleaning cylinder 131a, the second cleaning cylinder 132a and the third cleaning cylinder 133a are the same cleaning cylinder, and the first impurity removing and filtering device 131b, the second impurity removing and filtering device 132b and the third impurity removing and filtering device 133b are the same impurity removing and filtering device.

As shown in fig. 4, the first cleaning cylinder 131a, the second cleaning cylinder 132a, and the third cleaning cylinder 133a are different cleaning cylinders, and the first impurity-removing filtering device 131b, the second impurity-removing filtering device 132b, and the third impurity-removing filtering device 133b are different impurity-removing filtering devices. The first cleaning cylinder 131a, the first impurity removing and filtering device 131b, the second cleaning cylinder 132a, the second impurity removing and filtering device 132b, the third cleaning cylinder 133a and the third impurity removing and filtering device 133b are sequentially connected between the chopping device 12 and the extraction device group 14.

The cleaning apparatus group 13 further comprises a meter 134. The meter 134 is used to perform mass calculation on the second semi-finished product charged into the first washing cylinder 131a, on the second organic solvent charged into the first washing cylinder 131a, on the salt solution charged into the second washing cylinder 132a, and on the water charged into the third washing cylinder 133 a. The scale 134 may be a scale. In other embodiments, the meter 134 may also be used to perform a volume calculation for the second semi-finished product added to the first cleaning cylinder 131a, for the second organic solvent added to the first cleaning cylinder 131a, for the salt solution added to the second cleaning cylinder 132a, and for the water added to the third cleaning cylinder 133 a. The gauge 134 may be a graduated cylinder. It should be noted that the mass and the volume can be converted from the relationship between the mass and the volume based on the mass or the volume calculated by the meter 134.

Further, to meet the requirement of industrial automation production, the impurity removing equipment group 13 may further include a processor 135. The processor 135 is configured to control a mass ratio of the second organic solvent to the second semi-finished product, to control a mass ratio of the salt solution to the second semi-finished product, and to control a mass ratio of the water added to the impurity removing apparatus to the second semi-finished product. Specifically, the processor 135 is configured to control a mass ratio of the second organic solvent to the second semi-finished product to be 0.5-3.5: 1, controlling the mass ratio of the salt solution to the second semi-finished product to be 0.5-3.5: 1, and controlling the mass ratio of the water added into the impurity removing equipment to the second semi-finished product to be 2.5-7: 1.

the extraction equipment group 14 is used for performing pre-soaking, softening and enzymolysis reaction on the third semi-finished product, the enzymolysis liquid obtained through the enzymolysis reaction is a fourth semi-finished product, and the solution obtained by filtering the fourth semi-finished product is the active collagen extracting solution. Specifically, in some embodiments, the extraction equipment group 14 can be used to adopt HCl solution right the third semi-finished product is soaked in advance, obtains soaking mixture in advance, adds the softening solution and softens, and the temperature of softening process is 20 degrees centigrade-40 degrees centigrade, and pH is 2.5-4.5, adds first group protease or the protease of second group and carries out enzymolysis reaction, and the enzymolysis liquid that obtains through enzymolysis reaction is the fourth semi-finished product, right the solution that the fourth semi-finished product was filtered and is obtained does active collagen extract.

Specifically, the extraction apparatus group 14 includes an extraction tank 141, a stirrer 142, and an extraction liquid filtering apparatus 143.

The extracting tank 141 is used for adopting the HCl solution to pre-soak the third semi-finished product to obtain a pre-soaking mixture, adding the softening solution to soften, adding the first group of protease or the second group of protease to perform an enzymolysis reaction, and obtaining the enzymolysis liquid through the enzymolysis reaction as the fourth semi-finished product.

The stirrer 142 is used for stirring the mixture in the extraction tank 141.

The extracting solution filtering device 143 is configured to filter the fourth semi-finished product, and an obtained solution is the active collagen extracting solution.

The extraction equipment set 14 may also include a meter 144, a thermometer 145, and a pH meter 146.

The meter 144 is used to calculate the volume of the softening solution added to the extraction tank 141 and the mass of the first or second group of proteases. The scale 144 may be a measuring cylinder and a scale.

The thermometer 145 is used to detect the temperature of the mixture in the extraction tank 141.

The pH meter 146 is used for detecting the pH value of the mixture in the extraction tank 141.

Further, in order to meet the requirements of industrial automation production, the extraction equipment group 14 further comprises a processor 147. The processor 147 is configured to control the volume ratio of the softening solution to the pre-soak mixture, the mass ratio of the first group of proteases to the third intermediate product, the mass ratio of the second group of proteases to the third intermediate product, the pH of the softening process, and the temperature of the softening process. Specifically, in one embodiment, the processor 147 may be configured to control the volume ratio of the softening solution to the pre-soak mixture to be in a range of 20-60: 1, the mass ratio of the first group of protease to the third semi-finished product is 4-10 ‰: 1, the mass ratio of the second group of protease to the third semi-finished product is 2-6 ‰: 1, controlling the addition of the softening solution to maintain the pH value at 2.5-4.5, and controlling the temperature in the softening process at 20-40 ℃.

The set of extraction devices 14 may further comprise a display component 148, and the processor 147 is further configured to control the display component 148 to display the mass calculated by the meter 144, the temperature sensed by the thermometer 145, and the pH value sensed by the pH meter 146.

The extract filtering device 143 comprises a first sub-device 1431 and a second sub-device 1432. The filter bags of the first subset 1431 have a mesh number of 30-50. The filter bag of the second subset 1432 has a mesh size of 150-300. The first sub-device 1431 is configured to filter the fourth semi-finished product to obtain a semi-finished product of an extracting solution. The second sub-device 1432 is configured to filter the semi-finished product of the extracting solution to obtain the active collagen extracting solution.

The present application also provides a skin care product. The skin care product comprises active collagen extract, sodium hyaluronate, balancing agent, softening agent, formulation regulator, preservative and water. The active collagen extracting solution is the active collagen extracting solution. The skin care product can also comprise perfume, essential oil, pigment and the like. The active collagen extracting solution, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative and the water are in a mass part relationship of 2-10 parts of the active collagen extracting solution, 2-6 parts of the sodium hyaluronate, 1-4 parts of the balancing agent, 100-200 parts of the emollient, 2-20 parts of the formulation regulator, 1-4 parts of the preservative and 1760-1900 parts of water. The amounts of the components in the skin care product can be adjusted so that the skin care product has various product forms, such as skin care essence, skin care water, skin care milk, skin care gel, facial mask, skin care cream or skin care cream, and the like.

In some embodiments, the active collagen extraction solution, the sodium hyaluronate, the balancing agent, the emollient, the formulation modifier, the preservative, and the water may be in a mass part relationship of 5 parts of the active collagen extraction solution, 5 parts of the sodium hyaluronate, 2 parts of the balancing agent, 150 parts of the emollient, 10 parts of the formulation modifier, 2 parts of the preservative, and 1800 parts of water.

In some embodiments, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative, and the water may be in a relationship of parts by mass of 8 parts of the active collagen extraction solution, 6 parts of the sodium hyaluronate, 1 part of the balancing agent, 200 parts of the emollient, 5 parts of the formulation regulator, 1 part of the preservative, and 1900 parts of water.

In some embodiments, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative, and the water may be in a relationship of parts by mass of 3 parts of the active collagen extraction solution, 2 parts of the sodium hyaluronate, 4 parts of the balancing agent, 100 parts of the emollient, 15 parts of the formulation regulator, 3 parts of the preservative, and 1850 parts of water.

In some embodiments, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative, and the water may be related in parts by mass in the range of 6 parts of the active collagen extraction solution, 3 parts of the sodium hyaluronate, 1 part of the balancing agent, 180 parts of the emollient, 20 parts of the formulation regulator, 2 parts of the preservative, and 1760 parts of water.

In some embodiments, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative, and the water may be in a relationship of parts by mass of 4 parts of the active collagen extraction solution, 4 parts of the sodium hyaluronate, 4 parts of the balancing agent, 120 parts of the emollient, 10 parts of the formulation regulator, 2 parts of the preservative, and 1900 parts of water.

The sodium hyaluronate is one or a combination of several of sodium hyaluronate with molecular weight of 120-150 ten thousand, 80-100 ten thousand, 20-40 ten thousand and 1-10 ten thousand daltons.

The balancing agent comprises one or more of dipotassium glycyrrhizinate, sodium dihydrogen phosphate and disodium hydrogen phosphate. The balancing agent can also comprise one or more of triethanolamine, methyl propanol, potassium hydroxide and sodium hydroxide.

The dosage form regulator is one or the combination of two of hydroxyethyl cellulose and xanthan gum.

The preservative comprises one or the combination of two of caprylyl hydroximic acid and ethylhexyl glycerin. The preservative can also comprise one or more of methyl hydroxybenzoate, chlorphenesin, benzoic acid, sorbic acid, salicylate, hydroxybenzoate esters, nipagin, p-chloroxylenol, 2, 4, 4' -trichloro-N-carbanilide, alkyl trimethyl ammonium chloride, alkyl brominated quinoline, cetyl pyridinium chloride, ethanol, methyl dibromoglutaronitrile, phenoxyethanol, diazolidinyl urea and iodine.

The pH of the active collagen extracting solution is 3.5-5.5, and the pH of the skin care product is 4.5-7.

b21, providing an active collagen extracting solution, sodium hyaluronate, a balancing agent, a softening agent, a formulation regulator, a preservative and water, wherein the active collagen extracting solution is the active collagen extracting solution, and the mass parts of the active collagen extracting solution, the sodium hyaluronate, the balancing agent, the softening agent, the formulation regulator, the preservative and the water are in a relation of 2-10 parts of the active collagen extracting solution, 2-6 parts of the sodium hyaluronate, 1-4 parts of the balancing agent, 100-200 parts of the softening agent, 2-20 parts of the formulation regulator, 1-4 parts of the preservative and 1760-1900 parts of water;

in some embodiments, the active collagen extraction solution, the sodium hyaluronate, the balancing agent, the emollient, the formulation modifier, the preservative, and the water may be related in parts by mass in the range of 5 parts of the active collagen extraction solution, 5 parts of the sodium hyaluronate, 2 parts of the balancing agent, 150 parts of the emollient, 10 parts of the formulation modifier, 2 parts of the preservative, and 1800 parts of water.

In some embodiments, the sodium hyaluronate, the balancing agent, the emollient, the formulation modifier, the preservative, and the water may be in a mass parts relationship of 8 parts of the active collagen extract, 6 parts of the sodium hyaluronate, 1 part of the balancing agent, 200 parts of the emollient, 5 parts of the formulation modifier, 1 part of the preservative, and 1900 parts of water.

The balancing agent comprises one or more of dipotassium glycyrrhizinate, sodium dihydrogen phosphate and disodium hydrogen phosphate. The balancing agent can also comprise one or a combination of triethanolamine, methyl propanol, potassium hydroxide and sodium hydroxide.

The softening agent is one or more of glycerol, butanediol, propylene glycol, glyceryl caprylate, 1, 2-hexanediol and glyceryl polyether-26.

And B22, mixing and stirring a part of the water, the sodium hyaluronate, the balancing agent and the formulation regulator at 40-60 ℃ according to the mass part relation to obtain a first mixture.

And B23, adding the softening agent and the preservative into the first mixture according to the mass part relation to obtain a second mixture.

And B24, cooling the second mixture to room temperature, adding the active collagen extracting solution into the second mixture according to the mass part relation, stirring, adding the other part of the water, and stirring to obtain the skin care product.

Referring to table 5, table 5 shows the quality indexes of the skin care product prepared by the skin care product preparation method provided by the present application:

TABLE 5

Example B, using the method of preparation of the skin care product provided herein, was analyzed as follows:

example B:

according to the proportion, 50.0KG of purified water, 100.0g of sodium hyaluronate (50.0 g with the molecular weight of 120-150 ten thousand, 25.0g with the molecular weight of 20-40 ten thousand and 25.0g with the molecular weight of 1-10 ten thousand), 100.0g of dipotassium glycyrrhizinate, 100.0g of hydroxyethyl cellulose and 100.0g of xanthan gum are sequentially put into a stirring tank and stirred and homogenized for 60min at 50 ℃, then 3.5KG of glycerol, 1KG of butanediol, 0.5KG of propanediol, 0.6KG of 1, 2-hexanediol, 261.0 KG of glycerol polyether and 7.0g of glyceryl caprylate and 70.0g of caprylyl hydroxamic acid are sequentially put into the tank, stirred and uniformly and cooled to room temperature, 37.0KG of the active collagen extracting solution prepared in the embodiment A is added, the residual water is supplemented to 6.3KG to 100KG, and stirred uniformly, and the skin care product is obtained.

Wherein the balancing agent is dipotassium glycyrrhizinate, the dosage form regulator is the combination of hydroxyethyl cellulose and xanthan gum, the emollient is the combination of glycerol, 1kg of butanediol, propylene glycol, 1, 2-hexanediol, glyceryl caprylate and glyceryl polyether-26, and the preservative is caprylhydroxamic acid.

Referring to table 6, table 6 shows the quality test results of the skin care product of example B:

TABLE 6

Therefore, the skin care product prepared by the preparation method of the skin care product meets the quality standard.

Further, the skin care product of example B was tested for moisturizing effect:

to test the moisturizing effect of the skin care prepared in example B and the comparative example, a process of applying cream on human skin was simulated using a small glass plate and a medical air-permeable adhesive tape as biomaterials such as a simulated horny layer, an epidermis, etc., 0.2g of skin care solution was applied on a prepared 0.9cm × 5.5cm simulation device, the device was placed in an oven at 25 ℃, the mass of the simulation device was precisely measured at intervals of 5min, the mass m1 before the sample was placed and the mass m2 after the sample was placed were precisely measured, and the moisturizing rate after 30min was calculated.

Referring to table 7, table 7 shows the results of the skin care product of example B in the moisture retention effect test.

	Sample 1	Sample 2	Sample 3	Comparative example
					In vitro moisture retention rate	49.97％	55.83％	57.38％	28.12％

TABLE 7

As can be seen from the data in table 7, the skin lotion prepared in this example has a good moisturizing and hydrating effect, which indicates that the skin lotion components of the example have a good moisturizing and hydrating effect.

Further, the skin care product of example B was tested for antioxidant effect:

to test the antioxidant effect of the lotions prepared in example B and comparative example, skin care products having antioxidant effect were purchased on the market as a general control group.

Dissolving DPPH free radical in absolute ethyl alcohol to prepare 2 x 10^-8A solution of DPPH in mol/L. Respectively adding 2mL of skin care essence into the test tubes, and addingAnd (2) strongly oscillating the 2mLDPPH solution, standing the solution in a dark place for 30min, taking absolute ethyl alcohol as a reference, respectively measuring the absorbance value of each reaction solution at 517nm, and calculating the removal rate of each sample to be measured on DPPH free radicals.

Referring to Table 8, Table 8 shows the results of the skin care product of example B for antioxidant effect testing.

	Sample 1	Sample 2	Sample 3	Common control
					DPPH radical clearance	78.19％	82.16％	85.19％	71.92％

TABLE 8

As can be seen from the data in table 8, the skin care prepared in this embodiment has a good antioxidant effect, which indicates that the collagen and the sodium hyaluronate have a good antioxidant effect after being compounded, and the sodium hyaluronate with both small and large molecules has a better effect after being compounded.

Further, the skin care product of example B was tested for tyrosinase inhibition:

to test the tyrosinase inhibition effect of the skin lotions prepared in the examples and comparative examples, products on the market, which are declared to have skin color-lightening efficacy, were purchased as a general control group. The test method was carried out in accordance with the "cosmetic-tyrosinase activity inhibition test method" which is a group standard published by the society for daily chemicals of the Shanghai.

1mL of skin care lotion, 0.9mL of phosphate buffer with pH value of 6.8 and 1mL of 0.03% tyrosine are respectively put into a test tube, 0.1mL of tyrosinase aqueous solution (1070U/mL) is added, the solution is uniformly mixed and placed in a water bath with constant 37 ℃ for reaction for 10 minutes, 2mL of levodopa solution is added, the reaction time of each tube is controlled to be 5 minutes, and the absorbance value A1 at 475nm is measured by a spectrophotometer. The tyrosinase was replaced by an equal volume of phosphate buffer and its absorbance value A2 was measured. The skin lotion was replaced with an equal volume of phosphate buffer and the absorbance value A3 was measured. The tyrosinase and skin lotion were replaced by equal volume of phosphate buffer, and the absorbance value A4 was measured. The inhibition rate of the skin lotion on tyrosinase was calculated, and the inhibition rate (%) was 100% x [1- (a1-a2/A3-a4) ].

Referring to Table 9, Table 9 shows the results of tyrosinase inhibition testing of the skin care product of example B.

	Sample No. 1	Sample 2	Sample 3	Common control
					Tyrosinase inhibition rate	80.37％	81.48％	87.83％	60.31％

TABLE 9

As can be seen from the data in table 9, the skin lotion sample prepared in this example has a good tyrosinase inhibition effect, and can achieve the effects of whitening skin and brightening skin.

In the skin care product provided by the application, the macromolecular and micromolecular functional component systems of the collagen extracting solution and the sodium hyaluronate enable macromolecular active components to act on the skin surface and the epidermal layer, and micromolecular active components act on the dermis layer, so that the effects of internal and external concurrent modification are achieved. The skin care lotion has pH of 4.5-7, and can provide weak acid environment for skin to inhibit bacterial growth and improve secretion and excretion of sebaceous gland.

When the active collagen extracting solution is prepared, fresh leather is provided, a skin layer and a subcutaneous fat layer are removed, and a first semi-finished product is obtained by adopting a first organic solvent and water for cleaning; the first semi-finished product is crushed into muddy flesh to obtain a second semi-finished product; sequentially cleaning the second semi-finished product by adopting a second organic solvent, a salt solution and water to obtain a third semi-finished product; and adopting HCl solution to carry out presoaking on the third semi-finished product to obtain a presoaking mixture, adding softening solution to soften, wherein the temperature of the softening process is 20-40 ℃, the pH value is 2.5-4.5, adding a first group of protease or a second group of protease to carry out enzymolysis reaction, the enzymolysis liquid obtained through the enzymolysis reaction is the fourth semi-finished product, and the solution obtained by filtering the fourth semi-finished product is the active collagen extracting solution, so that the whole preparation process does not need low temperature conditions, the production difficulty of the active collagen extracting solution is reduced, and the active collagen extracting solution with low production difficulty, the preparation method and the system thereof, and the skin care product and the preparation method thereof are realized.

In addition, the extraction of macromolecular active collagen at present generally aims at the biomedical field, the extraction of a proto-tissue material source needs to meet the strict requirements of laws and regulations of an animal source, the extraction process is complex and tedious, the conditions are harsh, the extraction cost is high, the method can be only used in the biomedical field with high added value, has great limitations and is difficult to produce and apply in a large scale, and the active collagen prepared by the complex process in the cosmetic field is used as an effective component to be added into a product.

Most of collagen cosmetics/skin care products on the market at present are collagen polypeptide or hydrolyzed collagen, and few patent reports of products for care mainly based on active collagen are reported, particularly, the products are applied and developed on the basis of prepared active collagen extracting solution, the active collagen plays an important role in the aspect of endowing the products with multiple effects due to the special structure and functional characteristics of the active collagen, and the existing cosmetics/skin care products have complex and various components and complicated preparation processes.

Active collagen is poorly water soluble and needs to be solubilized under acidic conditions, while it carries negative charges under acidic conditions, which causes problems with the compatibility of collagen in cosmetic/skin care applications, such as: in a solution lower than the isoelectric point of the collagen, the collagen presents electropositivity, and the sodium hyaluronate presents electronegativity, so that the collagen and the sodium hyaluronate have stronger electrostatic attraction effect when being blended, flocculent polymer precipitation is generated, and the compatibility is poor. At present, strong electrolyte such as sodium chloride or phosphate is generally used as a balancing substance to solve the problem, but the addition amount of salt is more, so that the sticky feeling and the texture of the solution are obviously changed, water molecules in the solution can generate hydration with small molecular salt, and OH in the water^-Can form hydrogen bonds with collagen and hydroxyl in hyaluronic acid molecules, and destroy the influence of the hydrogen bonds among the original collagen molecules on the triple-helical structure of the collagen, thereby influencing the activity of the collagen.

The extraction process of the active collagen provided by the application is simple and efficient, and the cost of the active collagen raw material can be effectively controlled; the skin care product taking collagen as a core component has the advantages of simple components, reasonable compatibility of all components, good compatibility, uniformity and stability, simple preparation method and easy large-scale industrial production; the skin care product is mild and non-irritant, has a good skin repairing function, effectively promotes cell proliferation, and has the effects of repairing, nourishing skin, moisturizing, resisting oxidation, brightening skin and firming skin.

The above description is only for the purpose of illustrating embodiments of the present application and is not intended to limit the scope of the present application, and all modifications of equivalent structures and equivalent processes, which are made by the contents of the specification and the drawings of the present application or are directly or indirectly applied to other related technical fields, are also included in the scope of the present application.

Claims

1. An active collagen extracting solution, which is characterized by comprising an active collagen main body and a substrate, wherein the active collagen main body comprises triple-helix active collagen, collagen peptide and hydrolyzed glue, in the active collagen main body, the mass fraction of the triple-helix active collagen is greater than or equal to 90% and less than 100%, the sum of the mass fractions of the collagen peptide and the hydrolyzed glue is greater than 0 and less than or equal to 10%, and the substrate is water.

2. The active collagen extract according to claim 1, wherein the pH of said active collagen extract is 3.5 to 5.5.

3. A method for producing an active collagen extract according to claim 1 or 2, which comprises:

4. The method of producing an active collagen extract according to claim 3, wherein said first organic solvent is 75% ethanol.

5. The method of preparing an active collagen extract according to claim 3, wherein the second organic solvent is a combination of at least two of acetone, ethanol and petroleum ether, and the mass ratio of the second organic solvent to the second semi-finished product is 0.5 to 3.5: 1.

6. the method of preparing an active collagen extract according to claim 5, wherein said second organic solvent is a mixture of acetone and ethanol, wherein the volume ratio of acetone to ethanol is 1: 0.5-3.5, or the second organic solvent is a mixture of petroleum ether and ethanol, and the volume ratio of the petroleum ether to the ethanol is 1: 0.5-3.5.

7. The method for preparing an active collagen extracting solution according to claim 3, wherein the salt solution is a NaCl solution with a mass fraction of 3% to 15%, and the mass ratio of the salt solution to the second semi-finished product is 0.5 to 3.5: 1.

8. the method for preparing an active collagen extracting solution according to claim 3, wherein in the step of sequentially washing the second semi-finished product with a second organic solvent, a salt solution and water to obtain a third semi-finished product, the mass ratio of the water to the second semi-finished product is 2.5-7: 1.

9. the method for preparing active collagen extracting solution according to claim 3, wherein the concentration of HCl solution is 0.001mol/L-0.02mol/L, the concentration of softening solution is 0.01mol/L-0.3mol/L, the first group of protease is pepsin, and the second group of protease comprises papain and/or bromelain, and the softening solution is acetic acid solution, citric acid solution or a mixed solution of acetic acid and citric acid.

10. The method of claim 9, wherein the second group of proteases further comprises one or more of trypsin, ficin, and cysteine proteases.

11. The method for preparing an active collagen extract according to claim 9 or 10, wherein the volume ratio of said softening solution to said pre-soaking mixture is 20-60: 1, the mass ratio of the first group of protease to the third semi-finished product is 4-10 ‰: 1, the mass ratio of the second group of protease to the third semi-finished product is 2-6 ‰: 1.

12. a system for producing an active collagen extracting solution, for producing the active collagen extracting solution according to claim 1 or 2, comprising:

13. The system for preparing active collagen extracting solution according to claim 12, wherein the skin slicing apparatus set comprises a cutting machine, a skin slicing machine and a cleaning device, the cutting machine is configured to cut fresh skin to obtain strips of skin, the skin slicing machine is configured to remove the epidermis layer and subcutaneous fat layer of the strips of skin to obtain the first intermediate product, and the cleaning device is configured to clean the first intermediate product with the first organic solvent and the water to obtain the first semi-finished product.

14. The system for preparing active collagen extracting solution according to claim 13, wherein the cleaning device comprises a cleaning tank, a skin strip feeding port, a first organic solvent feeding port, a water feeding port and a skin strip discharging port, the skin strip feeding port, the first organic solvent feeding port, the water feeding port and the skin strip discharging port are all communicated with the cleaning tank, and the skin strip feeding port, the first organic solvent feeding port, the water feeding port and the skin strip discharging port are the same opening or different openings.

15. The system according to claim 12, wherein the impurity removing equipment set comprises a first cleaning tank, a first impurity removing and filtering equipment, a second cleaning tank, a second impurity removing and filtering equipment, a third cleaning tank and a third impurity removing and filtering equipment, the first cleaning tank is used for mixing the second organic solvent and the second semi-finished product, the first impurity removing and filtering equipment is used for filtering the mixture in the first cleaning tank to remove liquid and obtain a second intermediate product, the second cleaning tank is used for mixing the salt solution and the second intermediate product, the second impurity removing and filtering equipment is used for filtering the mixture in the second cleaning tank to remove liquid and obtain a third intermediate product, and the third cleaning tank is used for mixing the water and the third intermediate product, the third edulcoration filtration equipment is used for right the mixture in the third washing jar filters, strains liquid, obtains the third semi-manufactured goods, first washing jar second washing jar with the third washing jar is same washing jar or different washing jars, first edulcoration filtration equipment second edulcoration filtration equipment with third edulcoration filtration equipment is same edulcoration filtration equipment or different edulcoration filtration equipment.

16. The system for preparing an active collagen extracting solution according to claim 15, wherein the trash extraction equipment set further comprises a meter for performing mass or volume calculation of the second semi-finished product added to the first washing tank, the second organic solvent added to the first washing tank, the salt solution added to the second washing tank, and the water added to the third washing tank.

17. The system for preparing an active collagen extracting solution according to claim 16, wherein the impurity removing equipment set further comprises a processor for controlling a mass ratio of the second organic solvent to the second semi-finished product, for controlling a mass ratio of the salt solution to the second semi-finished product, and for controlling a mass ratio of the water added to the impurity removing equipment to the second semi-finished product.

18. The system for preparing active collagen extracting solution according to claim 12, wherein the extracting device set comprises an extracting tank, an agitator and an extracting solution filtering device, the extracting tank is configured to pre-soak the third semi-finished product with HCl solution to obtain a pre-soaked mixture, add a softening solution to soften the third semi-finished product, add the first group of proteases or the second group of proteases to perform the enzymatic hydrolysis reaction, the enzymatic hydrolysate obtained by the enzymatic hydrolysis reaction is the fourth semi-finished product, the agitator is configured to agitate the mixture in the extracting tank, the extracting solution filtering device is configured to filter the fourth semi-finished product, and the obtained solution is the active collagen extracting solution.

19. The system for preparing an active collagen extracting solution according to claim 18, wherein said extraction equipment set further comprises a meter for calculating the volume of the softening solution added to the extraction tank and the mass of the first group of proteases or the second group of proteases, a thermometer for detecting the temperature of the mixture in the extraction tank, and a pH meter for detecting the pH of the mixture in the extraction tank.

20. The system for preparing an active collagen extraction solution according to claim 19, wherein said set of extraction devices further comprises a processor for controlling the volume ratio of said softening solution to said pre-soaking mixture, controlling the mass ratio of said first group of proteases to said third semi-finished product, controlling the mass ratio of said second group of proteases to said third semi-finished product, controlling the pH of the softening process, and controlling the temperature of the softening process.

21. The system for preparing an active collagen extraction solution according to claim 20, wherein said set of extraction devices further comprises a display unit, and said processor is further configured to control said display unit to display the mass calculated by said meter, the temperature detected by said thermometer, and the pH detected by said pH meter.

22. The system for preparing active collagen extracting solution as claimed in claim 18, wherein the extracting solution filtering device comprises a first sub-device and a second sub-device, the mesh number of the filter bags of the first sub-device is 30-50, the mesh number of the filter bags of the second sub-device is 150-300, the first sub-device is used for filtering the fourth semi-finished product to obtain the extracting solution semi-finished product, and the second sub-device is used for filtering the extracting solution semi-finished product to obtain the active collagen extracting solution.

23. A skin care product characterized by comprising an active collagen extraction solution, sodium hyaluronate, a balancing agent, an emollient, a formulation regulator, a preservative and water, wherein the active collagen extraction solution is the active collagen extraction solution according to claim 1 or 2, and the mass parts of the active collagen extraction solution, the sodium hyaluronate, the balancing agent, the emollient, the formulation regulator, the preservative and the water are in a relationship of 2-10 parts of the active collagen extraction solution, 2-6 parts of the sodium hyaluronate, 1-4 parts of the balancing agent, 100-200 parts of the emollient, 2-20 parts of the formulation regulator, 1-4 parts of the preservative and 1760-1900 parts of the water.

24. The skin care product of claim 23 wherein the sodium hyaluronate is one or a combination of sodium hyaluronate with a molecular weight of 120 to 150, 80 to 100, 20 to 40 and 1 to 10 million daltons.

25. The skin care product of claim 23, wherein the balancing agent comprises one or a combination of dipotassium glycyrrhizinate, sodium dihydrogen phosphate and disodium hydrogen phosphate.

26. The skin care product of claim 23, wherein the emollient is one or more of glycerin, butylene glycol, propylene glycol, glyceryl caprylate, 1, 2-hexanediol, and glyceryl polyether-26.

27. The skin care product of claim 23 wherein said dosage form modifier is one or a combination of hydroxyethyl cellulose and xanthan gum.

28. The skin care product of claim 23 wherein the preservative comprises one or a combination of caprylhydroxamic acid and ethylhexyl glycerin.

29. The skin care product of claim 23, wherein the skin care product has a pH of from 4.5 to 7.

30. A method for preparing a skin care product, comprising:

providing an active collagen extracting solution, sodium hyaluronate, a balancing agent, a softening agent, a dosage form regulator, a preservative and water, wherein the active collagen extracting solution is the active collagen extracting solution as claimed in claim 1 or 2, and the mass parts of the active collagen extracting solution, the sodium hyaluronate, the balancing agent, the softening agent, the dosage form regulator, the preservative and the water are in a relation of 2-10 parts of the active collagen extracting solution, 2-6 parts of the sodium hyaluronate, 1-4 parts of the balancing agent, 100-200 parts of the softening agent, 2-20 parts of the dosage form regulator, 1-4 parts of the preservative and 1760-1900 parts of water;