CN110960441B

CN110960441B - SOD composition with stable activity and preparation process and application thereof

Info

Publication number: CN110960441B
Application number: CN201911274856.7A
Authority: CN
Inventors: 胡建锋; 吴军; 刘冰凌
Original assignee: Ningbo Jiurui Biological Engineering Co ltd
Current assignee: Ningbo Jiurui Biological Engineering Co ltd
Priority date: 2019-12-12
Filing date: 2019-12-12
Publication date: 2022-09-13
Anticipated expiration: 2039-12-12
Also published as: CN110960441A

Abstract

The invention discloses an SOD composition with stable activity, which is characterized in that: the material comprises the following substances in percentage by mass: 0.9% of sodium chloride, 3% -10% of collagen, 2% -5% of protein, 3% -6% of copper-blue peptide, 0.3% -0.8% of vitamin E, 0.2% of SOD, 0.2% -0.5% of thickening agent, 3% -5% of glycerol, 0.1% -0.5% of xanthan gum, 2% -5% of caprylic/capric triglyceride, 0.5% -2% of phytosterol oleate, 1% -3% of zinc gluconate, 0.3% -0.6% of polyacrylamide dimethyl ammonium taurinate, 0.5% -1% of ascorbic acid and the balance of water; also discloses the application of the SOD composition with stable activity in preparing cream cosmetic skin care products; the invention also discloses a preparation method of the SOD composition with stable activity.

Description

SOD composition with stable activity and preparation process and application thereof

Technical Field

The invention relates to the field of daily chemicals, in particular to an SOD composition with stable activity, a preparation process and application thereof.

Background

SOD is superoxide dismutase, is an antioxidant metalloenzyme existing in organisms, plays a vital role in the balance of organism oxidation and antioxidation, and is widely applied to the fields of food, daily chemical industry, pharmacy and the like. SOD has strong effects of scavenging free radicals, resisting aging, whitening, tendering skin and the like, but is a biological enzyme, so that the environmental requirement on survival is very strict, and particularly, the SOD has short shelf life (the activity of the SOD is reduced linearly at room temperature for 1-3 months) in cosmetics and is far from being suitable for the 3-year shelf life of the cosmetics.

Disclosure of Invention

The invention aims to provide an SOD composition with stable activity, a preparation process and application thereof, which have the advantages of stable SOD activity and long survival time.

The technical purpose of the invention is realized by the following technical scheme:

an active stable SOD composition characterized by: the material comprises the following substances in percentage by mass: 0.9 percent of sodium chloride, 3 to 10 percent of collagen, 2 to 5 percent of protein, 3 to 6 percent of blue copper peptide, 0.3 to 0.8 percent of vitamin E, 0.2 percent of SOD, 0.2 to 0.5 percent of thickening agent, 3 to 5 percent of glycerol, 0.1 to 0.5 percent of xanthan gum, 2 to 5 percent of stabilizing agent, 0.5 to 2 percent of antioxidant additive, 1 to 3 percent of zinc gluconate, 0.3 to 0.6 percent of polyacrylamide dimethyl taurinate, 0.5 to 1 percent of ascorbic acid and the balance of water.

By adopting the technical scheme, the SOD has stable activity, is easier to survive and has longer quality guarantee time. The blue copper peptide and the zinc gluconate are matched to further keep the activity of SOD, zinc ions can be separated from the zinc gluconate in the system, and the copper ions in the blue copper peptide and the zinc ions in the zinc gluconate form a coordination structure with the SOD so as to jointly stabilize the structure of the SOD. On the other hand, the blue copper peptide can react with ascorbic acid, and the blue copper peptide promotes the ascorbic acid to be dehydrogenated to generate dehydroascorbic acid, so as to destroy proteins and nucleic acids of bacteria, thereby achieving the effect of killing the bacteria. Furthermore, the ascorbic acid can increase the permeability of the cell membrane of the bacteria to copper ions in the copper-blue peptide, thereby playing a role in enhancing the sterilization, reducing the consumption and damage of the bacteria in the SOD composition to SOD after sterilization, and further improving the survival time of the SOD.

The SOD activity is stable and the SOD is more survivable by adding the components; the SOD composition has high-efficiency moisturizing effect by adding collagen and glycerol; the xanthan gum also has thickening effect, and can enable the SOD composition to have proper viscosity; the copper peptides can also enable the SOD composition to have the effect of helping skin generate collagen and elastin; the poly acryloyl dimethyl ammonium taurate has excellent emulsifying, dispersing and thickening capabilities; the vitamin E and the ascorbic acid enable the SOD composition to have the functions of resisting oxidation and free radicals, and achieve the effects of resisting aging, whitening and the like. The addition of 0.9% sodium chloride solution can simulate the ion concentration and osmotic pressure in blood plasma of mammal, so as to stabilize the spatial structure of SOD. The protein can provide essential nutrients for skin layer containing collagen, supplement amino acids beneficial to human body, enhance collagen activity in skin, maintain stratum corneum water and fiber structure integrity, improve skin cell living environment, promote skin tissue metabolism, promote circulation, and care skin

The invention is further configured to: the stabilizer is caprylic capric triglyceride.

By adopting the technical scheme, the caprylic capric triglyceride is high-purity grease formed by esterifying caprylic acid, capric acid and glycerol. It is an excellent moisturizing oil, has good spreadability, gives the skin a slippery but not greasy feeling, and is easily absorbed by the skin. Has good effect on the uniformity and fineness of cosmetics and can ensure that the skin is smooth and glossy.

The invention is further configured to: the protein is milk protein.

By adopting the technical scheme, the milk protein is taken as an efficient nutritional agent, and can promote the regeneration of cells after being coated on the skin, strengthen the moisturizing effect of the outer layer of the skin and provide moisture and comfort; meanwhile, the milk protein can inhibit the proliferation of melanocytes and the activity of tyrosinase, and has the whitening effect; the milk protein combines with vitamin C and vitamin E, and can protect skin from free radical damage and repair skin.

The invention is further configured to: the thickening agent is carbomer.

By adopting the technical scheme, carbomer is added as a thickening agent, so that the overall viscosity of the SOD composition can be regulated and controlled; meanwhile, carbomer can act together with ammonium polyacryl dimethyl taurate to emulsify glycerin, caprylic capric triglyceride and phytosterol oleate, so that the SOD composition is more uniform and fine.

The invention is further configured to: the antioxidant additive is phytosterol oleate.

By adopting the technical scheme, the phytosterol has good oxidation resistance, but the cyclopentanoperhydrophenanthrene structure in the molecule and the hydrocarbon chain on C-17 determine that the phytosterol is insoluble in water, and the hydroxyl on C-3 ensures that the solubility of the phytosterol in the oil phase is very low, so that the phytosterol oleate obtained through esterification modifies the hydroxyl of the phytosterol so as to improve the solubility of the phytosterol in the oil phase, and has good oxidation resistance. Meanwhile, the phytosterol can act synergistically with vitamin E, and chain oxidation reaction is interrupted by supplying H to oxygen free radicals or peroxy free radicals, so that the effect of resisting oxidation is achieved.

The invention is further configured to: the material comprises the following substances in percentage by mass: 0.9% of sodium chloride, 3% of collagen, 2% of milk protein, 6% of copper-blue peptide, 0.3% of vitamin E, 0.2% of SOD, 0.2% of carbomer, 3% of glycerol, 0.1% of xanthan gum, 3% of caprylic capric triglyceride, 2% of zinc gluconate, 0.5% of phytosterol oleate, 0.3% of ammonium polyacryloyldimethyl taurate, 0.5% of ascorbic acid and the balance of water.

By adopting the technical scheme, the components with stable activity are preferably selected as the mass percentage of the SOD composition, so that the activity of SOD enzyme can be stabilized to the maximum degree and the whitening effect of skin can be improved.

The invention also aims to provide the application of the SOD composition with stable activity in preparing a pasty skin care product.

The invention also provides a preparation method of the SOD composition with stable activity, which comprises the following steps:

(1) adding water, glycerol, xanthan gum and sodium chloride into a mixing pot, and stirring to dissolve uniformly.

(2) Adding caprylic capric triglyceride, phytosterol oleate, polyacrylamide dimethyl taurinate and carbomer into an oil phase pot, and stirring and dispersing uniformly.

(3) Pumping the oil phase materials into the mixing pot, starting the homogenizer, and stirring uniformly.

(4) Adding collagen, zinc gluconate, milk protein, copper cyanamide, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition.

By adopting the technical scheme, in the step (1), sodium chloride is added into water to be fully dissolved, and then glycerin is added, wherein the glycerin is used as a good solvent and can be mutually dissolved with the water in any proportion. Adding xanthan gum, stirring and dissolving uniformly, so that the components of the xanthan gum can be fully dispersed in water, and slightly thickening the xanthan gum to ensure that the composition obtained in the step (1) has better stability;

in the step (2), polyacrylamide dimethyl ammonium taurate is used as an excellent emulsifying and dispersing agent, caprylic capric triglyceride, phytosterol oleate, polyacrylamide dimethyl ammonium taurate and carbomer are added into an oil phase pot, and the mixture is stirred and dispersed uniformly, so that the oil phase can be fully emulsified, and the later mixing process is facilitated.

In the step (3), the materials in the oil phase pot are pumped into the mixing pot, the homogenizer is started and stirred uniformly, so that the stirring is more thorough, and the mixing effect is better and more uniform.

And (4) adding collagen, zinc gluconate, milk protein, copper cyanamide, vitamin E, ascorbic acid and SOD into the mixture, and uniformly stirring and dissolving the mixture to obtain the SOD composition, which is simple and convenient. The SOD composition obtained by the steps has stable property and uniform and fine texture.

Detailed Description

The present invention will be described in further detail below.

Example 1:

an SOD composition with stable activity comprises the following mass substances (g):

example 2:

example 3:

example 4:

example 5:

the activity-stable SOD compositions of examples 1-5 were prepared as follows:

(3) Pumping the oil phase pot material into a mixing pot, starting a homogenizer, and stirring uniformly.

(4) Adding collagen, zinc gluconate, milk protein, ceruloplasmin, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition.

Comparative example 1:

the active stable SOD composition of comparative example 1 was prepared as follows:

(4) Adding collagen, zinc gluconate, milk protein, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition in the comparative example 1.

Comparative example 2:

the active stable SOD composition of comparative example 2 was prepared as follows:

(4) Adding collagen, zinc gluconate, milk protein, blue copper peptide and vitamin E, SOD into the mixing pan, stirring and dissolving uniformly to obtain the SOD composition of comparative example 2.

Comparative example 3:

the active stable SOD composition of comparative example 3 was prepared as follows:

(2) Adding caprylic/capric triglyceride, phytosterol oleate, polyacrylamide dimethyl taurinate and carbomer into an oil phase pot, and stirring and dispersing uniformly.

(4) Adding collagen, milk protein, copper-blue peptide, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition in the comparative example 3.

Comparative example 4:

the active stable SOD composition of comparative example 4 was prepared as follows:

(1) adding water, glycerol and xanthan gum into a mixing pot, stirring and dissolving uniformly.

(4) Adding collagen, milk protein, blue copper peptide, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition in the comparative example 4.

Comparative example 5:

comparative example 6:

comparative example 7:

the active stable SOD compositions of comparative examples 5-7 were prepared as follows:

(4) Adding collagen, milk protein, copper-blue peptide, vitamin E, ascorbic acid and SOD into a mixing pot, and stirring to dissolve uniformly to obtain the SOD composition in the comparative examples 5-7.

The sources of the partial raw materials are as follows:

SOD, Shanghai Green plum Biotech Co., Ltd;

blueish peptide, american peptide, trade name CTPT-1;

milk protein, CLR, germany, under the trade name Lactokine Fluid PF;

phytosterol oleate, tradename CARDIOAID-SF, of the Aids Union, USA;

collagen, CLR, germany, under the trade name Collagen CLR.

And (3) experimental test:

1. reagent and apparatus

(1) Pyrogallol solution: 142mg of pyrogallol were dissolved in 10mmol/L HCl and brought to a volume of 25ml, and refrigerated in a brown flask.

(2) Tris-HCl-EDTA buffer: measuring 150ml of 0.5mol/L Tris and 91.7ml of 500mmol/L HCl, supplementing 15ml of 100mmol/L diethylenetriaminepentaacetic acid, finally diluting to 1.5L with deionized water, and adjusting the pH value to obtain 50mmol/L Tris-HCL buffer solution with pH of 8.2.

2. Sample(s)

Collecting SOD composition 10g, soaking in water or dissolving to 100ml, and filtering to obtain filtrate.

3. Principle of

Pyrogallol is stable in an acid environment, but can undergo an autoxidation reaction in a weak base environment, and only one electron is accepted by the pyrogallol during autoxidation to generate a superoxide anion free radical (O) ₂ +O ₂ ^- ) And in the autoxidation process of the same, a colored intermediate product is generated at a certain rate, and the SOD can convert O into ₂ ^- Disproportionating and decomposing into H ₂ O ₂ And O ₂ Thereby inhibiting the autoxidation rate of pyrogallol. Whereby O can be decomposed according to SOD ₂ ^- The activity of SOD (SOD activity unit is obtained when 50% of the reaction rate is inhibited by SOD in reaction solution per minute) is indirectly calculated.

4. Test method

(1) And (3) measuring the self-oxidation rate of the pyrogallol, namely taking 4.5ml of Tris-HCl-EDTA buffer solution with the pH value of 8.2 into a 10ml colorimetric tube, keeping the temperature at 25 ℃ for 10min, adding 10 mu L of 45mmol/L pyrogallol solution with the constant temperature at 25 ℃, uniformly mixing, and rapidly measuring the photometric density value at 325nm wavelength in a 1cm quartz cuvette. Measuring optical density value every 30s for 4min to obtain the self-oxidation rate OD of pyrogallol _A /min。

(2) SOD activity determination, taking 4.5ml of Tris-HCl-EDTA buffer solution with pH of 8.2 to put in a 10ml colorimetric tube at 25 DEG CKeeping the temperature for 10min, adding 10ml SOD composition sample solution with constant temperature of 25 deg.C, rapidly mixing, measuring density value at 325nm wavelength once every 30s for 4min, and calculating optical density value change rate OD _B /min。

(3) Enzyme activity was calculated according to the following formula:

V ₁ total volume of reaction solution, ml

V ₂ Determination of sample volume, ml

n- -sample dilution factor

OD _A Autoxidation rate of pyrogallol

OD _B Rate of change of optical density value of sample

The SOD activity is measured by adopting a test method of a trace pyrogallol method. According to the above method, multiple SOD compositions were prepared, sampled at regular intervals and tested for SOD activity by the method of micro pyrogallol (325nm), respectively, and the numerical change of unit enzyme activity was observed.

The unit enzyme activity (U/ml) test results are shown in tables 1 and 2 below:

TABLE 1 enzyme activity measurement tables of examples 1 to 5 and comparative examples 1 to 3 units (unit: U/ml)

As can be seen from Table 1, in examples 1-5, different amounts of ceruloplasmin, zinc gluconate and ascorbic acid were added, wherein the amount of zinc gluconate was changed in examples 3-5, and the experimental data show that the SOD enzyme in example 5 survived for the longest time and the increase in the amount of zinc gluconate has an effect of promoting the activity of the SOD enzyme.

In comparative example 3, no zinc gluconate was added as compared with the examples, and the experimental data showed that the enzymatic activity of comparative example 3 decreased rapidly as compared with the examples, and by the twelfth month, the SOD enzyme had been inactivated. This shows that the zinc ion provided by zinc gluconate plays an important role in the activity of SOD, and the zinc gluconate can improve the activity of SOD and prolong the survival time of SOD enzyme when used in SOD composition.

In comparative example 2, in which ascorbic acid was not added as compared with the examples, experimental data showed that the comparative example 2 had a rapid decrease in enzyme activity as compared with the examples, and by the twelfth month, the SOD enzyme had been inactivated. This indicates that the addition of ascorbic acid as a strong antioxidant reducing component can increase the activity of SOD and prolong the survival time of SOD.

In comparative example 1, in which no bluepatide was added as compared with the examples, experimental data showed that the enzymatic activity of comparative example 1 decreased rapidly as compared with the examples, and by the twelfth month, the SOD enzyme had been inactivated. This shows that the copper ions provided by the copper-coated peptides play an important role in the activity of SOD, and the copper-coated peptides used in the SOD composition can improve the activity of SOD and prolong the survival time of SOD enzyme.

TABLE 2 Unit enzyme activity assay table (unit: U/ml) at different NaCl concentrations

The sodium chloride solution in the components also plays an important role, the ion concentration and osmotic pressure in plasma of mammals can be simulated by adding 0.9% of sodium chloride, the purpose of stabilizing the spatial structure of SOD is achieved, but the enzyme activity of SOD can be inhibited when the concentration of sodium chloride is too high or too low. In summary, sodium chloride solutions, ceruloplasmin, zinc gluconate and ascorbic acid at appropriate concentrations are not acceptable in SOD compositions. The activity of SOD can be further maintained by the cooperation of the bluecopper peptide and the zinc gluconate, zinc ions can be separated from the zinc gluconate in the system, and the copper ions in the bluecopper peptide and the zinc ions in the zinc gluconate form a coordination structure with the SOD so as to jointly stabilize the structure of the SOD; the ascorbic acid can increase the permeability of the cell membrane of the bacteria to copper ions in the blue copper peptide, thereby playing a role in enhancing the sterilization, reducing the consumption and damage of the bacteria in the SOD composition to SOD after sterilization, and further improving the survival time of the SOD. Any of the components plays an important role in the stabilization of SOD and acts together with the activity of SOD.

The above-described embodiments do not limit the scope of the present invention. Any modification, equivalent replacement, and improvement made within the spirit and principle of the above-described embodiments should be included in the protection scope of the technical solution.

Claims

1. An active stable SOD composition characterized by: the material comprises the following substances in percentage by mass: 0.9% of sodium chloride, 3% -10% of collagen, 2% -5% of protein, 3% -6% of ceruloplasmin, 0.3% -0.8% of vitamin E, 0.2% of SOD, 0.2% -0.5% of thickening agent, 3% -5% of glycerol, 0.1% -0.5% of xanthan gum, 2% -5% of stabilizing agent, 0.5% -2% of antioxidant additive, 1% -3% of zinc gluconate, 0.3% -0.6% of ammonium polyacryl dimethyl taurate, 0.5% -1% of ascorbic acid and the balance of water, wherein the thickening agent is carbomer, and the antioxidant additive is phytosterol oleate.

2. The active stable SOD composition of claim 1, wherein: the stabilizer is caprylic capric triglyceride.

3. The active stable SOD composition of claim 2, wherein: the protein is milk protein.

4. The active stable SOD composition of claim 3, wherein: the material comprises the following substances in percentage by mass: 0.9% of sodium chloride, 3% of collagen, 2% of milk protein, 6% of ceruloplasmin, 0.3% of vitamin E, 0.2% of SOD, 0.2% of carbomer, 3% of glycerol, 0.1% of xanthan gum, 2% of caprylic capric triglyceride, 3% of zinc gluconate, 0.5% of phytosterol oleate, 0.3% of ammonium polyacryloyldimethyl taurate, 0.5% of ascorbic acid and the balance of water.

5. Use of the active stable SOD composition of any one of claims 1-4 for preparing a cream-like cosmetic skin care product.

6. A preparation method of SOD composition with stable activity is characterized in that: the preparation method comprises the following steps:

(1) adding water, glycerol, xanthan gum and sodium chloride into a mixing pot, stirring and dissolving uniformly,

(2) adding caprylic capric triglyceride, phytosterol oleate, polyacrylamide dimethyl taurinate and carbomer into an oil phase pot, stirring and dispersing uniformly,

(3) pumping the oil phase materials into a mixing pot, starting a homogenizer, stirring uniformly,