Background
The chitosan is obtained by treating shrimp and crab shells in a series of ways, is non-toxic and tasteless, is widely applied to the industries of food, medicine and the like, has immunoregulation activity, and can effectively promote wound healing. For example, the addition of a proper amount of chitosan into cream cosmetics can improve the immunity of human bodies to infection caused by bacteria and fungi, prevent the growth of original bacteria, not only can not cause infection to damaged skin, but also can promote the healing of the damaged skin and eliminate facial diseases. In addition, chitosan can also be mixed with formalin to prepare cosmetics containing formalin, and the prepared cosmetics have good bactericidal effect.
Salicylic acid, also known as o-hydroxybenzoic acid, is a fat-soluble organic acid. Salicylic acid has wide application, for example, can be used as a standard reagent for titration by an alkaline method and an iodometric method; in the rubber industry as scorch retarders and for the production of UV absorbers and foaming agents; the compound can be used as an additive in certain weak acid electrolyte and also can be used as a complexing agent for electroplating or chemical plating; can be used as cosmetic antiseptic; as a raw material of the pharmaceutical industry, is used for preparing aspirin, sodium salicylate, salicylamide, analgesic, phenyl salicylate, blood control-67 and other medicaments; can be used for reducing the risk of long-term heart disease of diabetic patients in clinical trials; salicylic acid is widely applied to external smearing medicines, is used in a plurality of skin care products, and has the effects of softening cutin, sterilizing, treating acne and the like. However, salicylic acid has poor water solubility, so that the application effect is limited, and salicylic acid has strong irritation and cannot be directly contacted with the skin.
Gamma-aminobutyric acid (GABA) is widely distributed in animals and plants such as green vegetables, rice, Chinese herbal medicines, beans and the like, is an important inhibitory neurotransmitter, participates in various metabolic activities, and has extremely high physiological activity. Besides the existing functions of lowering blood pressure, improving liver function, preventing arteriosclerosis and cerebral apoplexy, preventing and treating dreaminess and insomnia, promoting relaxation, relieving pain, improving anti-stress, eliminating fatigue, keeping the mood level, regulating the function of brain nerve cells, strengthening brain, soothing the nerves, promoting brain blood circulation, preventing senile dementia and life habit diseases, removing active oxygen, preventing blood vessel and cell aging, improving the absorption rate of nutrient components, especially hard substances, and keeping nutrition balance; in addition, the gamma-aminobutyric acid has the effects of resisting oxidation and aging, repairing skin suffering from external pressure, relieving pressure and relaxing nerves, can quickly penetrate deep into the horny layer of the skin, reforms fine lines and has a good effect on repairing concave-convex disordered muscle lines.
The ionic liquid is a salt which is composed of an organic cation and an inorganic (or organic) anion and is in a liquid state at room temperature (or near room temperature), and is generally called room-temperature ionic liquid or room-temperature molten salt, and the ionic liquid has good thermal stability and strong dissolving capacity, and can enhance the physiological activity of the ion per se, and has attracted much attention of researchers in recent years. Gamma-aminobutyric acid is a commonly used ionic liquid raw material, and if the gamma-aminobutyric acid ionic liquid can be prepared from organic acids such as salicylic acid or alkaloids such as matrine, active substances with strong physiological activity and good solubility can be expected to be obtained.
Therefore, if gamma-aminobutyric acid and salicylic acid are prepared into the ionic liquid, the solubility of the salicylic acid is expected to be enhanced, and the irritation caused by the salicylic acid can be further reduced by wrapping chitosan in the gamma-aminobutyric acid salicylic acid ionic liquid.
Disclosure of Invention
The first technical problem to be solved by the invention is as follows: provides a preparation method of chitosan-coated gamma-aminobutyric acid salicylic acid with good solubility and mild skin.
The second technical problem to be solved by the invention is: provides the chitosan coated gamma-aminobutyric acid salicylic acid prepared by the method.
The third technical problem to be solved by the invention is: provides an application of the chitosan coated gamma-aminobutyric acid salicylic acid.
In order to solve the first technical problem, the invention adopts the technical scheme that: a preparation method of chitosan-coated gamma-aminobutyric acid salicylic acid comprises the following steps:
s1, mixing the components according to the solid-liquid ratio of 1 mg: (1-5) adding chitosan into a glacial acetic acid solution according to the proportion of mL to prepare a chitosan aqueous solution, wherein the mass percentage concentration of glacial acetic acid in the glacial acetic acid solution is 1-3%;
and S2, dropwise adding the gamma-aminobutyric acid salicylic acid ionic liquid into the chitosan aqueous solution under stirring, standing at room temperature, and thus obtaining the chitosan-coated gamma-aminobutyric acid salicylic acid.
Further, the preparation method of the gamma-aminobutyric acid salicylic acid ionic liquid comprises the following steps:
s201, adding gamma-aminobutyric acid into a solvent under a low-oxygen and normal-temperature environment to prepare a gamma-aminobutyric acid solution;
s202, adding salicylic acid into the gamma-aminobutyric acid solution to react to obtain the gamma-aminobutyric acid salicylic acid ionic liquid.
Further, the solvent is water.
Further, in the step S202, the molar ratio of the gamma-aminobutyric acid to the salicylic acid is 1 (0.8-1.2); preferably, the molar ratio is 1: (0.9-1.1).
Further, in the step S202, the reaction temperature after salicylic acid is added is 15-30 ℃; preferably, the reaction temperature is 20-25 ℃; most preferably 20 deg.c.
Further, in the step S202, the reaction time is 9-16 h, preferably 12-13 h.
The invention has the beneficial effects that: the chitosan-coated gamma-aminobutyric acid salicylic acid prepared by the scheme of the invention has good solubility in oil system or water system solvents; the irritation caused by salicylic acid can be reduced by coating chitosan, and the sustained-release effect can be achieved, so that the effect is more durable, and meanwhile, the prepared chitosan-coated gamma-aminobutyric acid salicylic acid has physiological effects of chitosan, salicylic acid and gamma-aminobutyric acid, and has good application prospects in the fields of cosmetics and medicines; the solvents used in the scheme of the invention are common solvents, the reaction conditions are mild, the operation is simple and convenient, the synthesis steps are simple, the post-treatment is convenient, and the prepared chitosan-coated gamma-aminobutyric acid salicylic acid has high coating rate.
In order to solve the second technical problem, the invention adopts the technical scheme that: the chitosan coated gamma-aminobutyric acid salicylic acid prepared by the method.
In order to solve the third technical problem, the invention adopts the technical scheme that: an application of the chitosan-coated gamma-aminobutyric acid salicylic acid in cosmetics.
Further, the preparation raw materials of the cosmetic comprise the following components in percentage by weight:
deionized water: balance of
AVC:0.1-0.3wt%
HA:0.05-0.2wt%
PEG-26 glycerol ether: 1-5 wt.%
Glycerol: 1-5 wt.%
Betaine: 1-8 wt.%
Dipropylene glycol: 2-15 wt%
WSK-1:1-6wt%
Coating gamma-aminobutyric acid salicylic acid ionic liquid with chitosan: 1-10 wt%
Nicotinamide: 0.5-2 wt%
1, 2-hexanediol: 0.2 to 1 wt%
A631:0.1-1wt%
ENNACOMPLEX 261:1-5wt%
PEG-60 hydrogenated castor oil: 0.01-0.05 wt%;
wherein said EnNAOMPLEX 261 comprises Hamamelis virginiana leaf extract, Aesculus hippocastanum seed extract, and Arnica extract.
The technical scheme adopted by the invention also comprises the following steps: an application of the chitosan-coated gamma-aminobutyric acid salicylic acid in the preparation of cosmetics.
Further, the application comprises the steps of:
1) completely soaking AVC and HA in deionized water, homogenizing until completely dispersed, adding PEG-26 glyceryl ether, glycerol, betaine and dipropylene glycol, heating to 80-90 deg.C, homogenizing at medium speed for 3-5min, and stirring at constant temperature for 20-40 min;
2) cooling the product obtained in the step 1) to 35-45 ℃, and adding WSK-1, chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid, nicotinamide, 1, 2-hexanediol, A631 and 261 under stirring;
3) heating and stirring PEG-60 hydrogenated castor oil at 35-45 ℃ until the castor oil is transparent, adding the mixture into the mixture prepared in the step 2), and cooling to obtain the cosmetic containing the gamma-aminobutyric acid salicylic acid coated by chitosan.
Further, the stirring speed of the medium-speed homogenizing is 3000-5000 rpm.
The invention has the beneficial effects that: the gamma-aminobutyric acid salicylic acid coated with the chitosan has good oxidation resistance, lasting moisture retention, mild skin and no stimulation or small irritation.
Detailed Description
In order to explain the technical content, the objects and the effects of the present invention in detail, the following description will be given with reference to the embodiments.
The embodiment 1 of the invention is as follows: a preparation method of chitosan gamma-aminobutyric acid salicylic acid comprises the following steps:
(1) preparing gamma-aminobutyric acid salicylic acid ionic liquid: wrapping and weighing 1.03g of gamma-aminobutyric acid (10mmol), adding 20mL of deionized water, and introducing N2(inert gas can also be used) so that the whole reaction process is in N2The reaction is carried out in the atmosphere, the reaction container is placed in a normal-temperature water bath, the reaction temperature is kept at 25 ℃, the reaction container is shielded from light, and the reaction is carried out under the condition of being shielded from light. 1.38g of salicylic acid (10mmol) was weighed out, the molar ratio of gamma-aminobutyric acid to salicylic acid was 1:1, adding a small amount of salicylic acid into the reaction container for many times (usually, adding the salicylic acid into the solution within 2min for 3-5 times, in this embodiment, adding the salicylic acid into the solution for 4 times) to ensure that N is contained2The real-time temperature of the atmosphere and the reaction is 0-5 ℃. After the addition, the ice salt bath was removed and the reaction was carried out for 12h under ambient temperature and in the absence of light under inert gas.
(2) The preparation process of the salicylic acid gamma-aminobutyric acid coated by chitosan comprises the following steps:
according to the solid-liquid ratio of 1 mg: 4mL, adding chitosan into a glacial acetic acid solution with the mass concentration of 1% under magnetic stirring to obtain a chitosan aqueous solution. Dropwise adding the gamma-aminobutyric acid salicylic acid into the chitosan aqueous solution under magnetic stirring, wherein the stirring speed is 700r/min, and standing at room temperature for 24h to obtain the chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid. The encapsulation efficiency of the test sample was 85.12%.
The embodiment 2 of the invention is as follows: a cosmetic containing gamma-aminobutyric acid salicylic acid coated with chitosan is prepared from the following raw materials: deionized water: 76.25 wt%; AVC: 0.13 wt%; HA (middle molecule): 0.05 wt%; PEG-26 glycerol ether: 1.2 wt%; glycerol: 2.3 wt%; betaine: 3.2 wt%; dipropylene glycol: 4.5 wt%; WSK-1: 3.3 wt%; coating gamma-aminobutyric acid salicylic acid ionic liquid with chitosan: 4 wt%; nicotinamide: 0.78 wt%; 1, 2-hexanediol: 0.42 wt%; a631: 0.24 wt%; 261: 3.6 wt%; RH-60: 0.03 wt%.
The preparation process comprises the following steps:
1) completely soaking AVC and HA (middle molecule) in deionized water, homogenizing and dispersing in a homogenizer at 3000r/min, adding PEG-26 glyceryl ether, glycerol, betaine and dipropylene glycol, heating to 85 deg.C, homogenizing at medium speed for 3min at 4000r/min, and stirring at constant temperature for 30 min.
2) And (3) cooling the product obtained in the step (1) to 40 ℃, and adding WSK-1, chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid, nicotinamide, 1, 2-hexanediol, A631 and A261 under the condition of magnetic stirring.
3) Heating and stirring RH-60 at 40 ℃ until the mixture is transparent, adding the mixture into the mixture obtained in the step 2, and cooling to obtain the chitosan-coated gamma-aminobutyric acid salicylic acid cosmetic.
The embodiment 3 of the invention is as follows: a cosmetic containing gamma-aminobutyric acid salicylic acid coated with chitosan is prepared from the following raw materials: deionized water: 64.38 wt%; AVC: 0.22 wt%; molecules in HA: 0.14 wt%; PEG-26 glycerol ether: 2.1 wt%; glycerol: 3.5 wt%; betaine: 4.5 wt%; dipropylene glycol: 8.2 wt%; WSK-1: 4.7 wt%; coating gamma-aminobutyric acid salicylic acid ionic liquid with chitosan: 6 wt%; nicotinamide: 1.1 wt%; 1, 2-hexanediol: 0.54 wt%; a631: 0.6 wt%; 261: 4 wt%; RH-60: 0.02 wt%.
The preparation process comprises the following steps:
1) completely soaking molecules in AVC and HA in deionized water, homogenizing and dispersing completely in a homogenizer at 4000r/min, adding PEG-26 glyceryl ether, glycerol, betaine and dipropylene glycol, heating to 85 deg.C, homogenizing at medium speed for 3min, homogenizing at 5000r/min, and stirring at constant temperature for 30 min.
2) And (3) cooling the product obtained in the step (1) to 40 ℃, and adding WSK-1, chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid, nicotinamide, 1, 2-hexanediol, A631 and A261 under the condition of magnetic stirring.
3) Heating and stirring RH-60 at 40 ℃ until the mixture is transparent, adding the mixture into the mixture obtained in the step 2, and cooling to obtain the chitosan-coated gamma-aminobutyric acid salicylic acid cosmetic.
The embodiment 4 of the invention is as follows: a cosmetic containing gamma-aminobutyric acid salicylic acid coated with chitosan is prepared from the following raw materials: deionized water: 52.64 wt%; AVC: 3 wt%; molecules in HA: 0.18 wt%; PEG-26 glycerol ether: 4.4 wt%; glycerol: 4.2 wt%; betaine: 7.6 wt%; dipropylene glycol: 11 wt%; WSK-1: 5 wt%; coating gamma-aminobutyric acid salicylic acid ionic liquid with chitosan: 7.1 wt%; nicotinamide: 1.6 wt%; 1, 2-hexanediol: 0.67 wt%; a631: 0.58 wt%; 261: 2 wt%; RH-60: 0.03 wt%.
The preparation process comprises the following steps:
1) completely soaking molecules in AVC and HA in deionized water, homogenizing and dispersing completely in a homogenizer at 4000r/min, adding PEG-26 glyceryl ether, glycerol, betaine and dipropylene glycol, heating to 85 deg.C, homogenizing at medium speed for 3min, homogenizing at 6000r/min, and stirring at constant temperature for 30 min.
2) And (3) cooling the product obtained in the step (1) to 40 ℃, and adding WSK-1, chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid, nicotinamide, 1, 2-hexanediol, A631 and A261 under the condition of magnetic stirring.
3) Heating and stirring RH-60 at 40 ℃ until the mixture is transparent, adding the mixture into the mixture obtained in the step 2, and cooling to obtain the chitosan-coated gamma-aminobutyric acid salicylic acid cosmetic.
Comparative example 1:
the cosmetic preparation procedure was the same as in example 2, and "chitosan-coated γ -aminobutyric acid salicylic acid ionic liquid: 4 wt% ", modified to" gamma-aminobutyric acid salicylic acid ionic liquid: 4 wt% ".
Comparative example 2:
the cosmetic preparation procedure was the same as in example 2, and "chitosan-coated γ -aminobutyric acid salicylic acid ionic liquid: 4 wt% ", instead" chitosan: 4 wt% ".
The cosmetics prepared in examples 2 to 4 and comparative examples 1 to 2 were compared in various aspects such as oxidation resistance, irritation, moisturizing effect, etc.
1) And (3) oxidation resistance test:
the cosmetics obtained in examples 2 to 4 and comparative example 1 were subjected to DPPH radical scavenging test (procedure according to the conventional procedure known in the art), and the results are shown in Table 1:
TABLE 1 Effect of different formulations on DPPH radical scavenging
As can be seen from Table 1, the cosmetics of gamma-aminobutyric acid (GABA) coated by chitosan have higher DPPH free radical clearance rate (increased to more than 1.5 times) than the cosmetics without chitosan coating, and the inoxidizability of the cosmetics of gamma-aminobutyric acid (GABA) coated by chitosan is obviously enhanced.
2) And (3) irritation test:
the effect of inhibition of the response of cosmetics to superficial epithelial cells of the inflammatory state of the brand was evaluated by analyzing IL-1 α, LTB4 or PGE2 released by Normal Human Epidermal Keratinocytes (NHEK). The cells were human-derived keratinocyte cell line (HL60), which were cultured in RPMI 1640(Life Technologies) medium supplemented with 2mmol of L-glutamine, 50U/mL of penicillin, 50. mu.g/mL of streptomycin, and 10% Fetal Calf Serum (FCS), at 37 ℃ and 5% CO2Under the conditions of (1).
For IL-1 α, cells were preincubated for 24 hours in the presence of phorbol ester and calcium ionophore A23187 (Sigma No. C752).
For LTB4, cells were preincubated for 24 hours in the presence of arachidonic acid and calcium ionophore a23187 (sigma No. C752).
For PEG2, cells were preincubated for 24 hours in the presence of phorbol esters.
The culture medium was replaced after incubation with the same medium but containing the cosmetics tested (examples 2-4 and comparative example 1).
At the end of this new incubation period (15min, 37 ℃), the supernatant was collected and the amount of IL-1 α, LTB4 or PGE2 released into the medium was analyzed by ELISA (R and D system kit, No. DE 0275).
For each test, a control without the cosmetic to be tested was run, with the test results shown in table 2 below:
TABLE 2 irritation test results Table
|
IL-1α
|
LTB4
|
PGE2
|
Example 2
|
-22%
|
-26%
|
-40%
|
Example 3
|
-23%
|
-20%
|
-44%
|
Example 4
|
-17%
|
-30%
|
-50%
|
Comparative example 1
|
15%
|
12%
|
18% |
The data in table 2 are the rate of increase or decrease in production relative to control IL-1 α, LTB4 or PGE2 without the cosmetic to be tested.
Under the same experimental conditions, the chitosan-coated gamma-aminobutyric acid salicylic acid ionic liquid causes obvious reduction of the yield of IL-1 alpha, LTB4 or PGE2, while the non-coated gamma-aminobutyric acid salicylic acid ionic liquid increases to different degrees, thereby indicating that the irritation of the gamma-aminobutyric acid salicylic acid ionic liquid coated by the scheme of the invention is greatly reduced.
3) And (4) testing the moisturizing effect:
selecting 5 volunteers for testing, wherein the female is 30-50 in age, firstly measuring facial skin moisture value of the volunteer by adopting a Kanga SC-D8F-03 type skin moisture tester, using the cosmetics of the embodiment 2 and the comparative example 2, carrying out statistical analysis on data by using SPSS10.0 software to obtain an average skin moisture value (P <0.05), and then placing the volunteers in the same working environment, wherein the relative humidity is 20% and the temperature is 28 ℃; facial skin moisture values were measured for test and control volunteers at baseline, 24h and 48h, respectively, as shown in table 3.
Table 3 moisturizing effect test results table
As can be seen from the above table 3, the chitosan coated with the gamma-aminobutyric acid salicylic acid ionic liquid has better moisturizing effect compared with the pure chitosan, and the moisture retention rate after 48 hours is obviously better than that of the pure chitosan.
In conclusion, the gamma-aminobutyric acid salicylic acid ionic liquid is wrapped by the chitosan, and the chitosan is added into the cosmetics, so that the physiological activities of the prepared cosmetics in various aspects such as moisture retention, oxidation resistance and the like are greatly improved, and the cosmetics prepared by the scheme of the embodiment of the invention have smaller irritation to skin.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent modifications made by the present invention in the specification or directly or indirectly applied to the related technical field are included in the scope of the present invention.