CN113233976B

CN113233976B - Gallic acid and aminoacetic acid eutectic and whitening cream containing eutectic

Info

Publication number: CN113233976B
Application number: CN202110499029.9A
Authority: CN
Inventors: 涂禄清; 邓建科
Original assignee: Guangzhou Cuipu Biotechnology Co ltd
Current assignee: Guangzhou Cuipu Biotechnology Co ltd
Priority date: 2021-05-08
Filing date: 2021-05-08
Publication date: 2021-11-16
Anticipated expiration: 2041-05-08
Also published as: CN113233976A

Abstract

The invention discloses a gallic acid-aminoacetic acid eutectic crystal and a whitening cream containing the eutectic crystal.

Description

Gallic acid and aminoacetic acid eutectic and whitening cream containing eutectic

Technical Field

The invention relates to the field of cosmetics, in particular to a gallic acid and aminoacetic acid eutectic crystal, a whitening cream containing the eutectic crystal and a preparation method of the whitening cream.

Background

Gallic acid is also called gallic acid or gallic acid, chemical name is 3, 4, 5 trihydroxy benzoic acid, chemical formula C₆H₂(OH)₃COOH, needle-like crystals, melting point 235-. The results of the research on the in-vitro antioxidant capacity of the gallic acid by the Xianxiayan and the like also show that the gallic acid has a far greater clearance rate on the free radicals of the 2, 2-biphenyl-1-pichydrazide in the concentration range of 5-50mg/L than the vitamin C which is a recognized antioxidant, and the gallic acid has the advantages of higher clearance rate than the vitamin CAnd has a certain concentration dependence relationship; the removal effect of gallic acid on hydroxyl free radicals is obviously higher than that of vitamin C within the range of 50-400 mg/L, and gallic acid with different concentrations has the inhibition effect on lipid peroxidation, and the inhibition rate is higher than that of vitamin C.

Cocrystals are crystals formed by the combination of a pharmaceutically active ingredient (API) and a cocrystal former (CCF, or another API) via hydrogen bonding or other non-covalent bonds, wherein the pure states of API and CCF are both solid at room temperature and have a fixed stoichiometric ratio between the components. By forming a co-crystal, properties of the API, such as increased solubility, dissolution and transdermal absorption, improved release characteristics, etc., can be greatly improved. Common methods for preparing the co-crystals include: solvent evaporation, solvothermal, cooling crystallization, sublimation, melting, suspension crystallization, milling, ultrasound, and the like. The eutectic can improve the dissolution rate and the transdermal absorption rate of insoluble substances, and is a new technical development trend of cosmetic combination. Gallic acid and aminoacetic acid eutectic have been reported in the literature, but no research on the application of gallic acid and aminoacetic acid eutectic in cosmetics has been found so far.

Disclosure of Invention

The invention aims to provide a compound cosmetic containing gallic acid-aminoacetic acid eutectic and a preparation method thereof.

The gallic acid-aminoacetic acid eutectic exists in a stable eutectic form, the molar ratio of the gallic acid to the aminoacetic acid is 1:1, and the gallic acid-aminoacetic acid eutectic is prepared by adopting an absolute ethyl alcohol solvent assisted wet grinding method. Gallic acid was purchased from beijing carbofuran technologies ltd, content: 99.0 percent; aminoacetic acid was purchased from bailcarb technologies ltd, beijing.

Further, the preparation method of the gallic acid-glycine eutectic is an absolute ethyl alcohol solvent assisted wet grinding method, the gallic acid and the glycine are respectively mixed according to a molar ratio of 1:1, the mixture is placed in a mortar, 1mL of absolute ethyl alcohol is added for grinding, 1mL of absolute ethyl alcohol is dropwise added every 2 minutes for grinding, so that a sample is always kept in a slurry state in the whole grinding process, the grinding time is 30min, and the sample is dried in a vacuum drier for later use. The grinding method mixes different drug molecules uniformly by mechanical force, and the two drug molecules generate acting forces with different strengths such as hydrogen bond, dipole moment, van der waals force and the like according to the structural characteristics of the two drug molecules, so that a eutectic can be formed. Characterization of gallic acid-glycine co-crystal:

(1) x-ray powder diffraction (PXRD) analysis is used for evaluating the state of substances in the gallic acid-aminoacetic acid drug composition, and about 50mg of a sample to be tested is taken to perform X-ray powder diffraction at room temperature. Adopting a Cu-Kalpha target, filtering by a graphite crystal monochromator, and obtaining the following working voltage: 20kV, receiving slit: 0.3mm, 2 θ angle: 5-40 DEG, step length: 0.05 °, diffraction patterns and data were recorded directly by the computer. A new peak different from the peak of gallic acid-glycine crystals appeared at 26.3 ° 2 θ, indicating the formation of a eutectic of gallic acid and glycine. Gallic acid contains O-H at the 3, 4, and 5 positions, and can form C ═ O with C ═ O on glycine^…O-H hydrogen bonds, thereby forming a new gallic acid-aminoacetic acid eutectic. Please refer to fig. 1.

(2) And evaluating the substance state of the gallic acid-aminoacetic acid drug combination by differential scanning calorimetry. Thermal analysis is carried out on gallic acid monomer, aminoacetic acid monomer and gallic acid-aminoacetic acid eutectic (1: 1) by adopting differential scanning calorimetry. The experimental method comprises the following steps: precisely weighing 3-5 mg of a sample to be measured, placing the sample in an aluminum sample crucible, taking a reference object as an empty aluminum crucible, heating the sample from room temperature to 450 ℃ at the heating rate of 10 ℃/min, and taking nitrogen as the atmosphere. In the DSC curve, please refer to fig. 2, a new peak appears at 163 ℃ in the gallic acid-aminoacetic acid eutectic, indicating the formation of gallic acid-aminoacetic acid eutectic. Melting point peaks of gallic acid and aminoacetic acid are respectively at 272 ℃ and 268 ℃, when gallic acid-aminoacetic acid eutectic is 1:1, two peaks at 272 ℃ and 268 ℃ disappear, and simultaneously, a melting point peak of the eutectic appears at 163 ℃.

(3) Evaluation of dissolution test

The dissolution test is carried out by referring to the fourth general rule dissolution and release determination method II (paddle method) of the Chinese pharmacopoeia 2015 edition. Taking 50mg of gallic acid standard substance, wherein gallic acid and aminoacetic acid 1:1 eutectic are equivalent to 50mg of gallic acid, taking 900mL of 0.1mol/L hydrochloric acid solution as a dissolution medium, stirring at a constant speed of 50r/min, measuring in a circulating water bath at 37 +/-0.5 ℃, sampling at 1,3, 5, 7, 9, 11, 13, 15 and 20min respectively, sampling 5mL of the dissolution medium each time, filtering through a 0.45 mu m polytetrafluoroethylene membrane, collecting subsequent filtrate, placing the subsequent filtrate in a test tube, supplementing 5mL of dissolution medium preheated to 37 ℃ into a dissolution cup, measuring the absorbance of the sample in each test tube at 259nm, calculating the amount of free gallic acid according to a standard curve, calculating the dissolution rate of the gallic acid at each time point, and drawing an in-vitro dissolution curve of the gallic acid. Referring to fig. 3, the dissolution rate of gallic acid after the gallic acid-aminoacetic acid eutectic is prepared is significantly improved compared with the gallic acid-aminoacetic acid physical mixture. It can be found that the cumulative dissolution amount of gallic acid-aminoacetic acid in 3min is more than 85%.

(4) Transdermal absorption test

The culture conditions of Caco-2 cells are as follows: DMEM high-sugar medium containing 10% fetal calf serum, 1% double antibody and 1% nonessential amino acids is used for culture in a biological incubator at 37 ℃ and under the conditions of 95% relative humidity and 5% CO 2. Caco-2 simulated small intestine epithelial cell transfer experiment adopts Caco-2 cell strain growing and differentiating to 30 generations, after cell differentiation and fusion reach 80%, using pancreatin to digest, using complete culture medium to regulate cell suspension concentration to be 1.5X 10⁵cells/mL were inoculated into a 12-well Transwell cuvette, and 0.5mL of the cell fluid was added to the upper layer (AP side) of the Transwell, while 1.5mL of the medium was added to the lower layer (BL side) of the Transwell. The medium was changed every other day, cell morphology was observed, and after continuous culture for 21 days, permeation experiments were started. Appropriate amounts of gallic acid and gallic acid-glycine (molar ratio 1:1) were dispersed in pH7.4 (containing HBSS and 25mM HEPES buffer) to prepare a suspension solution of 20. mu.g/mL in terms of gallic acid, which was used as the group 2 administration groups. pH7.4 (containing HBSS, 25mM HEPES and 4% BSA) was used as a receiving solution. 0.4mL of a 20. mu.g/ML gallic acid-containing solution as a test solution was added to the AP side1.2mL of a culture medium having a pH of 7.4 was added to the BP side. Each group of 4 duplicate wells. The 12-well plate was incubated on a 37 ℃ temperature-controlled shaker at 100rpm to begin the transport experiment. Sampling every 20min, acting for 2h, pretreating the sample, and analyzing the content of gallic acid by HPLC liquid phase. The chromatographic conditions are as follows: with Diamonsil C₁₈A (250X 4.6mm, 5 μm) chromatographic column as a stationary phase, an acetonitrile-0.03% phosphoric acid solution (10:90) as a mobile phase, a detection wavelength of 259nm, a column temperature of 30 ℃, a flow rate: 0.6mL/min, injection volume: 25 μ l. The apparent permeability coefficient Papp is calculated. The calculation result of the apparent permeability coefficient Papp is shown in figure 4, which shows that the gallic acid-aminoacetic acid permeability is obviously improved.

The gallic acid-aminoacetic acid eutectic whitening cream is characterized in that the weight percentage of the gallic acid-aminoacetic acid eutectic is 0.2% -5%, when the content of the gallic acid-aminoacetic acid eutectic is lower than 0.2%, the whitening effect is not obvious, when the addition amount of the gallic acid-aminoacetic acid eutectic is higher than 5%, the formula cost is overhigh, and the skin discomfort caused by the heat sensation is caused. For example, the gallic acid-glycine co-crystal can be added in an amount of 0.2%, 0.3%, 0.5%, 1.0%, 2.0%, 3.5%, 5%, etc.

In some embodiments, the whitening cream further comprises one or more of an emulsifier, a thickener, a humectant, and a preservative;

preferably, the weight percentage of the emulsifier is 2% -15%, preferably 2% -10%;

preferably, the weight percentage of the thickening agent is 2% -10%, preferably 2% -8%;

preferably, the weight percentage of the humectant is 12-20%, preferably 12-17%;

preferably, the weight percentage of the preservative is 0.1-1%, preferably 0.1-0.5%.

In some embodiments, the emulsifier is formulated from behenyl alcohol, and arachidyl glucoside (Montanov202) and glyceryl stearate PEG-100 stearate (SIMULSOL 165). The selection of the emulsifier has an influence on the appearance, absorption effect, spreadability, size and uniformity of the dispersed particle size of the cream sample, especially stability. Therefore, the whitening cream is required to have the quality of modern cosmetics and to be stable and effective. The research finds that when the whitening cream uses a compound emulsifier of the docosanol, the eicosyl glucoside (Montanov202) and the glyceryl stearate PEG-100 stearate (SIMULSOL 165), the cream has excellent appearance, spreadability, absorption effect, viscosity, emulsified particle size and other indexes.

In some embodiments, the thickener is formulated from hydroxyethylcellulose and polyacrylic acid 13/polyisobutylene/polysorbate 20(Sepiplus 400). The effect of different thickening systems on the final sample, which is not only manifested in the stability and appearance of the sample, but also in the feel of use and the efficacy properties of the sample, will vary as well, being one of the important components of a thickening system in a cosmetic formulation. The experimental cream needs to be added with plant extracts with higher ion content, when a thickening system of a formula is designed, an ion-resistant thickening agent is selected, and the ion-resistant hydroxyethyl cellulose and SEPIPLUS400 are selected to be compounded with each other, so that the requirement of formula design is met.

In some embodiments, the humectant is selected from one or more of 1, 3-propanediol, propylene glycol, pentanediol, hexanediol, sorbitol, glycerol, dipropylene glycol, 1, 3-butanediol, trehalose, xylitol, erythritol, glyceryl polyether-26, β -glucan, methyl glucitol polyether-20, methyl glucitol polyether-10, erythrulose, biogan-1, betaine, rhamnose, oat β -glucan, acetyl chitosamine, polyethylene glycols, water-soluble fats and oils, water-soluble silicone oils, and sodium hyaluronate.

In some embodiments, the preservative is a combination of ZANTHOXYLUM BUNGEANUM (ZANTHOXYLUM BUNGEANUM) fruit extract, PULSATILLA CHINENSIS (PULSATILLA CHINENSIS) extract, and spanish moss (TILLANDSIA USNEOIDES) extract.

In some embodiments, the whitening cream comprises the following components in percentage by weight as shown in the table:

also provided in some embodiments is a method of preparing the whitening cream, the method comprising the steps of:

(1) adding deionized water into a water phase pot, adding docosyl alcohol, eicosyl glucoside, glyceryl stearate PEG-100 stearate, glycerol, 1, 3-propylene glycol and betaine under stirring, stirring uniformly, and heating to 85 deg.C for dissolving.

(2) Adding hydroxyethyl cellulose into an oil phase pot, heating to 80 ℃, and dissolving for later use.

(3) Preheating an emulsifying pot to 85 ℃, adding the phase (1), then adding the phase (2) and polyacrylic acid 13/polyisobutylene/polysorbate 20(Sepiplus400), homogenizing and emulsifying (3200R/min) for 5min, stirring (25R/min), vacuumizing to 0.4 atmospheric pressure, and slowly cooling with cooling water to 45 ℃ for later use.

(4) Adding gallic acid-aminoacetic acid eutectic and fructus Zanthoxyli (Zanthoxylum BUNGEANUM) fruit extract, radix Pulsatillae (Pulsatilla CHINENSIS) extract, and Spanish moss (TILLANDSIA USNEOIDES) extract into (3), stirring (15R/min) for 15 min, discharging at 38 deg.C, and storing and standing.

Compared with the prior art, the gallic acid-aminoacetic acid in the cosmetic exists in a cocrystallization state, the synergistic effect of the gallic acid and the aminoacetic acid is exerted, the dissolution rate of the gallic acid is improved, the transdermal absorption capacity of the gallic acid is increased, and the curative effect of the cosmetic is improved.

Drawings

FIG. 1 is an X-ray powder diffraction PXRD pattern.

Figure 2 is a differential scanning calorimetry, DSC, graph.

FIG. 3 is a graph showing the results of dissolution test.

FIG. 4 is a graph showing the results of apparent permeability coefficients of Caco-2 cell permeation experiments.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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 invention.

Examples 1 to 5: effect of emulsifiers on the sensory Effect of whitening creams

According to international standard ISO8589-2007 general guideline for design sensory analysis laboratory and national standard GB/T13868-2009 general knowledge principle for sensory analysis-establishment sensory analysis laboratory, etc., volunteers require: the number of 20 healthy people aged 20-30 years, the volunteers with unlimited characters fill in corresponding feeling evaluation tables according to the feelings during and after use, all the feelings are represented by numbers, the trial process adopts a double-blind method, namely, each volunteer does not know the real information of a sample and the feelings of other volunteers, and the research process is carried out independently. All data are summarized and subjected to statistical analysis to obtain the final evaluation of the tested sample. Preferably 9 to 10 minutes; preferably 7 to 8 minutes; generally 5 to 6 points; worse 3-4 points; very poor 1-2 points.

The effect of each emulsifier was examined with the basic formulation in table 1:

table 2 shows the results of the evaluation of the effect of the various emulsifiers when the basic formulation in Table 1 is unchanged: wherein the emulsifier is: example 1: isostearyl glucoside

Example 2: ceteareth-25

Example 3: tween-60 and span 60

Example 4: brij 72 and Briji 721 both: polyoxyethylene (2) stearyl alcohol ether and polyoxyethylene (21) stearyl alcohol ether

Example 5: montanov202 and SIMULSOL 165 both: behenyl alcohol and arachidyl glucoside and glyceryl stearate PEG-100 stearate

It can be seen from examples 1-5 that the moisturizing effect and the refreshing degree of the cream are not greatly affected by the emulsifier, mainly the moisturizing effect and the refreshing degree are more affected by the humectant and the grease, and the combination of different emulsifiers has little effect on the moisturizing effect and the refreshing degree under the condition that the basic formula is not changed. The cream appearance, spreading emulsifier example 3, 4, 5, worked better than the other example 1, 2. The five groups of emulsifiers have the greatest influence on the absorption effect and viscosity of the cream, wherein the groups of examples 3 and 5 are better than the groups of other examples 1, 2 and 4. Wherein the particle size, particle size uniformity (distribution), and particle size regularity of the examples 4 and 5 were observed by microscope, which is better than those of the other examples 1, 2, and 3. In summary of the results, the Montanov202 and SIMULSOL 165 of example 5, i.e., behenyl alcohol and arachidyl glucoside, and glyceryl stearate PEG-100 stearate emulsifiers, are better.

Examples 6 to 12: whitening effect test of whitening cream product

TABLE 3 whitening cream formulations of examples 6-12

Comparative examples 1 to 5 preparation

Comparative examples 1-5, the same weight percentages of emulsifier, humectant, thickener, preservative and deionized water as in example 6, except that comparative example 1 is a blank, comparative example 2 adds 5% gallic acid monomer, comparative example 3 adds 5% glycine monomer, comparative example 4 adds a gallic acid-glycine physical mixture, where the gallic acid content is equivalent to 5%, and comparative example 5 is 5% commercially available vitamin C.

Determination of melanin: the skin melanin tester MDD4-Mexameter MX18 is used for detection, during testing, the probe is vertically pressed on the skin surface of a testee, a spring for ensuring constant pressure during each test is arranged in the probe, and after each test, the skin melanin value is displayed on a display screen. The instrument is based on the principle of spectral absorption, and the content of melanin and heme in skin is determined by measuring the reflection quantity of light with specific wavelength irradiated on the skin of a human body. A smaller melanin number indicates a whiter skin, whereas a darker skin is indicated. The subject's requirements: selecting healthy people of 20-25 years old without skin disease history and cosmetic allergy history. The results are shown in Table 4.

As can be seen from table 4, the content of melanin of comparative example 1 has no significant difference (P >0.05) compared to the content of melanin before application on days 7, 14 and 28, respectively, indicating that the basic formula without gallic acid-glycine has no whitening effect. In example 6 (the added amount of gallic acid-glycine was 0.2%), the melanin content was reduced on the 7 th, 14 th and 28 th days after application, but the reduction was not significant, and was substantially equivalent to the effect of the 5% gallic acid monomer in comparative example 2. The whitening effect of the cream containing gallic acid-aminoacetic acid eutectic 0.2% is weaker. In example 7, the melanin content has no significant difference (P >0.05) between the melanin content before application on the 7 th day and the melanin content before use on the 14 th day, and the melanin content has a significant difference (P <0.05) between the melanin content before application on the 28 th day and the melanin content before use, so that the cream containing gallic acid-glycine eutectic 0.3% starts to have certain whitening effect on the 28 th day of application; in example 8, the melanin content was not significantly different (P >0.05) on the 7 th day of application compared to the melanin content before use, while the melanin content was not significantly different (P <0.05) on the 14 th and 28 th days compared to the melanin content before use, respectively, and the melanin content was not significantly different (P >0.05) on the 14 th and 28 th days, which indicates that 0.5% gallic acid-glycine co-crystal had the whitening effect on the 14 th day, and the whitening effect was gradually stabilized from the 14 th to 28 th days. In example 9, the melanin contents were significantly different (P <0.05) on the 7 th, 14 th and 28 th days of application and the melanin contents were significantly different (P <0.05) on the 7 th and 14 th days and were not significantly different (P >0.05) on the 14 th and 28 th days, indicating that 1.0% gallic acid-glycine co-crystal had a whitening effect on the 7 th day, and the whitening effect was still enhanced from the 7 th to 14 th days, and the whitening effect was gradually stabilized until the 14 th day. In examples 10 and 11, the melanin contents were significantly different (P <0.05) on the 7 th, 14 th and 28 th days of application and the melanin contents were significantly different (P <0.05) on the 7 th and 14 th days and the melanin contents were significantly different (P <0.05) on the 14 th day and the 28 th day, respectively, indicating that the skin whitening effect was exhibited from the 7 th day to the 14 th day when the gallic acid-glycine co-crystal was 2.0% and 3.5%, and the skin whitening effect was still enhanced from the 7 th to the 14 th days. In example 12, the melanin contents were significantly different (P <0.05) on the 7 th, 14 th and 28 th days of application and the melanin contents were significantly different (P <0.05) on the 7 th and 14 th days of application, and the melanin and heme contents were not significantly different (P >0.05) on the 14 th and 28 th days of application, which indicates that gallic acid-glycine eutectic 5.0% had the whitening effect on the 7 th day, and the whitening effect was still enhanced from the 7 th to 14 th days, and the whitening effect was gradually stabilized after the 14 th day. From examples 6-12, it can be seen that the whitening effect of the picric acid-glycine co-crystal is gradually increased with the increase of the content, but the increase to 5% tends to be smooth.

Example 13: preparation of whitening creams of examples 6 to 12

While the foregoing is directed to the preferred embodiment of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.

Claims

1. A gallic acid and aminoacetic acid eutectic, characterized by that, said gallic acid and aminoacetic acid exist in stable eutectic form, adopt Cu-Kalpha radiation, in the X-ray powder diffraction spectrum expressed by degree 2 theta, gallic acid and aminoacetic acid eutectic appear the new peak in 2 theta ═ 26.3 degrees, in DSC curve, gallic acid and aminoacetic acid eutectic appear the new peak in 163 degrees centigrade; the molar ratio of the gallic acid to the aminoacetic acid is 1:1, and the gallic acid is prepared by adopting an absolute ethyl alcohol solvent assisted wet grinding method.

2. The gallic acid and aminoacetic acid eutectic crystal of claim 1, wherein the anhydrous ethanol solvent assisted wet grinding method comprises mixing gallic acid and aminoacetic acid at a molar ratio of 1:1, placing in a mortar, adding anhydrous ethanol for grinding, adding anhydrous ethanol while grinding to keep the sample in a slurry state all the time during the whole grinding process, and drying in a vacuum drier for later use.

3. The gallic acid and aminoacetic acid eutectic of claim 2, wherein said anhydrous ethanol is added to 1m L, and 1m L anhydrous ethanol is added dropwise every 2min for grinding for 30 min.

4. A whitening cream characterized by comprising the gallic acid and aminoacetic acid eutectic crystal as claimed in any one of claims 1 to 3, wherein the weight percentage of the gallic acid and the aminoacetic acid eutectic crystal is 0.2 to 5 percent.

5. The whitening cream of claim 4, further comprising one or more of an emulsifier, a thickener, a humectant, and a preservative;

the weight percentage of the emulsifier is 2% -15%;

the weight percentage of the thickening agent is 2% -10%;

the weight percentage of the humectant is 12-20%;

the weight percentage of the preservative is 0.1-1%.

6. The whitening cream according to claim 5,

the weight percentage of the emulsifier is 2% -10%;

the weight percentage of the thickening agent is 2% -8%;

the weight percentage of the humectant is 12-17%;

the weight percentage of the preservative is 0.1-0.5%.

7. The whitening cream according to claim 6, wherein the emulsifier is compounded from Montanov202 and SIMULSOL 165.

8. The whitening cream according to claim 6, wherein the thickener is formulated from hydroxyethyl cellulose and Sepiplus 400.

9. The whitening cream according to claim 6, wherein the humectant is selected from one or more of 1, 3-propylene glycol, pentanediol, hexanediol, sorbitol, glycerin, dipropylene glycol, 1, 3-butylene glycol, trehalose, xylitol, erythritol, glyceryl polyether-26, β -glucan, methyl glucitol polyether-20, methyl glucitol polyether-10, erythrulose, biogan-1, betaine, rhamnose, oat β -glucan, acetyl chitosamine, polyethylene glycols, water-soluble oils, water-soluble silicone oils, and sodium hyaluronate;

the preservative is a compound of a zanthoxylum bungeanum maxim extract, a Chinese pulsatilla root extract and a Spanish moss extract.

10. The whitening cream according to any one of claims 6 to 9, comprising the following components in percentage by weight: