A deep eutectic solvent composition capable of forming reverse micelles and a method of making the same
By preparing a eutectic solvent composition consisting of a hydrogen bond donor, a hydrogen bond acceptor, and an oil phase carrier, the problems of instability and excessive hydrophilicity of reverse micelles were solved, achieving stability and skin permeability, and it can be applied to protein separation and purification.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- GUANGZHOU KEYING COSMETICS CO LTD
- Filing Date
- 2026-03-18
- Publication Date
- 2026-06-05
AI Technical Summary
In existing methods for preparing reverse micelles, the stability of reverse micelles is affected by factors such as light, heat, and oxygen, leading to instability. At the same time, the excessive hydrophilicity of water-soluble active ingredients prevents them from penetrating the skin.
Reverse micelles are prepared by using a eutectic solvent composition consisting of a hydrogen bond donor, a hydrogen bond acceptor, and an oil phase carrier, through mixing, evaporation, homogenization, and the addition of water-soluble active ingredients. The hydrogen bond donors are selected from glycyrrhizic acid, azelaic acid, C10~C23 saturated fatty acids, etc., and the oil phase carriers are selected from isononyl isononanoate, meadowfoam seed oil, etc.
It improves the stability of reverse micelles, solves the problems of instability due to light, heat and oxygen, and enables skin penetration of water-soluble active ingredients by improving hydrophilicity.
Smart Images
Figure CN122141289A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of reverse micelle technology, specifically to a eutectic solvent composition capable of forming reverse micelles and a method for preparing the same. Background Technology
[0002] Reversed micelles are nanoscale aggregates spontaneously formed by surfactants in organic solvents; they are transparent, thermodynamically stable systems. Reversed micelles possess unique solubility properties, capable of dissolving biomolecules such as amino acids, peptides, and proteins, providing a hydrophilic microenvironment for these molecules. Therefore, they are commonly used for the separation and purification of biomolecules, such as protein extraction and the separation of protein mixtures. Reversed micelle extraction is a liquid-liquid extraction method that utilizes various surfactants to separate organic and inorganic substances, and its unique advantages have led to its wide range of applications.
[0003] The reverse micelles prepared by existing reverse micelle preparation methods are unstable due to the instability of the water-soluble active substances in them to light, heat, oxygen, etc. Summary of the Invention
[0004] This invention provides a eutectic solvent composition capable of forming reverse micelles and a method for preparing the same, thereby improving the stability of reverse micelles.
[0005] To solve the above problems, the technical solution provided by the present invention is as follows: A eutectic solvent composition capable of forming reverse micelles, comprising a hydrogen bond donor, a hydrogen bond acceptor, and an oil phase carrier, wherein the hydrogen bond acceptor is a Sophora flavescens extract.
[0006] The hydrogen bond donor is any one of glycyrrhizic acid, azelaic acid, C10~C23 saturated fatty acids, and 10-hydroxydecanoic acid.
[0007] The hydrogen bond donors are C10-C16 saturated fatty acids.
[0008] The oil phase carrier is any one of isononyl isononanoate, meadowfoam seed oil, sunflower seed oil, triglyceride (ethylhexanoate), and jojoba seed oil.
[0009] The mass ratio of the hydrogen bond donor, hydrogen bond acceptor, and oil phase carrier is 1-5:2-6:4-8.
[0010] The method for preparing the above-mentioned eutectic solvent composition capable of forming reverse micelles includes the following steps: 1) Mix the hydrogen bond donor and hydrogen bond acceptor evenly in a solvent to obtain a mixture; 2) Evaporate the mixture from step 1) to remove the solvent, and obtain a concentrated solution; 3) Mix the concentrate from step 2) with the oil phase carrier evenly, then add the water-soluble active ingredient, mix evenly at 40-50℃, and homogenize to obtain the final product.
[0011] Preferably, the solvent in step 1) is one of ethanol and methanol.
[0012] Preferably, the temperature at which the hydrogen bond donor and hydrogen bond acceptor are mixed in the solvent in step 1) is 40-50°C.
[0013] Preferably, in step 3), the mass ratio of the concentrate to the oil phase carrier is 1-5:3-7, and more preferably 1-3:3-7.
[0014] Preferably, the water-soluble active ingredient is added dropwise in step 3).
[0015] Preferably, the water-soluble active ingredient is at least one selected from PDRN, cyclic peptides, chain polypeptides, and small molecule compounds, wherein the small molecule compound is at least one selected from vitamin C, nicotinamide, and tranexamic acid. The chain polypeptide is any one selected from dipeptide-15, nonapeptide-1, and acetyl hexapeptide-8.
[0016] The present invention provides the application of the eutectic solvent composition capable of forming reverse micelles in protein separation and purification.
[0017] The beneficial effects of this invention are: The eutectic solvent composition of the present invention can solve the problem in the prior art that the instability of the active material to light, heat, oxygen, etc., leads to the instability of the reverse micelles, and can also solve the problem that water-soluble active materials cannot penetrate the skin due to excessive hydrophilicity. Attached Figure Description
[0018] Figure 1 A graph showing the variation of water content in different oil phase carriers; Figure 2 The results of LUM stability analysis test for sunflower seed oil as the oil phase carrier; Figure 3 Image of a sample with 30% water content, using sunflower seed oil as the oil phase carrier; Figure 4 A graph showing the cumulative permeation of vitamin C using sunflower seed oil as an oil phase carrier. Figure 5 A graph showing the decay trend of vitamin C under high temperature conditions using sunflower seed oil as the oil phase carrier; Figure 6 A graph showing the cumulative permeation of PDRN using sunflower seed oil as the oil phase carrier. Detailed Implementation
[0019] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments and test examples. However, those skilled in the art should understand that the embodiments are only used to illustrate the technical solution of the present invention and should not be regarded as limiting the scope of protection of the present invention.
[0020] Unless otherwise specified, the raw materials, reagents, instruments, etc. used in the following embodiments are all commonly used in the field and are publicly available or commercially obtainable.
[0021] The extraction method for Sophora flavescens extract can be adopted using existing technologies. The method used in the example includes: selecting Sophora flavescens root, washing and cutting it into sections, extracting it by reflux with anhydrous ethanol, removing the solvent by vacuum evaporation of the extract, and then separating and purifying it by column chromatography.
[0022] Example 1 The eutectic solvent composition capable of forming reverse micelles in this embodiment is made of a hydrogen bond donor, a hydrogen bond acceptor, and an oil phase carrier, with a mass ratio of 2.4:0.6:7. The hydrogen bond donor is glycyrrhizic acid, the hydrogen bond acceptor is Sophora flavescens extract, and the oil phase carrier is sunflower seed oil.
[0023] The method for preparing the eutectic solvent composition capable of forming reverse micelles in this embodiment includes the following steps: 1) Take the hydrogen bond donor and hydrogen bond acceptor, add them to ethanol, and stir at 40°C to mix them evenly to obtain a mixture; the hydrogen bond donor is glycyrrhizic acid, and the hydrogen bond acceptor is Sophora flavescens extract; the mass ratio of hydrogen bond donor to hydrogen bond acceptor is 4:1. 2) The mixture from step 1) is subjected to rotary evaporation to remove ethanol, resulting in a concentrated solution; 3) Add the concentrate obtained in step 2) to the oil phase carrier and stir to mix evenly; the mass ratio of concentrate to oil phase carrier is 3:7; the oil phase carrier is sunflower seed oil; 4) Take a 2% PDRN aqueous solution (the amount used is 10% of the concentrated solution in step 2) and add it dropwise to the liquid that has been mixed evenly in step 3). Stir while adding the solution. After the addition is complete, stir at 40°C for 6 hours. Then pour the stirred liquid into a high-pressure microjet device, homogenize and circulate it 3 times, and then discharge the product.
[0024] Example 2 The eutectic solvent composition capable of forming reverse micelles in this embodiment is made of a hydrogen bond donor, a hydrogen bond acceptor, and an oil phase carrier, with a mass ratio of 2.1:0.9:7. The hydrogen bond donor is azelaic acid, the hydrogen bond acceptor is Sophora flavescens extract, and the oil phase carrier is sunflower seed oil.
[0025] The method for preparing the eutectic solvent composition capable of forming reverse micelles in this embodiment includes the following steps: 1) Take the hydrogen bond donor and hydrogen bond acceptor, add them to ethanol, and stir at 40°C to mix them evenly to obtain a mixture; the hydrogen bond donor is azelaic acid, and the hydrogen bond acceptor is Sophora flavescens extract; the mass ratio of hydrogen bond donor to hydrogen bond acceptor is 2.1:0.9. 2) The mixture from step 1) is subjected to rotary evaporation to remove ethanol, resulting in a concentrated solution; 3) Add the concentrate obtained in step 2) to the oil phase carrier and stir to mix evenly; the mass ratio of concentrate to oil phase carrier is 3:7; the oil phase carrier is sunflower seed oil; 4) Take a 2% PDRN aqueous solution (the amount used is 10% of the concentrated solution in step 2) and add it dropwise to the liquid that has been mixed evenly in step 3). Stir while adding the solution. After the addition is complete, stir at 40°C for 6 hours. Then pour the stirred liquid into a high-pressure microjet device, homogenize and circulate it 3 times, and then discharge the product.
[0026] Example 3 The eutectic solvent composition capable of forming reverse micelles in this embodiment and its preparation method differ from those in Example 1 in that the hydrogen bond donor is a C10 saturated fatty acid, while the others are the same as in Example 1.
[0027] Example 4 The difference between the eutectic solvent composition capable of forming reverse micelles and its preparation method in this embodiment and that in Example 1 is that the hydrogen bond donor is 10-hydroxydecanoic acid, while the others are the same as in Example 1.
[0028] Comparative Example 1 The difference between the eutectic solvent composition and its preparation method in this comparative example, which can form reverse micelles, and Example 3 is that the hydrogen bond donor is a C9 saturated fatty acid. After the eutectic solvent formed by this composition is added to the oil phase, phase separation occurs at low temperature, indicating that it is unstable.
[0029] Comparative Example 2 The difference between the eutectic solvent composition and its preparation method in this comparative example, which can form reverse micelles, and Example 3 is that the hydrogen bond donor is a C17 saturated fatty acid. After the eutectic solvent formed by this is added to the oil phase, a stable supramolecular reverse micelle oil solution is formed. Subsequently, after adding an aqueous solution containing active ingredients, phase separation occurs due to the inability to effectively encapsulate the micelles. It is speculated that the carbon chain is too long, resulting in excessive hydrophobicity, which leads to the water core being too small to effectively encapsulate the micelles.
[0030] Comparative Example 3 The eutectic solvent composition and its preparation method for forming reverse micelles in this comparative example differ from those in Example 1 in that the hydrogen bond acceptors used are other commonly used hydrogen bond acceptors, such as betaine, carnitine, and choline chloride. These eutectic solvents exhibit varying degrees of crystallization and precipitation, and are insoluble in the carrier oil.
[0031] Comparative Example 4 The difference between the eutectic solvent composition and its preparation method in this comparative example, which can form reverse micelles, and Example 2 is that the hydrogen bond donor is a short-chain dicarboxylic acid—adipic acid. The resulting eutectic solvent is immiscible with the oil phase carrier, is a hydrophilic eutectic solvent, and cannot form reverse micelles.
[0032] Experimental Example 1 Water-carrying capacity test: Following the steps of Example 3, the oil phase carrier was replaced with different oils, including: jojoba seed oil, squalane, sunflower seed oil, meadowfoam seed oil, isononyl isononanoate, caprylic / capric triglyceride, isohexadecane, ethylhexyl palmitate, and macadamia seed oil. After adding the oil phase carrier, water was slowly added dropwise, and the endpoint was considered when the oil phase solution began to separate or become turbid. The results showed that sunflower seed oil had a good water-carrying capacity, such as... Figure 1 As shown. Figure 3 Image of a sample with 30% water content using sunflower seed oil as the oil phase carrier.
[0033] Experiment Example 2 Following the steps in Example 3, the obtained sample underwent a LUM stability analysis test. The specific operation was as follows: the sample was continuously irradiated using a multi-light source system, centrifuged at 4000 rpm, tested for 12 hours, and data was acquired every 45 seconds. The results showed that the reverse micelle sample exhibited good stability, such as... Figure 2 As shown.
[0034] Experimental Example 3 Following the steps of Example 3, vitamin C was prepared into an aqueous solution and added dropwise to reverse micelle oil for encapsulation, resulting in a final total vitamin C content of 10,000 ppm. An in vitro transdermal absorption test was then performed: Ex vivo skin samples from undamaged Bama miniature pigs were cleaned with physiological saline, and the skin was fixed to a Franz diffusion cell with the keratin layer facing upwards. PBS was added to the receiving cell, and after equilibration for 30 minutes, air bubbles were removed by tapping with a rubber bulb. The water bath temperature was set to 32±1℃, and 1 g of the prepared sample was accurately weighed. The receiving cell stirring speed was set to 600 rpm. Samples were collected at 4h, 8h, 12h, 24h, and 48h, centrifuged at 12,000 rpm for 10 minutes, and the supernatant was collected. After filtering through a 0.22 μm filter membrane, liquid chromatography was performed, and the cumulative permeate was calculated, as shown in the attached figure. Figure 4 As shown, the diffusion cell has an opening diameter of 20 mm and an effective diffusion area of approximately 3.14 cm².2 .
[0035] The formula for calculating cumulative transmittance is as follows:
[0036] in, V represents the cumulative permeation rate, and V is the total volume of the receiving liquid. The concentration of the drug in the receiving solution during the nth sampling is given. Let be the mass concentration of the drug in the receiving solution during the i-th sampling. Where A is the sampling volume and A is the effective diffusion area.
[0037] Experiment Example 4 Following the steps of Example 3, vitamin C was prepared into an aqueous solution and added dropwise to the reverse micelle oil for encapsulation, resulting in a final total vitamin C content of 10,000 ppm. A vitamin C decay test was then performed using the following method: The prepared reverse micelle vitamin C and an aqueous solution of vitamin C of the same concentration were placed at 45°C, and samples were taken at 0, 7, 14, and 28 days. The vitamin C content was tested using ultraviolet spectrophotometry at a wavelength of 246 nm. (See attached...) Figure 5 As shown.
[0038] Experimental Example 5 Following the steps of Example 3, PDRN was prepared as an aqueous solution and added dropwise to reverse micelle oil for encapsulation, resulting in a final total PDRN content of 500 ppm. An in vitro transdermal absorption test was then performed: Ex vivo skin samples from undamaged Bama miniature pigs were cleaned with physiological saline, and the skin was fixed to a Franz diffusion cell with the stratum corneum facing upwards. PBS was added to the receiving cell, and after equilibration for 30 minutes, air bubbles were removed by tapping with a rubber bulb. The water bath temperature was set to 32 ± 1℃, and 1 g of the prepared sample was accurately weighed. The receiving cell stirring speed was set to 600 rpm. Samples were collected at 4 h, 8 h, 12 h, 24 h, and 48 h, centrifuged at 12000 rpm for 10 minutes, and the supernatant was collected. After filtering through a 0.22 μm filter membrane, liquid chromatography was performed, and the cumulative permeate was calculated as shown in the attached figure. Figure 6 As shown, the diffusion cell has an opening diameter of 20 mm and an effective diffusion area of approximately 3.14 cm². 2 .
[0039] The formula for calculating cumulative transmittance is as follows:
[0040] in, V represents the cumulative permeation rate, and V is the total volume of the receiving liquid. The concentration of the drug in the receiving solution during the nth sampling is given. Let be the mass concentration of the drug in the receiving solution during the i-th sampling. Where A is the sampling volume and A is the effective diffusion area.
Claims
1. A eutectic solvent composition capable of forming reverse micelles, characterized in that, It is made of hydrogen bond donor, hydrogen bond acceptor and oil phase carrier, wherein the hydrogen bond acceptor is Sophora flavescens extract.
2. The eutectic solvent composition capable of forming reverse micelles according to claim 1, characterized in that, The hydrogen bond donor is any one of glycyrrhizic acid, azelaic acid, C10~C23 saturated fatty acids, and 10-hydroxydecanoic acid.
3. The eutectic solvent composition capable of forming reverse micelles according to claim 1, characterized in that, The oil phase carrier is any one of isononyl isononanoate, meadowfoam seed oil, sunflower seed oil, triglyceride (ethylhexanoate), and jojoba seed oil.
4. The eutectic solvent composition capable of forming reverse micelles according to claim 1, characterized in that, The mass ratio of the hydrogen bond donor, hydrogen bond acceptor, and oil phase carrier is 1-5:2-6:4-8.
5. A method for preparing a eutectic solvent composition capable of forming reverse micelles as described in claim 1, characterized in that, Includes the following steps: 1) Mix the hydrogen bond donor and hydrogen bond acceptor evenly in a solvent to obtain a mixture; 2) Evaporate the mixture from step 1) to remove the solvent, and obtain a concentrated solution; 3) Mix the concentrate from step 2) with the oil phase carrier evenly, then add the water-soluble active ingredient, mix evenly at 40-50℃, and homogenize to obtain the final product.
6. The method for preparing the eutectic solvent composition capable of forming reverse micelles according to claim 5, characterized in that, The solvent mentioned in step 1) is either ethanol or methanol.
7. The method for preparing the eutectic solvent composition capable of forming reverse micelles according to claim 5, characterized in that, In step 1), the temperature at which the hydrogen bond donor and hydrogen bond acceptor are mixed in the solvent is 40-50℃.
8. The method for preparing the eutectic solvent composition capable of forming reverse micelles according to claim 5, characterized in that, In step 3), the mass ratio of the concentrate to the oil phase carrier is 1-5:3-7.
9. The method for preparing the eutectic solvent composition capable of forming reverse micelles according to claim 1, characterized in that, The water-soluble active ingredient is at least one of PDRN, cyclic peptide, chain polypeptide, and small molecule compound, wherein the small molecule compound is at least one of vitamin C, nicotinamide, and tranexamic acid.
10. The use of any of the eutectic solvent compositions capable of forming reverse micelles as described in claims 1 to 4 in the separation and purification of proteins.