Preparation method and application of escin for cosmetics

Abstract

The invention discloses a preparation method and application of escin for cosmetics, and belongs to the technical field of plant active ingredient extraction. The preparation method of the aescin comprises the following steps: adding aesculus chinensis seeds into an organic solvent, then adding alkaloid, carrying out heating reflux extraction, and then removing the organic solvent to obtain a crude extract; dissolving the crude extract in water, adding cyclic oligosaccharide, stirring for reaction, adding organic acid, heating for reaction, and purifying to obtain escin. The preparation method provided by the invention can improve the retention rate of aescin while improving the extraction rate of aescin.

Description

Technical Field

[0001] The invention relates to the technical field of plant active ingredient extraction, in particular to a method for preparing aescin for cosmetics and application thereof. Background Art

[0002] Horse chestnut (scientific name: Aesculus chinensis Bunge), also known as Solo tree, monkey chestnut, and Solo tree, is a deciduous tree of the Sapindaceae family. Its seeds are Solo seeds. Solo seeds are sweet and warm in nature. They have the effects of soothing the liver and regulating qi, harmonizing the stomach and relieving pain. They are often used to treat liver and stomach qi stagnation, chest and abdominal distension, and epigastric pain. Aescin is the main active ingredient of Solo seed extract, which has obvious pharmacological effects such as anti-inflammatory and detumescence, anti-oxidation, reducing nerve damage, and anti-tumor effects.

[0003] The current methods for extracting aescin mainly include ethanol reflux extraction, methanol ultrasonic extraction, organic solvent extraction and liquid chromatography. However, the ethanol reflux extraction method has the disadvantage of low extraction rate; the methanol ultrasonic extraction method has high noise and cannot be mass-produced, and methanol has certain toxicity. The aescin obtained by methanol extraction will have a trace of methanol residue, which is not suitable for use in cosmetics; the organic solvent extraction method requires a large amount of organic solvent, high energy consumption for recovering solvent, large environmental pollution, many process steps, long time consumption and high cost; the liquid chromatography method is inefficient and too expensive, and is not suitable for large-scale industrial production. In addition, in order to improve the extraction rate of aescin, the prior art usually adds sodium hydroxide that can increase the solubility of aescin during the extraction process, resulting in aescin being converted into sodium aescin, and it is impossible to obtain aescin with higher value, and the generated sodium aescin is not in the cosmetics use catalog and cannot be used as a cosmetic ingredient, thereby limiting its scope of application. How to improve the extraction rate and purity of aescin and overcome the problems of low extraction efficiency, multiple process steps and high cost in the prior art has become a technical problem to be solved by those skilled in the art. Summary of the invention

[0004] The purpose of the present invention is to provide a method for preparing aescin for cosmetics and its application, so as to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention: a method for preparing aescin, comprising the following steps:

[0007] The seeds of horse chestnut (Aesculus hippocastanum) are added to an organic solvent, and then an alkaloid is added, and the organic solvent is removed after heating and refluxing for extraction to obtain a crude extract;

[0008] Dissolve the crude extract in water, add cyclic oligosaccharide, stir and react, then add organic acid, heat and react, and purify to obtain the aescin.

[0009] Furthermore, the organic solvent includes an ethanol solution with a concentration of 70 vol.%.

[0010] Furthermore, the alkaloid includes matrine;

[0011] The dosage ratio of the aesculus seeds to the alkaloid is 1 kg: 8 - 10 g.

[0012] Matrine can increase the solubility of aescin and solve the problem of forming sodium aescin caused by using sodium hydroxide to increase the solubility of aescin.

[0013] Furthermore, the time for heating and reflux extraction is 3 - 5 h.

[0014] Furthermore, the cyclic oligosaccharide includes hydroxypropyl-β-cyclodextrin or hydroxypropyl-γ-cyclodextrin;

[0015] Cyclic oligosaccharide can protect the aescin component and reduce the loss of aescin during subsequent processing.

[0016] The dosage ratio of the aesculus seeds to the cyclic oligosaccharide is 1 kg: 300 - 500 g.

[0017] Furthermore, the organic acid includes ferulic acid;

[0018] The dosage ratio of the alkaloid to the organic acid is 8 - 10 g: 10 - 15 g.

[0019] Adding an excessive amount of organic acid can react completely with the alkaloid to form a water-soluble organic salt, which can remove the alkaloid in the system; and the organic acid is hardly soluble in cold water, and the excessive organic acid can be removed by lowering the temperature and filtering.

[0020] Furthermore, the purification includes: adding a decolorizing agent for decolorization (stirring and decolorizing at 70 °C for 4 - 6 h), and then passing through a cation exchange resin (adsorbing organic base cations) and an anion exchange resin (adsorbing organic acid radicals) in sequence to obtain an effluent;

[0021] Filter the effluent with an ultrafiltration membrane and then dry it to obtain the aescin.

[0022] Furthermore, the decolorizing agent is activated carbon and polyamide resin with a mass ratio of 3:1.

[0023] Furthermore, the filtration using the ultrafiltration membrane includes: first using the ultrafiltration membrane to remove macromolecular substances above 2500 Da, and then passing through a secondary ultrafiltration membrane to remove small molecular substances below 1000 Da.

[0024] Polyamide resin can adsorb impurity components such as flavonoids in the solution; activated carbon can adsorb some pigments, flavonoids, inorganic salts, etc. in the solution, and can remove some organic salts formed by organic acids and alkaloids.

[0025] The second technical solution of the present invention: Application of aescin prepared by the above preparation method in the preparation of cosmetics.

[0026] The present invention discloses the following technical effects:

[0027] (1) The preparation method of the present invention can improve the extraction rate of aescin while increasing the retention rate of aescin.

[0028] (2) The product obtained by the preparation method of the present invention is off-white in color, has good water solubility, and its solubility in water can reach 328.29 g / L.

[0029] (3) The preparation method of the present invention can avoid using sodium hydroxide, prevent the formation of aescin sodium from aescin, improve the extraction rate of aescin, and at the same time use cyclic oligosaccharides (such as cyclodextrin derivatives) to improve the recovery rate of aescin and enhance the stability of the final product.

[0030] (4) The method of the present invention has the advantages of high extraction efficiency, simple process, low cost, etc., and is suitable for large-scale industrial production. Description of the Drawings

[0031] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0032] Figure 1 It is the HPLC chromatogram of the aescin prepared in Example 1;

[0033] Figure 2 It is the picture before the stability test of the aescin prepared in Example 1;

[0034] Figure 3 It is the picture after the stability test of the aescin prepared in Example 1;

[0035] Figure 4 It is the physical picture of the aescin prepared in Example 1 and two commercially available aescin (commercially available sample 1 and commercially available sample 2);

[0036] Figure 5Prepare a solubility comparison chart of aescin prepared in Example 1 and two commercially available aescin (Commercially available sample 1 and Commercially available sample 2). Detailed Description of the Invention

[0037] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0038] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0039] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0040] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific embodiments of the present invention specification, which are obvious to those skilled in the art. Other embodiments obtained from the specification of the present invention are obvious to those skilled in the art. The specification and examples of the present invention are merely exemplary.

[0041] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0042] It should be noted that the aspects not described in detail in the present invention are all conventional operating means in the art and are not the focus of the present invention.

[0043] Example 1

[0044] A method for preparing aescin for cosmetics:

[0045] (1) Dry and crush the horse chestnut seeds, and sieve them (100 mesh) to obtain horse chestnut seed powder.

[0046] (2) Take 1 kg of the powder of Aesculus seeds, add 20 L of an ethanol aqueous solution with a concentration of 70 vol.%, then add 9 g of matrine, heat under reflux for 4 h, filter through a plate and frame filter press, and concentrate the filtrate under reduced pressure at 70 °C until there is no ethanol left to obtain the crude extract 1.

[0047] (3) Add deionized water to the crude extract 1 to make the total volume 3 L, cool it and then filter. Then add 400 g of hydroxypropyl-β-cyclodextrin to the filtrate, keep it warm (40 °C) and stir for 1 h (stirring speed is 50 r / min), then add 10 g of ferulic acid, heat to 70 °C and stir for 0.5 h (stirring speed is 50 r / min), quickly cool down the reaction solution, refrigerate for 8 h, and filter to obtain the crude extract 2.

[0048] (4) Dilute the crude extract 2 with deionized water to a solid content of 5%, add 30 g of a decolorizing agent (the decolorizing agent is activated carbon and polyamide resin with a mass ratio of 3:1), stir and decolorize at 70 °C for 5 h (stirring speed is 50 r / min), filter, quickly cool down the filtrate, pass the filtrate cooled to room temperature through an S1830 type cation exchange resin, and then through an S1200 type strong base anion exchange resin, repeat the conventional column passing operation twice, take the final effluent, remove macromolecular substances above 2500 Da through an ultrafiltration membrane, and then remove small molecular substances below 1000 Da through a secondary ultrafiltration membrane, and dry the filtrate to obtain aescin.

[0049] Example 2

[0050] A preparation method of aescin for cosmetics:

[0051] (1) Dry the Aesculus seeds, crush them and sieve them (100 mesh) to obtain the powder of Aesculus seeds.

[0052] (2) Take 1 kg of the powder of Aesculus seeds, add 20 L of an ethanol aqueous solution with a concentration of 70 vol.%, then add 10 g of matrine, heat under reflux for 3 h, filter through a plate and frame filter press, and concentrate the filtrate under reduced pressure at 70 °C until there is no ethanol left to obtain the crude extract 1.

[0053] (3) Add deionized water to the crude extract 1 to make the total volume 3 L, cool it and then filter. Then add 500 g of hydroxypropyl-γ-cyclodextrin to the filtrate, keep it warm (40 °C) and stir for 1 h (stirring speed is 50 r / min), then add 15 g of ferulic acid, heat to 70 °C and stir for 0.5 h (stirring speed is 50 r / min), quickly cool down the reaction solution, refrigerate for 8 h, and filter to obtain the crude extract 2.

[0054] (4) Add deionized water to the crude extract 2 to dilute it to a solid content of 5%, add 30 g of decolorizing agent (the decolorizing agent is activated carbon and polyamide resin with a mass ratio of 3:1), stir and decolorize at 70 °C for 5 h (stirring speed is 50 r / min), filter, quickly cool the filtrate, pass the filtrate cooled to room temperature through S1830 cation exchange resin, and then through S1200 strong base anion exchange resin. Repeat the conventional column operation twice, take the final effluent, remove macromolecular substances above 2500 Da through an ultrafiltration membrane, and then remove small molecular substances below 1000 Da through a secondary ultrafiltration membrane. Dry the filtrate to obtain aescin.

[0055] Comparative Example 1

[0056] Same as Example 1, except that matrine is replaced with an equal mass of sodium hydroxide.

[0057] Sodium hydroxide is a strong base, which will greatly increase the extraction rate of aescin, but it is easy to react with aescin to form sodium aescin, resulting in a decrease in the final retention rate of aescin. Secondly, because it is a strong base, it will increase the operation difficulty of the subsequent desalting process.

[0058] Comparative Example 2

[0059] Same as Example 1, except that matrine is replaced with an equal mass of L-carnitine.

[0060] L-carnitine has a fishy smell, and through testing, it is found that it does not greatly improve the extraction rate of aescin, and the final product has a slight fishy smell, which limits its use.

[0061] Comparative Example 3

[0062] Same as Example 1, except that ferulic acid is replaced with an equal mass of gallic acid.

[0063] Gallic acid is stronger in acidity than ferulic acid. An overly strong acidic substance will destroy the stable system of aescin and cyclic oligosaccharides, resulting in the loss of aescin during the subsequent desalting process.

[0064] Comparative Example 4

[0065] Same as Example 1, except that ferulic acid is replaced with an equal mass of succinic acid.

[0066] Succinic acid has a certain solubility in water, which will increase the difficulty of subsequent removal, and it has an irritating smell after heating, which will affect the smell of the final product.

[0067] Comparative Example 5

[0068] Same as Example 1, except that hydroxypropyl-β-cyclodextrin is replaced with an equal mass of chitosan (molecular weight is 10 kDa).

[0069] When the molecular weight of chitosan is too large, during the subsequent ultrafiltration membrane stage, it will cause part of the aescin it encapsulates to be filtered, affecting the final retention rate of aescin, and the obtained aescin has poor water solubility and stability.

[0070] Comparative Example 6

[0071] Same as Example 1, the difference is only that step (3) is specifically: deionized water is added to the crude extract to make the total volume 3L, cooled and then filtered, and then 10g of ferulic acid is added to the filtrate, heated to 70°C and stirred for 0.5h (stirring speed is 50r / min), the reaction solution is quickly cooled, refrigerated for 8h, and filtered to obtain crude extract 2.

[0072] No cyclic oligosaccharide was added in this comparative example, so it will cause excessive loss of aescin components, poor water solubility and stability in the subsequent decolorization and desalting processes.

[0073] Effect Example 1

[0074] Determine the content of aescin in the horse chestnut seed powder and aescin, and calculate the extraction rate of aescin. The aescin includes (aescin A, aescin B, aescin C, aescin D, and the HPLC chromatogram of the aescin prepared in Example 1 is shown in Figure 1 ), and the determination method refers to the content detection under the item of Semen Aesculi in Part I of the Pharmacopoeia 2020 Edition.

[0075] The extraction rate of aescin = (the content of aescin in crude extraction 1 × the mass of crude extraction 1) / (the content of aescin in the horse chestnut seed powder × the mass of the horse chestnut seed powder) × 100%.

[0076] Determine the retention rate of aescin in the aescin, and the calculation method is as follows:

[0077] The retention rate of aescin = (the content of aescin in the aescin × the mass of the aescin) / (the content of aescin in crude extraction 1 × the mass of crude extraction 1) × 100%.

[0078] Table 1 Extraction rate and retention rate of aescin

[0079] Grouping Extraction rate of aescin (%) Retention rate of aescin (%) Example 1 98.75 89.84 Example 2 98.38 89.64 Comparative Example 1 98.79 46.72 Comparative Example 2 85.62 75.39 Comparative Example 3 98.28 79.36 Comparative Example 4 98.16 76.92 Comparative Example 5 98.63 57.69 Comparative Example 6 98.25 40.73

[0080] Effect Example 2

[0081] Dissolve the aescin prepared in Example 1 in deionized water to make the concentration of aescin 1 wt.%, obtaining an aqueous solution of aescin with a concentration of 1 wt.%, which is in a clear and transparent state. Then, place it under the conditions of normal temperature, low temperature (4°C), high temperature (45°C), pH 5 - 6 (45°C), and pH 4 - 10 for 3 months to test the stability of aescin.

[0082] Dissolve the aescin prepared in Example 1 in a 5 wt.% butanediol solution to make the concentration of aescin 1 wt.%, obtaining a butanediol solution of aescin with a concentration of 1 wt.%. Place it under the conditions of normal temperature, high temperature (45°C), and pH 4 - 6 for 6 months to test the stability of aescin.

[0083] The figure before the test is shown in Figure 2 , and the figure after the test is shown in Figure 3 .

[0084] Figure 2 and Figure 3 From left to right in

[0085] are the stabilities of the aqueous solution of aescin with a concentration of 1 wt.% under the conditions of normal temperature, low temperature (4°C), high temperature (45°C), pH 5 (45°C), pH 6 (45°C), pH 4, pH 5, pH 6, pH 7, pH 8, pH 9, pH 10, and the stabilities of the butanediol solution of aescin with a concentration of 1 wt.% under the conditions of normal temperature, high temperature (45°C), pH 4, pH 5, pH 6.

[0086] The physical pictures of the aescin prepared in Example 1 and two commercially available aescin (commercially available sample 1 and commercially available sample 2) are shown in Figure 4 ;

[0087] Add the aescin prepared in Example 1 and two commercially available aescin (commercially available sample 1 and commercially available sample 2) into water respectively to prepare solutions with a concentration of 10 wt.%, and conduct a solubility comparison. The results are shown in Figure 5 .

[0088] It can be seen from Figures 2 - 5 that the aescin prepared in Example 1 of the present invention has good stability, is off - white in color, and has good water solubility.

[0089] The above - described embodiments are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A preparation method of aescin, characterized in that, It includes the following steps: Add the horse chestnut seeds into an organic solvent, then add an alkaloid, extract by heating under reflux, and remove the organic solvent to obtain a crude extract; Dissolve the crude extract in water, add a cyclic oligosaccharide, stir and react, then add an organic acid, heat and react, and purify to obtain the aescin; 2. The preparation method according to claim 1, characterized in that, The organic solvent includes an ethanol solution with a concentration of 70 vol.%; 3. The preparation method according to claim 1, characterized in that, The alkaloid includes matrine; The dosage ratio of the horse chestnut seeds to the alkaloid is 1 kg: 8 - 10 g; 4. The preparation method according to claim 1, characterized in that, The time for heating under reflux extraction is 3 - 5 h; 5. The preparation method according to claim 1, characterized in that The cyclic oligosaccharide includes hydroxypropyl-β-cyclodextrin or hydroxypropyl-γ-cyclodextrin; The dosage ratio of the horse chestnut seeds to the cyclic oligosaccharide is 1 kg: 300 - 500 g; 6. The preparation method according to claim 1, characterized in that, The organic acid includes ferulic acid; The dosage ratio of the alkaloid to the organic acid is 8 - 10 g: 10 - 15 g; 7. The preparation method according to claim 1, wherein The purification includes: adding a decolorizing agent for decolorization, and then passing through a cation exchange resin and an anion exchange resin in sequence to obtain an effluent; Filter the effluent with an ultrafiltration membrane and then dry it to obtain the aescin; 8. The preparation method according to claim 7, characterized in that, The decolorizing agent is activated carbon and polyamide resin with a mass ratio of 3:1; 9. The preparation method according to claim 7, characterized in that, The filtration with the ultrafiltration membrane includes: first removing macromolecular substances with a molecular weight above 2500 Da with the ultrafiltration membrane, and then removing small molecular substances with a molecular weight below 1000 Da through a secondary ultrafiltration membrane; 10. Use of aescin prepared by the preparation method according to any one of claims 1 - 9 in the preparation of cosmetics.

Images