A method for preparing and applying aescin for cosmetic use

Abstract

This invention discloses a method for preparing and applying aescin in cosmetics, belonging to the field of plant active ingredient extraction technology. The method for preparing aescin includes the following steps: adding horse chestnut seeds to an organic solvent, then adding an alkaloid, heating and refluxing to extract, and then removing the organic solvent to obtain a crude extract; dissolving the crude extract in water, adding a cyclic oligosaccharide, stirring and reacting, then adding an organic acid, heating and reacting, and finally purifying to obtain aescin. The preparation method of this invention can improve both the extraction rate and retention rate of aescin.

Description

Technical Field

[0001] This invention relates to the field of plant active ingredient extraction technology, and in particular to a method for preparing and applying aescin for cosmetic use. Background Technology

[0002] Horse chestnut (scientific name: *Aesculus chinensis* Bunge), also known as spinach, monkey chestnut, and sal tree, is a deciduous tree belonging to the genus *Aesculus* in the family Sapindaceae. Its seeds are called sal seeds. Sal seeds are sweet and warm in nature, and 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 stomach pain. Aescin is the main active ingredient in sal seed extract, and it exhibits significant pharmacological effects such as anti-inflammatory and swelling-reducing properties, antioxidant activity, nerve damage reduction, and anti-tumor activity.

[0003] Current methods for extracting aescin mainly include ethanol reflux extraction, methanol ultrasonic extraction, organic solvent extraction, and liquid chromatography. However, ethanol reflux extraction suffers from low extraction rates; methanol ultrasonic extraction is noisy, unsuitable for large-scale production, and methanol is toxic, leaving trace amounts of methanol residue, making it unsuitable for cosmetic use; organic solvent extraction requires large amounts of organic solvents, resulting in high energy consumption for solvent recovery, significant environmental pollution, numerous process steps, long processing time, and high costs; liquid chromatography is inefficient and too expensive, making it unsuitable for large-scale industrial production. Furthermore, existing technologies often add sodium hydroxide to increase aescin solubility during extraction to improve extraction rates, causing aescin to convert into sodium aescinate, failing to obtain the more valuable aescin. Moreover, sodium aescinate is not listed in the cosmetics usage catalog and cannot be used as a cosmetic ingredient, thus limiting its application scope. 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 existing technologies has become a technical problem that needs to be solved by those skilled in the art. Summary of the Invention

[0004] The purpose of this invention is to provide a method for preparing and applying aescin in cosmetics, so as to solve the problems existing in the prior art.

[0005] To achieve the above objectives, the present invention provides the following solution:

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

[0007] Horse chestnut seeds (Brahma seeds) were added to an organic solvent, followed by the addition of alkaloids. The mixture was heated under reflux and then the organic solvent was removed to obtain a crude extract.

[0008] The crude extract was dissolved in water, then cyclic oligosaccharides were added, the mixture was stirred and reacted, organic acids were added, the mixture was heated and reacted, and then purified to obtain the aesculin.

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

[0010] Furthermore, the alkaloids include matrine;

[0011] The ratio of horse chestnut seeds to alkaloids is 1 kg: 8-10 g.

[0012] Matrine can increase the solubility of aescin, thus solving the problem of sodium aescin formation caused by using sodium hydroxide to increase the solubility of aescin.

[0013] Furthermore, the heating and reflux extraction time is 3 to 5 hours.

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

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

[0016] The ratio of horse chestnut seeds to cyclic oligosaccharides is 1 kg: 300-500 g.

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

[0018] The ratio of alkaloids to organic acids is 8-10g:10-15g.

[0019] Adding excess organic acid can react completely with alkaloids to produce water-soluble organic salts, which can remove alkaloids from the system; and since organic acid is poorly soluble in cold water, excess organic acid can be removed by lowering the temperature and filtering.

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

[0021] The effluent was filtered using an ultrafiltration membrane and then dried to obtain the aesculin.

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

[0023] Furthermore, the ultrafiltration process includes: first removing macromolecular substances with a value of 2500 Da or higher using an ultrafiltration membrane, and then removing small molecular substances with a value of 1000 Da or lower using a secondary ultrafiltration membrane.

[0024] Polyamide resin can adsorb impurities 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: the application of aesculin 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 and the retention rate of aescin at the same time.

[0028] (2) The product obtained by the preparation method of the present invention is off-white, 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 the use of sodium hydroxide, prevent the formation of sodium aescinate from aescin, and improve the extraction rate of aescin. At the same time, the use of cyclic oligosaccharides (such as cyclodextrin derivatives) improves the recovery rate of aescin and enhances the stability of the final product.

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

[0031] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 The HPLC chromatogram of aescin prepared in Example 1;

[0033] Figure 2 A diagram showing the aescin prepared in Example 1 before stability testing;

[0034] Figure 3 The image shows the stability test results of the aescin prepared in Example 1.

[0035] Figure 4 The images show the physical samples of aescin prepared in Example 1 and two commercially available aescins (commercial sample 1 and commercial sample 2).

[0036] Figure 5The diagram shows a comparison of the solubility of aescin prepared in Example 1 and two commercially available aescins (commercial sample 1 and commercial sample 2). Detailed Implementation

[0037] 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 terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0039] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0040] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0041] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0042] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0043] Example 1

[0044] A method for preparing aescin for cosmetic use:

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

[0046] (2) Take 1 kg of horse chestnut seed powder, add 20 L of 70 vol.% ethanol aqueous solution, then add 9 g of matrine, heat and reflux for 4 h, filter through plate and frame filter, concentrate the filtrate under reduced pressure at 70 °C until no ethanol is present, and obtain crude extract 1.

[0047] (3) Add deionized water to crude extract 1 to a total volume of 3L, let cool and filter. Then add 400g of hydroxypropyl-β-cyclodextrin to the filtrate, keep warm (40℃) and stir for 1h (stirring speed 50r / min). Then add 10g of ferulic acid, heat to 70℃ and stir for 0.5h (stirring speed 50r / min). The reaction solution is rapidly cooled and refrigerated for 8h. Filter to obtain crude extract 2.

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

[0049] Example 2

[0050] A method for preparing aescin for cosmetic use:

[0051] (1) Dry the horse chestnut seeds, then crush and sieve them (100 mesh) to obtain horse chestnut seed powder.

[0052] (2) Take 1 kg of horse chestnut seed powder, add 20 L of 70 vol.% ethanol aqueous solution, then add 10 g of matrine, heat and reflux for 3 h, filter through plate and frame filter, concentrate the filtrate under reduced pressure at 70 °C until no ethanol is present, and obtain crude extract 1.

[0053] (3) Add deionized water to crude extract 1 to a total volume of 3L, let it cool and filter it. Then add 500g of hydroxypropyl-γ-cyclodextrin to the filtrate, keep it warm (40℃) and stir for 1h (stirring speed of 50r / min). Then add 15g of ferulic acid, heat to 70℃ and stir for 0.5h (stirring speed of 50r / min). The reaction solution is rapidly cooled and refrigerated for 8h. After filtration, crude extract 2 is obtained.

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

[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 alkali, which can significantly increase the extraction rate of aescin. However, it easily reacts with aescin to form sodium aescinate, resulting in a decrease in the final retention rate of aescin. Furthermore, because it is a strong alkali, it increases the difficulty of subsequent desalination processes.

[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 meaty smell, and tests have shown that it does not significantly improve the extraction rate of aescin, and the final product has a slight meaty 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 more acidic than ferulic acid. Excessively strong acidic substances can disrupt the stable system of aescin and cyclic oligosaccharides, leading to the loss of aescin during subsequent desalination.

[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 degree of solubility in water, which increases the difficulty of subsequent removal, and it has an irritating odor when heated, which will affect the odor 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 of 10 kDa).

[0069] If the molecular weight of chitosan is too large, it will cause some of the aescin to be filtered out during the subsequent ultrafiltration stage, affecting the final aescin retention rate. In addition, the obtained aescin has poor water solubility and poor stability.

[0070] Comparative Example 6

[0071] Same as Example 1, except that step (3) is as follows: add deionized water to the crude extract until the total volume is 3L, cool and filter, then add 10g ferulic acid to the filtrate, heat to 70℃ and stir for 0.5h (stirring speed is 50r / min), cool the reaction solution quickly, refrigerate for 8h, filter, and obtain crude extract 2.

[0072] This comparative sample did not contain cyclic oligosaccharides, which resulted in excessive loss of aescin components during subsequent decolorization and desalination processes, leading to poor water solubility and instability.

[0073] Example 1

[0074] The content of aescin in horse chestnut seed powder and aescin was determined, and the extraction rate of aescin was calculated. The aescin included aescin A, aescin B, aescin C, and aescin D. The HPLC chromatogram of the aescin prepared in Example 1 is shown in [reference needed]. Figure 1 The determination method refers to the content detection under the "Medicinal Materials and Processed Scutellaria baicalensis" section of Part I of the 2020 Pharmacopoeia. 。

[0075] The extraction rate of aescin is calculated as follows: (aescin content in crude extract 1 × mass of crude extract 1) / (aescin content in horse chestnut seed powder × mass of horse chestnut seed powder) × 100%.

[0076] The retention rate of aescin in aescin was determined using the following method:

[0077] Aescin retention rate = (Aescin content × Aescin mass) / (Aescin content of crude extract 1 × Mass of crude extract 1) × 100%.

[0078] Table 1 Extraction and retention rates 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] Example 2

[0081] The aescin prepared in Example 1 was dissolved in deionized water to obtain an aqueous solution of aescin with a concentration of 1 wt.%, which was clear and transparent. The solution was then placed under the conditions of room 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] The aescin prepared in Example 1 was dissolved in a 5 wt.% butanediol solution to obtain an aescin butanediol solution with a concentration of 1 wt.%. The stability of the aescin was tested by placing it under the conditions of room temperature, high temperature (45°C), and pH 4-6 for 6 months.

[0083] See the image before the test. Figure 2 See the image after the test. Figure 3 .

[0084] Figure 2 and Figure 3 From left to right, the table shows the stability of a 1 wt.% aescin aqueous solution under ambient temperature, low temperature (4℃), high temperature (45℃), pH 5 (45℃), pH 6 (45℃), pH 4, pH 5, pH 6, pH 7, pH 8, pH 9, and pH 10 conditions, as well as the stability of a 1 wt.% aescin butanediol solution under ambient temperature, high temperature (45℃), pH 4, pH 5, and pH 6 conditions.

[0085] Example 3

[0086] Images of the aescin prepared in Example 1 and two commercially available aescins (sample 1 and sample 2) are shown below. Figure 4 ;

[0087] The aescin prepared in Example 1 and two commercially available aescins (commercial sample 1 and commercial sample 2) were added to water to prepare solutions with a concentration of 10 wt.%, and their solubility was compared. The results are shown in [Figure 1]. Figure 5 .

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

[0089] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A process for the preparation of aescin, characterized in that, The method comprises the following steps: adding the horse chestnut seeds into an organic solvent, then adding alkaloids, removing the organic solvent after heating and reflux extraction, and obtaining a crude extract; adding the crude extract into water, dissolving, then adding cyclic oligosaccharides, stirring and reacting, then adding organic acids, heating and reacting, and purifying to obtain the horse chestnut saponin; the organic solvent is an ethanol solution with a concentration of 70 vol.%; the alkaloids are matrine; the cyclic oligosaccharides are hydroxypropyl-β-cyclodextrin or hydroxypropyl-γ-cyclodextrin; the organic acids are ferulic acid; the purifying comprises the following steps: adding a decolorizing agent to decolorize, then sequentially passing through a cation exchange resin and an anion exchange resin to obtain effluent, filtering the effluent by using an ultrafiltration membrane, and drying to obtain the horse chestnut saponin; the filtering by using an ultrafiltration membrane comprises the following steps: first removing macromolecular substances above 2500 Da by using an ultrafiltration membrane, and then removing small molecular substances below 1000 Da by using a secondary ultrafiltration membrane.

2. The preparation method according to claim 1, wherein the use amount ratio of the horse chestnut seeds and the alkaloids is 1 kg: 8-10 g.

3. The method of claim 1, wherein, the heating and reflux extraction is performed for 3-5 h.

4. The method of claim 1, wherein, the use amount ratio of the horse chestnut seeds and the cyclic oligosaccharides is 1 kg: 300-500 g.

5. The preparation method according to claim 1, characterized in that, the use amount ratio of the alkaloids and the organic acids is 8-10 g: 10-15 g.

6. The method of claim 1, wherein, the decolorizing agent is activated carbon and polyamide resin with a mass ratio of 3:1.

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