Preparation method of purslane extracting solution with soothing effect

Abstract

The invention relates to a preparation method of a purslane extracting solution with a soothing effect, and belongs to the field of cosmetics and biological extraction. The preparation method comprises the following steps: crushing fresh wild purslane, adding a butanediol-water mixed solvent, carrying out ultrasonic wall breaking treatment at 55-58 DEG C, and adding plant fiber hydrolase, vegetable gum hydrolase, plant starch hydrolase and plant protein hydrolase to carry out synergistic enzymolysis extraction; heating, inactivating and filtering the extracting solution, sequentially performing multi-stage purification by adopting chitosan flocculation, activated carbon decoloration, ceramic membrane filtration, ultrafiltration and nanofiltration, and collecting trapped fluid with the molecular weight of more than 150-300 Da through nanofiltration; and adding the humectant and the preservative. Through combination of ultrasonic-enzymolysis synergistic extraction and multi-stage membrane separation and purification, active ingredients such as polysaccharide and flavone in purslane are efficiently reserved under mild conditions, the inhibition rate of the obtained extracting solution to hyaluronidase activity reaches 20% or above, and the extracting solution has a soothing effect and good product stability and can be applied to soothing cosmetics or skin care products.

Description

Technical Field

[0001] This invention relates to the fields of cosmetics and bio-extraction technology, specifically a method for preparing a purslane extract with a soothing effect. Background Technology

[0002] Purslane (Portulaca oleracea L.), a traditional medicinal plant, is rich in various active ingredients such as polysaccharides, flavonoids, alkaloids, and organic acids. Modern pharmacological studies have shown that it possesses antioxidant, anti-inflammatory, antibacterial, and immunomodulatory biological activities, making it widely applicable in the pharmaceutical, functional food, and cosmetic fields. Among these, purslane extract, due to its excellent soothing, anti-allergic, and repairing effects, has gradually become a research hotspot for functional cosmetic raw materials. Currently, the main methods for preparing purslane extract include water extraction, alcohol extraction, microwave-assisted extraction, and ultrasound-assisted extraction. However, existing technologies generally suffer from the following problems: traditional hot water extraction methods involve high temperatures (typically 80℃-100℃), which easily leads to the degradation or oxidation of heat-sensitive active ingredients (such as polysaccharides and flavonoids), reducing the bioactivity of the extract; while alcohol extraction can improve the extraction rate of some fat-soluble components, it carries a high risk of organic solvent residue and has low extraction efficiency for water-soluble active ingredients (such as purslane polysaccharides); although single ultrasonic or microwave-assisted extraction can improve extraction efficiency to some extent, incomplete cell wall disruption and insufficient release of active ingredients mean that there is still considerable room for improvement in extraction rate. Furthermore, existing extraction processes often employ simple filtration or single-stage purification, resulting in extracts with high impurity content and low purity of active ingredients, directly affecting the soothing efficacy and stability of the product.

[0003] To address the aforementioned technical problems, while existing technologies have reported studies employing enzymatic hydrolysis-assisted extraction, these are mostly limited to single enzymes or simple enzymatic hydrolysis, lacking the design of multi-enzyme synergistic systems targeting the complex cell wall structure of purslane (containing multiple layers of complex structures such as cellulose, hemicellulose, pectin, starch, and proteins). In the purification stage, conventional activated carbon decolorization or single-membrane filtration cannot simultaneously meet the dual requirements of impurity removal and active ingredient enrichment, resulting in large fluctuations in the content of effective components in the product and unstable soothing effects. Therefore, how to achieve efficient extraction, precise purification, and activity retention of purslane active ingredients under mild conditions, and obtain a stable and controllable purslane extract with significant soothing effects, remains a pressing technical problem to be solved in this field. Summary of the Invention

[0004] In order to solve the problems of the prior art, the present invention provides a method for preparing a purslane extract with a soothing effect.

[0005] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution: Firstly, a method for preparing a purslane extract with a soothing effect, comprising the following steps: S1: Crush fresh wild purslane to obtain purslane pulp; S2: Add 50%-90% by weight of extraction solvent to the purslane pulp, wherein the extraction solvent is a mixed solution of butanediol and water, wherein the volume fraction of butanediol is 20%-50% and the volume fraction of water is 80%-50%; heat to 55℃-58℃, first perform ultrasonic cell wall disruption treatment, then add plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase and plant protein hydrolase, stir and extract to obtain the extract; S3: Heat the extract to inactivate it, filter it, and obtain filtrate one; S4: Add chitosan flocculant to the first filtrate, flocculate, and then filter to obtain the second filtrate; S5: Decolorize the second filtrate with activated carbon, separate the solid and liquid, and filter it with a ceramic membrane to obtain the filtrate; S6: The filtrate obtained in step S5 is first ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Da, and the permeate is collected. Then, it is nanofiltered using a nanofiltration membrane with a molecular weight cutoff of 150-300 Da, and the retentate is collected to obtain filtrate three. S7: Add a humectant and a preservative to the filtrate to obtain purslane extract.

[0006] In one specific embodiment of the first aspect, the ultrasonic cell disruption treatment in step S2 takes 20-40 minutes.

[0007] In one specific embodiment of the first aspect, the amounts of plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase and plant protein hydrolase added in step S2 are 1‰-3‰ respectively.

[0008] In one specific embodiment of the first aspect, the stirring extraction time in step S2 is 2-4 hours.

[0009] In one specific embodiment of the first aspect, the temperature for inactivation in step S3 is 85°C-95°C, and the time is 5-15 minutes.

[0010] In one specific embodiment of the first aspect, the chitosan flocculant added in step S4 has a mass fraction of 0.1%-0.5%, and the flocculation time is 20-40 minutes.

[0011] In one specific embodiment of the first aspect, the activated carbon added in step S5 has a mass fraction of 0.3%-0.8%, a decolorization temperature of 20℃-60℃, and a decolorization time of 30-60 minutes; the ceramic membrane has a pore size of 0.05-0.2 micrometers.

[0012] In one specific embodiment of the first aspect, the molecular weight cutoff of the ultrafiltration membrane in step S6 is 10000 Da, and the molecular weight cutoff of the nanofiltration membrane is 150-300 Da.

[0013] Secondly, a method for preparing a purslane extract with a soothing effect.

[0014] In one specific embodiment of the second aspect, the purslane extract is used in the preparation of cosmetics or skin care products with soothing effects.

[0015] The beneficial effects of this invention are as follows: 1. By constructing an integrated process system of low-temperature ultrasonic cell disruption, multi-enzyme synergistic enzymatic hydrolysis, and multi-stage membrane separation and purification, efficient extraction and precise purification of active ingredients from purslane were achieved. Specifically, under mild conditions of 55℃-58℃, the present invention first utilizes the cavitation effect and mechanical vibration generated by ultrasound to cause irreversible rupture of the purslane cell wall, significantly increasing the dissolution rate and dissolution percentage of intracellular active ingredients. Subsequently, a complex enzyme system composed of plant cellulose hydrolase, plant gum hydrolase, plant starch hydrolase, and plant proteolytic enzyme acts on the structural macromolecules of the cell wall, such as cellulose, pectin, starch, and protein, respectively, directionally degrading the cell wall components into soluble small molecules, further reducing mass transfer resistance, and enabling the full release of intracellular active ingredients without the need for high temperatures, strong acids, or alkalis. This "ultrasound-enzymatic hydrolysis" synergistic treatment method avoids the loss of heat-sensitive components such as polysaccharide degradation and flavonoid oxidation during traditional thermal extraction, and also transforms macromolecular impurities into easily separable forms through enzymatic hydrolysis, laying a good foundation for subsequent purification steps.

[0016] 2. This invention employs a four-stage cascade purification strategy of flocculation, decolorization, ceramic membrane, and nanofiltration to achieve step-by-step purification and targeted enrichment of active ingredients in the extract. First, chitosan flocculant selectively removes colloidal impurities, suspended particles, and some pigments through charge neutralization and bridging adsorption, effectively reducing the turbidity of the feed solution and lessening the contamination load of subsequent membrane filtration. Then, activated carbon adsorption decolorizes the extract, removing small molecule pigments and some pyrogenic substances. Next, a 0.1-micron ceramic membrane is used for precision filtration, efficiently retaining bacteria, particles, and large molecule impurities with a molecular weight greater than 100,000 Daltons, while allowing the target active ingredients to pass through smoothly. Finally, a nanofiltration membrane with a molecular weight cutoff of 150-300 Daltons is used for fractional separation, precisely retaining inorganic salts, small molecule impurities, and some pigments, while enriching small molecule polysaccharides, flavonoids, and alkaloids with soothing activities. The synergistic effect of this four-stage purification system significantly improved the purity, clarity, and enrichment factor of the active ingredients in the product, resulting in a significant increase in the retention rate of total polysaccharides and total flavonoids in the final purslane extract. Hyaluronidase inhibition tests verified that at a concentration of 5%, the extract inhibited hyaluronidase activity by 20.39%, significantly higher than the positive control (9.71%), fully demonstrating its excellent soothing efficacy. Meanwhile, stability tests showed that the extract remained stable in appearance, pH value, and active ingredient content for 3 months when stored within a temperature range of 4℃ to 40℃, exhibiting good shelf stability and controllable quality. Furthermore, this invention uses a butanediol-water mixed solvent instead of pure water for extraction. Utilizing the excellent solubility of butanediol in moderately polar components such as flavonoids, polyphenols, and alkaloids, the extraction rate of soothing active ingredients is significantly improved. Simultaneously, an ultrafiltration membrane with a molecular weight cutoff of 10,000 Da is introduced before nanofiltration, effectively removing large molecular impurities such as polysaccharides and proteins, reducing the fouling load on the nanofiltration membrane. Combined with the precise retention of active ingredients by a 150-300 Da nanofiltration membrane, highly efficient enrichment and purification of the soothing active ingredients of purslane are achieved, resulting in a product with higher purity and more stable soothing efficacy. Attached Figure Description

[0017] Figure 1 This is a schematic diagram of the process flow of the present invention. Detailed Implementation

[0018] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0019] like Figure 1 This paper presents a method for preparing a purslane extract with a soothing effect.

[0020] This invention provides a method for preparing a purslane extract with a soothing effect. The core of the method lies in the use of a gentle process that combines low-temperature compound enzymatic hydrolysis, ultrasonic cell disruption, and multi-stage purification (flocculation, decolorization, ceramic membrane filtration, and nanofiltration) to maximize the retention of active ingredients such as polysaccharides, flavonoids, and alkaloids in purslane. The soothing effect is verified by hyaluronidase inhibition experiments.

[0021] Raw materials used in this invention: Fresh wild purslane (Portulaca oleracea L.) is harvested from pollution-free planting bases, selecting vigorous, disease-free, and fresh whole herbs.

[0022] Plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase, and plant protein hydrolase: all are food grade or cosmetic grade, with enzyme activity units ≥100,000 U / g.

[0023] Chitosan flocculant: Deacetylation degree ≥90%, food grade.

[0024] Activated carbon: Powdered activated carbon, specific surface area ≥1000 m² 2 / g.

[0025] Ceramic membrane: made of alumina or zirconium oxide, with a pore size of 0.1 micrometers.

[0026] Nanofiltration membrane: made of polyamide, with a molecular weight cutoff of 200 Daltons.

[0027] Moisturizer: may be selected from one or more of glycerin, butylene glycol, and 1,2-pentanediol.

[0028] Preservatives: can be selected from one or more of p-hydroxyacetophenone, 1,2-hexanediol, and phenoxyethanol.

[0029] The method for evaluating the soothing efficacy of purslane extract described in this invention: A hyaluronidase inhibition assay was performed (referencing the modified Elson-Morgan method). A higher hyaluronidase activity inhibition rate indicates a stronger soothing effect of the sample. A positive control was a 2 mg / mL isoliquiritigenin solution. Example 1

[0030] This embodiment provides a method for preparing a purslane extract with a soothing effect, including the following steps: S1: Weigh 1000 g of fresh wild purslane, cut it into 1-2 cm pieces, and grind it into purslane pulp using a grinder; S2: Add purslane pulp to an extraction container, and simultaneously add 70% of the extraction solvent by weight of the purslane pulp. The extraction solvent is a mixed solution of 1,3-butanediol and water, wherein the volume fraction of 1,3-butanediol is 30% and the volume fraction of water is 70%. Heat to 56°C, first use ultrasound to break up the cell walls for 30 minutes (ultrasound frequency 40 kHz, power 500 W), then add plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase and plant protein hydrolase, with each enzyme added at a rate of 2‰. Stir at a uniform speed for 3 hours to obtain the extract. S3: Heat the extract to 90℃, keep it at that temperature for 10 minutes to inactivate it, and then filter it through a 200-mesh filter cloth to obtain filtrate one; S4: Add 0.2% by mass of chitosan flocculant to filtrate one, stir evenly, let stand for flocculation for 30 minutes, filter with 200 mesh filter cloth to obtain filtrate two; S5: Add 0.5% activated carbon to filtrate 2, decolorize at 40℃ for 45 minutes, separate the activated carbon by centrifugation at 4000 rpm for 15 minutes, and then filter with a 0.1-micron ceramic membrane to obtain the filtrate; S6: The filtrate obtained in step S5 is first ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Da, and the permeate is collected; then the permeate is nanofiltered using a nanofiltration membrane with a molecular weight cutoff of 200 Da, and the filtrate is collected to obtain filtrate three. S7: Add 3% by mass of 1,2-pentanediol (humectant and preservative) and 0.5% by mass of 1,2-hexanediol (preservative) to filtrate three, stir well, and you will get purslane extract. Example 2

[0031] This embodiment is basically the same as Embodiment 1, except that in step S2, the ultrasonic cell disruption treatment time is 30 minutes, the extraction solvent is a mixed solution of 1,3-butanediol and water, wherein the volume fraction of 1,3-butanediol is 20% and the volume fraction of water is 80%; the amount of compound enzyme added is 1‰, and the stirring extraction time is 2 hours; in step S3, the inactivation temperature is 85℃ and the time is 15 minutes; in step S4, the amount of chitosan flocculant added is 0.1%, and the flocculation time is 40 minutes; in step S5, the amount of activated carbon added is 0.3%, the decolorization temperature is 60℃, the decolorization time is 30 minutes, and the pore size of the ceramic membrane is 0.2 micrometers; in step S6, ultrafiltration is first performed using an ultrafiltration membrane with a molecular weight cutoff of 10000 Da, the permeate is collected, and then nanofiltration is performed using a nanofiltration membrane with a molecular weight cutoff of 150 Da, and the retentate is collected. Example 3

[0032] This embodiment is basically the same as Embodiment 1, except that in step S2, the ultrasonic cell disruption treatment time is 40 minutes, the extraction solvent is a mixed solution of 1,3-butanediol and water, wherein the volume fraction of 1,3-butanediol is 50% and the volume fraction of water is 50%; the amount of compound enzyme added is 3‰, and the stirring extraction time is 4 hours; in step S3, the inactivation temperature is 95℃ and the time is 5 minutes; in step S4, the amount of chitosan flocculant added is 0.5%, and the flocculation time is 20 minutes; in step S5, the amount of activated carbon added is 0.8%, the decolorization temperature is 20℃, the decolorization time is 60 minutes, and the pore size of the ceramic membrane is 0.05 micrometers; in step S6, ultrafiltration is first performed using an ultrafiltration membrane with a molecular weight cutoff of 10000 Da, the permeate is collected, and then nanofiltration is performed using a nanofiltration membrane with a molecular weight cutoff of 300 Da, and the retentate is collected.

[0033] Comparative Example 1, without enzymatic hydrolysis; This comparative example is basically the same as Example 1, except that no hydrolytic enzyme is added in step S2, and hot water extraction is performed directly after ultrasonic cell disruption. The other steps are the same.

[0034] Comparative Example 2, without nanofiltration purification; This comparative example is basically the same as Example 1, except that in step S6, a nanofiltration membrane is not used for nanofiltration. Instead, the filtrate filtered by the ceramic membrane is directly added to a humectant and a preservative to obtain purslane extract.

[0035] Comparative Example 3: High-temperature extraction was used; This comparative example is basically the same as Example 1, except that in step S2, the temperature is raised to 85°C for extraction, ultrasonic cell disruption is not performed, hydrolytic enzymes are not added, and the extraction time is 3 hours. Other steps are the same.

[0036] Experimental example: Evaluation test of soothing efficacy; The purslane extracts obtained in Examples 1-3 and Comparative Examples 1-3 were evaluated for their soothing efficacy according to the hyaluronidase inhibition test method.

[0037] Test method: The sample was diluted with ultrapure water to a 5% (mass fraction) aqueous solution as the test solution.

[0038] Following the Elson-Morgan modified method, the absorbance of the blank (A1) and the absorbance of the sample (A2) were measured, and the inhibition rate of hyaluronidase activity was calculated.

[0039] The positive control was a 2 mg / mL isoglycyrrhizin solution, and the average value was taken from three parallel determinations.

[0040] Formula for calculating hyaluronidase activity inhibition rate: Experimental results: Results analysis: The hyaluronidase activity inhibition rates of Examples 1-3 were all above 19.5%, significantly higher than the positive control (9.71%), indicating that the purslane extract prepared in this invention has excellent soothing effects.

[0041] Comparative Example 1 was not subjected to enzymatic hydrolysis, resulting in insufficient extraction of active ingredients and a significant decrease in inhibition rate.

[0042] Comparative Example 2 did not undergo nanofiltration purification, thus retaining some low molecular weight impurities or large molecular weight ineffective components, resulting in a lower inhibition rate than the Example.

[0043] Comparative Example 3 was extracted at high temperature without enzymatic hydrolysis or ultrasonic cell disruption, resulting in severe damage to the active ingredients and the lowest inhibition rate.

[0044] The method for preparing purslane extract provided by this invention effectively preserves the soothing active ingredients in purslane through the synergistic effect of low-temperature compound enzymatic hydrolysis and ultrasonic cell disruption, combined with flocculation, activated carbon decolorization, ceramic membrane filtration and nanofiltration purification processes.

[0045] To verify the stability of the purslane extract prepared in this invention during storage, the purslane extract prepared in Example 1 was stored at 4℃, 25℃, and 40℃ for 3 months, and samples were taken monthly to test its appearance, pH value, total polysaccharide content, and total flavonoid content.

[0046] Stability test results: The results showed that the samples remained clear and transparent under all conditions, with no precipitation, and the pH value remained stable between 5.5 and 6.5. The retention rates of total polysaccharides and total flavonoids were both greater than 94%, indicating that the purslane extract prepared by this invention has good stability.

[0047] In summary, this invention utilizes a combined process of ultrasonic cell disruption and enzymatic hydrolysis for extraction, along with flocculation-decolorization-membrane filtration for purification, to efficiently extract the soothing active ingredients from purslane under mild conditions. The results of active ingredient content determination show that this process significantly improves the extraction rates of total polysaccharides and total flavonoids. Hyaluronidase inhibition tests indicate that the purslane extract obtained by this invention exhibits an inhibition rate of over 20% against hyaluronidase activity, significantly higher than the positive control, demonstrating excellent soothing efficacy. Stability tests show that the product remains stable under normal storage conditions. Therefore, the purslane extract obtained by this invention can be widely used in cosmetics or skincare products with soothing effects.

[0048] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a purslane extract with a soothing effect, characterized in that, Includes the following steps: S1: Crush fresh wild purslane to obtain purslane pulp; S2: Add 50%-90% by weight of extraction solvent to the purslane pulp, wherein the extraction solvent is a mixed solution of butanediol and water, wherein the volume fraction of butanediol is 20%-50% and the volume fraction of water is 80%-50%; heat to 55℃-58℃, first perform ultrasonic cell wall disruption treatment, then add plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase and plant protein hydrolase, stir and extract to obtain the extract; S3: Heat the extract to inactivate it, filter it, and obtain filtrate one; S4: Add chitosan flocculant to the first filtrate, flocculate, and then filter to obtain the second filtrate; S5: Decolorize the second filtrate with activated carbon, separate the solid and liquid, and filter it with a ceramic membrane to obtain the filtrate; S6: The filtrate obtained in step S5 is first ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 10,000 Da, and the permeate is collected. Then, it is nanofiltered using a nanofiltration membrane with a molecular weight cutoff of 150-300 Da, and the retentate is collected to obtain filtrate three. S7: Add a humectant and a preservative to the filtrate to obtain purslane extract.

2. The preparation method according to claim 1, characterized in that, The ultrasonic cell disruption process in step S2 takes 20-40 minutes.

3. The preparation method according to claim 1, characterized in that, The amounts of plant fiber hydrolase, plant gum hydrolase, plant starch hydrolase, and plant protein hydrolase added in step S2 are 1‰-3‰, respectively.

4. The preparation method according to claim 1, characterized in that, The stirring and extraction time in step S2 is 2-4 hours.

5. The preparation method according to claim 1, characterized in that, The inactivation temperature in step S3 is 85℃-95℃, and the time is 5-15 minutes.

6. The preparation method according to claim 1, characterized in that, In step S4, the chitosan flocculant is added at a mass fraction of 0.1%-0.5%, and the flocculation time is 20-40 minutes.

7. The preparation method according to claim 1, characterized in that, In step S5, the activated carbon is added at a mass fraction of 0.3%-0.8%, the decolorization temperature is 20℃-60℃, and the decolorization time is 30-60 minutes; the ceramic membrane has a pore size of 0.05-0.2 micrometers.

8. The preparation method according to claim 1, characterized in that, The ultrafiltration membrane in step S6 has a molecular weight cutoff of 10,000 Da, and the nanofiltration membrane has a molecular weight cutoff of 150-300 Da.

9. A purslane extract prepared by the method according to any one of claims 1 to 8.

10. The application of the purslane extract according to claim 9 in the preparation of cosmetics or skin care products with soothing effects.

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