Cactus polysaccharide composition as well as preparation method and application thereof

Abstract

The invention relates to the technical field of functional cosmetic raw materials, in particular to a cactus polysaccharide composition as well as a preparation method and application thereof. According to the cactus polysaccharide composition disclosed by the invention, the cactus polysaccharides with specific number-average molecular weight are combined according to a specific proportion, so that the cactus polysaccharide composition has a more comprehensive anti-irritation effect and has the characteristics of high stability and better skin feeling and solubility. The cactus polysaccharide can be prepared through a simple process, exogenous components do not need to be introduced, after the cactus aqueous extract is subjected to enzymolysis through amylase, the cactus polysaccharide with the specific number-average molecular weight is screened and combined according to the specific proportion, the anti-irritation effect requirement can be met, and safety is better. The cactus polysaccharide composition can be applied to cosmetics / pharmaceuticals as an anti-irritation raw material.

Description

Technical Field

[0001] This invention relates to the field of cosmetic raw material technology, and in particular to a cactus polysaccharide composition, its preparation method and application. Background Technology

[0002] Adverse skin reactions caused by cosmetics are becoming increasingly common. Many products, despite passing safety tests, still cause irritation for consumers. Consumers' concerns about cosmetic-induced irritation and sensitivity have reached unprecedented levels, with cosmetic safety ranking first among the four key characteristics of cosmetics. Currently, the cosmetic industry requires anti-irritant ingredients to simultaneously meet the core requirements of "strong efficacy, pleasant skin feel, high stability, and good safety."

[0003] Polysaccharides, as an important class of natural macromolecules, are widely found in plants, animals, and microorganisms. Due to their excellent biocompatibility, non-toxicity, and diverse biological activities, they have been widely used in food, medicine, and cosmetics. Cactus polysaccharides, rich in active groups such as hydroxyl and carboxyl groups, can repair the skin barrier and inhibit the release of inflammatory factors, and are therefore widely used in anti-irritant skincare products.

[0004] Some examples of using polysaccharides as cosmetic raw materials have been disclosed in related technologies. Low molecular weight polysaccharides (LMW) are fast-acting but easily degraded, while high molecular weight polysaccharides (HMW) have excellent stability but poor permeability and a poor skin feel, often with a noticeable stickiness. Furthermore, they may pose further irritation problems, limiting the application of polysaccharide products in certain scenarios. To achieve anti-irritation effects, polysaccharide raw materials often need to be combined with other exogenous ingredients. However, introducing exogenous ingredients may not meet the needs of subsequent formulation applications and can also negatively impact stability. Related technologies use polysaccharides of different molecular weights in combination, but this can easily lead to antagonistic and other instability issues, failing to guarantee the full utilization of the efficacy characteristics of polysaccharides of different molecular weights. Irritation problems are also difficult to avoid, often requiring the addition of other ingredients.

[0005] In view of this, the present invention is hereby proposed. Summary of the Invention

[0006] One object of the present invention is to provide a cactus polysaccharide composition that has a more comprehensive anti-irritant effect and is characterized by high stability, better skin feel and solubility.

[0007] Another objective of this invention is to provide a method for preparing a cactus polysaccharide composition, which is simple in process, does not require the introduction of exogenous components, and can achieve the desired anti-irritant efficacy with superior safety.

[0008] A third objective of this invention is to provide an application of a cactus polysaccharide composition, which can be used as an anti-irritant ingredient in cosmetics / pharmaceuticals.

[0009] To achieve the above-mentioned objective of the present invention, the present invention provides a cactus polysaccharide composition comprising cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C; Among them, the number average molecular weight of cactus polysaccharide A is above 100,000 Da, the number average molecular weight of cactus polysaccharide B is 10,000-100,000 Da, and the number average molecular weight of cactus polysaccharide C is below 10,000 Da. The mass ratio of cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C in the cactus polysaccharide composition is cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15): (50-70): (10-30).

[0010] The cactus polysaccharide composition uses cactus polysaccharides with specific number-average molecular weights combined in a specific ratio, exhibiting more comprehensive anti-irritant effects, and also possessing the characteristics of high stability, better skin feel, and superior solubility.

[0011] Another aspect of the present invention provides a method for preparing the above-mentioned cactus polysaccharide composition, comprising enzymatically hydrolyzing a water extract of cactus with amylase, and then screening out cactus polysaccharide A with a number average molecular weight of more than 100,000 Da, cactus polysaccharide B with a number average molecular weight of 10,000-100,000 Da and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da, and mixing cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C in a mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15):(50-70):(10-30) to obtain the final product.

[0012] This method is simple and does not require the introduction of exogenous components. After enzymatically hydrolyzing the water extract of cactus with amylase, cactus polysaccharides with specific number-average molecular weights are screened and combined in a specific ratio to achieve the desired anti-irritant effect with better safety.

[0013] Another aspect of the present invention provides the application of the above-mentioned cactus composition, which can be used as an anti-irritant ingredient in cosmetics / pharmaceuticals.

[0014] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention relates to a cactus polysaccharide composition, which uses cactus polysaccharides of specific number-average molecular weights combined in a specific ratio to exhibit more comprehensive anti-irritant effects, while also possessing high stability, superior skin feel, and better solubility. It can be prepared through a simple process without the introduction of exogenous components. After enzymatically hydrolyzing the water extract of cactus with amylase, cactus polysaccharides of specific number-average molecular weights are selected and combined in a specific ratio to achieve the desired anti-irritant efficacy with superior safety. This cactus polysaccharide composition can be used as an anti-irritant raw material in cosmetics and pharmaceuticals. Attached Figure Description

[0015] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0016] Figure 1 This is a transmission electron microscope image of the cactus polysaccharide composition obtained in Example 1 of the present invention, wherein... Figure 1 a and Figure 1 b are electron micrographs of the cactus polysaccharide compositions obtained in Examples 1 and 2, respectively; Figure 2 The figures show the test results of TRPV1 inhibition experiments in various embodiments and comparative examples of the present invention; Figure 3 The figures show the DNA protection test results of various embodiments and comparative examples of the present invention; Figure 4 The figures show the test results of the ABTS free radical scavenging test for various embodiments and comparative examples of the present invention; Figure 5 The figures show the test results of various embodiments and comparative examples of the present invention in anti-stimulation tests. Detailed Implementation

[0017] The technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings and specific embodiments. However, those skilled in the art will understand that the embodiments described below are some embodiments of the present invention, but not all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall be followed. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0018] The present invention provides a cactus polysaccharide composition comprising cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C; Among them, the number average molecular weight of cactus polysaccharide A is above 100,000 Da, the number average molecular weight of cactus polysaccharide B is 10,000-100,000 Da, and the number average molecular weight of cactus polysaccharide C is below 10,000 Da. The mass ratio of cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C in the cactus polysaccharide composition is cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15): (50-70): (10-30).

[0019] The described cactus polysaccharide composition, using cactus polysaccharides of specific number-average molecular weights in a specific ratio, exhibits more comprehensive anti-irritant effects, along with high stability, superior skin feel, and better solubility. This cactus polysaccharide composition demonstrates a multi-dimensional anti-irritant mechanism, exhibiting a uniform spherical structure under microscopic observation (electron microscopy). The cactus polysaccharides of different number-average molecular weights are recombine rather than existing independently, thus demonstrating both the individual characteristics of each number-average molecular weight cactus polysaccharide and their synergistic effects in terms of anti-irritant efficacy.

[0020] In one specific embodiment of the present invention, the mass ratio of cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C in the cactus polysaccharide composition is cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15):(50-70):(10-30), which can be any numerical ratio within this range, such as 5:70:10, 10:60:20, 15:50:30, etc. The mass fraction of cactus polysaccharide A in this mass ratio can be any mass fraction between 5 and 15 parts, such as 5 parts, 6 parts, 7.5 parts, 9 parts, 10 parts, 11.4 parts, 12.8 parts, 14 parts, 14.3 parts, or 15 parts. The mass fraction of cactus polysaccharide B can be any number of parts between 50 and 70, such as 50, 52.6, 55, 60, 63, 65, 67.4, 69, or 70 parts. The mass fraction of cactus polysaccharide C can be any number of parts between 10 and 30, such as 10, 12.1, 15, 16, 20, 24.7, 25, 27, or 30 parts.

[0021] In one specific embodiment of the present invention, cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C are prepared by enzymatic hydrolysis of cactus water extract with amylase.

[0022] Another aspect of the present invention provides a method for preparing the above-mentioned cactus polysaccharide composition, comprising enzymatically hydrolyzing a water extract of cactus with amylase, and then screening out cactus polysaccharide A with a number average molecular weight of more than 100,000 Da, cactus polysaccharide B with a number average molecular weight of 10,000-100,000 Da and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da, and mixing cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C in a mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15):(50-70):(10-30) to obtain the final product.

[0023] This method is simple and does not require the introduction of exogenous components. After enzymatically hydrolyzing the cactus water extract with amylase, cactus polysaccharides with specific number-average molecular weights are screened and combined in a specific ratio to achieve the desired anti-irritant efficacy with superior safety. The applicant discovered that after hydrolyzing the cactus water extract with amylase, the resulting cactus polysaccharide materials with different number-average molecular weights exhibit excellent compatibility, facilitating the recombination of cactus polysaccharides with different number-average molecular weights into a homogeneous whole.

[0024] In one specific embodiment of the present invention, the mass ratio of cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C in the cactus polysaccharide composition is cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15):(50-70):(10-30), which can be any numerical ratio within this range, such as 5:70:10, 10:60:20, 15:50:30, etc. The mass fraction of cactus polysaccharide A in this mass ratio can be any mass fraction between 5 and 15 parts, such as 5 parts, 6 parts, 7.5 parts, 9 parts, 10 parts, 11.4 parts, 12.8 parts, 14 parts, 14.3 parts, or 15 parts. The mass fraction of cactus polysaccharide B can be any number of parts between 50 and 70, such as 50, 52.6, 55, 60, 63, 65, 67.4, 69, or 70 parts. The mass fraction of cactus polysaccharide C can be any number of parts between 10 and 30, such as 10, 12.1, 15, 16, 20, 24.7, 25, 27, or 30 parts.

[0025] In one specific embodiment of the present invention, the method for preparing the cactus water extract includes extracting the cactus stem with water at 40-70℃ for 1-3 hours.

[0026] In one specific embodiment of the present invention, the cactus stem can be first crushed and then extracted with water. The extraction temperature can be selected between 40-70℃, such as 40℃, 42℃, 48℃, 50℃, 55℃, 60℃, 63.6℃, ​​67.2℃ or 70℃. The extraction time can be 1-3 hours, such as 1 hour, 1.2 hours, 1.5 hours, 2 hours, 2.3 hours, 2.8 hours or 3 hours, depending on the extraction temperature, to ensure sufficient extraction.

[0027] In one specific embodiment of the present invention, the mass ratio of the cactus stem to water is cactus:water = 1:(10-40), such as 1:10, 1:12, 1:15, 1:20, 1:23, 1:26.5, 1:30, 1:31, 1:35 or 1:40. The amount of water used is sufficient to fully extract the cactus stem.

[0028] In one specific embodiment of the present invention, after extraction, the material is left to stand for 12-18 hours to allow the extract residue to precipitate.

[0029] In one specific embodiment of the present invention, the cactus water extract is decolorized and filtered before being enzymatically hydrolyzed.

[0030] In one specific embodiment of the present invention, the cactus water extract is decolorized using activated carbon. The amount of activated carbon can be determined according to the general decolorization process. For example, the amount of activated carbon is 0.5-2.0% of the material mass, such as 0.5%, 0.6%, 0.74%, 0.9%, 1.0%, 1.2%, 1.25%, 1.4% or 1.5% of the material mass.

[0031] In one specific embodiment of the present invention, after decolorization, filtration is performed, for example, by using a paperboard filter to remove activated carbon and some impurities.

[0032] In one specific embodiment of the present invention, the amylase includes α-amylase.

[0033] In one specific embodiment of the present invention, the amount of amylase used is preferably sufficient to fully decompose the starch in the material. For example, the amount of amylase used in the enzymatic hydrolysis is 0.005%-0.02% of the material mass, such as 0.005%, 0.007%, 0.01%, 0.012%, 0.014%, 0.015%, 0.016%, 0.018%, 0.019%, or 0.02%.

[0034] In one specific embodiment of the present invention, the enzymatic hydrolysis temperature is preferably such that the amylase activity is ensured and the enzymatic hydrolysis reaction is fully carried out, for example, the enzymatic hydrolysis temperature is 50-55℃, such as 50℃, 51℃, 51.4℃, 52℃, 53℃, 54℃ or 55℃.

[0035] The enzymatic hydrolysis time is based on the standard that the enzymatic hydrolysis reaction is fully completed. The completion of the enzymatic hydrolysis reaction can be determined by sampling and detecting the starch content. For example, under the above conditions, the enzymatic hydrolysis time can be more than 30 minutes.

[0036] In one specific embodiment of the present invention, after enzymatic hydrolysis, the obtained material is desalted and then screened to obtain cactus polysaccharide A with a number average molecular weight of more than 100,000 Da, cactus polysaccharide B with a number average molecular weight of 10,000-100,000 Da, and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da.

[0037] In one specific embodiment of the present invention, the desalination is carried out using cationic resin and anionic resin. The material can be desalinated by sequentially passing through cationic resin and anionic resin.

[0038] In one specific embodiment of the present invention, the screening is performed using membrane separation. Materials with a number-average molecular weight (NM) of 100,000 or higher are selected as cactus polysaccharide A, materials with a NM of 10,000-100,000 Da are selected as cactus polysaccharide B, and materials with a NM of less than 10,000 Da are selected as cactus polysaccharide C. Filter membranes with NMs of 100,000 Da and 10,000 Da can be selected. Materials collected on the 100,000 Da filter membrane are collected as cactus polysaccharide A, materials between the 100,000 Da and 10,000 Da filter membranes are collected as cactus polysaccharide B, and materials passing through the 10,000 Da filter membrane are collected as cactus polysaccharide C.

[0039] In one specific embodiment of the present invention, after the mixture is evenly mixed, an adsorbent is added for filtration, and the fine filtrate is collected. Impurities contained therein can be further removed by the adsorbent, and the turbidity of the polysaccharide solution can be reduced as needed. Since polysaccharides have good solubility, the adsorbent is adsorbed and then the adsorbent is washed, so generally no loss of polysaccharides will be caused.

[0040] In one specific embodiment of the present invention, the adsorbent may be diatomaceous earth.

[0041] In one specific embodiment of the present invention, the mixture is compounded after being thoroughly mixed.

[0042] In one specific embodiment of the present invention, glycerol is selected for compounding, and glycerol-water is used as the solvent system, which is more conducive to ensuring the stability of the material and can promote the recombination of cactus polysaccharides with different number average molecular weights. The mass ratio of material to glycerol is material:glycerol = 1:(0.5-1.5), such as 1:0.5, 1:0.6, 1:0.72, 1:0.9, 1:1, 1:1.1, 1:1.3 or 1:1.5.

[0043] In one specific embodiment of the present invention, the materials may be appropriately concentrated before compounding to reduce the amount of glycerin used; for example, concentrated to 1 / 2 of the original mass.

[0044] In one specific embodiment of the present invention, a preservative is added to the compounded material. The amount of the preservative should be determined according to the type of preservative and relevant national regulations and standards. For example, the amount of preservative is 1%-3% of the total mass of the material, such as 1%, 1.2%, 1.8%, 2%, 2.25%, 2.5%, 2.7% or 3%.

[0045] In one specific embodiment of the present invention, the preservative includes pentylene glycol / hexanediol.

[0046] In one specific embodiment of the present invention, the final material is sterilized to obtain a cactus polysaccharide composition.

[0047] In one specific embodiment of the present invention, the sterilization includes sterilization at 95-100℃ for more than 40 minutes.

[0048] Another aspect of the present invention provides the application of the above-mentioned cactus composition, which can be used as an anti-irritant ingredient in cosmetics / pharmaceuticals.

[0049] Example 1 A method for preparing a cactus polysaccharide composition, comprising: (1) Extraction: Weigh the cactus stem powder and add it to pure water at a ratio of 1:20 (m / m) of material to liquid mass; heat to 50℃, stir and extract for 2 hours, and let stand for 16 hours to allow the extract residue to precipitate.

[0050] (2) Decolorization: Extract the supernatant, add 1.0% activated carbon by weight of the supernatant, and keep warm at 80℃ for 1 hour.

[0051] (3) Filtration: H70 paperboard is used for filtration, and the filtrate is collected.

[0052] (4) Enzymatic hydrolysis: Add 0.01% of α-amylase by weight of filtrate, incubate at 50℃ for 30 min, take a sample to test starch, and end the enzymatic hydrolysis after the starch is completely decomposed.

[0053] (5) Desalting: The enzymatic hydrolysate is passed through cation exchange resin 001*7 and anion exchange resin D309 respectively to obtain a desalting solution.

[0054] (6) Membrane separation: The desalination liquid was separated by filter membranes with a number average molecular weight of 100,000 Da and 10,000 Da, respectively. The material with a number average molecular weight of more than 100,000 Da was collected as cactus polysaccharide A, the material between the filter membrane with a number average molecular weight of 100,000 Da and the filter membrane with a number average molecular weight of 10,000 Da was collected as cactus polysaccharide B, and the material with a number average molecular weight of less than 10,000 Da was collected as cactus polysaccharide C.

[0055] (7) Compound preparation: The mass ratio of cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C is 10:60:20. Mix them evenly.

[0056] (8) Compounding: The mass ratio of material mass to glycerol dosage is material:glycerol = 1:1. Add 2% hexanediol of the total mass of the obtained material and mix evenly.

[0057] (9) Sterilization: Sterilize at 98℃ for 40 min to obtain a cactus polysaccharide composition.

[0058] Example 2 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: In step 7, the following mixtures are prepared: cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C are mixed in a mass ratio of 5:50:30.

[0059] Example 3 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: In step 7, the following mixtures are prepared: cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C are mixed in a mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = 15:70:10.

[0060] Examples 4-5 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the differences shown in Table 1:

[0061] Comparative Example 1 A cactus polysaccharide extract was prepared using the same method as in Example 1, with the only difference being: Steps (6) and (7) are omitted to obtain a cactus polysaccharide extract.

[0062] Comparative Example 2 A cactus polysaccharide extract was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide A with a number-average molecular weight of 100,000 Da or higher is collected; 2) Omit step (7).

[0063] Comparative Example 3; A cactus polysaccharide extract was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide B with a number average molecular weight between 10,000 and 100,000 Da is collected; 2) Omit step (7).

[0064] Comparative Example 4 A cactus polysaccharide extract was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide C with a number average molecular weight of less than 10,000 Da is collected.

[0065] 2) Omit step (7).

[0066] Comparative Example 5 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide A with a number average molecular weight of more than 100,000 Da and cactus polysaccharide B with a number average molecular weight between 10,000 and 100,000 Da are collected; 2) Step (7) Mix the cactus polysaccharide A and cactus polysaccharide B evenly according to the mass ratio of 10:60.

[0067] Comparative Example 6 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide A with a number average molecular weight of more than 100,000 Da and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da are collected; 2) Step (7) Mix the cactus polysaccharide A and cactus polysaccharide C evenly according to the mass ratio of 10:20.

[0068] Comparative Example 7 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: 1) In step (6), only cactus polysaccharide B with a number average molecular weight between 10,000 and 100,000 Da and cactus polysaccharide C with a number average molecular weight below 10,000 Da are collected; 2) Step (7) Mix the cactus polysaccharide B and cactus polysaccharide C evenly according to the mass ratio of 60:20.

[0069] Comparative Example 8 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: Step (7) Mix the cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C evenly according to the mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = 20:40:5.

[0070] Comparative Example 9 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: Step (7) Mix the cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C evenly according to the mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = 3:40:40.

[0071] Comparative Example 10 A cactus polysaccharide composition was prepared using the same method as in Example 1, with the only difference being: Step (7) Mix the cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C evenly according to the mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = 3:80:8.

[0072] Comparative Example 11 A method for preparing a cactus polysaccharide extract, comprising: (1) Extraction: Weigh the cactus stem powder and add it to pure water at a ratio of 1:20 (m / m); heat to 50℃, stir and extract for 2 hours, and let stand for 16 hours to allow the extract residue to precipitate.

[0073] (2) Decolorization: Extract the supernatant, add 1.0% activated carbon by weight of the supernatant, and keep warm at 80℃ for 1 hour.

[0074] (3) Filtration: H70 paperboard is used for filtration, and the filtrate is collected.

[0075] (4) Desalination: The filtrate is passed through cation exchange resin 001*7 and anion exchange resin D309 respectively to obtain desalted solution.

[0076] (5) Alcohol precipitation: Add 95% ethanol solution at a mass-volume ratio of 1:9 (g / mL) of material to alcohol, stir at room temperature for 30 min, refrigerate at 4℃ for 16 h, discard the supernatant and collect the precipitate.

[0077] (6) Redissolution: Add pure water at a mass ratio of precipitate:water = 1:2 (m / m) and stir until dissolved.

[0078] (7) Compounding: Compound solution: glycerol = 1:1 by mass ratio, add 2% hexanediol of the total mass of the resulting material, and mix well.

[0079] (8) Sterilization: Sterilize at 98℃ for 40 min, a cactus polysaccharide extract Comparative Example 12 A method for preparing a cactus polysaccharide extract, comprising: A cactus polysaccharide extract was prepared using the same method as in Example 1, with the only difference being: Step (4) Add 0.2% of the filtrate by weight of neutral protease, keep warm at 82℃ for 1 hour, and then heat to 100℃ and keep warm for 30 minutes.

[0080] Experimental Example 1: Characterization of Cactus Polysaccharide Structure The morphology of cactus polysaccharides was observed using low-pressure TEM transmission electron microscopy. For electron micrographs of the cactus polysaccharide compositions prepared in Examples 1 and 2, please refer to [link to example image]. Figure 1 .

[0081] Depend on Figure 1 It can be seen that the cactus polysaccharide solution contains clearly and distinctly arranged, loosely spaced, and uniformly sized spherical molecules. This may be because the polysaccharide molecules have a spherical chain conformation. The boundaries between cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C are relatively blurred, which may be because they have recombinated to form a uniform cactus polysaccharide.

[0082] Experimental Example 2: TRPV1 Inhibition Test Test method: (1) Inoculation: The cells were seeded into 24-well plates and incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0083] (2) Capsaicin stimulation: Capsaicin was prepared according to the experimental groups in Table 2. After cells were cultured in 24-well plates for 24 hours, the cells were administered to the groups, with 3 replicates per group. Cell culture medium was added to the blank control group, and cell culture medium containing 0.01% capsaicin was added to the negative control group, positive control group, and sample group. The cells were cultured for another 24 hours at 37°C in an incubator with 5% CO2.

[0084] (3) Sample feeding: After the cells in the 24-well plate have grown for 24 hours, samples are fed to the groups, with 3 replicates per group. Cell culture medium is added to both the blank control and the negative control group. Cell culture medium containing 0.001% 4-tert-butylcyclohexanol is added to the positive control group. Cell culture medium containing 0.5% of the samples from each example and comparative example is added to the sample groups. The samples are then cultured for another 24 hours at 37°C in an incubator with 5% CO2.

[0085] (4) TRPV1 detection: On the fourth day, immunofluorescence staining was performed. The sample was washed three times with PBS, fixed with 4% paraformaldehyde for 30 min, washed three times with PBS, permeabilized with 0.5% Triton X-100 for 20 min, washed three times with PBS, blocked with 5% BSA at room temperature for 60 min, discarded after blocking, and incubated overnight at 4°C with the prepared TRPV1 primary antibody (purchased from Abcam). On the fifth day, the sample was washed three times with PBS, incubated with the prepared fluorescent secondary antibody (purchased from Abcam) at room temperature in the dark for 1 h, washed three times with PBS, and an anti-fluorescence quencher was added. The sample was then observed and photographed using a fluorescence microscope.

[0086] (5) Data processing: Perform fluorescence photo processing to calculate IOD value.

[0087] Experimental results: Test results are available Figure 2 .Depend on Figure 2 It was found that, compared with medium and low molecular weight polysaccharides, high molecular weight polysaccharides showed the best inhibitory effect on TRPV1, indicating that high molecular weight polysaccharides form a protective film on the cell surface, reducing the binding of capsaicin to TRPV1. The effect was enhanced when polysaccharides of different number-average molecular weights were combined in a certain proportion. This is partly because the high molecular weight polysaccharides retained their efficacy characteristics, and also indicates that the combination of these polysaccharides in a certain proportion has a synergistic effect.

[0088] Table 2 Experimental Groups

[0089] Experiment Example 3: DNA Protection Test 8-hydroxy-2′deoxyguanosine (8-OHdG) is widely recognized as a biomarker for DNA oxidative damage. This experiment, based on an H2O2-induced keratinocyte damage model, evaluated the protective effect of samples against DNA oxidative damage by detecting the effect of sample addition on intracellular 8-OHdG levels.

[0090] Test method: (1) Inoculation: The cells were seeded into 24-well plates and incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0091] (2) H2O2 induction: After culturing cells for 24 h, wash once with PBS and add H2O2 induction solution. Incubate for 20 min and wash twice with PBS. The blank control group does not add H2O2 induction solution, but adds the same volume of PBS solution.

[0092] (3) Sample feeding: After induction, cell culture medium was added to both the blank control and the negative control group, cell culture medium containing 10 μg / mL vitamin E was added to the positive control, and cell culture medium containing 0.5% of each example and comparative sample was added to the sample group. The samples were then cultured for 18-24 h at 37°C in an incubator with 5% CO2.

[0093] (4) 8-OHdG detection: Immunofluorescence staining was performed, followed by washing three times with PBS, fixation with 4% paraformaldehyde for 30 min, washing three times with PBS, permeabilization with 0.5% Triton X-100 for 20 min, washing three times with PBS, blocking with 5% BSA at room temperature for 60 min, and incubation overnight at 4°C with 8-OHdG primary antibody (purchased from Abcam). The next day, the sample was washed three times with PBS, incubated with fluorescent secondary antibody (purchased from Abcam) at room temperature for 1 h, washed three times with PBS, and observed and photographed under a fluorescence microscope.

[0094] (5) Data processing: Perform fluorescence photo processing to calculate IOD value.

[0095] Test results are available Figure 3 .Depend on Figure 3 It is known that cactus polysaccharides have a certain protective effect against DNA damage. When polysaccharides of different number-average molecular weights are combined in a specific ratio, a synergistic effect can be further demonstrated, effectively enhancing the protective effect against DNA damage.

[0096] Experimental Example 4: ABTS Free Radical Scavenging Test Test method: 5 mL of 7 mmol / L 2,2'-adiazonium-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt (abbreviated as ABTS) and 88 μL of 140 mmol / L potassium persulfate solution were mixed and allowed to stand for 16 h at room temperature in the dark to form an ABTS free radical stock solution. The ABTS stock solution was diluted with 50% ethanol to achieve an absorbance of 0.8 at 734 nm. Following the reaction system in Table 3, three replicates were performed for each group. The test sample and ABTS solution were added, mixed thoroughly, and reacted for 2 h. The absorbance was then measured at 734 nm.

[0097] The test samples were cactus polysaccharide products prepared according to the methods of Example 1 and Comparative Examples 1-10, and the test results are shown in [reference to relevant documentation]. Figure 4 The test samples in the sample group were calculated according to the mass concentration of the cactus polysaccharide products obtained in each example or comparative example, and the sample concentration was set to 5%. The free radical scavenging rate was tested according to the reaction system described below.

[0098] Table 3 ABTS Free Radical Scavenging Test Reaction System

[0099] The calculation formula is as follows:

[0100] In the formula, A is the absorbance value of the mixed solution of ABTS and the test sample, B is the absorbance value of the ABTS solution after mixing 50% ethanol by volume, and C is the absorbance value of the mixed solution of 50% ethanol by volume and the sample.

[0101] Test results are available Figure 4 .Depend on Figure 4 It is known that cactus polysaccharides have the effect of scavenging ABTS free radicals and have antioxidant effects. When polysaccharides of different number average molecular weights are combined in a specific ratio, they can further show a synergistic effect and have a stronger antioxidant effect.

[0102] Experimental Example 5: Anti-stimulation Test Test methods Sodium dodecyl sulfate (SDS) is an anionic surfactant that, when applied to the skin, disrupts the skin barrier function and causes skin irritation. SDS acts on red blood cells (RBCs), altering cell membrane permeability and causing hemoglobin leakage, leading to hemolysis. Preliminary experiments were conducted to adjust red blood cell density and SDS dosage, ensuring the hemolysis rate in the negative control group was between 60% and 90%. Centrifuge tubes were prepared according to the reaction system shown in Table 4, with the addition of the test sample, PBS, RBC suspension, and SDS, and mixed thoroughly. The sample mass fraction was 5%. The tubes were incubated on a shaker for 10 minutes, then centrifuged. The observed phenomena were observed, and the supernatant was collected for optical density (OD530).

[0103] Table 4. Response system for anti-stimulation test

[0104] Note: In the table, "+" represents "added" and "-" represents "not added". The sample in the model control is deionized water.

[0105] Formula for calculating red blood cell hemolysis rate:

[0106] Formula for calculating the hemolysis inhibition rate of erythrocytes:

[0107] Experimental results Test results are available Figure 5 .Depend on Figure 5 It is known that cactus polysaccharides have certain anti-irritant effects. When polysaccharides of different number average molecular weights are combined in a specific ratio, they can further exhibit synergistic effects, with stronger anti-irritant activity and more significant cell-protective effects.

[0108] Experiment Example 6: Stability Test The cactus polysaccharide products obtained in Example 1, Comparative Examples 2-4, Comparative Example 8 and Comparative Example 11 were freeze-dried and then prepared into 1% aqueous solutions. The stability of the samples was then investigated.

[0109] (1) Heat resistance test The aqueous solutions of cactus polysaccharide products from each example and comparative example were kept at 50±2℃ for 5 days, 10 days, and 1 month, respectively. After returning to room temperature, the turbidity was observed compared with that before the experiment. ○ indicates no turbidity; × indicates turbidity.

[0110] (2) Cold resistance test The aqueous solutions of cactus polysaccharide products from each example and comparative example were kept at -17±2℃ for 5 days, 10 days, and 1 month, respectively. After returning to room temperature, the samples were observed to see if there was any obvious precipitation compared with before the experiment. ○ indicates no precipitation; × indicates precipitation.

[0111] Table 5 Results of heat resistance and cold resistance stability tests

[0112] The experimental results above show that cactus polysaccharides may become unstable during long-term storage. When polysaccharides of different number average molecular weights are combined in a specific ratio, the tertiary molecular weight polysaccharides form an "interlocked stable system," exhibiting superior stability. Combining polysaccharides of different number average molecular weights in a specific ratio has a synergistic effect on improving stability.

[0113] Experiment Example 7: Skin Feel Test The cactus polysaccharide products of Example 1 and Comparative Examples 2-4 were prepared into aqueous solutions with a concentration of 5 wt% (solvent: deionized water). Eight testers were selected to rate all the samples according to "smoothness (1-5 points, 5 points is the smoothest), stickiness (1-5 points, 5 points is the stickiest), and absorption speed (1-5 points, 5 points is the fastest)" and ranked them by preference (1 is the highest). The test results are shown in Table 6.

[0114] Table 6 Skin feel test results

[0115] Experimental Example 9: Solubility After freeze-drying the cactus polysaccharide products obtained in Example 1 and Comparative Examples 2-4, deionized water was added at a concentration of 5 wt% and dissolved at 25°C with stirring. The time to complete dissolution (without visible particles) was recorded.

[0116] Table 6 shows that the skin feel of the cactus polysaccharide composition with different number-average molecular weight polysaccharides combined in a specific ratio is better than that of the comparative examples. This is because the combination of different number-average molecular weights balances the relationship between "smoothness, stickiness, and absorption." Generally, high molecular weight polysaccharides provide a smooth feel, medium molecular weight polysaccharides reduce the stickiness of high molecular weight polysaccharides, and low molecular weight polysaccharides accelerate the overall absorption rate. Comparative example 4, which screened small molecular weight polysaccharides, theoretically has the best absorption effect and the lowest stickiness. However, by combining different number-average molecular weight polysaccharides in a specific ratio, the absorption effect is close to that of simple small molecular weight polysaccharides. The addition of medium and high molecular weight polysaccharides did not significantly affect absorption, and the stickiness did not increase significantly. The smoothness was significantly improved, demonstrating the uniformity of different number-average molecular weight cactus polysaccharides in the obtained composition. This further illustrates that while maintaining their own efficacy characteristics, they are combined into a whole, showing a better synergistic effect.

[0117] Table 7 Solubility Test Results

[0118] As can be seen from the results in Table 7, high molecular weight polysaccharides tend to aggregate, resulting in poor solubility and long dissolution time in aqueous solution. When polysaccharides of different number average molecular weights are combined in a certain proportion, the aggregation of molecular weight polysaccharides can be reduced by forming hydrogen bonds between molecules. At the same time, the overall dissolution resistance is reduced by synergistic dispersion, thereby improving the overall solubility. Its solubility is close to that of the best-soluble small molecule polysaccharide (Comparative Example 4).

[0119] This invention combines cactus polysaccharides of different number-average molecular weights in a specific ratio, maintaining the unique efficacy of each polysaccharide while creating a homogeneous whole. This results in a complementary and synergistic effect, further enhancing the overall performance of the cactus polysaccharide composition. Simultaneously, the recombination of these polysaccharides forms an "interlocked stable system," maintaining high stability even under extreme conditions, meeting the industrial requirements for long-term storage of skincare products. By controlling the number-average molecular weight and its proportions, the skin feel and solubility are further improved, enhancing the user experience and reducing the processing difficulty in skincare product manufacturing. This invention achieves synergistic efficacy solely through the combination of different number-average molecular weight ranges of cactus polysaccharides, eliminating the need for exogenous active ingredients. This results in a simpler composition, reducing the probability of skin sensitization and better meeting the core requirement of "safe and non-irritating" skincare products for sensitive skin.

[0120] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A cactus polysaccharide composition, characterized in that, The cactus polysaccharide composition includes cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C; Among them, the number average molecular weight of cactus polysaccharide A is above 100,000 Da, the number average molecular weight of cactus polysaccharide B is 10,000-100,000 Da, and the number average molecular weight of cactus polysaccharide C is below 10,000 Da. The mass ratio of cactus polysaccharide A, cactus polysaccharide B and cactus polysaccharide C in the cactus polysaccharide composition is cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15): (50-70): (10-30).

2. The cactus polysaccharide composition according to claim 1, characterized in that, The cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C were prepared by enzymatic hydrolysis of cactus water extract with amylase.

3. The method for preparing a cactus polysaccharide composition as described in claim 1 or 2, characterized in that, The process involves enzymatically hydrolyzing cactus water extract with amylase, then screening out cactus polysaccharide A with a number average molecular weight of over 100,000 Da, cactus polysaccharide B with a number average molecular weight of 10,000-100,000 Da, and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da. The cactus polysaccharide A, cactus polysaccharide B, and cactus polysaccharide C are then mixed evenly in a mass ratio of cactus polysaccharide A: cactus polysaccharide B: cactus polysaccharide C = (5-15):(50-70):(10-30).

4. The method for preparing the cactus polysaccharide composition according to claim 3, characterized in that, The method for preparing the cactus water extract includes extracting the cactus stem with water at 40-70℃ for 1-3 hours. Preferably, the mass ratio of the cactus stem to water is cactus:water = 1:(10-40). Preferably, the sample is allowed to stand for 12-18 hours after extraction; Preferably, the cactus water extract is decolorized and filtered before being enzymatically hydrolyzed; Preferably, the cactus water extract is decolorized using activated carbon at a dosage of 0.5-2.0% of the material mass. Preferably, the amylase includes α-amylase; Preferably, the amount of amylase used in the enzymatic hydrolysis is 0.005%-0.02% of the material mass; Preferably, the enzymatic hydrolysis temperature is 50-55℃; Preferably, after enzymatic hydrolysis, the obtained material is desalted and then screened to obtain cactus polysaccharide A with a number average molecular weight of more than 100,000 Da, cactus polysaccharide B with a number average molecular weight of 10,000-100,000 Da, and cactus polysaccharide C with a number average molecular weight of less than 10,000 Da. Preferably, the desalination is carried out using cationic resin and anionic resin; Preferably, the screening is carried out by membrane separation, and materials with a number average molecular weight of more than 100,000 Da are selected as cactus polysaccharide A, materials with a number average molecular weight of 10,000-100,000 Da are selected as cactus polysaccharide B, and materials with a number average molecular weight of less than 10,000 Da are selected as cactus polysaccharide C. Preferably, the mixture is compounded after being thoroughly mixed; Preferably, the compound is made of glycerol, and the mass ratio of the material to the amount of glycerol is material:glycerol = 1:(0.5-1.5). Preferably, a preservative is added after compounding, and the amount of preservative is 1%-3% of the total mass of the materials; Preferably, the preservative includes pentylene glycol / hexanediol. Preferably, the final material is sterilized to obtain a cactus polysaccharide composition; Preferably, the sterilization includes sterilization at 95-100°C for more than 40 minutes.

5. The cactus polysaccharide composition as described in claim 1 or 2, or the cactus polysaccharide composition prepared by the method described in claim 3 or 4, for use in the preparation of cosmetic / pharmaceutical products.

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