A malve polysaccharide skin cream and a preparation method thereof

Abstract

The application discloses a malve nut polysaccharide skin care cream and a preparation method thereof, and belongs to the technical field of skin care products. According to the mass fraction, the components of the malve nut polysaccharide skin care cream comprise 60-85 parts of water, 3-15 parts of a moisturizing agent, 5-15 parts of an emollient, 6-10 parts of an emulsifier, 0.1-0.5 parts of a preservative and 1-3 parts of a malve nut polysaccharide extract, and a pH regulator is used to adjust the pH of the system to 4.5-6.5. The malve nut polysaccharide skin care cream is prepared by adding the active ingredient malve nut polysaccharide extract to the aqueous skin cream, and the malve nut polysaccharide extract has the characteristics of anti-inflammation, anti-oxidation, anti-allergy, viscoelasticity and film-forming property. By controlling the added amount, the prepared malve nut polysaccharide skin care cream not only retains the refreshing skin feeling and fast absorption characteristics of the aqueous skin cream, but also has the functions of long-acting water locking and barrier repair.

Description

Technical Field

[0001] This invention belongs to the field of skin care technology, specifically relating to a golden sunflower polysaccharide skin care cream and its preparation method. Background Technology

[0002] Skin creams, as common topical preparations, are usually classified into two categories based on their base characteristics: water-based and oil-based.

[0003] 1. Water-based skin cream (O / W type lotion)

[0004] With water as the continuous phase and oil dispersed within it, its core advantage lies in:

[0005] (1) Lightweight feel: Easy to spread, absorbs quickly, and leaves no sticky residue after use. Suitable for oily skin or hot and humid environments.

[0006] (2) Compatible with hydrophilic active ingredients: such as vitamin C derivatives, sodium hyaluronate, etc., which can enhance moisturizing and antioxidant effects;

[0007] (3) Excellent breathability: reduces the risk of clogged pores.

[0008] Its inherent defects include:

[0009] (1) Limited moisturizing effect: The water evaporates easily and the long-term water-locking ability is insufficient, especially in dry environments where the moisturizing effect is significantly reduced;

[0010] (2) Weak barrier repair: Lacking sufficient lipid components, it is difficult to effectively repair the skin barrier and has limited improvement on dry / damaged skin;

[0011] (3) Stability challenges: The system relies on emulsifiers / preservatives to maintain stability, which may increase the complexity of the formulation and the risk of irritation.

[0012] 2. Oil-based skin cream (W / O type or pure oil ointment)

[0013] With oils and fats as the continuous phase, its significant characteristics are:

[0014] (1) Powerful water-locking and sealing: The oil film can significantly reduce transepidermal water loss (TEWL), making it suitable for extremely dry environments or skin with a damaged barrier;

[0015] (2) High permeability: Promotes the delivery of fat-soluble active ingredients (such as retinol and vitamin E);

[0016] (3) High formula stability: It is not easy for microorganisms to grow, and the need for preservatives is low.

[0017] However, its main drawbacks are:

[0018] (1) Heavy skin feel: poor spreadability, easy to produce greasy feeling, may cause acne or discomfort, poor user experience;

[0019] (2) Limited application scenarios: It may aggravate shine on oily / combination skin and has low applicability in hot climates;

[0020] (3) Narrow component compatibility: It is difficult to effectively carry high concentrations of hydrophilic active substances.

[0021] In comparison, water-based skin creams are more favored by consumers due to their easy spreadability, rapid absorption, and non-sticky feel after use. However, their inherent defects limit their effectiveness to some extent. How to overcome the defects as much as possible while retaining the advantages of water-based skin creams, so that water-based skin creams can retain the refreshing feel and fast absorption characteristics, while also having long-lasting moisturizing and barrier repair functions, is the current research direction of those skilled in the art. Summary of the Invention

[0022] The purpose of this invention is to provide a golden sunflower polysaccharide skin cream and its preparation method. By adding the active ingredient golden sunflower polysaccharide extract to a water-based skin cream, and utilizing the anti-inflammatory, antioxidant, anti-allergic, viscoelastic, and film-forming properties of the golden sunflower polysaccharide extract, and by controlling the amount added, the resulting golden sunflower polysaccharide skin cream not only retains the refreshing feel and rapid absorption characteristics of water-based skin creams, but also possesses long-lasting moisturizing and barrier repair functions.

[0023] To achieve the above objectives, the present invention provides the following technical solution:

[0024] One of the technical solutions of this invention is to provide a golden sunflower polysaccharide skin cream, which, by mass parts, comprises: 60-85 parts water, 3-15 parts moisturizer, 5-15 parts emulsifier, 6-10 parts emulsifier, 0.1-0.5 parts preservative and 1-3 parts golden sunflower polysaccharide extract, and uses a pH adjuster to adjust the pH of the system to 4.5-6.5.

[0025] The anti-inflammatory effect of the golden hibiscus polysaccharide extract in this invention can reduce skin inflammation, soothe secondary damage to the skin caused by external stimuli, and help repair the skin barrier. Its antioxidant effect can resist oxidative damage to the skin caused by free radicals, delay aging, and improve skin condition. The small amount of flavonoids contained in the extract also contributes to anti-inflammation, anti-oxidation, and anti-allergy effects. Furthermore, the viscoelasticity and film-forming properties of golden hibiscus polysaccharide allow it to replace conventional thickeners, improving the application feel while increasing the product's viscosity and stability, preventing separation, and effectively reducing transepidermal water loss, thus enhancing the product's water-locking function. In addition, skincare products containing thickeners may cause problems such as clogged pores and acne. Thickeners form a thick "glue" layer on the skin surface, which not only affects makeup application but may also increase the burden on the skin, potentially clogging pores, damaging the skin's natural barrier, accelerating skin aging, and causing skin sensitivity. The golden sunflower polysaccharide extract used as a substitute for thickeners in this invention does not have the above problems. The large molecular weight golden sunflower polysaccharide extract can lock in skin moisture while maintaining good breathability, making it an excellent component to replace thickeners.

[0026] Preferably, the moisturizer includes one or more of the following: glycerin, sodium hyaluronate, propylene glycol, butylene glycol, amino acid moisturizers, sorbitol, and trehalose.

[0027] Optionally, the amino acid-based moisturizer includes one or more of arginine, serine, and glycine.

[0028] Preferably, the emollient comprises one or more of the following: light synthetic esters, silicone oil derivatives, natural oils, and fatty alcohols.

[0029] Optionally, the light synthetic esters include one or more of the following: caprylic triglyceride, caprylic triglyceride, ethylhexyl palmitate, and dioctyl carbonate; the silicone oil derivatives include one or more of the following: cyclopentasiloxane, polydimethylsiloxane, and cyclohexylsiloxane; the natural oils include one or more of the following: squalane (plant-derived), jojoba oil, and almond oil; and the fatty alcohols include one or more of the following: cetyl alcohol, stearyl alcohol, and cetearyl alcohol.

[0030] Preferably, the emulsifier includes one or more of the following: polysorbate emulsifiers, lecithin, glyceryl stearate, and acrylic emulsifiers.

[0031] Optionally, the polysorbate emulsifier includes one or more of polysorbate-20, polysorbate-60, and polysorbate-80; the acrylic emulsifier includes one or more of sodium dodecyl sulfate, sodium dodecyl sulfonate, and sodium dodecylbenzene sulfonate.

[0032] Preferably, the preservative includes one or more of the following: phenoxyethanol, sodium benzoate, potassium sorbate, a composition of p-hydroxyacetophenone and capryloyl hydroxamic acid, polyol preservatives, isothiazolinone preservatives, and paraben preservatives.

[0033] Optionally, the polyol preservative includes pentanediol and / or hexanediol; the isothiazolinone preservative includes methylisothiazolinone and / or methylchloroisothiazolinone; and the paraben preservative includes methylparaben and / or propylparaben.

[0034] Preferably, the extraction solution used when extracting the golden sunflower polysaccharide extract is a weakly acidic aqueous solution with a pH of 4.5 to 6.5.

[0035] The main reason why this invention uses an aqueous solution with a specific pH value as the extraction solution is that this pH value is within the acidic range of the skin surface. The golden sunflower polysaccharide extract obtained by extracting with an extract solution within this pH range has better anti-inflammatory and antioxidant effects.

[0036] Preferably, the pH adjuster includes: citric acid, lactic acid, triethanolamine, sodium hydroxide, or potassium hydroxide.

[0037] The second technical solution of the present invention provides a method for preparing the above-mentioned golden sunflower polysaccharide skin cream, comprising the following steps:

[0038] Moisturizer, emollient, and golden sunflower polysaccharide extract were added to water accounting for 50-70% of the total water volume. After mixing evenly, the mixture was homogenized once. Then, emulsifier was added, and the mixture was homogenized twice. Preservative and the remaining water were added, and the mixture was mixed evenly. The pH value was adjusted to 4.5-6.5 using a pH adjuster. The mixture was homogenized three times to obtain the golden sunflower polysaccharide skin cream.

[0039] Preferably, the conditions for the first, second, and third homogenizations are all 2500–3500 rpm for 5–10 min.

[0040] The beneficial technical effects of the present invention are as follows:

[0041] This invention uses a water-based skin cream as a base and adds the active ingredient, golden sunflower polysaccharide extract. By utilizing the anti-inflammatory, antioxidant, anti-allergic, viscoelastic, and film-forming properties of golden sunflower polysaccharide extract and controlling the amount added, the resulting golden sunflower polysaccharide skin cream not only retains the refreshing feel and fast absorption characteristics of water-based skin creams, but also has long-lasting moisturizing and barrier repair functions. Detailed Implementation

[0042] Hibiscus manihot L., hailed as the "giant panda of the plant kingdom," is a gem of the Malvaceae family. Not only are its flowers vibrant, but the polysaccharides found deep within its roots, stems, leaves, and flowers are a focal point of ongoing exploration in modern life sciences. Hibiscus manihot L. Polysaccharides (HMLP), one of its core active ingredients, demonstrates remarkable application potential in health maintenance, disease prevention, and even cutting-edge biomaterials due to its complex structural characteristics and broad physiological activities.

[0043] Golden sunflower is an annual herbaceous plant with a tall, upright stem, palmately lobed leaves, and large, bright yellow flowers with a deep red center, giving it a unique appearance. Its capsule is pyramidal, and its seeds are kidney-shaped.

[0044] Golden sunflower is not a new discovery of modern science. In the traditional Chinese medicine system, its roots, leaves, flowers, and seeds are all used medicinally. Traditional Chinese medicine considers it to be sweet and cold in nature, possessing the effects of clearing heat and detoxifying, promoting diuresis and relieving strangury, cooling the blood and stopping bleeding, and moistening dryness and lubricating the intestines. It is often used to treat sore throat, dysentery, strangury, carbuncles and boils, constipation, and external bleeding. Its tender leaves, young pods, and petals can also be eaten as vegetables, are rich in nutrients, and are a typical example of a plant with both medicinal and edible properties. Modern research is a deeper exploration of the material basis of its traditional medicinal effects.

[0045] Golden hibiscus polysaccharides are a mixture of high molecular weight heteropolysaccharides extracted from different parts of the golden hibiscus (especially the flowers and leaves). Their chemical nature and properties are fundamental to understanding their biological activity.

[0046] The mainstream extraction method remains hot water extraction, which utilizes the water solubility of polysaccharides. To improve efficiency, it is often supplemented with ultrasound-assisted extraction (using cavitation to break cells), microwave-assisted extraction (rapid molecular motion generating heat), or enzymatic hydrolysis (using cellulase, pectinase, etc., to selectively destroy cell walls). Then, impurities are removed and purified to obtain relatively pure polysaccharide components. Ethanol fractionation is the most commonly used method, utilizing the differences in solubility of polysaccharides with different molecular weights or structures at different ethanol concentrations for separation.

[0047] Golden sunflower polysaccharides possess multidimensional biological activities:

[0048] (1) The "intelligent regulator" of the immune system:

[0049] Activation of Immune Cells: HMLP significantly activates macrophages, enhancing their ability to phagocytose foreign substances (phagocytic activity) and promoting the secretion of various key immune signaling molecules (cytokines), such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and interferon (IFN) with antiviral activity. These cytokines constitute a complex signaling network, recruiting and activating more immune cells (such as neutrophils and natural killer (NK) cells), initiating inflammatory defense responses. HMLP can also promote the proliferation and differentiation of T lymphocytes and B lymphocytes, enhancing cellular immunity (such as killing infected cells) and humoral immunity (producing more antibodies) responses.

[0050] (2) "Free radical scavengers" on the antioxidant front:

[0051] Neutralizing reactive oxygen species (ROS): Human metabolism and environmental stress constantly generate free radicals (ROS). Excessive ROS attack lipids, proteins, and DNA, leading to cell damage, aging, and various chronic diseases (arteriosclerosis, neurodegenerative diseases, cancer, etc.). HMLP can effectively remove superoxide anions (O3). 2·- It contains various ROS such as hydroxyl radicals (·OH), hydrogen peroxide (H2O2), etc.

[0052] (3) Potential interveners in glucose metabolism disorders:

[0053] Improving insulin resistance and lowering blood sugar: Animal models and some cell experiments show that HMLP can improve insulin sensitivity and reduce insulin resistance, thereby helping to lower blood sugar levels.

[0054] (4) “Ecological Guardians” and “Physical Barrier Workers” of Gut Health:

[0055] Prebiotic effect: As a type of dietary fiber, HMLP is not easily broken down by enzymes in the upper digestive tract and can reach the colon intact. There, it becomes the exclusive "food" (prebiotic) for beneficial gut bacteria (such as Bifidobacteria and Lactobacillus). These beneficial bacteria ferment HMLP to produce short-chain fatty acids (SCFAs, such as acetic acid, propionic acid, and butyric acid). SCFAs are the main energy source for colon cells, can lower the intestinal pH (inhibiting the growth of harmful bacteria), enhance the intestinal barrier function, regulate host immunity, and have systemic anti-inflammatory and metabolic regulatory effects.

[0056] Physical barrier protection: HMLP absorbs water and swells to form a gel-like substance, increasing stool volume and water content, promoting intestinal peristalsis, and relieving constipation. At the same time, this mucus-like gel layer can form a physical protective film on the intestinal mucosa surface, reducing the direct stimulation and damage of harmful substances to the intestinal wall, and helping to maintain the integrity of the intestinal barrier.

[0057] (5) "Multi-faceted Cooperative Warriors" on the Anti-tumor Battlefield:

[0058] Indirect immune-mediated: HMLP's powerful immune-enhancing effect is a crucial foundation for its anti-tumor potential. It enhances the immune system's ability to recognize and eliminate tumor cells by activating macrophages, NK cells, and T cells.

[0059] (6) The "loyal guardian" of liver health:

[0060] Protection against chemically induced liver injury: In animal models of liver injury induced by toxins such as alcohol, carbon tetrachloride (CCl4), and acetaminophen (APAP), HMLP showed significant protective effects. It reduced serum transaminase (ALT, AST, markers of hepatocellular injury) and bilirubin levels, and improved pathological changes in liver tissue (such as steatosis, necrosis, and inflammatory infiltration).

[0061] Based on the excellent bioactivity and high safety of golden sunflower polysaccharides, golden sunflower polysaccharides have shown promising application prospects in many fields, such as functional foods and health foods, and the pharmaceutical field.

[0062] Golden sunflower has been approved by the National Health Commission of China as a new resource food (new food raw material), which has laid the legal foundation for its application in the food field.

[0063] Currently, the application of golden sunflower polysaccharides in the skincare field is limited. How to fully utilize their properties to develop skincare products that are more easily accepted by consumers is a new research direction in this field.

[0064] This invention effectively overcomes the adverse effects of thickeners on skincare products by replacing thickeners in skincare products with golden sunflower polysaccharide extract. Skincare products prepared with golden sunflower polysaccharide extract as the active ingredient not only retain the refreshing feel and fast absorption characteristics of water-based skincare creams, but also have long-lasting moisturizing and barrier repair functions.

[0065] In addition, the inventors also discovered that the polysaccharide extract of golden sunflower obtained with a specific extract solution has a more prominent effect on anti-inflammatory and antioxidant properties in skin care products.

[0066] The components of the golden sunflower polysaccharide skin cream provided by this invention include: water, moisturizer, emollient, emulsifier, preservative, golden sunflower polysaccharide extract, and pH adjuster.

[0067] Besides the golden sunflower polysaccharide extract, the functions of the other components in this invention are as follows:

[0068] Water: Primarily used as a solvent to dissolve other water-soluble ingredients and form the basic texture of the product. It also provides instant hydration and a cooling sensation.

[0069] Moisturizers: They absorb moisture from deep within the skin and the environment, and bind it to the stratum corneum, increasing the skin's hydration level.

[0070] Emollients: fill the gaps between keratinocytes, smooth skin texture, provide a soft touch, and reduce transepidermal moisture loss. Emollients in water-based creams are usually chosen to be lighter in texture.

[0071] Emulsifiers: They stably mix the aqueous and oil phases of a product together to form a uniform cream texture. Without emulsifiers, the water and oil will separate into layers.

[0072] Preservatives: Prevent the growth of microorganisms (bacteria, fungi), ensuring product safety and extending shelf life. Water-based products especially require preservatives.

[0073] 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. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0074] 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.

[0075] Furthermore, regarding the 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. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0076] 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 to or equivalent to those described herein may be used in the implementation or testing of this invention.

[0077] 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.

[0078] Example 1

[0079] Preparation of Golden Sunflower Polysaccharide Skin Cream:

[0080] (1) The dried petals of the golden sunflower were crushed and then soaked in a 10wt.% citric acid anhydrous ethanol solution for 20 min to defatted the petals. The solution was filtered and washed with water. The residue was added to a deionized water solution with pH adjusted to 5.0 using citric acid at a ratio of 1:20. The solution was refluxed at 90℃ for 1 h and then filtered. After the filtrate was cooled, anhydrous ethanol was added until the alcohol concentration in the system was 70wt.%. At this point, the golden sunflower polysaccharide precipitated out. The solution was filtered and freeze-dried to obtain the crude golden sunflower polysaccharide.

[0081] (2) The obtained crude polysaccharide from *Hippophae rhamnoides* was loaded onto a DEAE-Sepharose FF column and connected to a fraction collector and a peristaltic pump. Deionized water was used for elution, and one tube of eluent was collected every 5 minutes. The absorbance of the eluent was measured in real time at 490 nm using the phenol-sulfuric acid method. The absorbance peak and the eluent before and after 5 minutes were collected, concentrated, dialyzed with a 3500 Da dialysis bag, and freeze-dried to obtain purified *Hippophae rhamnoides* polysaccharide extract with a polysaccharide purity of 98.1%.

[0082] (3) Prepare each raw material according to the components in Table 1. Add the moisturizer, emollient and purified golden sunflower polysaccharide extract to 50 parts of deionized water, stir at 800 rpm for 10 min to mix the materials evenly, and then homogenize at 2800 rpm for 10 min. Continue to add emulsifier, stir at 800 rpm for 10 min to mix the materials evenly, and then homogenize at 2800 rpm for 10 min. Add preservative and the remaining 25 parts of deionized water, stir at 800 rpm for 10 min to mix the materials evenly, adjust the pH of the system to 5.0 using citric acid value, and then homogenize at 2800 rpm for 10 min to obtain golden sunflower polysaccharide skin cream.

[0083] Table 1. Dosage of each component

[0084]

[0085] Example 2

[0086] Preparation of Golden Sunflower Polysaccharide Skin Cream:

[0087] Compared with Example 1, the only difference is that in the extraction step of the crude polysaccharide of *Hippophae rhamnoides*, the extraction solution used is deionized water, and the purity of the purified *Hippophae rhamnoides* polysaccharide extract is 97.9%.

[0088] Comparative Example 1

[0089] Preparation of regular skin cream:

[0090] Compared with Example 1, the difference is that 2.5 parts of purified golden sunflower polysaccharide extract were replaced with 1.5 parts of carbomer and 1 part of xanthan gum.

[0091] The skin irritation of each group of skin creams prepared in Examples 1-2 and Comparative Example 1 was investigated: the test was conducted according to the skin irritation test in Section 2.3.3 of the "Disinfection Technical Specifications" (2002 edition).

[0092] Animal preparation: New Zealand rabbits are pre-fed in a laboratory animal facility for 3 days to acclimatize. 24 hours before applying the skincare product, use electric clippers to remove the fur along both sides of the rabbit's spine on its back, taking care not to damage the skin. The removed area should be at least 5cm x 5cm on each side.

[0093] Skincare product application method: Using a self-comparison method of left and right sides of the same body, 0.5g of skincare cream was evenly applied to a 3cm×3cm area of ​​hair-removed, intact skin on the left side, then covered with a layer of non-irritating cellophane, and then secured with non-irritating adhesive tape. The right side of the skin was treated with the same method as a negative control group.

[0094] Symptom observation: Local skin reactions were observed at 1 h, 24 h, and 48 h after removal of the test substance. Scoring was performed according to the skin irritation reaction scoring criteria in Table 2-11 of the "Disinfection Technical Specifications" (2002 edition), and the skin irritation intensity grading was determined in Table 2-12. Specific scoring results are shown in Table 2.

[0095] Table 2. Scoring results of a single complete skin irritation test.

[0096]

[0097] Table 2 shows that the skin care products prepared in Examples 1 and 2 of this invention did not cause any skin irritation. The skin care product prepared in Comparative Example 1 showed barely visible erythema after 24 hours, indicating that using the golden sunflower polysaccharide extract provided by this invention to replace conventional thickeners can avoid the irritation caused by thickeners.

[0098] The moisturizing effects of the skin creams prepared in Examples 1-2 and Comparative Example 1 were examined:

[0099] Detection method:

[0100] The subjects' faces were cleansed with facial cleanser and the skin was dried with a dry facial tissue. They then sat quietly for 30 minutes in a laboratory at a temperature of 21±1℃ and 50±10%RH.

[0101] One side of the subject's cheek was randomly selected as the skin cream 1 area, and the other side was selected as the skin cream 2 area.

[0102] Researchers used the Tewameter TM300 skin moisture loss test probe to measure the transepidermal water loss (TEWL) value of the skin at 0h, 2h, and 6h after applying the skin cream. The results are shown in Table 3.

[0103] Table 3. Results of Moisturizing Efficacy Test

[0104]

[0105] As can be seen from Table 3, the golden sunflower polysaccharide skin cream prepared in Examples 1 and 2 of this invention can lock in moisture for a long time, while ordinary water-soluble skin creams generally have a lower water-locking ability.

[0106] The purified golden hibiscus polysaccharide extracts obtained in Examples 1 and 2 were subjected to an inhibition assay of the expression level of the cellular inflammatory factor IL-6.

[0107] Using the standard ELISA method:

[0108] The experiment was conducted using HDF-a cells in the logarithmic growth phase, with cells grown at a rate of 2 × 10⁶ cells / year. 5 The cells were seeded in 12-well plates and cultured at 37°C and 5% CO2 until 70% growth was achieved. Groups were set up: a blank stimulation control group, a lipopolysaccharide (LPS) stimulation group, and a *Hippophae rhamnoides* polysaccharide group (final concentrations of 20, 40, 60, 100, 150, and 200 μM, respectively). The culture medium was carefully removed. Fresh complete culture medium containing different concentrations of *Hippophae rhamnoides* polysaccharide extract was added to the *Hippophae rhamnoides* polysaccharide groups, while an equal volume of water was added to the blank stimulation control group and the LPS stimulation group. After 1 hour, except for the blank stimulation control group, 1 μg / mL LPS was added to all wells for 4 hours to induce cell inflammation. The plates were then removed, and ELISA experiments were performed according to the kit instructions.

[0109] Collect the culture medium supernatant: Collect the supernatant into a 1.5 mL EP tube, centrifuge at 1000 rpm for 10 minutes, and use the supernatant for ELISA detection of cytokines.

[0110] The secretion of IL-6 in the supernatant of the culture medium was detected using an enzyme-linked immunosorbent assay (ELISA) kit (Beyotime, catalog number: IL-6:PI236). The procedure was performed according to the kit's instructions, as follows:

[0111] (1) Reagent preparation

[0112] ① Remove from the refrigerator and allow to equilibrate to room temperature for 20 minutes. ② Dilute the washing buffer (20×) with double-distilled water to 1× to prepare the required washing buffer. ③ Add the standard diluent to one bottle of standard according to the volume indicated on the standard label and incubate at room temperature for 15 minutes. ④ Take five clean 1.5 mL centrifuge tubes and pre-add 250 μL of standard diluent to each tube to perform serial dilutions of the standard, ultimately obtaining six standard concentrations: 1000, 500, 250, 125, 62.5, and 31.25 pg / mL. Finally, add the diluted standard to the wells of the pre-coated plate sequentially, adding the standard diluent directly as the 0 pg / mL concentration, for a total of seven standard concentrations. ⑤ Add 300 μL to each well, and repeat after approximately 15-30 seconds, washing the plate five times in total. Pat the plate dry on paper.

[0113] (2) Operation steps

[0114] ① Calculate the number of pre-coated strips required for one experiment, and take out the required strips and place them in a 96-well frame.

[0115] ② Add the sample or standard of different concentrations to the corresponding well at a rate of 100 μL / well, seal the reaction well with a sealing film (transparent), and incubate at room temperature for 120 min.

[0116] ③ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0117] ④ Add 100 μL of biotinylated antibody per well, seal the wells with a clear sealing film, and incubate at room temperature for 60 minutes.

[0118] ⑤ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0119] ⑥ Add 100 μL of horseradish peroxidase-labeled streptavidin per well. Seal the reaction wells with white sealing film and incubate at room temperature in the dark for 20 minutes.

[0120] ⑦ Wash the board 5 times, and pat it dry on thick absorbent paper on the last wash.

[0121] ⑧ Add 100 μL of TMB colorimetric reagent solution per well, seal the reaction wells with white sealing film, and incubate at room temperature in the dark for 20 minutes.

[0122] ⑨ Add 50 μL of stop solution per well, mix well, and immediately measure the expression level of IL-6.

[0123] The test results showed that, compared with the LPS group, the purified *Hippophae rhamnoides* polysaccharide extracts from Examples 1 and 2 could downregulate the secretion level of the inflammatory factor IL-6. The IC50 of the purified *Hippophae rhamnoides* polysaccharide extract from Example 1 was [missing information]. 50 =75.62 μM, IC50 of the purified golden sunflower polysaccharide extract in Example 250 =135.62 μM, and all were downregulated in a dose-dependent manner. Compared with conventional water-extracted polysaccharides, the golden sunflower polysaccharide extract obtained by the present invention using a weakly acidic solution with pH=5.0 exhibits better anti-inflammatory activity against skin cells.

[0124] The antioxidant capacity of the purified golden hibiscus polysaccharide extracts obtained in Examples 1 and 2 was investigated:

[0125] DPPH radical scavenging capacity assay: 0.5 mg of purified Hibiscus mutabilis polysaccharide extract from each group was dissolved in 0.5 mL of water, and 0.5 mmol·L⁻¹ was added. -1 Mix 3.5 mL of DPPH solution thoroughly and let stand at room temperature in the dark for 30 min; zero the instrument with anhydrous ethanol and measure the absorbance at 517 nm, denoted as A. 测定 Distilled water is used as a substitute for the extract, denoted as A. 空白 Anhydrous ethanol replaces DPPH, denoted as A. 对照 ; Calculate DPPH clearance rate according to formula (1).

[0126] Hydroxyl radical (·OH) scavenging capacity determination: 0.5 mg of purified Hibiscus mutabilis polysaccharide extract from each group was dissolved in 0.5 mL of water, and 6 mmol·L⁻¹ water was added. -1 1 mL of ferrous sulfate, 6 mmol·L -1 2 mL of hydrogen peroxide, 6 mmol·L⁻¹ -1 Mix 1 mL of salicylic acid, place in a 37°C water bath for 30 min; zero the apparatus with distilled water, and measure the absorbance at 510 nm, denoted as A. 测定 Distilled water is used as a substitute for the extract, denoted as A. 空白 Distilled water replacing hydrogen peroxide is denoted as A. 对照 The OH scavenging rate is calculated according to formula (1).

[0127] Clearance rate (%) = [A 空白 -(A 测定 -A 对照 )] / A 空白 ×100%(1)

[0128] The calculated DPPH· scavenging rate and ·OH scavenging rate are shown in Table 4.

[0129] Table 4. Scavenging rates of DPPH· and ·OH

[0130] DPPH clearance rate (%) OH removal rate (%) Example 1 <![CDATA[89.5 a ]]> <![CDATA[96.4 a ]]> Example 2 <![CDATA[76.9 b ]]> <![CDATA[87.5 b ]]>

[0131] Note: Different lowercase letters on the same column in Table 4 indicate significant differences.

[0132] As can be seen from Table 4, compared with conventional water-extracted polysaccharides, the golden sunflower polysaccharide extract obtained by the present invention using a weakly acidic solution with pH=5.0 has better antioxidant activity.

[0133] 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 skin cream containing anti-inflammatory and antioxidant golden sunflower polysaccharides, characterized in that, The components, by mass parts, are: 60-85 parts deionized water, 3-15 parts moisturizer, 5-15 parts skin moisturizer, 6-10 parts emulsifier, 0.1-0.5 parts preservative and 1-3 parts golden sunflower polysaccharide extract, and the pH of the system is adjusted to 4.5-6.5 using a pH adjuster. The extraction solution used for extracting the polysaccharide from the golden sunflower is a weakly acidic aqueous solution with a pH of 4.5 to 5.

0.

2. The anti-inflammatory and antioxidant sunflower polysaccharide skin cream according to claim 1, characterized in that, The moisturizer includes one or more of the following: glycerin, sodium hyaluronate, propylene glycol, butylene glycol, amino acid moisturizers, sorbitol, and trehalose.

3. The anti-inflammatory and antioxidant sunflower polysaccharide skin cream according to claim 1, characterized in that, The emollients include one or more of the following: light synthetic esters, silicone oil derivatives, natural oils, and fatty alcohols.

4. The anti-inflammatory and antioxidant sunflower polysaccharide skin cream according to claim 1, characterized in that, The emulsifier includes one or more of the following: polysorbate emulsifiers, lecithin, glyceryl stearate, and acrylic emulsifiers.

5. The anti-inflammatory and antioxidant sunflower polysaccharide skin cream according to claim 1, characterized in that, The preservatives include one or more of the following: phenoxyethanol, sodium benzoate, potassium sorbate, a combination of p-hydroxyacetophenone and capryloyl hydroxamic acid, polyol preservatives, isothiazolinone preservatives, and paraben preservatives.

6. The anti-inflammatory and antioxidant sunflower polysaccharide skin cream according to claim 1, characterized in that, The pH adjuster includes: citric acid, lactic acid, triethanolamine, sodium hydroxide, or potassium hydroxide.

7. A method for preparing the anti-inflammatory and antioxidant golden sunflower polysaccharide skin cream according to any one of claims 1 to 6, characterized in that, Includes the following steps: Moisturizer, emollient, and golden sunflower polysaccharide extract were added to water accounting for 50-70% of the total water volume. After mixing evenly, the mixture was homogenized once. Then, emulsifier was added, and the mixture was homogenized a second time. Preservative and the remaining water were added, and the mixture was mixed evenly. The pH value was adjusted to 4.5-6.5 using a pH adjuster. The mixture was homogenized a third time to obtain the anti-inflammatory and antioxidant golden sunflower polysaccharide skin cream.

8. The preparation method of the anti-inflammatory and antioxidant golden sunflower polysaccharide skin cream according to claim 7, characterized in that, The conditions for the first, second, and third homogenizations are all 2500-3500 rpm for 5-10 minutes.

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