Compound jasmine hydrolat, preparation method and application thereof

Abstract

The application discloses a compound jasmine hydrolat as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine cosmetics, wherein the active ingredients include jasmine hydrolat and a traditional Chinese medicine extract, and the raw materials of the traditional Chinese medicine extract include traditional Chinese medicines such as gynostemma pentaphyllum, aloe, radix scrophulariae and giant knotweed rhizome. The application provides a natural and safe traditional Chinese medicine cosmetic, the extraction efficiency of the active ingredients in the compound jasmine hydrolat is remarkably improved through optimization of an extraction process, and efficient utilization of the active ingredients is ensured. By adding appropriate amounts of pH, humectants and preservatives, the color and aroma of the compound jasmine hydrolat are remarkably improved, and the compound jasmine hydrolat is more easily accepted by consumers. Through UV modeling experiments, it is verified that the compound jasmine hydrolat can remarkably improve the appearance of skin, repair the structure of epidermal thickness, collagen fibers and elastic fibers, reduce the levels of IL-1, IL-6 and TNF-alpha, reduce the level of MDA, and improve the levels of SOD and T-AOC, and has clear antioxidant and anti-inflammatory functions.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine cosmetics technology, and in particular relates to a compound jasmine hydrosol, its preparation method and application. Background Technology

[0002] Jasmine flowers possess properties that regulate qi and relieve pain, warm the stomach and spleen, reduce swelling and detoxify, and enhance the immune system. They exhibit significant anti-inflammatory and detoxifying effects on diseases such as dysentery, abdominal pain, conjunctivitis, and sores. The *Ben Cao Zai Xin* records that it "clears heat, removes cold accumulation, treats sores and boils, and eliminates carbuncles and tumors." Recent studies have found that ethanol extracts of jasmine leaves can promote wound healing in rat models of surgical incisions; petroleum ether extracts of jasmine leaves have a significant inhibitory effect on paw edema in mice, and their anti-inflammatory effect is comparable to diclofenac sodium, indicating that extracts from jasmine plants have potential application value in the treatment of acute inflammation.

[0003] Long-term UV radiation causes molecular-level damage that accumulates over time, eventually manifesting as skin pigmentation, oxidative damage, and other superficial changes. More seriously, this persistent state of oxidative stress can activate inflammatory cascades and may induce light-induced carcinogenesis, leading to complex pathophysiological effects. Therefore, it is crucial to scientifically and rationally prevent and alleviate the damage caused by UV radiation to the skin, as well as to research products with proven protective effects.

[0004] As chlorofluorocarbons (CFCs) continue to be released into the atmosphere, ozone layer depletion is becoming increasingly severe, leading to a significant increase in the intensity of ultraviolet (UV) radiation received by the Earth's surface. This enhanced UV exposure not only damages human skin tissue but also contributes to a sustained rise in the incidence of malignant skin tumors. Currently, most UV protection and repair products on the market are chemically synthesized drugs, which have problems such as significant side effects and high costs, making it difficult to meet consumers' demand for natural, safe, and functional foods.

[0005] Hydrosols are byproducts of steam distillation of aromatic plants. They contain not only trace amounts of essential oils but are also rich in various plant-based active substances, such as tannins, flavonoids, and other mineral nutrients. Traditional Chinese medicines like Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum have skin barrier-repairing effects; combining these ingredients can potentially provide anti-inflammatory and soothing benefits. However, current technologies show low utilization rates for jasmine hydrosols, low extraction efficiency of their active ingredients, and a lack of mature techniques for effectively integrating them with traditional Chinese medicine in herbal cosmetics. Furthermore, the strong odors and dark colors of herbal extracts make them difficult to apply directly to cosmetics, limiting their market promotion and application.

[0006] Therefore, how to improve the odor and color of Chinese herbal extracts, enhance their extraction efficiency and the stability of active ingredients, and realize their good application in cosmetics are urgent problems to be solved. At the same time, verifying the anti-inflammatory and soothing effects of extracts and finished products, and clarifying their safety and functional effectiveness are also technical challenges that need to be overcome in this field. Summary of the Invention

[0007] In view of this, the purpose of this invention is to provide a novel anti-inflammatory and soothing natural medicine and its preparation method, as well as a traditional Chinese medicine cosmetic and its preparation method.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A compound jasmine hydrosol, the active ingredients of which include jasmine hydrosol and traditional Chinese medicine extract, wherein the raw materials of the traditional Chinese medicine extract include: Centella asiatica, aloe vera, Scutellaria baicalensis and Polygonum cuspidatum.

[0009] Jasmine hydrosol is the aqueous distillate separated by condensation during the extraction of essential oil from jasmine flowers using steam distillation. In processes where the primary goal is to obtain essential oil, it is usually a byproduct and is also known as jasmine flower dew. It contains trace amounts of jasmine essential oil and water-soluble active ingredients, and is naturally mild with a sweet aroma.

[0010] The compound jasmine hydrosol mentioned above uses the following raw materials for its herbal extract: Centella asiatica (3-12 parts), Aloe vera (0.5-2 parts), Scutellaria baicalensis (1-4 parts), and Polygonum cuspidatum (1-4 parts).

[0011] The compound jasmine hydrosol described above has a volume ratio of jasmine hydrosol to traditional Chinese medicine extract of 1:15~20, preferably 1:17.

[0012] The compound jasmine hydrosol described above also includes pH adjusters, humectants, and preservatives.

[0013] The preparation method of the compound jasmine hydrosol and the Chinese herbal extract as described above is as follows: pulverize the Chinese herbs Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum, add an appropriate amount of organic solvent, stir and mix to obtain a Chinese herbal mixture, then use ultrasonic extraction multiple times, combine the extracts, concentrate, and obtain the Chinese herbal extract.

[0014] The specific method for preparing traditional Chinese medicine extracts includes the following steps: S1 Pretreatment: Select high-quality Chinese medicinal herbs such as Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum for later use.

[0015] S2 Grinding: Put the Chinese medicinal herbs Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum into a grinder for grinding; S3 Ultrasonic Extraction Method: Mix the powders of Chinese medicinal herbs Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum in a certain proportion, add an ethanol aqueous solution and stir to obtain a Chinese medicinal mixture, then sonicate. S4 filtration: First, use a filter cloth to coarsely filter the extract, then use a centrifuge to centrifuge, take the supernatant to evaporate the ethanol, add water, filter and concentrate to obtain the Chinese herbal extract.

[0016] As described above, in step S3, the volume fraction of the ethanol-water solution used is 50%-70%, the extraction time is 30-90 min, the extraction temperature is 50-70℃, the mass-to-volume ratio of the Chinese herbal medicine to the ethanol-water solution is 1:30~50 g / mL, and the mass-to-volume ratio of the amount of Chinese herbal medicine fed to the concentrated Chinese herbal extract is 0.20~0.24 g / mL.

[0017] Preferably, the volume fraction of the ethanol-water solution is 60%, the extraction time is 60 min, the extraction temperature is 50℃, the extraction is performed twice, and the mass-to-volume ratio of the Chinese herbal medicine to the ethanol-water solution is 1:50 g / mL.

[0018] Preferably, the ratio of the amount of Chinese herbal medicine added to the mass-volume ratio of the concentrated Chinese herbal medicine extract is 0.22 g / mL.

[0019] This invention provides a method for preparing compound jasmine hydrosol, comprising the following steps: R1. Mix the herbal extract with pH adjuster, humectant, and preservative evenly, and then decolorize. R2. Add jasmine hydrosol and mix well; R3. Sterilize to obtain compound jasmine hydrosol.

[0020] The preparation method described above is as follows: the pH adjuster is triethanolamine, the humectant is glycerin, the preservative is phenoxyethanol, the ratio of glycerin to traditional Chinese medicine extract is 1:1 to 3:1, the pH is adjusted to 6 to 7, and macroporous resin is used for decolorization.

[0021] The present invention also provides an application of compound jasmine hydrosol in the preparation of cosmetics, wherein the active ingredients of the cosmetics include the above-mentioned compound jasmine hydrosol.

[0022] In the application described above, the mass addition amount of the compound jasmine hydrosol is 50-100%.

[0023] As described above, the cosmetic is formulated as an aqueous solution, emulsion, spray, or mask.

[0024] This invention proposes a traditional Chinese medicine cosmetic based on jasmine hydrosol and herbal extracts. By optimizing the extraction and preparation processes, the anti-inflammatory and soothing components of jasmine hydrosol and herbal extracts are efficiently utilized, while also exhibiting good color, aroma, and stability.

[0025] This product is natural, safe, and has no side effects. It uses jasmine hydrosol, aloe vera, centella asiatica, polygonum cuspidatum, and scutellaria baicalensis extracts as raw materials, fully utilizing the hydrosol, a byproduct of essential oils, while also incorporating the effects of traditional Chinese medicine, providing a natural and safe herbal cosmetic. By optimizing the extraction process, the extraction efficiency of active ingredients in the compound jasmine hydrosol is significantly improved, ensuring efficient utilization of these ingredients. Through orthogonal experiments to optimize the hydrosol formula, and by adding appropriate amounts of pH, humectants, and preservatives, the color and aroma of the compound jasmine hydrosol are significantly improved, making it more acceptable to consumers. UV modeling experiments verify that the compound jasmine hydrosol of this invention can significantly improve skin appearance, repair epidermal thickness and collagen and elastic fiber structures; reduce IL-1, IL-6, and TNF-α levels, reduce MDA levels, and increase SOD and T-AOC levels, exhibiting clear antioxidant and anti-inflammatory functions.

[0026] This invention enables the high-value utilization of jasmine hydrosol, promotes the sustainable development of by-products, and has significant economic and environmental benefits.

[0027] The preparation method of the present invention is simple and low in cost, suitable for large-scale industrial production, and has good market application prospects. Attached Figure Description

[0028] Figure 1 This is a flow chart of the preparation process of the compound jasmine hydrosol in this application; Figure 2 These are images showing the skin condition of rabbits after applying compound jasmine hydrosol and distilled water for fourteen consecutive days. Each rabbit had compound jasmine hydrosol applied to its left side and distilled water applied to its right side. Figure 3 These are observation diagrams of the ocular irritation test results of animal formulations, where (A) no drug was administered on the blank side; (B) eye condition 1 hour after drug administration; (C) no drug was administered on the blank side; (D) eye condition 24 hours after drug administration; (E) no drug was administered on the blank side; (F) eye condition 48 hours after drug administration; (G) no drug was administered on the blank side; and (H) eye condition 72 hours after drug administration. Figure 4 These are experimental data graphs showing the effects of compound jasmine hydrosol on the general behavior and appearance of mice with photodamaged skin, and the appearance of mouse skin during the first week of the experiment. Figure 5 These are experimental data graphs showing the effects of compound jasmine hydrosol on the general behavior and appearance of mice with photodamaged skin, and the appearance of mouse skin in the fourth week of the experiment. Figure 6 It is a score for the appearance of skin damage in mice; Figure 7 It refers to the thickness of the epidermis; Figure 8 This describes the changes in subcutaneous blood vessels in mice. Figure 9The effect of compound jasmine hydrosol on the pathological changes of mouse skin tissue (HE staining ×100). Figure 10 The effect of compound jasmine hydrosol on the pathological changes of mouse skin tissue (VG staining ×100). Figure 11 It is Victoria blue dye (×100). Figure 12 This is a biochemical colorimetric method for detecting the content of tissue oxidation indicators in the skin of experimental mice, where (A) is T-AOC, (B) is MDA, and (C) is SOD; Figure 13 The effect of compound jasmine hydrosol on the expression levels of inflammatory cytokines in mouse skin, where (A) is TNF-α, (B) is IL-1β, and (C) is IL-6. Detailed Implementation

[0029] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below. The specific embodiments listed below are merely descriptions of the principles and features of the present invention, and the examples are only for explaining the present invention and are not intended to limit 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.

[0030] I. Preparation of Compound Jasmine Hydrosol 1.1 Preparation of Traditional Chinese Medicine Extracts: 1) Pretreatment: Select high-quality Chinese medicinal herbs such as Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum for later use.

[0031] 2) Grinding: Put the Chinese medicinal herbs Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum into a grinder and grind them.

[0032] 3) Ultrasonic extraction: Weigh the powders of Chinese herbs Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum according to the specified ratio, and add them to an ethanol-water solution according to the material-liquid ratio. Stir and mix to obtain a Chinese herbal mixture, and then extract by ultrasound and heating.

[0033] 4) Filtration: First, use a filter cloth to coarsely filter the extract, then use a centrifuge to centrifuge and take the supernatant for later use. Evaporate the ethanol from the supernatant, add water, filter and concentrate to obtain the Chinese herbal extract.

[0034] Orthogonal experiments were used to optimize extraction conditions, including determining the concentration and volume of ethanol solution, extraction temperature, and extraction time.

[0035] 1.2 Preparation of Compound Jasmine Hydrosol: Hydrosol components include jasmine hydrosol, Centella asiatica (a traditional Chinese medicine), aloe vera, Scutellaria baicalensis, Polygonum cuspidatum extract, pH adjuster, humectant, preservative, etc.

[0036] R1. Mix the herbal extract with pH adjuster, humectant, and preservative until uniform, adjust the pH of the solution to 6-7, and decolorize. R2. Add jasmine hydrosol and mix well; R3. Sterilization.

[0037] The hydrosol formula was optimized through orthogonal experiments, and appropriate amounts of pH adjusters, humectants and preservatives were added to ensure the color, aroma and stability of the hydrosol.

[0038] II. Investigation of Preparation Process 2.1 Process Investigation of Traditional Chinese Medicine Extracts Extraction time, water volume, extraction temperature, and ethanol volume fraction were used as factors of investigation. Total flavonoids, total polysaccharides, and total saponins were used as indicators (weighting coefficients for total flavonoids were 0.4, total polysaccharides were 0.2, and total saponins were 0.4). A four-factor, three-level orthogonal experiment was conducted, with the factor and level design shown in Table 8. A comprehensive weighted scoring method was used, with the maximum value of total flavonoids, total polysaccharides, and total saponins in the orthogonal experiment set at 100 points. The total amount of each indicator component was converted into a score, and a comprehensive score was calculated. The orthogonal experiment results were then calculated based on the comprehensive score, and variance analysis was performed to determine the optimal extraction process for compound jasmine hydrosol.

[0039] Table 1 Orthogonal Experiment Arrangement Table

[0040] Table 2. Orthogonal Experiment Data

[0041] Table 3. Results of orthogonal experimental variance analysis of alcohol extraction process

[0042] Orthogonal experimental analysis showed that the influence of the four main factors on the ethanol extraction effect of the medicinal materials was in the following order: A (extraction time) > B (extraction temperature) > D (ethanol volume fraction) > C (solid-liquid ratio). The degree of influence of each factor at different levels was in the following order: A2 > A1 > A3, B1 > B3 > B2, C2 > C3 > C2, D2 > D1 > D3. Further analysis using analysis of variance showed that factors A (extraction time), B (extraction temperature), C (ethanol volume fraction), and D (solid-liquid ratio) had no significant impact on the experimental results, with no statistically significant differences (P>0.05). The comprehensive score indicated that the optimal ethanol extraction process was A2B1C2D3. Combining the results of single-factor experiments and considering factors such as the enterprise's production cycle and production costs, the final optimal process was A2B1C2D3, involving two extractions: extraction with 60% ethanol, two extractions, each lasting 60 minutes, at a temperature of 50℃, with a solid-liquid ratio of 1:50.

[0043] 2.2 Process Validation Experiment According to the prescription ratio (1g aloe vera, 2g scutellaria baicalensis, 2g polygonum cuspidatum, 6g centella asiatica), three parallel portions of the medicinal materials were weighed and extracted separately according to the optimized process. After the extracts were combined, the extract was first coarsely filtered using a filter cloth, then centrifuged. The supernatant was evaporated to dryness, water was added, filtered, and concentrated to 50ml. The content of its active ingredients was then determined. The process validation results are shown in Table 4.

[0044] Table 4 Process Validation Test Table

[0045] 2.3 Process Investigation of Compound Hydrosol Formulation Based on the single-factor experiments, glycerol dosage (A), extract dosage (B), and decolorizing agent dosage (C) were selected as the factors to be investigated. An orthogonal experimental design was conducted using a three-factor, three-level orthogonal experiment (Table 5) to screen the optimal formulation and proportion of compound jasmine hydrosol. The results are shown in Table 6. In experiments 3, 7, and 8, the hydrosol was darker in color, slightly cloudy, had a weaker aroma, and low uniformity. In experiment 4, the hydrosol had good uniformity and pleasantness, but a slightly darker color. In experiments 2, 6, and 9, the hydrosol had good uniformity, but an off-odor. In experiments 1 and 5, the hydrosol was slightly darker in color and had low uniformity. The K value is the mean of the experimental results at the corresponding level for each factor; the R value is the difference between the maximum and minimum values ​​of K for each factor, plus the range. A larger R value indicates greater variability and a greater influence of the factor, therefore A>C>B. A larger K value indicates a greater influence of the level on the results, therefore A1>C1>B2. The best overall score was A1B2C1, which is 10mL glycerol, 10mL extract, using decolorizing agent macroporous resin, and employing D101 macroporous adsorption resin (D101, polystyrene-type non-polar adsorption resin). It has good comfort, strong stability, and the product is a pale yellow transparent liquid.

[0046] Table 5. Orthogonal Experimental Factor Level Design Table

[0047] Table 6. Results of Orthogonal Experiments

[0048] Sensory evaluation of compound jasmine hydrosol preparation In the above experiment, for the sensory evaluation of compound jasmine hydrosol, this application combined the characteristics of compound jasmine hydrosol itself and formulated a scoring standard for compound jasmine hydrosol based on influencing factors such as color, odor and stability of the product (Table 7). A 20-person evaluation group consisting of 10 males and 10 females scored the hydrosol on items such as aroma, color, centrifugation, cold resistance and heat resistance according to sensory evaluation and experimental data standards.

[0049] Table 7 Scoring Criteria

[0050] Example 1 Preparation of Chinese herbal extracts Step 1: Crush the Chinese herbs Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum into powder. The powder should contain 6 parts Centella asiatica, 1 part aloe vera, 2 parts Scutellaria baicalensis, and 2 parts Polygonum cuspidatum. Step 2: Add 60% ethanol solution at a material-to-liquid ratio of 1:40; Step 3: Extract at 50℃, 1 hour each time, 2 times; Step 4: First, use a filter cloth to coarsely filter the extract, then use a centrifuge to centrifuge, take the supernatant, evaporate the ethanol, add water, filter and concentrate to obtain the Chinese herbal extract. The mass-volume ratio of the amount of Chinese herbal medicine added to the concentrated Chinese herbal extract is 0.22 g / mL.

[0051] Formula for compound jasmine hydrosol Element: Traditional Chinese medicine concentrate: 10mL; pH adjuster (triethanolamine): 0.4mL Moisturizer (glycerin): 10mL; Preservative (phenoxyethanol): 0.6g Jasmine hydrosol: 170mL Preparation method: The concentrated Chinese herbal medicine solution was mixed evenly with pH adjuster, humectant, and preservative. It was then decolorized using macroporous resin and jasmine hydrosol was added and mixed evenly.

[0052] Sterilization. The hydrosol is filled into glass bottles sterilized by high-pressure steam to obtain a Chinese herbal cosmetic that can be used directly, or it can be used as a main ingredient in the preparation of other cosmetics.

[0053] III. Efficacy Verification of Compound Jasmine Hydrosol 3.1 Multiple skin irritation experiments of compound jasmine hydrosol on New Zealand rabbits Six clean-grade New Zealand White rabbits, half male and half female, weighing between 2 and 2.5 kg, were purchased from Hunan Taiping Biotechnology Co., Ltd., animal production license number SCXK (Gui) 2017-0001. They were housed at the Experimental Animal Center of Guangxi University of Traditional Chinese Medicine, with a room temperature of 20-25℃ and humidity of 60%-80%. Standard formulated feed was used, and water was unlimited. They underwent acclimatization feeding for 7 days prior to the experiment in the experimental animal facility.

[0054] Before the experiment, the hair on both sides of the spine on the back of the experimental animals was shaved off, and the downy hair was removed with depilatory cream. The hair removal area was 3cm × 3cm on each side, and the application area was 2.5cm × 2.5cm. Approximately 0.5mL of hydrosol was applied to one side of the skin once a day for 14 consecutive days. Starting from the second day, the hair was shaved off before each application, and any residual test substance was removed with water. The results were observed after 1 hour, and scored according to Table 8. The control area and the test area were treated in the same way. The average score per animal per day was calculated using the following formula, and the skin irritation intensity was determined according to Table 9.

[0055] Average score per animal per day = (∑Erythema and edema score / Number of animals tested) / 14 Table 8 Skin Irritation Response Scores

[0056] Table 9 Skin Irritation Intensity Grading

[0057] After applying hydrosol and distilled water to intact skin on both sides of rabbits respectively, a control group was created by comparing the left and right sides of the same animal. The effects of compound jasmine hydrosol on the skin irritation of New Zealand rabbits were observed, such as... Figure 2 As shown in Table 10, after 14 days of continuous application of the hydrosol, the hair on both sides of the animals was removed. The average score for both sides of the animals in this group was 0. Based on the evaluation criteria, the average score of the animals indicates that the compound jasmine hydrosol is non-irritating to the skin of New Zealand rabbits. In terms of behavior, the animals did not exhibit any abnormal behavior after administration, and their mental state was good, with normal food and water intake and formed feces. No adverse reactions were observed.

[0058] Table 10 Results of Skin Safety Tests

[0059] 3.2 Acute eye irritation experiment of compound jasmine hydrosol on New Zealand rabbits Four clean - grade New Zealand white rabbits, with half males and half females, weighing between 2 - 2.5 kg, were purchased from Hunan Taiping Biotechnology Co., Ltd. The animal production license number is SCXK (Gui) 2017 - 0001. They were raised in the Experimental Animal Center of Guangxi University of Chinese Medicine. The room temperature was 20 - 25 °C and the humidity was 60% - 80%. The experimental animals and the experimental environment met the corresponding national regulations. Standard formulated feed was used, and drinking water was unlimited. They were adaptively fed in the experimental environment for 7 days before the experiment.

[0060] Gently pull down the lower eyelid of one side of the rabbit's eye, and drop (or smear) 0.1 mL of the test substance into the conjunctival sac, and make the upper and lower eyelids passively close for 1 s to prevent the loss of the test substance. The other eye was not treated as a self - control. Do not rinse the eyes within 24 h after dropping the test substance. Clinical examination and scoring: Examine the animal's eyes at 1, 24, 48, 72 h, and on the 4th and 7th days after dropping the test substance. If no irritation reaction appears within 72 h, the test can be terminated. Except for observing the cornea, iris, and conjunctiva, other damage effects should be recorded and reported. The integral of the eye irritation reaction should be recorded according to the scoring criteria for eye damage in Table 11 during each examination.

[0061] Table 11 Scoring criteria for eye damage

[0062] According to the evaluation criteria, by calculating the average score of the evaluation of the animals, it can be determined that the hydrosol has no irritation to the eyes of New Zealand white rabbits. As can be seen from the above - mentioned scoring criteria for eye damage, the scoring for eye damage within 1 - 72 h is shown in Table 12. Figure 3 as shown below.

[0063] Table 12 Scoring results of eye damage

[0064] 3.3 Verification of anti - inflammatory and soothing effects: Experimental method: 1. Use a self - made ultraviolet modeling box to establish a mouse skin photo - damage model, and the modeling is in the form of ultraviolet irradiation.

[0065] SPF - grade KM female mice, weighing between 18 - 22 g, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. The animal production license number is SCXK (Xiang) 2021 - 0002. They were raised in the SPF - grade Experimental Animal Center of Guangxi University of Chinese Medicine. The breeding environment temperature was (22 ± 2) °C and the humidity was (55 ± 5)%. The lighting was natural day and night, and they had free access to drinking water and food. The bedding, feed, cages, water bottles, etc. used for animal breeding were all sterilized by high - pressure steam, and the breeding room was disinfected daily. The KM mice were adaptively fed for 7 days before the experiment.

[0066] Based on mouse weight, mice were randomly assigned to six groups using Microsoft Excel: a control group, a positive control group (vitamin E), a model group, and low-, medium-, and high-dose compound jasmine hydrosol groups, with eight mice in each group. The compound jasmine hydrosol product prepared in Example 1 was used as the high-dose group, diluted 10 times as the medium-dose group, and diluted 100 times as the low-dose group. One day before the experiment, the hair on the backs of the mice was removed using a shaving tool and depilatory cream, leaving a bare area of ​​approximately 4cm × 2cm. The hair was then trimmed appropriately according to the growth of the mouse hair. Except for the control group, all mice were given UV irradiation for 10 minutes to establish the model. Thirty minutes before irradiation, the control group and the model group received no medication. Vitamin E was applied to the bare skin of the mice in the positive control group, and the corresponding medications were applied to the low-, medium-, and high-dose compound jasmine hydrosol groups, with a dosage of 0.5 mL per mouse.

[0067] Extensive pigmentation, erythema, and wrinkles appeared on the irradiated areas of the mice's backs, with localized ulceration, scabs, and a leathery texture, indicating that the photodamage model of the mouse skin was successfully established. Irradiation dose and experimental groups are shown in Tables 13 and 14. During the feeding period, the experimental animals were provided with sufficient water and feed daily for four consecutive weeks. During the experiment, the mice's water and food intake, mental state, and skin appearance were observed and recorded daily after the model was established and after the hydrosol was applied.

[0068] Table 13 Weekly Irradiation Dose

[0069] Table 14 Experimental Groups and Drug Administration

[0070] Figure 4 , Figure 5 These are experimental data graphs showing the effects of compound jasmine hydrosol on the general behavior and appearance of mice with photosensitive skin, specifically the skin appearance of mice in the first and fourth weeks of the experiment.

[0071] See Table 15 for the scoring data and graphs of skin appearance damage. Figure 6 .

[0072] Table 15 Scoring of skin appearance damage in mice ( (x±s, n=8)

[0073] Note: Compared with the blank group # P <0.05, ## P <0.01; compared with the model group * P <0.05, **P <0.01 Table 16 Epidermal thickness ( (x±s, n=8)

[0074] Note: Compared with the blank group # P <0.05, ## P <0.01; compared with the model group * P <0.05, ** P <0.01 The experimental mice were fasted for 12 hours before sacrifice, but allowed free access to water. Their weight was measured, and they were euthanized after anesthesia with an appropriate amount of sodium pentobarbital. The skin on their backs was peeled off and divided into three sections for the following procedures: First, the skin was cut along the edge of the hairless area, and the thickness at the same midpoint was measured using calipers. Experimental data on the relative thickness of the epidermis and related figures are shown in Table 16 and [Figure number missing]. Figure 7 Excess fat and connective tissue were removed to ensure a smooth surface. Surface bloodstains were gently rinsed with saline solution. The tissue was then laid flat against a black background and photographed using a high-resolution camera under uniform lighting conditions. The experimental results are shown in [link to experimental results]. Figure 8 The image shows experimental data on changes in subcutaneous blood vessels in mice.

[0075] One sample of skin tissue was cut into appropriate sizes, fixed in 4% paraformaldehyde solution, embedded in paraffin, sectioned, stained with hematoxylin and eosin (HE), and mounted. Another sample of tissue was cut into small pieces and homogenized in a homogenizing tube at a ratio of 1g (tissue): 9mL (physiological saline) to prepare a 10% tissue homogenate, ensuring complete cell disruption, and then subjected to biochemical assays. The remaining sample was flash-frozen in liquid nitrogen and then transferred to -80℃ for storage until later use.

[0076] 2. Detection of biochemical indicators in skin tissue According to the instructions of the corresponding kits (commercially available malondialdehyde (MDA) kit, superoxide dismutase (SOD) kit, total antioxidant capacity (T-AOC) kit, and BCA protein concentration assay kit), the activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA), and the level of total antioxidant capacity (T-AOC) in skin tissue were measured to assess oxidative stress and antioxidant capacity. See Table 17 and... Figure 12 The data and graphs for anti-skin photodamage in the fourth week (comparison of T-AOC, MDA, and SOD levels) are shown.

[0077] Table 17 Effects of Compound Jasmine Hydrosol on Mouse Skin Tissue The effects of T-AOC, MDA and SOD levels ( (x±s, n=8)

[0078] Note: Compared with the blank group # P <0.05, ## P <0.01; compared with the model group* P <0.05,** P <0.01 3. Detection of inflammatory factor markers in skin tissue The levels of IL-1β, IL-6, and TNF-α in rat skin tissue were measured separately, following the instructions of the ELISA kits (commercially available interleukin-6 ELISA kit, interleukin-1β ELISA kit, and tumor necrosis factor-α ELISA kit). This study further investigated how the hydrosol exerted a relative therapeutic effect on mouse skin inflammation by influencing oxidative stress and the expression levels of inflammatory factors and tumor necrosis factor in the skin. (See Table 18 and...) Figure 13 The data and graphs for anti-skin photodamage at week four (comparison of IL-1β, IL-6, and TNF-α levels) are shown. Table 18 Effects of Compound Jasmine Hydrosol on the Expression Levels of Inflammatory Cytokines in Mouse Skin ( (x±s, n=8)

[0079] Note: Compared with the blank group # P <0.05, ## P <0.01; compared with the model group* P <0.05,** P <0.01 Compared with the blank group, the MDA level in the model group was significantly higher ( P <0.05), while SOD and T-AOC levels were significantly reduced ( P <0.05). Compared with the model group, the MDA levels in the low, medium, and high dose groups of compound jasmine hydrosol were significantly reduced ( P <0.05), while SOD and T-AOC levels were significantly elevated ( P <0.05); compared with the blank group, the levels of IL-1β, IL-6 and TNF-α in the model group were significantly increased ( P <0.01, P <0.05). Compared with the model group, the levels of IL-1β, IL-6, and TNF-α in the vitamin E group were significantly reduced ( P <0.01, P<0.05), while the levels of IL-1β, IL-6, and TNF-α in the low, medium, and high dose groups of compound jasmine hydrosol were also significantly reduced ( P <0.05, P <0.01). Among them, the high-dose group showed a particularly significant effect in reducing IL-1β and TNF-α ( P <0.01).

[0080] Therefore, it can be seen that compound jasmine hydrosol can significantly improve the appearance of the skin, repair the thickness of the epidermis and the structure of collagen / elastic fibers. Compound jasmine hydrosol has a significant protective effect against skin photodamage, which indicates that compound jasmine hydrosol has significant anti-inflammatory and antioxidant effects and can effectively alleviate skin photodamage.

[0081] 4. Pathological and histomorphological observation 4.1 HE staining Step 1: Mouse skin tissue soaked in paraformaldehyde is prepared into paraffin blocks and sectioned. The sections are then immersed in xylene bottles (No. I and No. II) for 20 minutes each, followed by immersion in anhydrous ethanol bottles (No. I and No. II) for 5 minutes each. Next, the sections are transferred to 75% ethanol bottles and immersed for 5 minutes each. Finally, they are washed with tap water to complete dewaxing. Step 2: The dewaxed sections are immersed in hematoxylin staining solution for 3-5 minutes, then carefully rinsed with tap water. Differentiation solution is then applied, followed by rinsing with tap water. The sections are then immersed in blueing solution for blueing, and finally rinsed with running water to complete hematoxylin staining. Step 3: The stained sections are immersed in 85% and 95% ethanol for 5 minutes each, followed by eosin staining solution for 5 minutes to complete eosin staining. Step 4: Place the hematoxylin-eosin stained sections sequentially into bottles I, II, and III of anhydrous ethanol and let them stand for 5 minutes each. Then, place the sections sequentially into bottles I and II containing xylene and let them stand for 5 minutes each. Finally, mount the sections with neutral resin. Step 5: Place the dehydrated and mounted samples under a microscope for observation and image acquisition and analysis.

[0082] The epidermis, acting as a physical barrier between the body and the external environment, effectively protects the skin from external disturbances. Morphological and histological studies indicate that the main characteristic of photodamage to the skin is the destruction of collagen structure and changes in epidermal thickness caused by excessive ultraviolet radiation. Ultraviolet radiation can lead to epidermal hyperplasia, a process achieved by activating epidermal growth factor receptors, thereby inducing keratinocyte proliferation. In this experiment, the pathological and histological changes in mouse skin were mainly observed using epidermal thickening, crusting and abscess formation, collagen fiber degeneration, and inflammatory cell infiltration as indicators for comprehensive analysis and evaluation. HE staining clearly revealed the general morphological structure of mouse skin. See also Figure 9The effect of compound jasmine hydrosol on histopathological changes in mouse skin tissue (HE staining ×100).

[0083] The skin structure of the control group mice was normal, and no pathological changes such as inflammatory cell infiltration were observed. In contrast, the epidermis of the model group mice was significantly thickened, with obvious crusted abscesses visible in the epidermal layer, accompanied by varying degrees of hyperkeratosis, significantly thickened spinous cells, elongated and undulating spinous processes, and inflammatory cell infiltration and erythrocyte exudation. Compared with the model group, the pathological changes in the skin of mice in the vitamin E group and the low, medium, and high dose groups of compound jasmine hydrosol were all alleviated, with the skin structure of the high dose group basically returning to normal.

[0084] 4.2 VG staining Collagen fibers are mainly distributed in the dermis of the skin. In normal tissues, the production and degradation of collagen fibers are in dynamic equilibrium. However, when the body suffers certain injuries, collagen fiber production exceeds degradation, leading to fibrosis and sclerosis of the skin tissue. VG staining is mainly used to distinguish collagen fibers from muscle fibers, thereby assessing the degree of damage and sclerosis in tissues and organs. After staining, collagen fibers appear red, while muscle fibers, nerve cell cytoplasm, and erythrocytes appear yellow. Figure 10 The effect of compound jasmine hydrosol on the histopathological changes of mouse skin tissue (VG staining ×100).

[0085] In the control group, collagen fibers in the dermis were arranged in parallel, parallel to the epidermis, and exhibited normal structure. In contrast, in the model group, collagen fibers in the superficial dermis were disordered and unevenly distributed, showing obvious hyperplasia, with collagen fibers agglomerated into bundles and abnormally deposited. Compared with the model group, the arrangement and distribution of collagen fibers in the vitamin E group and the low, medium, and high dose groups of compound jasmine hydrosol were all improved to some extent, and the improvement effect showed a dose-dependent relationship. Among them, the skin structure in the high-dose group was basically restored to normal, indicating that compound jasmine hydrosol has a significant protective effect against photodamage to the skin.

[0086] 4.3 Victoria Blue Staining When the skin is exposed to excessive ultraviolet radiation, the elastic fibers in the dermis will suffer pathological damage, gradually changing from the originally neat and orderly network structure to irregular lumps, and then symptoms such as disordered arrangement, thickening, and breakage will appear

[39] . Victoria blue staining can be used to observe the proliferation or breakage of elastic fibers in skin tissue. After staining, the elastic fibers appear blue. Figure 11 The effect of compound jasmine hydrosol on the histopathological changes of mouse skin tissue was determined by Victoria blue staining (×100).

[0087] In the control group, the mice had normal skin structure, with thin, sparsely distributed elastic fibers in the dermis, roughly parallel to the epidermis, maintaining skin elasticity. In contrast, the model group showed a significant increase and thickening of elastic fibers, disordered arrangement, and breakage, exhibiting a crisscrossed and disordered state, leading to loss of elastic fiber function and even skin hardening, indicating that ultraviolet radiation caused elastic fiber degeneration. Compared to the model group, the medium and high dose groups of compound jasmine hydrosol showed sparse, orderly, and well-organized elastic fibers, with significantly reduced damage, indicating that compound jasmine hydrosol has a significant protective and repairing effect on skin photodamage.

[0088] The above experimental results show that the compound jasmine hydrosol can significantly improve skin appearance, repair epidermal thickness and collagen and elastic fiber structure; reduce IL-1, IL-6, and TNF-α levels, reduce MDA levels, and increase SOD and T-AOC levels.

Claims

1. A compound jasmine hydrosol, characterized in that: Its active ingredients include jasmine hydrosol and traditional Chinese medicine extracts. The raw materials of the traditional Chinese medicine extracts include: Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum.

2. The compound jasmine hydrosol according to claim 1, characterized in that: The raw materials for the Chinese herbal extract include 3-12 parts of Centella asiatica, 0.5-2 parts of Aloe vera, 1-4 parts of Scutellaria baicalensis, and 1-4 parts of Polygonum cuspidatum.

3. The compound jasmine hydrosol according to claim 1, characterized in that: The volume ratio of jasmine hydrosol to traditional Chinese medicine extract is 1:15~20.

4. The compound jasmine hydrosol according to claim 1, characterized in that: It also includes pH adjusters, humectants, and preservatives.

5. A compound jasmine hydrosol according to claim 4, characterized in that, The preparation method of traditional Chinese medicine extract includes the following steps: S1 Pretreatment: Select high-quality Chinese medicinal herbs such as Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum for later use; S2 Grinding: Put the Chinese medicinal herbs Centella asiatica, aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum into a grinder for grinding; S3 Ultrasonic Extraction Method: Mix the powders of Chinese medicinal herbs Centella asiatica, Aloe vera, Scutellaria baicalensis, and Polygonum cuspidatum in a certain proportion, add an ethanol aqueous solution and stir to obtain a Chinese medicinal mixture, then sonicate. S4 filtration extract, centrifuged, supernatant collected, ethanol evaporated, water added, filtered and concentrated to obtain traditional Chinese medicine extract.

6. A compound jasmine hydrosol according to claim 5, characterized in that: The volume fraction of the ethanol-water solution used in step S3 is 50%-70%, the mass-to-volume ratio of the Chinese herbal medicine to the ethanol-water solution is 1:30~50 g / mL, the extraction time is 30-90 min, the extraction temperature is 50-70℃, and the mass-to-volume ratio of the amount of Chinese herbal medicine fed to the concentrated Chinese herbal extract is 0.20~0.24 g / mL.

7. A method for preparing a compound jasmine hydrosol according to any one of claims 1-4, characterized in that, Includes the following steps: R1. Mix the herbal extract with pH adjuster, humectant, and preservative evenly, and then decolorize. R2. Add jasmine hydrosol and mix well; R3. Sterilize to obtain compound jasmine hydrosol.

8. The method for preparing a compound jasmine hydrosol according to claim 7, characterized in that: The pH adjuster is triethanolamine, the humectant is glycerin, the preservative is phenoxyethanol, the ratio of glycerin to traditional Chinese medicine extract is 1:1 to 3:1, the pH is adjusted to 6 to 7, and macroporous resin is used for decolorization.

9. The application of a compound jasmine hydrosol in the preparation of cosmetics, characterized in that: The active ingredient of the cosmetic includes the compound jasmine hydrosol as described in any one of claims 1-4.

10. The application according to claim 9, characterized in that: The cosmetic is formulated as an aqueous solution, emulsion, spray, or mask, and the compound jasmine hydrosol is added at a mass ratio of 50-100%.

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