A plant-enhancing essence and its preparation method

Abstract

This invention discloses a preparation process and quality evaluation method for a plant essence. The essence formula was optimized using single-factor and orthogonal experiments. The dosage of Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract was screened using the hydroxyl radical scavenging rate as an indicator. Furthermore, the dosage of excipients such as carbomer resin 940 and sodium hyaluronate was screened using viscosity as an indicator. This essence has three functions: moisturizing, anti-oxidation, and whitening, and can improve skin condition safely and without irritation.

Description

Technical Field

[0001] This invention relates to the field of natural cosmetics technology, specifically to an essence of a natural plant cosmetic and its preparation process and quality evaluation. Background Technology

[0002] The skincare market has evolved to its current state, offering a wide variety of products. The most important aspect of skincare is the unity of appearance and efficacy. Today, with the rapid advancements in the skincare industry, the pace of product updates and replacements is accelerating, and more and more raw materials are being used in skincare products. However, consumers are increasingly favoring natural products as raw materials. This is mainly because natural products possess the unique properties of being low-irritant, safe, and pollution-free, aligning with the current pursuit of high-quality, healthy, and environmentally friendly skincare. Therefore, natural products are increasingly being incorporated into skincare products. Traditional Chinese medicine has a history of thousands of years in my country, and its application to skincare dates back over 2000 years. According to relevant data, ancient Chinese people utilized natural plant-based herbs for skincare, providing assistance and reference for broader research into plant-based skincare products.

[0003] Bletilla striata is a plant belonging to the genus Bletilla in the Orchidaceae family. It is a traditional Chinese herbal medicine. Bletilla striata gum is a substance that can be used as a suspending agent and emulsifier. Bletilla striata has a good effect on scavenging free radicals and can be used to whiten and lighten spots, delay skin aging, and nourish the skin.

[0004] Dictamni root bark is the dried root bark of a plant in the Rutaceae family, known for its anti-inflammatory, antibacterial, and antioxidant properties. Clinically, it is used to treat various skin diseases. Extracts of Dictamni root bark can combat Candida albicans in vitro. In ancient times, Dictamni root bark was often combined with different traditional Chinese medicines to treat skin conditions.

[0005] Tremella fuciformis, also known as snow fungus or white fungus, has polysaccharides that have antioxidant and skin-improving effects. Tremella polysaccharides are also known as "plant hyaluronic acid".

[0006] Bletilla striata herbal hand cream, Dictamnus dasycarpus antibacterial cream, and Tremella fuciformis pearl nourishing cream—these pure natural herbal extracts are gentle, have few side effects, and are loved by consumers. Because natural plant polysaccharides have a viscous, moisturizing effect, natural plants are often added to skincare products in the form of plant polysaccharides.

[0007] Lin Min et al. (A Traditional Chinese Medicine Composition, Anti-inflammatory Acne-removing Essence and Its Application [P]. Fujian Province: CN115813982A, 2023-03-21) developed an anti-inflammatory acne-removing essence using patchouli, coptis, burdock, and notoginseng as raw materials. The essence contains 0.1-10% of the aforementioned traditional Chinese medicine composition, 0.1-2% sodium hyaluronate, and the remainder is a cosmetic base. The four components of patchouli ferment, coptis extract, burdock extract, and notoginseng extract work synergistically to effectively remove acne, eliminate skin inflammation, repair damaged skin, and make the skin smooth and elastic.

[0008] Luo Jiachao et al. (An oil-controlling and moisturizing essence and its preparation method [P]. Sichuan Province: CN115778842A, 2023-03-14). Using licorice, scutellaria, lavender and peppermint as raw materials, an oil-controlling and moisturizing essence was prepared. The oil-controlling and moisturizing essence includes the following ingredients: sodium hyaluronate, butylene glycol, propylene glycol, glycerin, ethanol, carbomer, tridecyl alcohol polyether, niacinamide, plant extracts, sodium hydroxide, and water as the balance. The essence moisturizes and controls oil in the skin and prevents impurities from entering the skin through the pores and causing damage. It does not affect the skin while controlling oil and moisturizing.

[0009] Ou Chunfeng et al. (Ou Chunfeng, Zou Hengfang, Chen Yurong. A skin whitening and brightening essence and its preparation method [P]. Guangdong Province: CN115671001A, 2023-02-03) disclosed a skin whitening and brightening essence using peony, sophora flavescens, and sage as raw materials. It includes the following components: solubilizer, moisturizer, niacinamide, p-hydroxyacetophenone, skin conditioning agent, 1,2-hexanediol, peony extract, sophora flavescens extract, sage extract, tranexamic acid, pH adjuster, antioxidant, 1,3-butanediol, and water. The peony extract, sophora flavescens extract, and sage extract have a synergistic effect in whitening. The essence of this invention also has excellent antioxidant, anti-aging, nourishing, repairing, and skin elasticity-improving effects.

[0010] "Empowering" cosmetics, as the name suggests, endow cosmetics with functionality. Common skincare products typically offer functions such as moisturizing, anti-aging, and whitening, with moisturizing being a basic requirement. Currently, serums on the market can effectively remove acne, moisturize and control oil, provide antioxidants, and combat aging, but their functions are limited. Summary of the Invention

[0011] The purpose of this invention is to overcome the shortcomings of existing technologies and provide an essence that is easily absorbed by the human skin, while simultaneously providing cosmetics with three functions: moisturizing, anti-oxidation, and whitening. It is also safe and non-irritating, with a promising market prospect. After 30 days of use, skin blemishes, pigmentation, and acne were significantly reduced, and skin moisture increased. There was no significant difference in pore size and acne (P>0.01), but significant differences were observed in blemishes, roughness, wrinkles, and pigmentation (P<0.05). Moisture content showed a highly significant difference (P<0.01). These results indicate that the essence can effectively hydrate and improve skin condition.

[0012] According to a first aspect of the present invention, the present invention provides a plant-enhancing essence comprising carbomer resin 940, sodium hyaluronate, glycerin, 1,2-propanediol, triethanolamine, Bletilla striata extract, Dictamnus dasycarpus root bark extract, Tremella fuciformis extract, and water; wherein the triethanolamine is used to adjust the pH of the system to a range acceptable to the skin.

[0013] Preferably, the preparation of the Bletilla striata extract is as follows: 40g of Bletilla striata is pulverized, soaked in distilled water for 0.5h, and extracted three times at 80℃. The first extraction has a material-to-liquid ratio of 1:10 (g / mL) and an extraction time of 1.5h; the second extraction has a material-to-liquid ratio of 1:6 (g / mL) and an extraction time of 1h; the third extraction has a material-to-liquid ratio of 1:6 (g / mL) and an extraction time of 0.5h. The extract is filtered through three layers of gauze, and the three filtrates are combined. The solution is concentrated to 500mL by heating in a water bath, centrifuged at 4800rpm for 10min, and the uppermost clear liquid is collected and stored in a refrigerator for later use. The polysaccharide content in the Bletilla striata extract is 65.78±3.7%.

[0014] Preferably, the preparation of the Dictamnus dasycarpus extract is as follows: 50g of Dictamnus dasycarpus is crushed, the material-to-liquid ratio is 1:25 (g / mL), and the extract is heated at 100℃ for 2 hours. The extract is filtered through 3 layers of gauze, centrifuged at 4800rpm for 10 minutes, and the uppermost clear liquid is collected and stored in a refrigerator for later use. The polysaccharide content in the Dictamnus dasycarpus extract is 6.47±1.1%.

[0015] Preferably, the preparation of the Tremella fuciformis extract is as follows: 40g of Tremella fuciformis is pulverized and extracted twice at 80℃. The first extraction has a material-to-liquid ratio of 1:25 (g / mL) and an extraction time of 1 hour; the second extraction has a material-to-liquid ratio of 1:10 (g / mL) and an extraction time of 1 hour. The extract is filtered through three layers of gauze, and the two filtrates are combined and concentrated to 400mL by heating in a water bath. The solution is then centrifuged at 4800 rpm for 10 minutes, and the uppermost clear liquid is collected and stored in a refrigerator for later use. The polysaccharide content in the Tremella fuciformis extract is 43.60±2.2%.

[0016] Preferably, the sum of Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract accounts for 30% ± 5% of the total amount of the essence, calculated by weight percentage.

[0017] Preferably, the following components are included by weight percentage: 0.3±0.02% carbomer resin 940, 0.04±0.01% sodium hyaluronate, 7±0.9% glycerin, 3±1.0% 1,2-propanediol, appropriate amount of triethanolamine, 7.47±1.2% Bletilla striata extract, 3.74±1.0% Dictamnus dasycarpus root bark extract, 18.79±2.3% Tremella fuciformis extract, and ultrapure water to bring the total to 100%.

[0018] According to a second aspect of the present invention, the present invention provides a method for preparing a plant-enhancing essence, comprising the following steps:

[0019] Add 1,2-propanediol, carbomer resin 940, sodium hyaluronate, and glycerin to ultrapure water at 80–85°C, stir until completely dissolved, and maintain for 20 minutes. Then cool to 40–45°C, add triethanolamine, and stir until transparent. Finally, add Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract, and stir until homogeneous and free of insoluble matter. Encapsulate to obtain the compound essence.

[0020] The present invention describes the use of the plant-enhancing essence for improving human skin condition and moisturizing the skin.

[0021] Preferably, improving human skin condition refers to improving one or more skin problems such as skin blemishes, pigmentation, acne, pores, roughness, and wrinkles. More preferably, it refers to improving one or more skin problems such as skin blemishes, pigmentation, acne, roughness, and wrinkles.

[0022] The essence described in this article is a pale yellow, slightly viscous liquid with a faint herbal fragrance, which meets the fragrance requirements of the Chinese Cosmetic Standard (GB / T 21171-2018).

[0023] The essence described in this invention has a pH of 6.0±0.02, which is consistent with the pH of human skin. Viscosity: 400±10.8 mPa·s, moderate viscosity; Centrifugation: Centrifugation at different speeds showed no water droplets or layering; Heat resistance: After 24 hours at 40±1℃, there was no significant difference compared to before the experiment; Cold resistance: After 24 hours at -15±1℃, there was no significant difference compared to before the experiment; Alternating hot and cold test: The essence did not become thinner, change color, layer, or precipitate, thus not altering its state; Combustion test: No splashing occurred during combustion, only a small amount of black smoke, indicating a low oil content. A lower lipid-water partition coefficient and weaker hydrophobicity are beneficial for skin absorption. This demonstrates the essence's good stability.

[0024] The total bacterial count of the essence described in this invention showed that Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli were not detected. The total number of molds and yeasts was ≤10, and the total number of colonies was <20. The microbial index test results met the requirements of the cosmetic industry standard (GB / T 7918-1987).

[0025] The essence described in this invention has a moisturizing rate of 99.38±3.7%, indicating the best moisturizing effect. The combination of three plant polysaccharides achieves antioxidant and anti-aging effects by scavenging hydroxyl free radicals at a rate of 65±2.7%. The essence's tyrosinase inhibition rate is 33.07±1.6%, demonstrating that the essence can significantly inhibit tyrosinase activity and has a whitening effect.

[0026] Compared with the prior art, the significant advantages of this invention are:

[0027] (1) In this invention, Bletilla striata, Dictamnus dasycarpus and Tremella fuciformis are added to the essence as three independent units. Through the transdermal absorption of the essence, the effects of moisturizing, anti-oxidation and whitening are achieved.

[0028] (2) By using a low material-liquid ratio and multiple hot water soaking methods, the polysaccharide components can be retained to the maximum extent, and the hot water extraction can largely preserve the polysaccharide structure without damage.

[0029] (3) The serum was compared with commercially available serums in terms of moisturizing, anti-oxidation and whitening. The results showed that the serum was superior to commercially available serums in terms of moisturizing, anti-oxidation and whitening effects. Attached Figure Description

[0030] Figure 1 Comparison of the ability of different amounts of extract to scavenge hydroxyl radicals.

[0031] Figure 2 Effect curve.

[0032] Figure 3 Flowchart of serum preparation process.

[0033] Figure 4 The relationship between the moisture absorption rate of each sample and the change over time.

[0034] Figure 5 The relationship between the moisture retention rate of each sample and the change over time.

[0035] Figure 6 Vitamin C hydroxyl radical scavenging rate curve.

[0036] Figure 7 Vitamin C dose-response curve.

[0037] Figure 8 Curve showing the inhibition of tyrosinase activity by arbutin.

[0038] Figure 9 Arbutin dose-response curve.

[0039] Figure 10 Comparative data analysis before and 30 days after product use.

[0040] Figure 11 Statistical difference graph. Detailed Implementation

[0041] Example 1: Determination of polysaccharide content

[0042] The phenol-sulfuric acid method was used. Using 1 mL of 5% phenol solution and 3.5 mL of 98% concentrated sulfuric acid as blank references, the absorbance was measured at 490 nm. A glucose standard curve was plotted with glucose concentration C on the x-axis and absorbance A on the y-axis.

[0043] The standard curve is y = 28.163x + 0.04, R0. 2 =0.9991.

[0044] Example 2: Preparation of plant extract

[0045] Preparation of Bletilla striata extract: 40g of Bletilla striata was pulverized and soaked in distilled water for 0.5h. Extraction was performed three times at 80℃. The first extraction had a solid-liquid ratio of 1:10 (g / mL) and an extraction time of 1.5h; the second extraction had a solid-liquid ratio of 1:6 (g / mL) and an extraction time of 1h; the third extraction had a solid-liquid ratio of 1:6 (g / mL) and an extraction time of 0.5h. The extract was filtered through three layers of gauze, and the three filtrates were combined and concentrated to 500mL by heating in a water bath. The solution was centrifuged at 4800rpm for 10min, and the supernatant was collected and stored in a refrigerator for later use. The polysaccharide content in the Bletilla striata extract was 65.78±3.7%.

[0046] Preparation of Dictamnus dasycarpus root bark extract: 50g of Dictamnus dasycarpus root bark was crushed, and the material-to-liquid ratio was 1:25 (g / mL). Extraction was carried out at 100℃ for 2 hours. The extract was filtered through three layers of gauze, centrifuged at 4800 rpm for 10 minutes, and the supernatant was collected and stored in a refrigerator for later use. The polysaccharide content in the Dictamnus dasycarpus root bark extract was 6.47±1.1%.

[0047] Preparation of Tremella fuciformis extract: 40g of Tremella fuciformis was pulverized and extracted twice at 80℃. The first extraction had a material-to-liquid ratio of 1:25 (g / mL) and an extraction time of 1 hour; the second extraction had a material-to-liquid ratio of 1:10 (g / mL) and an extraction time of 1 hour. The extract was filtered through three layers of gauze, and the two filtrates were combined and concentrated to 400mL by heating in a water bath. The solution was then centrifuged at 4800 rpm for 10 minutes, and the supernatant was collected and stored in a refrigerator for later use. The polysaccharide content in the Tremella fuciformis extract was 43.60±2.2%.

[0048] The polysaccharide obtained by the hot water extraction method in this invention has a high content and can be directly added to the formulation, saving costs and facilitating industrial production.

[0049] Example 3 Functional Evaluation Method

[0050] (1) Moisturizing experiment

[0051] Moisture absorption rate determination: Weigh 10g of the sample to be tested and place it in a desiccator with a temperature of 20±2℃ and a constant humidity of 81%. Weigh it every 2 hours using a balance and calculate the moisture absorption rate.

[0052] Moisture absorption rate (%) = (m2 - m1) ÷ m1 × 100%

[0053] Moisture retention rate determination: Weigh 10g of the sample to be tested and place it in a weighing bottle that has been dried to constant weight. Place the weighing bottle in a desiccator containing silica gel (humidity 40%), weigh it every 2 hours using a balance, and calculate the moisture retention rate.

[0054] Moisturizing rate (%) = m2 ÷ m1 × 100%

[0055] In the formula: m1 is the mass of the sample before drying; m2 is the mass of the sample after drying.

[0056] (2) Antioxidant experiment

[0057] Salicylic acid-ferrous sulfate method: (1) A1: Mix 0.8 mL of 9 mmol / L FeSO4 solution and 0.60 mL of 9 mmol / L salicylic acid-ethanol solution, and dilute to 10.0 mL with distilled water; (2) A2: Mix 0.80 mL of 9 mmol / L FeSO4 solution, 0.60 mL of 9 mmol / L salicylic acid-ethanol solution, 1.0 mL of 1% H2O2 and plant extracts of different concentrations, and dilute to 10.0 mL with distilled water; (3) A3: Mix 0.80 mL of 9 mmol / L FeSO4 solution, 0.60 mL of 9 mmol / L salicylic acid-ethanol solution and plant extracts of different concentrations, and dilute to 10.0 mL with distilled water.

[0058] Take 1 mL of each of the diluted A1, A2, and A3 solutions and measure their absorbance at 510 nm. Using distilled water as a reference, calculate the clearance rate in the sample using the following formula:

[0059]

[0060] (3) Tyrosinase inhibition rate

[0061] Phosphate buffer solution (PBS): Weigh 7.16g of sodium dihydrogen phosphate and dilute to 100mL with distilled water to obtain solution A; weigh 3.12g of disodium hydrogen phosphate and dilute to 100mL with distilled water to obtain solution B; take 51mL of solution A and 49mL of solution B, and mix them to obtain the phosphate buffer solution.

[0062] L-Tyrosine Solution: Accurately weigh 50.0 mg L-tyrosine into a 50 mL volumetric flask and dissolve in phosphate buffer. Accurately weigh 1.0 mg tyrosinase, dissolve in phosphate buffer and bring the volume to 10 mL. Store at 4°C for later use.

[0063] Table 1 Reaction Solution Preparation Scheme

[0064]

[0065] Using tyrosinase inhibition rate as the indicator: Four solutions were added according to Table 1, with three parallel experiments set up for each group. The mixture was incubated in a 37℃ water bath for 10 min, then the tyrosinase solution was added, stirred thoroughly, and heated in a 37℃ water bath for another 20 min. After removing from the water bath, 200 μL of each reaction solution was added to a 96-well plate, and the absorbance was measured at 475 nm, recorded as A1, A2, A3, and A4. Arbutin was used as a control. A 1 mg / mL arbutin solution was prepared, and then diluted to concentrations of 0.8 mg / mL, 0.6 mg / mL, 0.4 mg / mL, and 0.2 mg / mL, respectively. The absorbance was measured at 475 nm using a microplate reader. The tyrosinase inhibition rate was calculated, a standard curve was plotted, a dose-response curve was plotted, and the IC50 was calculated. 50 .

[0066]

[0067] In the formula: A1: absorbance value containing only the substrate and tyrosinase system;

[0068] A2: Absorbance value containing only the substrate;

[0069] A3: Absorbance value of a system containing only substrate, tyrosinase, and inhibitor;

[0070] A4: Absorbance value of systems containing only substrate and inhibitor.

[0071] Example 4: Single-factor experiment on plant extracts and excipients for essence.

[0072] The example provides a method for single-factor optimization of plant extracts and excipients, the steps of which are as follows:

[0073] (1) Single-factor experiment of plant extract

[0074] The ability of different concentrations of Bletilla striata extract, Dictamnus dasycarpus root bark extract and Tremella fuciformis extract to scavenge hydroxyl radicals was determined by the salicylic acid-ferrous sulfate method in Example 3(2), and the experiment was repeated three times.

[0075] The fixed factors were 1 mL of Dictamnus dasycarpus root bark extract and 1 mL of Tremella fuciformis extract. The variables for Bletilla striata extract were 0.5 mL, 1 mL, 1.5 mL, 2 mL, and 2.5 mL. When the amount of Bletilla striata extract added was 1.5 mL, the plant extract showed the highest scavenging ability of ·OH to reach 94.50%.

[0076] The fixed factors were 1 mL of Bletilla striata extract and 1 mL of Tremella fuciformis extract. The variables for Dictamnus dasycarpus extract were 0.5 mL, 1 mL, 1.5 mL, 2 mL, and 2.5 mL. When the amount of Dictamnus dasycarpus extract added was 1.0 mL, the plant extract showed the highest scavenging ability of ·OH to reach 84.20%.

[0077] The fixed factors were 1 mL of Bletilla striata extract and 1 mL of Dictamnus dasycarpus extract. The variables for the Tremella fuciformis extract were 0.5 mL, 1 mL, 1.5 mL, 2 mL, and 2.5 mL. When the amount of Tremella fuciformis extract added was 3.0 mL, the plant extract showed the highest scavenging ability of ·OH to reach 95.59%.

[0078] according to Figure 1 In the screening of antioxidant results, the amount of Bletilla striata extract used was 1.5 mL, the amount of Dictamnus dasycarpus extract used was 1.0 mL, and the amount of Tremella fuciformis extract used was 3.0 mL.

[0079] In summary, the amount of Dictamnus dasycarpus extract should be 1.0 mL, the amount of Bletilla striata extract should be 1.5 mL, and the amount of Tremella fuciformis extract should be 3.0 mL, accounting for 30% of the total extract, with a ratio of 2:3:6. The extracts should be added according to the actual required amount.

[0080] (2) Single-factor experiment of excipients

[0081] Viscosity was determined according to the national standard "Method for Determining the Viscosity of Liquid Products by Rotation Viscometer for Surfactants and Detergents" (GB / T15357). Viscosity was used as an indicator (the viscosity of the essence was 400±10.8 Pa·s) to screen the dosage of carbomer resin 940 and sodium hyaluronate. Fixed factors included 1.5 mL of Bletilla striata extract, 1.0 mL of Dictamnus dasycarpus root bark extract, 3.0 mL of Tremella fuciformis extract, 10 g of glycerin, and 3 g of 1,2-propanediol. The experimental variables were: carbomer resin 940 dosage of 0.1 g, 0.2 g, 0.3 g, 0.4 g, and 0.5 g (sodium hyaluronate dosage of 0.04 g); and sodium hyaluronate dosage of 0.02 g, 0.03 g, 0.04 g, 0.05 g, and 0.06 g (carbomer resin 940 dosage of 0.3 g). Water was used to bring the total dosage to 100 g in both experiments.

[0082] Using the moisturizing rate in Example 3-(1) as an indicator, the amounts of glycerin and 1,2-propanediol were screened.

[0083] Fixed factors included 1.5 mL of Bletilla striata extract, 1.0 mL of Dictamnus dasycarpus root bark extract, 3.0 mL of Tremella fuciformis extract, 0.3 g of Carbomer resin 940, and 0.04 g of sodium hyaluronate. The research variables were the amount of glycerol added (5 g, 7 g, 9 g, 11 g, 13 g, with 3 g of 1,2-propanediol added) and the amount of 1,2-propanediol added (1 g, 2 g, 3 g, 4 g, 5 g, with 10 g of glycerol added). Water was used to bring the total amount to 100 g in both experiments.

[0084] Results on excipient dosage: Viscosity affects the sensory evaluation of a serum and reduces the user experience. Considering the viscosity of serums in daily life, the preferred dosage is 0.3 ± 0.02 g of carbomer resin 940 and 0.04 ± 0.01 g of sodium hyaluronate. When the amount of glycerin added is 7 ± 0.9 g, the water loss is lowest and the moisturizing rate is highest; however, a higher amount of glycerin increases the serum's viscosity, lowering the sensory score. Therefore, the preferred amount of glycerin added is 7 g. When the amount of 1,2-propanediol added is 3 ± 1.0 g, the water loss is lowest; when the amount of 1,2-propanediol added is 3 ± 1.0 g, the moisturizing rate is highest.

[0085] (3) Orthogonal experiment

[0086] Seven factors were selected to further optimize the dosage: sodium hyaluronate, carbomer resin 940, glycerin, 1,2-propanediol, Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract. Three levels were selected for each factor (see Table 2) according to L18(3) 7An orthogonal array (see Table 3) was used for the experiment. Viscosity, pH, moisture retention rate, tyrosinase inhibition rate, and hydroxyl radical scavenging rate were used as indicators. A comprehensive scoring method was used for evaluation. The ranking scoring method described in the literature was adopted to rank and give the corresponding score values, which were used as unit indicators for data analysis and processing to select the optimal combination. (The orthogonal experiment was scaled down by a factor of 10, and water was added to make up to 10g at the end.)

[0087] Table 2. Orthogonal Factors and Levels

[0088]

[0089] Note: 1.5 mL of Bletilla striata extract = 1.02 g, 1.0 mL of Dictamnus dasycarpus root bark = 0.68 g, and 3.0 mL of Tremella fuciformis extract = 2.05 g.

[0090] Table 3 Orthogonal Experiment Plan

[0091]

[0092]

[0093] The methods for determining viscosity and moisture retention are as described above; pH is determined in accordance with GB / T13531.1-2008.

[0094] This invention first investigated the addition amounts of Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract through single-factor experiments, and then further optimized the addition amounts of plant extracts and excipients using orthogonal experiments.

[0095] Viscosity, moisturizing rate, hydroxyl radical scavenging rate, and tyrosinase inhibition rate were used as indicators, and a ranking scoring method was employed for comprehensive evaluation. Viscosity scores are shown in Table 4, moisturizing rate scores in Table 5, hydroxyl radical scavenging rate scores in Table 6, and tyrosinase inhibition rate scores in Table 7. The intuitive analysis and variance analysis results of the experimental data are shown in Table 8.

[0096] Table 4. Serum Viscosity Score Data Table

[0097]

[0098]

[0099] Table 5. Serum Moisturizing Rate Score Data Table

[0100]

[0101] Table 6. Data on Hydroxyl Free Radical Scavenging Rate of Serum

[0102]

[0103]

[0104] Table 7. Data on Tyrosinase Inhibition Rate of Serum

[0105]

[0106]

[0107] Table 8. Visual Analysis of Orthogonal Experiment Results

[0108]

[0109]

[0110] Based on the intuitive analysis of Table 8, the degree of influence of each factor on the experiment was determined. If the seven factors—Bletilla striata extract, Dictamnus dasycarpus root bark extract, Tremella fuciformis extract, Carbomer resin 940, sodium hyaluronate, glycerin, and 1,2-propanediol—are labeled A, B, C, D, E, F, and G respectively, then the order of primary and secondary factors is: B > C > F > A > D > E > G. In this experiment, the factor with the highest comprehensive score needs to be selected. The optimal combination of levels for each factor is determined based on the experimental index values ​​to select the optimal experimental scheme. Simultaneously, the comprehensive effect curve (…) Figure 2 The correct comprehensive analysis results were obtained. Based on the comprehensive and intuitive analysis table and effect curve, the optimal solution B1C1F2A1D2E2G1 was determined, which is: 0.68g of Bletilla striata extract, 0.34g of Dictamnus dasycarpus root bark extract, 1.71g of Tremella fuciformis extract, 0.03g of Carbomer resin 940, 0.004g of sodium hyaluronate, 0.7g of glycerin, and 0.03g of 1,2-propanediol. The percentages are: 6.8g of Bletilla striata extract, 3.4g of Dictamnus dasycarpus root bark extract, 17.1g of Tremella fuciformis extract, 0.3g of Carbomer resin 940, 0.04g of sodium hyaluronate, 7g of glycerin, and 0.3g of 1,2-propanediol.

[0111] Based on a 100% formula ratio, the sum of Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract accounts for 30% of the total essence, with the ratio of the three in the order 2:3:6. According to the orthogonal experimental results, the total added amount of the three is calculated to be 27.3%, which, when converted to 30%, yields Bletilla striata extract at 7.47%, Dictamnus dasycarpus root bark extract at 3.74%, and Tremella fuciformis extract at 18.79%. The final specific formula is shown in Table 9.

[0112] Table 9 Final Formula of Compound Plant Extract Liquid

[0113]

[0114] Note: The dosage of triethanolamine should be based on the final compound plant extract system pH = 6.0 ± 0.02, which meets the requirements of the human body.

[0115] Example 5: Preparation method of serum

[0116] Carbomer resin 940 0.3±0.02%, sodium hyaluronate 0.04±0.01%, glycerin 7±0.9%, 1,2-propanediol 3±1.0%, Bletilla striata extract 7.47±1.2%, Dictamnus dasycarpus root bark extract 3.74±1.0%, Tremella fuciformis extract 18.79±2.3%, ultrapure water to 100%.

[0117] 1,2-Propanediol, carbomer resin 940, sodium hyaluronate, and glycerin were added to ultrapure water at 80–85°C and stirred until completely dissolved, maintaining this temperature for 20 minutes. The solution was then cooled to 40–45°C, and triethanolamine was added and stirred until transparent. Finally, extracts of Bletilla striata, Dictamnus dasycarpus, and Tremella fuciformis were added and stirred until homogeneous and free of insoluble matter. The solution was then packaged to obtain the compound essence. Figure 3 ); and used in the tests of Examples 6-10.

[0118] Example 6: Functional Evaluation Results

[0119] (1) Moisturizing experiment

[0120] The hygroscopic and moisturizing effects of glycerin, 1,2-propanediol, tremella extract and bletilla extract, essence sample (prepared in Example 5) and control group (Meiji Hydrating Essence - SuG Makeup Network Preparation No. 2019003654) were determined according to the method in Example 3 (1).

[0121] The results showed that at 6 hours, the moisturizing ability ranked as follows: serum > control group > glycerin > tremella extract > Bletilla striata extract > 1,2-propanediol. The moisturizing rate of the experimental group (serum sample) was 99.38±3.7%, while that of the control group was 89.37±2.9%. The experimental group showed the best moisturizing effect. Figure 4 and 5 As shown in the figure.

[0122] (2) Antioxidant experiment

[0123] The ability of the essence sample to scavenge hydroxyl radicals was determined using the salicylic acid-ferrous sulfate method in Example 3 (2).

[0124] The results showed that the serum sample prepared in Example 5 had a hydroxyl radical scavenging rate of 65±2.7%, achieving antioxidant and anti-aging effects, while the hydroxyl radical scavenging rate of the control group serum was 30.14±2.6%. This indicates that the serum sample had a better antioxidant effect.

[0125] Figure 6 This is the hydroxyl radical scavenging rate curve of vitamin C. The regression equation for vitamin C is y = 95.765x + 3.919, and its linear coefficient is R0.2 =0.9975. (Based on the vitamin C dose-response curve) Figure 7 ), to obtain the IC of vitamin C hydroxyl free radical scavenging 50 It is 0.5744 mg / mL.

[0126] (3) Whitening experiment

[0127] The whitening effect of the serum sample and the control group was determined according to the method in Example 3 (3). The results showed that the tyrosinase inhibition rate of the serum sample prepared in Example 5 was 33.07±1.6%, and the tyrosinase inhibition rate of the control group was 30.72±1.3%.

[0128] Figure 8 This is a graph showing the inhibitory effect of arbutin on tyrosinase activity. The regression equation for arbutin is y = 53.806x + 22.35, with a linear coefficient R0. 2 =0.9951. (Based on the dose-response curve of arbutin) Figure 9 The IC50 of arbutin inhibiting tyrosinase activity was obtained. 50 It is 0.5837 mg / mL.

[0129] Tyrosinase inhibition rate is an important indicator for evaluating the activity of whitening agents. The higher the inhibition rate, the stronger the ability to inhibit tyrosinase activity, and the better the whitening effect. Therefore, this serum can significantly inhibit tyrosinase activity and has a whitening effect.

[0130] Example 7 Sensory Evaluation

[0131] Fifty healthy individuals aged 20-50 with the ability to correctly identify the cause were selected as volunteers. A 4cm sample was taken from the volunteers' arms. 2 2g of sample was evenly applied to the arm of the volunteer, who then used their other hand to massage in circular motions until the sample was completely absorbed. Sensory evaluation was conducted during the application process. Scores were categorized as Excellent (90 points), Good (80 points), and Pass (60 points), and a questionnaire was used to collect and analyze the volunteers' scores.

[0132] A questionnaire survey was conducted to collect data on the use of woody products by people with different skin types during and after use. The results were summarized and are shown in Table 10 below.

[0133] Table 10 Sensory Evaluation Results

[0134]

[0135] Example 8: Evaluation of Physicochemical Properties

[0136] Appearance: The color and luster of the essence are observed to determine if the product's color and luster are comfortable and aesthetically pleasing, and whether it is uniform without layering. The product is also tested for any irritating odor using the sense of smell. Results: The product is a pale yellow, slightly viscous liquid with a faint herbal fragrance, meeting the specified aroma requirements.

[0137] pH Measurement: The pH value of the serum sample was measured at room temperature (25℃) according to the national standard General Test Method for Cosmetics (GB / T13531.1-2008). The pH meter was calibrated according to the instrument's instructions. The electrode was inserted into the serum, and the reading was recorded. The experiment was performed in triplicate. The result is expressed as 6.0 ± 0.02, which is consistent with the pH value of human skin.

[0138] Viscosity: The viscosity was determined using a rotational viscometer according to the national standard "Method for Determining the Viscosity of Liquid Products by Rotational Viscometer for Surfactants and Detergents" (GB / T15357). The serum was allowed to stand at room temperature for 24 hours to remove air bubbles. The serum was then poured into a beaker, and the temperature was adjusted to 23°C in a constant-temperature water bath. A suitable rotor was immersed in the solution to prevent air bubble formation. Once the rotational viscometer reading stabilized, the data was recorded in triplicate. The result is expressed as 400 ± 10.8 mPa·s, indicating a moderate viscosity that is beneficial for skin absorption.

[0139] Centrifugation test: Following the experimental method described in the People's Republic of China National Standard for Skin Care Lotion (GBT29665-2013), 5g of essence was taken and centrifuged at 2000r / min, 3000r / min, 4000r / min, and 5000r / min for 30min respectively. The results showed that no water droplets or layering occurred in the essence after centrifugation at different speeds.

[0140] Heat resistance test: Following the experimental method described in the National Standard of the People's Republic of China for Skin Care Lotion (GBT29665-2013), two 2g portions of the essence were taken. One portion of the essence was placed in a constant temperature incubator at 40±1℃ for 24 hours, then removed and cooled to room temperature before being compared with the other portion. The presence of any thinning, discoloration, layering, or sedimentation was observed to determine the heat resistance of the essence. Results showed that after 24 hours at 40±1℃, there was no significant difference compared to before the experiment.

[0141] Cold resistance test: Following the experimental method described in the National Standard of the People's Republic of China for Skin Care Lotion (GBT29665-2013), two 2g samples of the serum were taken. One sample was placed in a refrigerator at -15±1℃ for 24 hours, then removed and cooled to room temperature before being compared with the other sample. The presence of any thinning, discoloration, layering, or sedimentation was observed to determine the cold resistance of the serum. Results showed that after 24 hours at -15±1℃, there was no significant difference compared to before the experiment.

[0142] Alternating heating and cooling test: Following the experimental method described in the People's Republic of China National Standard for Determination of Storage Stability of Water-in-Oil Emulsions of Surfactants (GB / T 16497—2007), 2g of the essence was placed in a constant temperature incubator at 40±1℃ for 24 hours, then at room temperature for 24 hours, and finally in a refrigerator at -15±1℃ for 24 hours. The presence of stratification, precipitation, and color changes was observed. Results indicated that the essence maintained its original state.

[0143] Combustion Test: 2g of serum was placed in a crucible, an asbestos mesh was placed on an induction cooker, and the mixture was heated. The state of the serum was observed, and any changes in odor were noted. Results showed that no splattering occurred during combustion, and only a small amount of black smoke was produced. This indicates a low oil content, a lower lipid-water partition coefficient, and weaker hydrophobicity, which can negatively impact skin absorption.

[0144] Example 9: Evaluation of Health Indicators

[0145] 1g of essence was added to 99mL of sterile physiological saline using a sterile pipette. After mixing, a 1:100 test solution was prepared. After incubation under specific conditions for 24 hours, the total bacterial count of 0.5mL sample was determined. According to the standard test method for microbial contamination in cosmetics in the 2015 edition of the "Cosmetic Safety Technical Specifications", the total bacterial count, total mold and yeast count, thermotolerant coliform bacteria, Staphylococcus aureus, and Pseudomonas aeruginosa in the essence were determined. The results are shown in Table 11.

[0146] Table 11 Evaluation Results of Microbiological Indicators

[0147]

[0148] Example 10: Human Experiment Determination

[0149] Sixty volunteers with healthy skin were selected and divided into six groups. A 4cm section was selected on the inner side of the arm. 2 Weigh 2g of serum and apply it evenly to the inner arm. Use a skin moisture and oil tester to measure the skin's moisture and oil content before application. Then measure the skin's moisture and oil content 5, 10, 15, 30, and 60 minutes after applying the serum and compare the results to determine the serum's moisturizing effect. Figure 4and 5 It can be concluded that the moisture absorption rate of glycerin, 1,2-propanediol, tremella and Bletilla striata extracts gradually increased and the moisturizing rate gradually decreased. The moisturizing rate of a single component decreased with time, while the moisturizing effect of the essence in the experimental group and the control group increased significantly with time.

[0150] However, the moisture absorption and moisturizing rates of both the serum and the control group gradually increased, and the rate of decrease in moisturizing rate reflected the quality of the substances' moisturizing performance. At 6 hours, the moisturizing ability ranked as follows: serum > control group > glycerin > tremella extract > Bletilla striata extract > 1,2-propanediol. The moisturizing rate of the serum was 99.38±3.7%, while that of the control group was 89.37±2.9%, indicating that the serum had the best moisturizing effect.

[0151] Combining the methods described above, the skin's oil and water levels were measured using a skin oil and water analyzer, as shown in Table 12. The results indicate that the serum can reduce skin oil and increase skin moisture, thus moisturizing and regulating the skin's oil-water balance.

[0152] Table 12 Results of Skin Water-Oil Test

[0153]

[0154] Example 11 Instrument Evaluation

[0155] Thirty participants with no history of allergies were selected. 2g of the serum was applied to a fixed area of ​​the skin. The skin condition of the participants' faces before using the product was detected using a magic mirror device and recorded. 2g of the serum was then applied evenly to the volunteers' faces until absorbed. The facial condition of the volunteers was detected again one month after using the product. By comparing the skin condition before and after use, the effect of the serum on human skin can be determined.

[0156] Based on the above experimental methods, before using this product, the Magic Mirror device was used to measure and analyze data such as age spots, pigmentation, wrinkles, acne, and moisture on the subject's face. Then, based on the data measured by the Magic Mirror device after 30 days of product use, the results were analyzed as follows: Figure 10 As shown.

[0157] Combine the data obtained by the Magic Mirror device before and after the subjects used the product with, for example Figure 10As shown, after using the essence for 30 days, skin spots, pigments, acne, etc. were significantly reduced, and skin moisture increased. There was no significant difference in pores and acne with P>0.01, and significant differences in spots, roughness, wrinkles and pigments with P<0.05, and a highly significant difference in moisture with P<0.01. The results showed that the essence could be used to moisturize and improve the skin condition. This result was because polysaccharide from Bletilla striata was a sticky polysaccharide that could form a flexible film on the skin surface to prevent the skin from losing moisture; tremella polysaccharide could improve the skin's conductivity and prevent toxic and harmful substances from entering the skin and causing damage, prevent the evaporation of skin moisture, increase the water content of the skin epidermis, improve skin roughness, and remove facial freckles. Tremella polysaccharide had the effect of promoting cell proliferation, division and reducing the number of senescent cells. Sodium hyaluronate ingested from the outside would be decomposed into oligosaccharides by bacteria in the cecum, absorbed through the large intestine, and enter the whole body through the blood or lymph, promoting the secretion of hyaluronic acid by dermal cells and maintaining the relative stability of skin water content. Sodium hyaluronate could improve the survival environment of skin cells, induce cell proliferation and differentiation, penetrate into the skin, help metabolism, accelerate cell renewal, improve skin immunity, and achieve the effect of beauty. Since the structure of tremella polysaccharide was extremely similar to that of hyaluronic acid and both could improve the skin condition, it was speculated that tremella polysaccharide might have the same transdermal absorption effect as hyaluronic acid. The experimental results showed that by compounding different kinds of plant extracts in a certain proportion, it could play a good role in moisturizing and hydrating, and had the effect of improving skin quality.

[0158] The statistical differences between the essence and the control essence in terms of moisturizing, antioxidant and whitening are as Figure 11 shown. The moisturizing rate was 0.01<P<0.05, with a significant difference; the P value of the hydroxyl radical scavenging rate was <0.01, with a highly significant difference, and there was no significant difference in the tyrosinase inhibition rate. <​​​​​​​​Clean the affected area with an alcohol-soaked cotton swab, keeping the area dry and clean at all times, and avoiding contact with other skin preparations (such as other skincare cosmetics or pharmaceutical preparations). Apply the serum evenly to the test area twice daily, to a thickness of approximately 1 mm. During the experiment, the test area must be kept dry and clean. Observe the skin at 2 hours, 4 hours, 8 hours, 16 hours, 32 hours, and 48 hours for any abnormal phenomena such as erythema, edema, or allergic reactions.

[0162] The natural plant itself is safe, and the extraction method is hot water extraction without the addition of organic solvents. The excipients used to prepare the essence are glycerin, 1,2-propanediol, sodium hyaluronate and other ingredients commonly added to skin care products. No chemical preservatives are added. The irritation evaluation results showed that no allergic reactions such as erythema or edema occurred after volunteers applied the essence, and the rating was 0.

[0163] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A plant-enhancing essence, comprising carbomer resin 940, sodium hyaluronate, glycerin, 1,2-propanediol, triethanolamine, aqueous extracts of Bletilla striata, Dictamnus dasycarpus, and Tremella fuciformis, and water; wherein the triethanolamine is used to adjust the pH of the system to a range acceptable to the skin; The polysaccharide content in the aqueous extract of Bletilla striata was 65.78±3.7%; the polysaccharide content in the aqueous extract of Dictamnus dasycarpus was 6.47±1.1%; and the polysaccharide content in the aqueous extract of Tremella fuciformis was 43.60±2.2%. The Bletilla striata aqueous extract was prepared as follows: 40g of Bletilla striata was pulverized, soaked in distilled water for 0.5h, and extracted three times at 80℃. The first extraction had a material-to-liquid ratio of 1:10 (g / mL) and an extraction time of 1.5h; the second extraction had a material-to-liquid ratio of 1:6 (g / mL) and an extraction time of 1h; and the third extraction had a material-to-liquid ratio of 1:6 (g / mL) and an extraction time of 0.5h. The extract was filtered through three layers of gauze, and the three filtrates were combined. The solution was concentrated to 500mL by heating in a water bath, centrifuged at 4800rpm for 10min, and the supernatant was collected and stored in a refrigerator for later use. The polysaccharide content in the Bletilla striata extract was 65.78±3.7%. Preparation of the aqueous extract of Dictamnus dasycarpus: 50g of Dictamnus dasycarpus was crushed, the material-to-liquid ratio was 1:25 (g / mL), and the extract was heated at 100℃ for 2 hours. The extract was filtered through 3 layers of gauze, centrifuged at 4800rpm for 10 minutes, and the uppermost clear liquid was collected and stored in a refrigerator for later use. The polysaccharide content in the extract of Dictamnus dasycarpus was 6.47±1.1%. Preparation of the Tremella fuciformis aqueous extract: 40g of Tremella fuciformis was pulverized and extracted twice at 80℃. The first extraction had a material-to-liquid ratio of 1:25 (g / mL) and an extraction time of 1h; the second extraction had a material-to-liquid ratio of 1:10 (g / mL) and an extraction time of 1h. The extract was filtered through three layers of gauze, and the two filtrates were combined and concentrated to 400mL by heating in a water bath. The extract was then centrifuged at 4800r / min for 10min. The supernatant was collected and stored in a refrigerator for later use. The polysaccharide content in the Tremella fuciformis aqueous extract was 43.60±2.2%. Calculated by weight percentage: 0.3±0.02% Carbomer resin 940, 0.04±0.01% Sodium hyaluronate, 7±0.9% Glycerin, 3±1.0% 1,2-Propanediol, appropriate amount of triethanolamine, 7.47±1.2% Bletilla striata extract, 3.74±1.0% Dictamnus dasycarpus root bark extract, 18.79±2.3% Tremella fuciformis extract, and ultrapure water to make up to 100%.

2. According to claim 1, the total weight percentage of the plant-enhancing essence is 30% ± 5% of the total essence, calculated as follows: Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract.

3. A method for preparing the plant-enhancing essence of claim 1, comprising the following steps: Add 1,2-propanediol, carbomer resin 940, sodium hyaluronate, and glycerin to hot ultrapure water and stir until completely dissolved; then cool to a boil, add triethanolamine and stir until transparent, finally add Bletilla striata extract, Dictamnus dasycarpus root bark extract, and Tremella fuciformis extract, stir until homogeneous and free of insoluble matter, and encapsulate to obtain the compound essence.

4. The use of the plant-enhancing essence as described in claim 1, in the preparation of a plant-enhancing essence that improves human skin condition and moisturizes the skin.

5. According to the use described in claim 4, the improvement of human skin condition refers to improving any one or more skin problems such as skin spots, pigmentation, acne, pores, roughness, and wrinkles.

6. The use according to claim 4, wherein improving human skin condition refers to improving any one or more skin problems such as blemishes, pigmentation, acne, roughness, and wrinkles.

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