Essential oil composition having whitening and spot-fading effects and method for preparing the same

Abstract

The present application relates to a kind of essential oil composition with whitening and freckle lightening effect and its preparation method, belong to the technical field of cosmetics.The essential oil composition with whitening and freckle lightening effect provided by the present application includes Italian helichrysum oil, rice germ oil, sandalwood oil, white spring chamomile flower oil, rust red rose seed oil and pink passion fruit seed oil treated by specific fermentation process, all ingredients are derived from natural plants, no chemical synthetic substance is added, irritability is low, safety is high, has good whitening, freckle lightening effect, also has anti-inflammatory, antioxidant and barrier repair effect, suitable for various skin.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology and relates to an essential oil composition with whitening and spot-fading effects and its preparation method. Background Technology

[0002] From a pathological perspective, melanin production and deposition is a complex process involving multiple steps. Melanin is synthesized in melanosomes within melanocytes and then transported to adjacent keratinocytes via melanosomes. As cells differentiate and migrate upwards, melanosomes gradually move towards the outer layer of the epidermis, leading to pigment deposition. Studies have shown that pigmentary disorders can significantly reduce patients' quality of life and cause varying degrees of psychological burden. Currently, widely used chemical whitening agents, such as hydroquinone and arbutin, while having some whitening effects, are often accompanied by safety issues. For example, hydroquinone is cytotoxic, and long-term use may lead to exogenous okra; kojic acid has poor stability and poses a potential carcinogenic risk. Meanwhile, with the popularization of skincare knowledge and the improvement of consumers' safety awareness, the market demand for naturally derived, low-irritant whitening and spot-fading products is also continuously growing. Summary of the Invention

[0003] The purpose of this invention is to provide an essential oil composition with whitening and spot-fading effects and its preparation method. This invention scientifically combines Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rosehip seed oil, and pink passionflower seed oil obtained through a specific fermentation process. All ingredients are derived from natural plants, and it does not contain traditional chemical whitening agents such as hydroquinone and arbutin. It has no added chemical synthetics, low irritation, and high safety. It has good whitening and spot-fading effects, and also has anti-inflammatory, antioxidant, and barrier repair effects, making it suitable for all skin types.

[0004] The objective of this invention can be achieved through the following technical solutions: In a first aspect, the present invention provides an essential oil composition with whitening and spot-fading effects, the essential oil composition comprising the following raw materials: Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil, wherein the weight ratio of Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil is (1-3):(4-8):(2-4):(1-3):(0.6-2):(2-6); the pink passionflower seed oil is obtained by supercritical CO2 extraction after fermentation of pink passionflower seeds by Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae.

[0005] Italian immortelle oil is rich in terpenoids and polyphenols, which can scavenge free radicals, inhibit the release of inflammatory factors, reduce oxidative stress damage to the skin, and alleviate skin inflammation, thereby delaying pigmentation and skin aging and evening out skin tone. In addition, Italian immortelle oil can also accelerate epidermal cell regeneration and help melanin-containing keratinocytes shed faster by promoting basal cell proliferation and epidermal renewal.

[0006] Rice germ oil is rich in vitamin E and phytosterols, which have a strong antioxidant effect. It can protect the skin from sun damage during the day and reduce the activation of melanin induced by ultraviolet rays. The unsaturated fatty acids and squalane contained in rice germ oil also help maintain the skin barrier function, promote epidermal cell regeneration, and effectively lock in moisture to improve dry skin, thereby improving pigmentation and making the skin tone more even and radiant.

[0007] Sandalwood oil contains components such as α-santalol, β-santalol, and santalone. These components have antioxidant and anti-inflammatory effects that can effectively relieve skin aging, reduce skin inflammation, and prevent the abnormal inflammation from increasing the activity of melanocytes, thus preventing pigmentation and age spots. At the same time, it can also promote blood circulation and accelerate metabolism, which helps to lighten age spots.

[0008] White chrysanthemum oil contains various anti-inflammatory components (such as triterpenoids), which can effectively reduce skin inflammation and pigmentation caused by inflammation, thereby fading dark spots. Its flavonoids and phenols can scavenge free radicals, reduce oxidative damage, delay skin aging, and repair damaged skin barriers. By inhibiting inflammatory factors, resisting oxidative stress, and repairing damaged barriers, it effectively exerts whitening and spot-fading effects.

[0009] Rosehip seed oil is rich in unsaturated fatty acids, vitamins, minerals and other active ingredients. It has anti-inflammatory and antioxidant properties, which can effectively reduce the damage of oxidative stress to the skin, alleviate skin inflammation, nourish the skin, promote the repair of the skin barrier, enhance skin vitality, effectively improve skin condition, make the skin tone more even and radiant, and thus exert whitening and spot-fading effects.

[0010] Preferably, the weight ratio of the Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil is (1.5-2.5):(5.5-6.5):(2.5-3.5):(1.5-2.5):(1-1.4):(3.5-4.5).

[0011] Specifically, the preparation of the pink passionflower seed oil includes the following steps: S1. Crush the pink passion fruit seeds, pass them through a 40-60 mesh sieve to obtain pink passion fruit seed powder, add deionized water, mix well, sterilize, and obtain the basic liquid. S2. Add compound bacterial solution to the base liquid and ferment aerobicly at 29-37℃ for 24-36 hours to obtain fermentation product; S3. Dry the fermentation product at 60-70℃ until the moisture content is 10-20wt%, pulverize it and pass it through a 40-60 mesh sieve. Place it in a supercritical CO2 extraction vessel, pump in an entrainer for extraction, collect the extract, and obtain pink passion fruit seed oil.

[0012] Preferably, in step S1, the ratio of pink passion fruit seed powder to deionized water is 1g:5-15mL; the sterilization method is radiation sterilization, specifically using... 60 Sterilization is performed using Co-γ rays with a radiation dose of 25 kGy.

[0013] Preferably, the inoculation amount of the compound bacterial solution in step S2 is 5.5% v / v-6.5% v / v of the base solution, and the effective viable count of the compound bacterial solution is 4 × 10⁻⁶. 9 CFU / mL - 5 × 10 9 The CFU / mL ratio of viable Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae in the compound bacterial solution is 0.05-0.15:1:0.3-0.4.

[0014] Preferably, in step S3, the supercritical CO2 extraction is performed under the following conditions: extraction pressure of 30-40 MPa, extraction temperature of 45-55℃, and CO2 flow rate of 20-30 L / h for 60-80 min. The entrainer is an ethanol solution with a mass concentration of 75%-85%, and the flow rate ratio of the entrainer to the supercritical CO2 is 1:(25-35).

[0015] Pink passion fruit seed oil contains unsaturated fatty acids (such as linoleic acid), vitamins, and minerals. These components can effectively moisturize the skin, enhance the skin barrier function, and have certain soothing and antioxidant effects. They also promote metabolism, fade dark spots, and brighten skin tone.

[0016] This invention extracts pink passion fruit seed oil through multi-strain synergistic fermentation followed by supercritical CO2 extraction. The main advantage is that fermentation pretreatment enhances the dissolution rate of the target components, and supercritical extraction is then performed on this basis. This not only increases the contact area between the target components and the supercritical fluid, improving the extraction efficiency of supercritical extraction, but also effectively avoids the damage of heat-sensitive fat-soluble components (such as unsaturated fatty acids) caused by high temperatures, thus maximizing the preservation of their biological activity.

[0017] In a second aspect, the present invention provides a method for preparing the essential oil composition with whitening and spot-fading effects described in the first aspect, comprising the following steps: Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil were mixed in a weight ratio and homogenized at 10,000 rpm to 20,000 rpm to obtain an essential oil composition with whitening and spot-fading effects.

[0018] Preferably, the essential oil composition can be used to prepare skin care products, including toners, lotions, creams, masks, serums, and sprays, and the amount of the essential oil composition added is 0.5%-3% of the total weight of the skin care products.

[0019] Thirdly, the present invention provides an essence comprising the following ingredients by weight percentage: 0.5%-3% of the essential oil composition with whitening and spot-fading effects described in the first aspect, 0.05%-0.3% of thickener, 3%-10% of moisturizer, 0.5%-5% of preservative, 0.01%-0.3% of pH adjuster, and the balance being deionized water.

[0020] Preferably, the thickener comprises at least one of xanthan gum, carbomer, ammonium acryloyl dimethyl taurate / VP copolymer, hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, sodium polyacrylate, hydroxyethyl cellulose, and sclerotium gum; Preferably, the moisturizer includes at least one of allantoin, panthenol, dipropylene glycol, low molecular weight sodium hyaluronate, glycerin, 1,3-butanediol, and 1,2-propanediol. Preferably, the preservative includes at least one of p-hydroxyacetophenone, ethylhexylglycerin, and phenoxyethanol; Preferably, the pH adjuster includes at least one of triethanolamine, citric acid, and arginine.

[0021] The beneficial effects of this invention are: (1) This invention scientifically combines Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rosehip seed oil, and pink passionflower seed oil processed with a specific fermentation process to construct a four-fold synergistic mechanism of "triple tyrosinase inhibition, blocking melanin transport, accelerating keratin metabolism, and anti-inflammatory repair". Among them, Italian immortelle oil and white chrysanthemum oil play an anti-inflammatory and antioxidant role, rosehip seed oil promotes the repair of the skin barrier, rice germ oil and sandalwood oil strengthen barrier repair and promote circulation, and pink passionflower seed oil has significantly increased active ingredients after fermentation, which has a good promoting effect on metabolism, lightening dark spots, and brightening skin tone. The six ingredients work synergistically from multiple aspects such as melanin production, transfer, metabolism and inflammation regulation, and the effect is significantly better than single ingredients or traditional simple compound systems, with good whitening and spot-lightening effects.

[0022] (2) This invention uses a combination of multi-strain (Lactobacillus rhamnosus, Saccharomyces cerevisiae, Aspergillus oryzae) synergistic fermentation and supercritical CO2 extraction technology to extract pink passion fruit seed oil. This process destroys the plant cell wall structure through fermentation pretreatment, and then performs supercritical extraction on this basis. This not only increases the contact area between the target components and the supercritical fluid and improves the extraction efficiency of supercritical extraction, but also effectively avoids the damage of heat-sensitive fat-soluble components (such as unsaturated fatty acids) to high temperatures, thus maximizing the preservation of their biological activity.

[0023] (3) The essential oil composition provided by the present invention is entirely derived from natural plants, does not contain traditional chemical whitening agents such as hydroquinone and arbutin, has no added chemical synthetics, is low in irritation and high in safety, has good whitening and spot-fading effects, and also has anti-inflammatory, antioxidant and barrier repair effects, and is suitable for all skin types. Detailed Implementation

[0024] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with embodiments, is provided below.

[0025] Italian immortelle oil and sandalwood oil were purchased from Guangzhou Keran Trading Co., Ltd. Rice germ oil was purchased from Hengyang Zimin Agricultural Technology Development Co., Ltd. White chrysanthemum oil was purchased from Wuhan Yuqing Jiaheng Pharmaceutical Co., Ltd. The rust-red rosehip seed oil was purchased from Shanghai Huien International Trade Co., Ltd. Lactobacillus rhamnosus was purchased from the China General Microbiological Culture Collection Center, with accession number CGMCC No. 18639; The brewing yeast was purchased from the China Industrial Microbial Culture Collection Center, with accession number CICC 1210; Aspergillus oryzae was purchased from the China Industrial Microbial Culture Collection Center, with accession number CICC 2336.

[0026] Unless otherwise specified, all other materials, reagents, etc. used in all embodiments and comparative examples of this invention are commercially available.

[0027] The raw materials and their weights of the essential oil compositions with whitening and spot-fading effects in Examples 1-5 and Comparative Examples 1-18 are shown in Table 1. The total weight of each essential oil composition is equal. The preparation of the essential oil compositions with whitening and spot-fading effects in Examples 1-5 and Comparative Examples 1-18 includes the following steps: The raw material components were mixed according to the weight ratio and homogenized at 15,000 rpm to obtain an essential oil composition with whitening and spot-fading effects.

[0028] The preparation of pink passion fruit seed oil in Examples 1-5, Comparative Examples 1-5, and Comparative Examples 7-8 includes the following steps: S1. Crush the pink passion fruit seeds and pass them through a 50-mesh sieve to obtain pink passion fruit seed powder. Add deionized water at a material-to-liquid ratio of 1g:10mL, mix well, and apply a radiation dose of 25kGy. 60 The base solution was obtained by radiation sterilization with Co-γ rays. S2. Add a compound bacterial solution consisting of *Lactobacillus rhamnosus*, *Saccharomyces cerevisiae*, and *Aspergillus oryzae* to the base liquid, and ferment aerobically at 33°C for 30 hours to obtain the fermentation product; wherein, the inoculum amount of the compound bacterial solution is 6% v / v of the base liquid, and the effective viable count of the compound bacterial solution is 4.5 × 10⁻⁶. 9 CFU / mL, the viable count ratio of Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae in the compound bacterial solution is 0.1:1:0.35; S3. The fermentation product is dried at 65℃ until the moisture content is 10wt%, pulverized and passed through a 50-mesh sieve, and placed in a supercritical CO2 extraction vessel. An entrainer is pumped in for extraction. The extraction pressure is set to 35MPa, the extraction temperature to 50℃, the CO2 flow rate to 25L / h, the entrainer to be an 80% ethanol aqueous solution, the flow ratio of the entrainer to CO2 to be 1:30, and the extraction time to be 70min. The extract is collected to obtain pink passion fruit seed oil.

[0029] Table 1. Raw materials and their weight ratios of essential oil compositions with whitening and spot-fading effects.

[0030] Comparative Example 9 Compared with Example 1, the difference is that the compound bacterial solution used in Comparative Example 9 to prepare pink passion fruit seed oil consisted of Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae in a live bacteria ratio of 1:0.35:0.1, while other conditions remained the same as in Example 1.

[0031] Comparative Example 10 Compared with Example 1, the difference is that the compound bacterial solution used in Comparative Example 10 to prepare pink passion fruit seed oil consisted of Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae in a live bacteria ratio of 0.35:0.1:1, while other conditions remained the same as in Example 1.

[0032] Comparative Example 11 Compared with Example 1, the difference is that the compound bacterial solution used in Comparative Example 11 to prepare pink passion fruit seed oil consisted of Lactobacillus rhamnosus and Saccharomyces cerevisiae with a live bacteria ratio of 0.1:1, while other conditions remained the same as in Example 1.

[0033] Comparative Example 12 Compared with Example 1, the difference is that the compound bacterial solution used in Comparative Example 12 to prepare pink passion fruit seed oil consisted of Saccharomyces cerevisiae and Aspergillus oryzae with a live bacteria ratio of 1:0.35, while other conditions remained the same as in Example 1.

[0034] Comparative Example 13 Compared with Example 1, the difference is that the compound bacterial solution used in Comparative Example 13 to prepare pink passion fruit seed oil consisted of Lactobacillus rhamnosus and Aspergillus oryzae with a live bacteria ratio of 0.1:0.35, while other conditions remained the same as in Example 1.

[0035] Comparative Example 14 Compared with Example 1, the difference is that in the preparation of pink passion fruit seed oil, Comparative Example 14 used Lactobacillus rhamnosus with preservation number CGMCC No.20110 instead of Lactobacillus rhamnosus in Example 1, while other conditions remained the same as in Example 1.

[0036] Comparative Example 15 Compared with Example 1, the difference is that Comparative Example 15 used Lactobacillus plantarum with accession number CICC 20242 instead of Lactobacillus rhamnosus in Example 1 when preparing pink passion fruit seed oil, while other conditions remained the same as in Example 1.

[0037] Comparative Example 16 Compared with Example 1, the difference is that Comparative Example 16 used Saccharomyces cerevisiae with preservation number CICC 1252 instead of Saccharomyces cerevisiae in Example 1 when preparing pink passion fruit seed oil, while other conditions remained the same as in Example 1.

[0038] Comparative Example 17 Compared with Example 1, the difference is that Comparative Example 17 used Aspergillus oryzae with preservation number CICC 2005 instead of Aspergillus oryzae in Example 1 when preparing pink passion fruit seed oil, while other conditions were the same as in Example 1.

[0039] Comparative Example 18 The difference compared to Example 1 is that the preparation of the pink passion fruit seed oil in Comparative Example 18 includes the following steps: S1. Crush the pink passion fruit seeds and pass them through a 50-mesh sieve to obtain pink passion fruit seed powder. Add deionized water at a material-to-liquid ratio of 1g:10mL, mix well, and apply a radiation dose of 25kGy. 60 The base solution was obtained by radiation sterilization with Co-γ rays. S2. Place the base liquid into a high-pressure tank and pressurize it to 3 kgf / cm³. 2 After the pressure is uniform, maintain it for 20 minutes, then return to normal pressure to obtain a paste-like base material; S3. Place the paste-like base material in a supercritical CO2 extraction vessel, pump in an entrainer for extraction, and set the extraction pressure to 35 MPa, extraction temperature to 50℃, CO2 flow rate to 25 L / h, the entrainer to be an 80% ethanol aqueous solution, the flow rate ratio of the entrainer to CO2 to be 1:30, and the extraction time to be 70 min. Collect the extract to obtain pink passion fruit seed oil.

[0040] Test Example 1: Tyrosinase Activity Inhibition Experiment Using the essential oil compositions with whitening and spot-fading effects prepared in Examples 1-5 and Comparative Examples 1-18 as samples, the compositions were dissolved and diluted with PBS solution at pH 6.8 to prepare sample solutions with a concentration of 3 mg / mL. 1.0 mL of the sample solution was added to each sample tube and sample control tube, and 1.0 mL of PBS solution was added to each negative control tube and blank control tube. 0.5 mL of 100 U / mL tyrosinase solution (purchased from Sigma) was added to each sample tube and negative control tube, and 0.5 mL of PBS solution was added to each sample control tube and blank control tube. The mixtures were shaken to mix, incubated in a water bath at 30°C for 10 min, and then 2.0 mL of 5 × 10⁻⁶ PBS solution was added to each tube. -4 Incubate with g / mL L-DOPA (from Sigma) for 5 min, then immediately measure the absorbance at 475 nm and calculate the tyrosinase inhibition rate using the following formula: Tyrosinase inhibition rate (%) = [1 - (T - T0) / (C - CO)] × 100%; In the formula, T is the absorbance of the sample tube, T0 is the absorbance of the sample control tube, C is the absorbance of the negative control tube, and C0 is the absorbance of the blank control tube. Each group was set up with three parallel groups, and the results were averaged and recorded as shown in Table 2 below.

[0041] Table 2 Example 1 96.8 Example 2 91.5 Example 3 94.6 Example 4 83.9 Example 5 87.4 Comparative Example 1 48.1 Comparative Example 2 52.7 Comparative Example 3 53.9 Comparative Example 4 55.3 Comparative Example 5 56.2 Comparative Example 6 50.5 Comparative Example 7 64.1 Comparative Example 8 65.7 Comparative Example 9 69.5 Comparative Example 10 67.8 Comparative Example 11 58.9 Comparative Example 12 61.2 Comparative Example 13 57.3 Comparative Example 14 63.0 Comparative Example 15 59.6 Comparative Example 16 60.7 Comparative Example 17 62.1 Comparative Example 18 63.8 As shown in Table 2, compared with Comparative Examples 1-18, the essential oil compositions prepared in Examples 1-5 can effectively inhibit tyrosinase activity and have good whitening and spot-fading effects.

[0042] Among them, by comparing the data of Examples 1-5 and Comparative Examples 1-8, it can be seen that the six components of the essential oil composition of the present invention—Italian immortelle oil, rice germ oil, sandalwood oil, white chamomile oil, rust-red rose seed oil, and pink passionflower seed oil—are indispensable. Furthermore, when the weight ratio of Italian immortelle oil, rice germ oil, sandalwood oil, white chamomile oil, rust-red rose seed oil, and pink passionflower seed oil is (1-3):(4-8):(2-4):(1-3):(0.6-2):(2-6), the composition exhibits better tyrosinase inhibition, especially Example 1. Through comparative examples... As shown in Examples 1 and 9-18, supercritical CO2 extraction after fermentation yields better pink passion fruit seed oil. Furthermore, the compound microbial strains selected during the preparation of pink passion fruit seed oil have a significant impact on the tyrosinase inhibition effect of the final essential oil composition. When Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae are fermented in a ratio of (0.05-0.15):1:(0.3-0.4) for compound fermentation of pink passion fruit seeds, the tyrosinase inhibition effect of the composition reaches its optimal level. Any substitution or omission of a certain microbial strain may result in the final tyrosinase inhibition effect failing to meet the expectations of this invention.

[0043] Test Example 2: DPPH Free Radical Scavenging Experiment Using the essential oil compositions with whitening and spot-fading effects prepared in Examples 1-5 and Comparative Examples 1-18 as samples, they were dissolved and diluted with 60% v / v ethanol aqueous solution to prepare sample solutions with a concentration of 5 mg / mL. Sample groups and control groups were set up. In the sample group, 100 μL of sample solution and 100 μL of DPPH solution were added to each well of a 96-well plate. In the control group, 100 μL of 60% v / v ethanol aqueous solution and 100 μL of DPPH solution were added to each well of a 96-well plate. The plates were incubated at room temperature in the dark for 30 min, and the absorbance was measured at a wavelength of 517 nm. The DPPH free radical scavenging rate was calculated according to the following formula: DPPH radical scavenging rate (%) = (1 - absorbance of sample group / absorbance of control group) × 100%; Each group was set up with three parallel groups, and the results were averaged and recorded as shown in Table 3 below.

[0044] Table 3 Example 1 92.4 Example 2 88.1 Example 3 90.7 Example 4 82.3 Example 5 85.9 Comparative Example 1 42.2 Comparative Example 2 46.0 Comparative Example 3 48.5 Comparative Example 4 50.9 Comparative Example 5 52.3 Comparative Example 6 44.7 Comparative Example 7 62.4 Comparative Example 8 65.1 Comparative Example 9 67.8 Comparative Example 10 69.5 Comparative Example 11 55.3 Comparative Example 12 55.7 Comparative Example 13 53.9 Comparative Example 14 60.2 Comparative Example 15 56.8 Comparative Example 16 57.6 Comparative Example 17 59.4 Comparative Example 18 61.9 As can be seen from the data of Example 1 and Comparative Examples 1-6 in Table 3, in the essential oil composition of the present invention, Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil work synergistically to exert excellent antioxidant effects when used in combination.

[0045] Comparing the data from Examples 1-5 and Comparative Examples 7-8, it can be seen that the ratio of Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil affects the DPPH removal effect of the composition. When the weight ratio of Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil is (1-3):(4-8):(2-4):(1-3):(0.6-2):(2-6), the DPPH removal effect of the composition is at a better level. Comparing Example 1 and Comparative Examples 9-18, it can be seen that the preparation process of pink passionflower seed oil affects the DPPH removal effect of the composition.

[0046] Test Example 3: Cell Scratch Repair Experiment HaCaT cells (Shanghai Guyan Industrial Co., Ltd.) were seeded into 12-well plates, with inserts placed in each well. The plates were cultured at 37°C in a 5% CO2 incubator with maintenance medium for 24 hours. Cells were divided into a normal control group (serum-free DMEM medium), a positive control group (serum-free DMEM medium containing 10000 IU / mL epidermal growth factor (EGF), and a test substance treatment group (serum-free DMEM medium containing 1 wt% of the essential oil compositions from Examples 1-5 or Comparative Examples 1-18). Each group had three replicates. Inserts were removed and the cells were cultured for another 24 hours. Cell growth at the scratch sites was observed under a microscope at 0 hours and 24 hours, and photographs were taken. The healing area was calculated using ImageJ software. The repair ability of each group was evaluated by the healing rate. The healing rate was calculated using the following formula: Healing rate % = (Scratch area at 0h - Scratch area at 24h) / Scratch area at 0h × 100%; The results, after taking the average, are recorded in Table 4 below.

[0047] Table 4 normal control group 35.2 Positive control group 100.0 Example 1 78.6 Example 2 74.9 Example 3 77.2 Example 4 68.8 Example 5 72.4 Comparative Example 1 41.6 Comparative Example 2 43.5 Comparative Example 3 44.3 Comparative Example 4 45.8 Comparative Example 5 46.7 Comparative Example 6 42.9 Comparative Example 7 54.3 Comparative Example 8 55.1 Comparative Example 9 57.2 Comparative Example 10 56.5 Comparative Example 11 48.4 Comparative Example 12 49.0 Comparative Example 13 47.3 Comparative Example 14 53.2 Comparative Example 15 49.8 Comparative Example 16 50.7 Comparative Example 17 51.9 Comparative Example 18 53.1 As shown in Table 4, compared with Comparative Examples 1-18, the essential oil compositions with whitening and spot-fading effects provided in Examples 1-5 can effectively promote the healing of HaCaT cells and have a good repair effect.

[0048] As can be seen from Comparative Example 1 and Comparative Examples 1-8, omitting or replacing any component in the composition resulted in a decrease in the healing rate of the composition. As can be seen from Comparative Example 1 and Comparative Examples 9-18, the preparation process of pink passion fruit seed oil also affected the final healing effect of the composition on HaCaT cells. It is evident that the six components in the essential oil composition of the present invention are indispensable in promoting cell healing and have a synergistic effect.

[0049] Test Example 4: Anti-inflammatory Effect Experiment RAW264.7 mouse monocytes / macrophages (Shanghai Yaji Biotechnology Co., Ltd.) that have grown to the logarithmic growth phase were divided into 1.5 × 10⁻⁶ cells. 5 Cells were seeded at a density of 800 μL / mL in 24-well plates and cultured in a cell incubator at 37°C and 5% CO2 for 24 h. When the cells were nearly confluent, they were administered at a concentration of 10 mg / L (50 μL) of the sample (the whitening and spot-fading essence composition prepared in Examples 1-5 or Comparative Examples 1-18). Each group had 3 replicates. The positive control was 10 μg / mL (50 μL) of dexamethasone. After 1 h, the cells were stimulated with 1 μg / mL of lipopolysaccharide (LPS). After incubation in a cell culture incubator for 24 hours, the cell culture medium was transferred to centrifuge tubes and centrifuged at 5000 r / min for 10 min to collect the cell supernatant. The levels of inflammatory factors TNF-α and IL-1β in RAW264.7 cells were detected using a mouse tumor necrosis factor α (TNF-α) kit (Shanghai Baiyi Biotechnology Co., Ltd., catalog number BY-M01996) and a mouse interleukin 1β (IL-1β) ELISA kit (Shanghai Baiyi Biotechnology Co., Ltd., catalog number BY-M03521), respectively. The stronger the inhibitory effect on inflammatory factors TNF-α and IL-1β, the better the anti-inflammatory effect of the composition. The anti-inflammatory effect of the composition is expressed by the inhibition rate of TNF-α and IL-1β, as shown in the following formula: Inhibition rate = [blank group - (sample group - positive group)] / blank group × 100%; The results, after being averaged, are recorded in Table 5 below.

[0050] Table 5. Results of anti-inflammatory effect test Blank group 0 0 positive group 100 100 Example 1 71.9 70.3 Example 2 70.5 68.9 Example 3 68.3 66.5 Example 4 62.8 60.7 Example 5 65.4 63.8 Comparative Example 1 31.2 30.6 Comparative Example 2 35.3 33.9 Comparative Example 3 35.7 34.3 Comparative Example 4 36.6 35.1 Comparative Example 5 37.9 36.4 Comparative Example 6 33.1 31.5 Comparative Example 7 47.7 45.6 Comparative Example 8 49.4 47.0 Comparative Example 9 52.5 49.8 Comparative Example 10 51.0 48.5 Comparative Example 11 39.4 37.6 Comparative Example 12 40.8 38.7 Comparative Example 13 38.7 36.9 Comparative Example 14 45.2 43.4 Comparative Example 15 41.9 39.8 Comparative Example 16 42.8 40.6 Comparative Example 17 44.1 42.2 Comparative Example 18 46.3 44.7 As shown in Table 5, the essential oil composition of the present invention with whitening and spot-fading effects can effectively inhibit the expression of TNF-α and IL-1β inflammatory factors and has good anti-inflammatory properties. When the weight ratio of Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil and pink passionflower seed oil is (1-3):(4-8):(2-4):(1-3):(0.6-2):(2-6), the composition has better anti-inflammatory effect.

[0051] Comparing the TNF-α inhibition rate and IL-1β inhibition rate of Example 1 and Comparative Examples 1-18, it can be seen that all six components in the essential oil composition of the present invention are indispensable. Any substitution, omission, or change in the preparation process of the components will lead to a decrease in the anti-inflammatory effect of the composition, indicating that the raw material components have a synergistic effect in anti-inflammatory aspects.

[0052] Application Example 1 and Comparative Application Example 1-18 The whitening and spot-fading essential oil compositions of Example 1 and Comparative Examples 1-18 were added to the serum at a concentration of 2 wt% to obtain the serums of Application Example 1 and Comparative Application Examples 1-18, respectively. In addition, a serum without the added essential oil composition was prepared as a blank application example. The formula of the serum is shown in Table 6.

[0053] Table 6. Serum Components and Dosage

[0054] The preparation method of the serum described in Application Example 1 and Comparative Application Examples 1-18 specifically includes the following steps: (1) At 80°C, the humectant, thickener and deionized water are mixed and stirred evenly at 1500 rpm. The mixture is kept warm for later use to obtain the first material. (2) Mix the preservatives and heat them to 60°C to melt them, to obtain the second material; (3) After cooling the first material to 60°C, add it to the second material at 200 rpm and stir evenly. Cool it to 45°C, add the essential oil composition with whitening and spot-fading effects, and continue to stir evenly at 200 rpm. Then add the pH adjuster and stir evenly at 200 rpm. Discharge the material to obtain the essence.

[0055] The preparation method of the serum described in the blank application example specifically includes the following steps: (1) At 80°C, the humectant, thickener and deionized water are mixed and stirred evenly at 1500 rpm. The mixture is kept warm for later use to obtain the first material. (2) Mix the preservatives and heat them to 60°C to melt them, to obtain the second material; (3) After cooling the first material to 60°C, add it to the second material at 200 rpm, stir evenly, cool it to 45°C, then add the pH adjuster, stir evenly at 200 rpm, and discharge the material to obtain the essence.

[0056] Example 1 of efficacy test: Human skin patch test Fifty participants (healthy men or women aged 25-55) were selected for testing according to the inclusion criteria, and the selected area was no more than 50 mm². 2A qualified spot test apparatus with a depth of approximately 1 mm was used. The sample was placed inside the small chamber of the apparatus, and 0.025 mL of the test sample (the essence prepared in Application Example 1, Control Application Examples 1-18, and the blank application example, and distilled water) was added. Twenty-one test areas of the same size were selected on both arms of the volunteer (1-2 cm apart), and each area was covered with hypoallergenic adhesive tape. The tape was gently pressed with the palm of the hand to ensure even application to the skin. The spot test apparatus was removed after 24 hours. Skin reactions were observed according to the standards in Table 7 at 30 min (after the indentation disappeared), 24 h, and 48 h after removal of the test apparatus, and the results were recorded. If the average irritation score of the application examples was not higher than that of the blank control group, and no case of grade 2 or higher adverse reaction occurred, the product was considered safe and non-irritating.

[0057] Table 7 Skin Reaction Grading Standards 0 negative reaction 1 Suspicious reaction, only slight erythema 2 Weak positive reaction (erythema reaction): erythema, infiltration, edema, and papules may be present. 3 Strong positive reaction (herpes reaction): erythema, infiltration, edema, papules; the reaction may extend beyond the test area. 4 Extremely strong positive reaction (confluent herpes simplex reaction): obvious erythema, severe infiltration, edema, confluent herpes simplex; reaction extends beyond the test area. After testing, the serums provided in Application Example 1, Comparative Application Examples 1-18, and Blank Application Example of this invention showed that their average irritation scores were not higher than those of the blank control group after human patch testing, and no adverse reactions of grade 2 or above occurred. This indicates that the lotion provided by this invention is safe and non-irritating to human skin and that no adverse reactions occurred during use.

[0058] Example 2 of efficacy test: Human efficacy test Based on the principles of human efficacy evaluation in the "Cosmetic Safety Technical Specifications (2015 Edition)," 60 participants (healthy men or women aged 25-55) were selected according to the subject inclusion criteria. The participants were randomly divided into 20 groups of 3 people each. The test product sample groups (the serum prepared by application example 1, comparative application examples 1-18, and blank application example) were used to evaluate the melanin removal function, skin barrier repair function, anti-inflammatory function, and pigmentation improvement function of the test product. The testing method was as follows: Subjects were instructed to apply the test product to their face once in the morning and once in the evening, at a dosage of (2.00±0.05) mg / cm³. 2 The test lasted for 21 days. All tests were conducted after the participants were cleaned and sat still for 30 minutes in the experimental environment (temperature 20±1℃, humidity 50±10%). Follow-up visits were conducted on day 1 (initial values) and day 21. Melanin content (MI) was measured using an MX18 melanin analyzer (CK, Germany); transepidermal water loss (TEWL) values ​​were measured using a Tewameter™ Hex probe; and the corresponding a*, b*, and L* values ​​were measured using a Colorimeter CL400 (CK, Germany). The ITA° was then calculated to evaluate the product's effectiveness in improving melanin content (MI), skin barrier damage, skin inflammation, and pigmentation. In this application example, the improvement in melanin content (MI) was expressed as the MI improvement rate; the repair effect on skin barrier damage was expressed as the TEWL improvement rate; the improvement effect on skin inflammation was expressed as the a* improvement rate; and the improvement effect on pigmentation was expressed as the ITA° improvement rate. The specific calculation formulas are as follows: ITA°=[Arc tan(L*-50) / b*]×180 / π; MI value improvement rate (%) / TEWL value improvement rate (%) / a* value improvement rate (%) / ITA° value improvement rate (%) = (value before use - value after use) / value before use × 100%; The results, after taking the average, are recorded in Table 8 below.

[0059] Table 8 Results of Human Efficacy Tests Blank control group 2.9 3.1 4.9 3.2 Application Example 1 39.1 34.3 32.9 55.4 Comparative Application Example 1 11.5 10.2 11.0 16.1 Comparative Application Example 2 12.8 10.9 12.4 17.8 Comparative Application Example 3 13.6 11.3 12.9 18.6 Comparative Application Example 4 14.3 11.8 13.7 19.2 Comparative Application Example 5 14.7 12.3 14.2 19.7 Comparative Application Example 6 12.2 10.7 11.9 16.5 Comparative Application Example 7 20.4 17.4 18.5 25.3 Comparative Application Example 8 21.9 18.3 19.4 28.4 Comparative Application Example 9 24.1 19.8 20.6 32.0 Comparative Application Example 10 23.3 19.2 20.1 30.7 Comparative Application Example 11 16.2 13.4 15.3 21.4 Comparative Application Example 12 16.9 14.6 15.9 22.1 Comparative Application Example 13 15.8 12.9 14.8 20.5 Comparative Application Example 14 19.1 16.4 17.6 24.6 Comparative Application Example 15 17.5 15.2 16.4 22.7 Comparative Application Example 16 18.0 15.7 16.8 23.2 Comparative Application Example 17 18.7 16.1 17.3 23.9 Comparative Application Example 18 20.1 17.0 17.9 24.8 As shown in Table 8, the essence containing the essential oil composition of the present invention with whitening and spot-fading effects can effectively reduce the melanin content of the skin, repair the skin barrier, soothe facial inflammation, brighten and even out skin tone, and work together to exert whitening effects.

[0060] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any indirect modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. An essential oil composition with whitening and spot-fading effects, characterized in that, The essential oil composition with whitening and spot-fading effects is composed of Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil. The weight ratio of the Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil is (1-3):(4-8):(2-4):(1-3):(0.6-2):(2-6). The pink passion fruit seed oil is obtained by supercritical CO2 extraction from pink passion fruit seeds after compound fermentation with Lactobacillus rhamnosus, Saccharomyces cerevisiae, and Aspergillus oryzae, including the following steps: S1. Crush the pink passion fruit seeds, pass them through a 40-60 mesh sieve to obtain pink passion fruit seed powder, add deionized water, mix well, sterilize, and obtain the basic liquid. S2. Add compound bacterial solution to the base liquid and ferment aerobicly at 29-37℃ for 24-36 hours to obtain fermentation product; S3. Dry the fermentation product at 60-70℃ until the moisture content is 10-20wt%, pulverize it and pass it through a 40-60 mesh sieve. Place it in a supercritical CO2 extraction vessel, pump in an entrainer for extraction, collect the extract, and obtain pink passion fruit seed oil. The viable count ratio of *Lactobacillus rhamnosus*, *Saccharomyces cerevisiae*, and *Aspergillus oryzae* in the compound bacterial solution is 0.05-0.15:1:0.3-0.

4. *Lactobacillus rhamnosus* was purchased from the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 18639; *Saccharomyces cerevisiae* was purchased from the China Industrial Microbiological Culture Collection Center (CICC 1210); and *Aspergillus oryzae* was purchased from the China Industrial Microbiological Culture Collection Center (CICC 2336).

2. The essential oil composition with whitening and spot-fading effects according to claim 1, characterized in that, The weight ratio of the Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil is (1.5-2.5): (5.5-6.5): (2.5-3.5): (1.5-2.5): (1-1.4): (3.5-4.5).

3. The essential oil composition with whitening and spot-fading effects according to claim 1, characterized in that, In step S1, the ratio of pink passion fruit seed powder to deionized water is 1g:5-15mL, and the sterilization method is radiation sterilization; the radiation sterilization is performed using... 60 Sterilization is performed using Co-γ rays with a radiation dose of 25 kGy.

4. The essential oil composition with whitening and spot-fading effects according to claim 1, characterized in that, In step S2, the inoculation amount of the compound bacterial solution is 5.5% v / v - 6.5% v / v of the base solution, and the effective viable count of the compound bacterial solution is 4 × 10⁻⁶. 9 CFU / mL - 5 × 10 9 CFU / mL.

5. The essential oil composition with whitening and spot-fading effects according to claim 1, characterized in that, In step S3, the supercritical CO2 extraction is performed at an extraction pressure of 30-40 MPa, an extraction temperature of 45-55℃, a CO2 flow rate of 20-30 L / h, and an extraction time of 60-80 min. The entrainer is an aqueous ethanol solution with a mass concentration of 75%-85%, and the flow rate ratio of the entrainer to CO2 is 1:(25-35).

6. A method for preparing the essential oil composition with whitening and spot-fading effects according to any one of claims 1-5, characterized in that, Includes the following steps: Italian immortelle oil, rice germ oil, sandalwood oil, white chrysanthemum oil, rust-red rose seed oil, and pink passionflower seed oil were mixed in a weight ratio and homogenized at 10,000 rpm to 20,000 rpm to obtain an essential oil composition with whitening and spot-fading effects.

7. The application of the essential oil composition with whitening and spot-fading effects prepared by the method according to claim 6 in the preparation of skin care products, characterized in that, The skincare products include toners, lotions, creams, masks, serums, and sprays, and the amount of the essential oil composition added is 0.5%-3% of the total weight of the skincare products.

8. An essence, characterized in that, The serum comprises the following ingredients by weight percentage: 0.5%-3% of the essential oil composition with whitening and spot-fading effects according to any one of claims 1-5, 0.05%-0.3% thickener, 3%-10% moisturizer, 0.5%-5% preservative, 0.01%-0.3% pH adjuster, and the balance being deionized water.

9. The essence according to claim 8, characterized in that, The thickener includes at least one of xanthan gum, carbomer, ammonium acryloyl dimethyl taurate / VP copolymer, hydroxyethyl acrylate / sodium acryloyl dimethyl taurate copolymer, sodium polyacrylate, hydroxyethyl cellulose, and sclerotium gum; The moisturizer includes at least one of allantoin, panthenol, dipropylene glycol, low molecular weight sodium hyaluronate, glycerin, 1,3-butanediol, and 1,2-propanediol. The preservative includes at least one of p-hydroxyacetophenone, ethylhexylglycerin, and phenoxyethanol; The pH adjuster includes at least one of triethanolamine, citric acid, and arginine.

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