Composition, application and cosmetics containing withanolide

Abstract

The invention belongs to the field of daily chemicals, and discloses a composition containing withanolide, wherein the composition comprises meadowfoam seed oil and withanolide; the mass content of withanolide in the meadowfoam seed oil is 5 to 45 ppm. The composition contains withanolide, and can effectively improve the application performance of meadowfoam seed oil. At the same time, the invention also provides a method for improving the performance of meadowfoam seed oil, a use thereof, and a cosmetic.

Description

Technical Field

[0001] The invention relates to the field of daily chemicals, and in particular to a composition containing withanolide, an application and a cosmetic. Background Art

[0002] Meadowfoam seed oil is one of the most stable vegetable oils in the world because it contains more than 98% long-chain fatty acids with antioxidant effects. The average oxidative stability index OSI of meadowfoam seed oil is the highest among all vegetable oils. It not only has oxidative stability, but also has good antioxidant activity. Meadowfoam seed oil can promote the skin's reabsorption of water. At the same time, because it can form certain long-chain fatty alcohols, it can form a layer of hydrate in the epidermis to achieve the purpose of long-term water locking and moisturizing. Therefore, it is widely used to increase the stability of other oils and as an emollient.

[0003] Other properties of meadowfoam seed oil, such as anti-wrinkle and soothing properties, are not obvious. Therefore, improving the relevant application performance of meadowfoam seed oil is an important measure to extend the application scope of meadowfoam seed oil.

[0004] The technical problem solved in this case is: how to improve the application performance of white meadowfoam seed oil. Summary of the invention

[0005] The purpose of the present invention is to provide a composition, which contains withanolide and can effectively improve the application performance of meadowfoam seed oil.

[0006] Meanwhile, the invention also provides a method for improving the performance of meadowfoam seed oil, application and cosmetics.

[0007] The specific scheme of the present invention is:

[0008] A composition comprises meadowfoam seed oil and withanolide; the mass content of withanolide in the meadowfoam seed oil is 5 to 45 ppm.

[0009] Withania somnifera is derived from Withania somnifera, which is recognized to have significant antioxidant and immune-enhancing properties.

[0010] The main antioxidant active substance of South African somniferum is withanolide;

[0011] Withanolide is a naturally occurring steroid lactone with C28 ergostane as the skeleton. Some scholars also call it C28-steroid lactone triterpenoids. Generally speaking, those with α-β-unsaturated ketone groups have characteristic absorption at 240nm, and those with conjugated double bonds have absorption at 235nm. Due to the contribution of the steroid nucleus mother and the unsaturated lactone ring, withanolide compounds have maximum absorption around 220-240nm.

[0012] In the experimental verification process of the present invention, we verified that white meadowfoam seed oil can be dissolved into withanolide as much as possible through various ways. After verification, its integration limit is about 45ppm;

[0013] The contribution of Withania somnifera root plant as raw material to the performance of Limnanthes alba seed oil is not as good as that of the product obtained by directly adding Withania somnifera root extract to Limnanthes alba seed oil for mixing. The possible reason is that direct oil extraction from Withania somnifera root may mix in other oil-soluble impurities and affect the performance. However, no matter which method is used to make Withania somnifera lactones enter Limnanthes alba seed oil, the performance of Limnanthes alba seed oil can be improved.

[0014] In addition, we have also verified the efficacy of the composition of the present invention, such as firming, anti-wrinkle, and soothing; after verification, we surprisingly found that the combination of the two can effectively improve the performance.

[0015] In the above composition, the mass content of withanolide in the meadowfoam seed oil is 9 to 35 ppm, more preferably 10.7 to 31.7 ppm, and more preferably 18 to 35 ppm.

[0016] In the above composition, the Withania somnifera root extract is dispersed in Limnanthes alba seed oil, so that at least part of withanolide is dissolved in Limnanthes alba seed oil.

[0017] At the same time, the invention also discloses a method for improving the performance of the meadowfoam seed oil, and the performance of the meadowfoam seed oil is improved by controlling the mass content of withanolide in the meadowfoam seed oil to be 5-45 ppm.

[0018] In the above method, the mass content of withanolide in the white meadowfoam seed oil is controlled to be 9 to 35 ppm.

[0019] In the above method, the performance refers to one or more of firming, anti-wrinkle, and soothing.

[0020] In addition, the present invention also discloses the use of the composition for preparing cosmetics.

[0021] In the above-mentioned use, the dosage form of the cosmetic is essence oil, emulsion or cream.

[0022] Finally, the present invention also discloses a cosmetic comprising the composition as described above.

[0023] In the above-mentioned cosmetics, the dosage form of the cosmetics is essence oil, emulsion or cream.

[0024] The beneficial effects of this application are:

[0025] Through the experimental verification of the present invention, when a trace amount of withanolide is dissolved in the white meadowfoam seed oil, its anti-wrinkle and anti-inflammatory properties are significantly improved. Compared with using rice bran oil and macadamia seed oil as a carrier, the application performance of the white meadowfoam seed oil containing withanolide of the present invention is more obvious, which has positive significance and commercial promotion value for broadening the application scope of white meadowfoam seed oil. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the spectrum of the liquid phase of Limnanthes alba seed oil;

[0027] Figure 2 This is the spectrum of Limnanthes alba seed oil + Withania somnifera root plant liquid phase;

[0028] Figure 3 This is the liquid chromatography spectrum of Limnanthes alba seed oil + Withania somnifera root extract;

[0029] Figure 4 This is the liquid chromatography spectrum of withanolide A. DETAILED DESCRIPTION

[0030] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that, if no specific conditions are specified in the embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0031] Part I

[0032] Materials and reagents refer to Table 1;

[0033] Table 1 Sources of materials and reagents

[0034] Serial number name factory batch number 1 Withania somnifera root extract Shaanxi Wanyuan Biotechnology Co., Ltd. TNFZQ231125 2 Withania somnifera root extract Shaanxi New Horizon Biotechnology Co., Ltd. XTY20240325 3 Withania somnifera root extract Shaanxi Hongda Plant Chemical Co., Ltd. EP24020703 4 Withania somnifera root extract Shaanxi Bolin Biotechnology Co., Ltd. 20240524 5 Withania somnifera root extract Xi'an Zhengdong Biotechnology Co., Ltd. 20231218

[0035] Meadowfoam seed oil is of plant origin and the manufacturer is Guangzhou Huashi Cosmetics Technology Co., Ltd.

[0036] Part 2

[0037] Example 1

[0038] Extraction of Withanolides from Withania somnifera Roots by Limnanthes alba Seed Oil

[0039] The extraction method is:

[0040] Step 1: extraction, the solid-liquid ratio of Withania somnifera root to Limnanthes alba seed oil was 1:5 (w / w), and ultrasonic extraction was performed at 50°C for 40 min;

[0041] Step 2: Centrifugation: After the ultrasound, the Withania somnifera root oil mixture was centrifuged at 7000 rpm for 10 min, and the supernatant was collected;

[0042] Step 3: Filtration: the supernatant was filtered through a 0.45 um nylon 66 filter membrane to obtain the Withania somnifera root oil extract.

[0043] Example 2

[0044] Using Withania somnifera extracts from different manufacturers (raw material information can be found above) as raw materials to prepare the composition;

[0045] The preparation method is:

[0046] 0.3 wt% of the Withania somnifera root extract was added to the white meadowfoam seed oil, and ultrasonic extraction was performed at 50°C for 40 minutes. After the ultrasonication, the Withania somnifera root oil mixture was centrifuged at 7000 rpm for 10 minutes, and the supernatant was collected. The supernatant was filtered through a 0.45 um nylon 66 filter membrane to obtain a Withania somnifera oil composition.

[0047] Comparative Example 1

[0048] The composition is prepared by using Withania somnifera root extract and rice bran oil as raw materials;

[0049] The preparation method is:

[0050] 0.3 wt% of the Withania somnifera root extract was added to the rice bran oil and ultrasonicated at 50°C for 40 min. After the ultrasonication, the Withania somnifera root oil mixture was centrifuged at 7000 rpm for 10 min to obtain the supernatant. The supernatant was filtered through a 0.45 um nylon 66 filter membrane to obtain a rice bran oil composition.

[0051] Comparative Example 2

[0052] A composition is prepared by using Withania somnifera root extract and macadamia nut seed oil as raw materials;

[0053] The preparation method is:

[0054] 0.3 wt% of the Withania somnifera root extract was added to the macadamia seed oil and ultrasonicated at 50°C for 40 min. After the ultrasonication, the Withania somnifera root oil mixture was centrifuged at 7000 rpm for 10 min, and the supernatant was collected. The supernatant was filtered through a 0.45 um nylon 66 filter membrane to obtain a macadamia seed oil composition.

[0055] Part 3

[0056] 3.1 The method for detecting the content of withanolide in white meadowfoam seed oil is:

[0057] HPLC was used to detect the withanolide content;

[0058] Detector: DAD detector;

[0059] Chromatographic column: Agilent ZORBAX C18 (250mm*4.6mm*5um);

[0060] Mobile phase: methanol-water, gradient elution, elution program see Table 2;

[0061] Table 2 Elution gradient

[0062] Time (min) Chromatography Methanol Ultrapure water 0 20 80 6 100 0 10 100 0

[0063] Injection volume: 10 μL;

[0064] Column temperature: 25°C;

[0065] Detection wavelength: 230nm;

[0066] Flow rate: 1.0 mL / min;

[0067] Combined with UV spectroscopy analysis, the withanolide content was calculated using the withanolide A standard curve at a wavelength of 230nm.

[0068] The standard curve of withanolide A obtained from the experiment is:

[0069] y = 19.247x +8.0085 R = 0.999;

[0070] Figure 1 This is the spectrum of the liquid phase of Limnanthes alba seed oil;

[0071] Figure 2 This is the spectrum of Limnanthes alba seed oil + Withania somnifera root plant liquid phase;

[0072] Figure 3 This is the liquid chromatography spectrum of Limnanthes alba seed oil + Withania somnifera root extract;

[0073] Figure 4 is the liquid phase spectrum of withanolide A;

[0074] It can be seen from the above spectra that the spectrum of white meadowfoam seed oil + withania somnifera root extract has many more material peaks than the spectrum of white meadowfoam seed oil, most of which are withanolide peaks, and the peak time of withania somnifera A is 8.402min. The absorbance value of withania somnifera seed oil + withania somnifera root extract withania somnifera lactone is higher than that of white meadowfoam seed oil + withania somnifera root plant withania somnifera lactone.

[0075] The test results can be seen in Table 3;

[0076] Table 3 Statistics of withanolide content in compositions obtained by different extraction methods and raw materials

[0077] Test samples Components Withanolide content / ppm Example 1 Limnanthes Alba Seed Oil + Withania Somnifera Root Plant 23.64 Example 2.1 Limnanthes Alba Seed Oil + Withania Somnifera Root Extract (Wanyuan) 31.77 Example 2.2 Limnanthes Alba Seed Oil + Withania Somnifera Root Extract (New Horizons) 10.71 Example 2.3 Limnanthes Alba Seed Oil + Withania Somnifera Root Extract (Hongda) 18.31 Example 2.4 Limnanthes Alba Seed Oil + Withania Somnifera Root Extract (Bolin) 13.71 Example 2.5 Limnanthes Alba Seed Oil + Withania Somnifera Root Extract (Zhengdong) 19.73 Comparative Example 1 Rice bran oil + Withania somnifera root extract (Wanyuan) 18.56 Comparative Example 2 Macadamia Seed Oil + Withania Somnifera Root Extract (Wanyuan) 8.63

[0078] By using different vegetable oils to extract or dissolve the Withania somnifera root extract, it can be found that withanolide has very poor solubility in vegetable oils due to its high polarity.

[0079] There is a certain difference in the solubility of withanolide in the Withania somnifera root extracts provided by different suppliers, ranging from 10 to 32 ppm. The solubility of withanolide in the meadowfoam seed oil obtained by the extraction method is 23 ppm.

[0080] 3.2 Application Performance Testing

[0081] Determination of sample experimental concentration: Different dose concentrations of the example and comparative example samples were set to act on fibroblasts, and the cytotoxicity of each sample was determined by the CCK-8 method. The determination method was the same as the above-mentioned cell survival rate test method, and the final test sample was selected at a concentration when the cell survival rate was above 85% for subsequent cell experiments.

[0082] Recovery cell viability assay

[0083] The samples were tested for their ability to restore cell viability in a hydrogen peroxide-damaged fibroblast model, and the cell survival rate was determined using the CCK8 method.

[0084] The experiment set up a blank group (culture medium), a model group (H2O2), and a sample administration group (a mixture of the example and the comparative example with H2O2 solution in a ratio of 1:1). The sample administration group was set up with 1 concentration gradient, 3 replicates, and 3 parallel experiments.

[0085] Determination method: Fibroblasts were cultured to the logarithmic growth phase, and the cells were resuspended in DMEM medium (containing sodium penicillin 200 kU / L, streptomycin 100 mg / L, pH 7.4) containing 15% fetal bovine serum to adjust the cell concentration to 1×10 5 100 μL of cell solution was added to each 96-well plate and cultured in a 37°C CO2 incubator for 1 day. The blank group, model group, and sample administration group were added to stimulate the cells and cultured for 24 hours. The supernatant was carefully aspirated, and the CCK-8 solution and serum-free DMEM medium were mixed at a ratio of 1:10, and 110 μL of the mixture was added to each well. After incubation at 37°C in the dark for 1 hour, the absorbance of each well at a wavelength of 450 nm was measured using an ELISA instrument, i.e., the OD value.

[0086] Detection of key protein component expression

[0087] The ELISA method was used to determine the expression levels of the key protein component COL1 and the proinflammatory cytokine IL-1β, and the operation steps were carried out according to the instructions of the kit.

[0088] Data processing

[0089] GraphPad Prism 9 was used to process the data, analyze them and visualize them.

[0090] The test results can be seen in Table 4:

[0091] Table 4

[0092] Test samples Sample experimental concentration Cell viability Type I collagen COL1 promotion rate Proinflammatory cytokine IL-1β inhibition rate Example 1 0.5% 93.06% 36.77% 26.03% Example 2.1 0.5% 94.42% 63.80% 45.17% Example 2.2 0.5% 92.41% 21.51% 15.23% Example 2.3 0.5% 93.89% 47.47% 33.61% Example 2.4 0.5% 92.61% 27.53% 19.49% Example 2.5 0.5% 93.17% 39.62% 28.05% Comparative Example 1 0.5% 88.58% 2.10% 4.33% Comparative Example 2 0.5% 86.32% 1.82% 3.66% Lignum Meadowfoam Seed Oil 0.5% 95.46% 8.81% 5.67% Rice bran oil 0.5% 88.59% 1.98% 4.08% Macadamia Seed Oil 0.5% 86.21% 1.49% 2.54%

[0093] It can be seen from the results of Example 1 and Example 2.1 to Example 2.5 in Table 4 that Example 1 and Example 2.1 to Example 2.5 all have a good effect of promoting the production of type I collagen, helping the skin to improve elasticity and firmness and reduce wrinkles; Example 1 and Example 2.1 to Example 2.5 all have a good effect of inhibiting the production of pro-inflammatory cytokine IL-1β, soothing skin inflammatory reactions, reducing aging problems caused by inflammation, and are very helpful for anti-aging of sensitive skin;

[0094] At the same time, the results of Example 2.1, Example 2.2, Example 2.4 and Example 2.5 can prove that the effects of promoting the production of type I collagen and inhibiting the production of the proinflammatory cytokine IL-1β are related to the solubility of withanolide in white meadowfoam seed oil, and the higher the concentration of withanolide, the better its performance.

[0095] It can be seen from Comparative Examples 1 and 2 that the promoting effect on the production of type I collagen is weak; the effects of promoting the production of type I collagen and inhibiting the production of the proinflammatory cytokine IL-1β by white meadowfoam seed oil, rice bran oil and macadamia nut seed oil are not obvious; through the comparison of Comparative Example 1 and Example 2.3, it can be seen that under the condition of similar concentrations of withanolide, the effect of the white meadowfoam seed oil containing withanolide in Example 2.3 on promoting the production of type I collagen and inhibiting the production of the proinflammatory cytokine IL-1β is significantly better than that in Comparative Example 1; through the comparison of Comparative Example 2 and Example 2.2, it can be seen that under the condition of similar concentrations of withanolide, the effect of the white meadowfoam seed oil containing withanolide in Example 2.2 on promoting the production of type I collagen and inhibiting the production of the proinflammatory cytokine IL-1β is significantly better than that in Comparative Example 2; it indicates that a very small amount of withanolide dissolved in white meadowfoam seed oil will significantly promote the application performance of white meadowfoam seed oil. Another reason for the above results is that although withanolide has excellent functionality, it is difficult to independently promote the improvement of the relevant properties of rice bran oil and macadamia seed oil when used in extremely small amounts. In addition, rice bran oil and macadamia seed oil themselves do not have the effect of promoting the production of type I collagen or inhibiting the production of pro-inflammatory cytokine IL-1β. Therefore, even if a small amount of withanolide is used, the corresponding performance is difficult to improve.

[0096] Part 4

[0097] 4.1 Application formula

[0098] Formula 1: Anti-aging and soothing essential oil;

[0099] The preparation method is as follows: add the components in Table 5 into an emulsifying pot in sequence, stir evenly, and filter out the material;

[0100] Table 5 Essential oil formula

[0101]

[0102] Formula 2: Anti-aging and soothing serum

[0103] Its preparation comprises the following steps:

[0104] ① Add the components of phase A in Table 6 into the emulsifying pot in sequence, start stirring and heat to 75-85 degrees;

[0105] ② Add the B phase component to the oil phase pot and heat it to 70-75 degrees, add it to the emulsifying pot and start homogenization;

[0106] ③ Add the C phase components into the emulsifying pot;

[0107] ④Dissolve the phase D components and add them into the emulsifying pot;

[0108] ⑤Add low-temperature materials below 45 degrees, stir evenly, and filter out the materials.

[0109] Table 6 Essence milk formula

[0110]

[0111] Recipe 3: Anti-aging and soothing face cream

[0112] Its preparation comprises the following steps:

[0113] ① Add the components of phase A in Table 7 into the emulsifying pot in sequence, start stirring and heat to 75-85 degrees;

[0114] ② Add the B phase component to the oil phase pot and heat it to 70-75 degrees, add it to the emulsifying pot and start homogenization;

[0115] ③ Add the C phase components into the emulsifying pot;

[0116] ④Dissolve the phase D components and add them into the emulsifying pot;

[0117] ⑤Add low-temperature materials below 45 degrees, stir evenly, and filter out the materials.

[0118] Table 7 Essence milk formula

[0119]

[0120] 4.2 Essential Oil Application Performance Test

[0121] Thirty-two healthy Chinese women with dry and sensitive skin and troubled by wrinkles such as nasolabial folds and crow's feet were selected. They were aged 20-55 years old, with an average age of 42.52±9.18 years old. They had no other chronic diseases or diseases under treatment, did not use hormone drugs or immunosuppressants, did not participate in other clinical trials, and did not apply topical preparations.

[0122] After cleaning their skin, volunteers took an appropriate amount of essence oil and rubbed it in their palms to warm it up. They evenly pressed it on their facial skin to fully moisturize it, and gently massaged it to penetrate and help soothe and relax the skin. They used it once in the morning and evening every day for 28 consecutive days, and then took the following measurements.

[0123] Skin elasticity Q1 value and skin firmness F4 value

[0124] Cutometer dual MPA580 was used to test the Q1 value of facial skin elasticity and the F4 value of skin firmness. The larger the Q1 value, the better the skin elasticity; the smaller the F4 value, the better the skin firmness. The facial skin data values ​​were tested before use, after 7 days of use, and after 28 days of use.

[0125] Experts assess wrinkle levels

[0126] Experts evaluated the wrinkle levels of crow's feet and nasolabial folds. The smaller the value, the shallower the crow's feet; the smaller the value, the shallower the nasolabial folds. The wrinkle levels were evaluated before use, after 7 days of use, and after 28 days of use.

[0127] The test results can be found in Table 8;

[0128] Table 8 Effects of essential oils on skin elasticity, firmness and wrinkles

[0129] (Sample size = 31, 1 of the original 32 people dropped out of the test due to personal reasons)

[0130]

[0131] The above experimental data show that the composition of the present invention can effectively improve skin firmness and skin elasticity when used in essence oil, and has anti-wrinkle effect.

Claims

1. A composition containing withanolide, characterized in that: The composition consists of meadowfoam seed oil and withanolide; the mass content of withanolide in the meadowfoam seed oil is 10.7-31.7 ppm; The composition is used to improve the performance of meadowfoam seed oil; the performance refers to one or more of skin firming, anti-wrinkle and soothing.

2. The method for preparing the composition according to claim 1, characterized in that: The Withania somnifera extract is dispersed in Limnanthes alba seed oil so that at least a portion of withanolides are dissolved in the Limnanthes alba seed oil.

3. A method for improving the performance of meadowfoam seed oil, characterized in that: The performance of the meadowfoam seed oil is improved by controlling the mass content of withanolide in the meadowfoam seed oil to 10.7-31.7 ppm, wherein the performance refers to one or more of skin firming, anti-wrinkle and soothing.

4. Use of the composition as claimed in claim 1 in preparing cosmetics.

5. The use according to claim 4, characterized in that The dosage form of the cosmetic is essential oil, emulsion or cream.

6. A cosmetic, characterized in that: Contains the composition as claimed in claim 1.

7. The cosmetic according to claim 6, characterized in that: The dosage form of the cosmetic is essential oil, emulsion or cream.

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