Essence oil for promoting skin regeneration and method for preparing the same

Abstract

This invention belongs to the field of cosmetic technology, specifically relating to an essential oil for promoting skin regeneration and its preparation method. By weight, it comprises the following components: 50-60 parts rosehip oil, 6-8 parts lavender essential oil, 8-10 parts rose essential oil, 4-6 parts modified polyaspartic acid penetration enhancer, and 1-3 parts stearyl alcohol-modified recombinant collagen flexible liposomes. In this invention, recombinant collagen is encapsulated in liposomes, and sodium deoxycholate is added during the liposome preparation process. The liposomes encapsulating the recombinant collagen deform as they pass through the intercellular channels of skin cells, adapting to the size of the intercellular channels, thereby improving the absorption of the recombinant collagen and enabling it to reach the dermis, promoting collagen production in the dermis and thus promoting skin regeneration.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to an essential oil that promotes skin regeneration and its preparation method. Background Technology

[0002] As people age, most will experience signs of skin aging. Blood circulation in the skin slows down, and the subcutaneous fat layer becomes loose and loses elasticity. Skin laxity and subcutaneous fat loss can also lead to changes in facial contours such as sagging cheeks and sunken temples. Therefore, skin laxity is one of the major signs of aging.

[0003] In recent years, the thinning of the ozone layer has led to more ultraviolet radiation (especially UVB) reaching the Earth's surface, making the skin more susceptible to photoaging damage. Although the energy of high-energy blue light (HEV) emitted by electronic devices such as mobile phones and computers is lower than that of ultraviolet radiation, long-term exposure may still promote the generation of free radicals, accelerate collagen degradation, and lead to dull skin and aging.

[0004] CN115998649A discloses a firming composition, its preparation method, and its application. This composition utilizes human-derived type III collagen with a core functional region exhibiting a 164.88° bend, a typical triple helix structure, excellent skin affinity, and the ability to accelerate skin regeneration and restore skin barrier function. Furthermore, this invention combines red algae extract and novel copper peptides with human-derived collagen to promote collagen synthesis and firm the skin from multiple angles.

[0005] However, collagen molecules with a triple helix structure are too large and are not easily absorbed after being applied to the skin, making it difficult for them to reach the dermis and exert their effects. Summary of the Invention

[0006] The purpose of this invention is to provide an essential oil that promotes skin regeneration and its preparation method, so as to solve the above-mentioned technical problems.

[0007] To achieve the above-mentioned technical objectives, the technical solution of the present invention is as follows:

[0008] An essential oil for promoting skin regeneration, comprising, by weight, the following components: 50-60 parts rosehip oil, 6-8 parts lavender essential oil, 8-10 parts rose essential oil, 4-6 parts modified polyaspartic acid penetration enhancer, and 1-3 parts stearyl alcohol modified recombinant collagen flexible liposomes.

[0009] The modified polyaspartic acid penetration enhancer is prepared by modifying polyaspartic acid under acidic conditions using stearyl alcohol as a modifier.

[0010] This invention also provides a method for preparing an essential oil that promotes skin regeneration, comprising the following steps:

[0011] S1. Preparation of modified polyaspartic acid penetration enhancer;

[0012] Polyaspartic acid was dissolved in N,N-dimethylformamide and stirred at 60°C until completely dissolved. Stearyl alcohol and 0.1 mol / L concentrated sulfuric acid were added to the solution. The mixture was reacted at 80-100°C for 6-12 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered, dialyzed, and vacuum dried to obtain the modified polyaspartic acid penetration enhancer.

[0013] S2, Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0014] Modified soybean lecithin was dissolved in anhydrous ethanol, and then cholesterol was added to obtain an organic phase. Sodium deoxycholate and recombinant collagen were dissolved in a phosphate solution with a pH of 6.8 to obtain an aqueous phase. The organic and aqueous phases were mixed and sonicated. Then, the mixture was evaporated under reduced pressure in a water bath at 40°C to obtain a gel. After hydration with phosphate solution at 40°C for 20 min, the mixture was immediately cooled and filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0015] Stearyl alcohol was dissolved in anhydrous ethanol and stirred at 60°C for 30 min to obtain a modified solution. The modified solution was slowly added dropwise to a suspension of recombinant collagen flexible liposomes, incubated at 60°C for 1 h, dialyzed for 48 h, and then vacuum dried to obtain stearic acid modified recombinant collagen flexible liposomes.

[0016] S3. Preparation of essential oils that promote skin regeneration;

[0017] Under stirring conditions, rosehip oil, lavender essential oil, and rose essential oil are mixed and heated to 35-40℃. The mixture is stirred at 150 rpm for 30-40 minutes. While stirring, stearyl alcohol is added to modify recombinant collagen flexible liposomes, and stirring is continued for 30 minutes. Finally, modified polyaspartic acid penetration enhancer is added, and the mixture is stirred for 30 minutes before being cooled to room temperature to obtain an essential oil that promotes skin regeneration.

[0018] As a further improvement, in step S1, the preparation method of the polyaspartic acid is as follows: preheat L-aspartic acid in an oven at 100°C for 5 minutes, add 85% by mass of phosphoric acid, stir evenly, and then place it under vacuum drying at 200°C and 0.08MPa for 2 hours. After taking it out, pulverize it to obtain crude polysuccinimide, and then purify it to obtain polysuccinimide.

[0019] Polysuccinimide was mixed with 2.5 mol / L sodium hydroxide solution and stirred at 60 °C for 24 h. After the reaction was completed, the pH was adjusted to 7-8, and the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed, and the precipitate was dried under vacuum to obtain polyaspartic acid.

[0020] As a further improvement, in step S2, the modified soybean lecithin is prepared by mixing hexane, palmitic acid and soybean lecithin, then adding lipase, reacting at 50-60°C for 50 hours, centrifuging and filtering to obtain a mixture, removing the organic solvent by rotary evaporation under reduced pressure, adding cold acetone, centrifuging, taking the precipitate and repeatedly adding cold acetone and centrifuging, repeating 5 times, and then vacuum drying the precipitate to obtain modified soybean lecithin.

[0021] As a further improvement, the ratio of soybean lecithin to n-hexane in the liquid-to-solid mixture is 0.2 g: 1 mL, and the mass ratio of soybean lecithin, palmitic acid, and lipase is 1:2 to 2.2:0.2.

[0022] As a further improvement, in step S2, the mass concentration of modified soybean lecithin in the organic phase is 2.5%, the mass ratio of modified soybean lecithin, cholesterol and sodium deoxycholate is 28:7:4, the volume ratio of the organic phase to the aqueous phase is 3~5:1, and the mass ratio of recombinant collagen to modified soybean lecithin is 1:3~5.

[0023] As a further improvement, in step S2, the concentration of stearyl alcohol in the modification solution is 0.1 g / L, and the volume ratio of the modification solution to the recombinant collagen flexible liposome suspension is 10~12:1.

[0024] As a further improvement, in step S1, the mass ratio of polyaspartic acid to stearyl alcohol is 1:2.5~3.

[0025] As a further improvement, the ratio of L-aspartic acid to phosphoric acid is 5-6 g:1 mL, and the ratio of polysuccinimide to sodium hydroxide solution is 1 g:5-6 mL.

[0026] Due to the adoption of the above technical solution, the beneficial effects of the present invention are as follows:

[0027] This invention provides an essential oil for promoting skin regeneration and its preparation method. The method involves encapsulating recombinant collagen in liposomes and adding sodium deoxycholate during liposome preparation. As the liposomes encapsulate the recombinant collagen, they deform to adapt to the size of the intercellular channels, improving the absorption of the recombinant collagen and enabling it to reach the dermis. This promotes collagen production in the dermis and skin regeneration. The oil is also formulated with lavender and rose essential oils. The citronellol and geraniol in rose essential oil promote collagen synthesis, while the terpenoids in lavender essential oil neutralize free radicals and improve skin problems caused by UV damage.

[0028] This invention adds a modified polyaspartic acid penetration enhancer in combination with stearyl alcohol-modified recombinant collagen flexible liposomes. The polyaspartic acid absorbs moisture from the skin surface, reducing the skin surface water content and creating a water content gradient, which further promotes the absorption of stearyl alcohol-modified recombinant collagen flexible liposomes. The modified polyaspartic acid penetration enhancer remains on the skin surface after absorbing moisture, thus improving the moisturizing performance of the essential oil.

[0029] This invention uses modified soybean lecithin to prepare flexible liposomes. In the preparation process of modified soybean lecithin, lipase is used as a catalyst to increase the content of saturated fatty acids in soybean lecithin by exchanging with palmitate, thereby improving the stability of flexible liposomes.

[0030] This invention modifies recombinant collagen flexible liposomes with stearyl alcohol. By grafting stearyl alcohol onto the surface of the flexible liposomes, the long carbon chains in stearyl alcohol weaken the polarity of the recombinant collagen flexible liposome surface, improve its compatibility with rosehip oil, and prevent the liposomes prepared by palmitic acid-modified soybean lecithin from having strong polarity and poor compatibility with the base oil rosehip oil, thereby improving the stability of the essential oil.

[0031] This invention modifies polyaspartic acid with stearyl alcohol, thereby improving the compatibility of polyaspartic acid with rosehip oil and enhancing the stability of the essential oil. Detailed Implementation

[0032] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. However, those skilled in the art will understand that the embodiments described below are some embodiments of the present invention, but not all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall be followed. Where the manufacturers of reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0033] Example 1: A method for preparing an essential oil that promotes skin regeneration, comprising the following steps:

[0034] Preparation of S1, polyaspartic acid;

[0035] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0036] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0037] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0038] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0039] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0040] S3, Preparation of modified soybean lecithin;

[0041] 10 mL of n-hexane, 4 g of palmitic acid, and 2 g of soybean lecithin were mixed, and then 0.4 g of lipase (Novozym435 enzyme) was added. The mixture was reacted at 50 °C for 50 h. After the reaction, the mixture was centrifuged and filtered to obtain a mixed solution. The n-hexane was removed by rotary evaporation under reduced pressure at 30 °C and 300 mbar. Then, 50 mL of cold acetone was added to precipitate the mixture. The precipitate was centrifuged and the addition of cold acetone was repeated. The process was repeated 5 times. The precipitate was then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0042] S4. Preparation of recombinant collagen flexible liposome suspension;

[0043] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol (density 0.789 g / cm³). 3 The organic phase was obtained by adding 0.7g of cholesterol to the organic phase. 0.4g of sodium deoxycholate and 0.93g of recombinant collagen were dissolved in 47.32mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250W for 10s. Then, the mixture was evaporated under reduced pressure in a water bath at 40℃ and 300mbar to obtain a gel. The gel was hydrated in 50mL of phosphate solution with pH 6.8 at 40℃ for 20min and then immediately cooled. The mixture was filtered through a 0.22μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0044] S5. Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0045] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 100 mL of recombinant collagen flexible liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen flexible liposomes.

[0046] S6. Preparation of essential oils that promote skin regeneration;

[0047] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part stearyl alcohol was added to modify recombinant collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0048] Example 2: A method for preparing an essential oil that promotes skin regeneration, comprising the following steps:

[0049] Preparation of S1, polyaspartic acid;

[0050] 240g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0051] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0052] 20g of polysuccinimide and 120mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 8 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0053] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0054] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 30g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 100℃ for 6h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The modified polyaspartic acid was then vacuum dried at 40℃ and 0.08MPa to obtain the penetration enhancer.

[0055] S3, Preparation of modified soybean lecithin;

[0056] 10 mL of n-hexane, 4.4 g of palmitic acid and 2 g of soybean lecithin were mixed, and then 0.4 g of lipase (Novozym435 enzyme) was added. The mixture was reacted at 60 °C for 50 h. After the reaction, the mixture was centrifuged and filtered to obtain a mixed solution. The n-hexane was removed by rotary evaporation under reduced pressure at 30 °C and 300 mbar. Then, 50 mL of cold acetone was added to precipitate the mixture. The precipitate was centrifuged and the addition of cold acetone was repeated. The process was repeated 5 times. The precipitate was then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0057] S4. Preparation of recombinant collagen flexible liposome suspension;

[0058] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.4 g of sodium deoxycholate and 0.56 g of recombinant collagen were dissolved in 28.39 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0059] S5. Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0060] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 83.3 mL of recombinant collagen flexible liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen flexible liposomes.

[0061] S6. Preparation of essential oils that promote skin regeneration;

[0062] Under stirring conditions, 60 parts of rosehip oil, 8 parts of lavender essential oil, and 10 parts of rose essential oil were mixed, heated to 35°C, and stirred at 150 rpm for 40 minutes. While stirring, 3 parts of stearyl alcohol were added to modify recombinant collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 6 parts of modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0063] Example 3: A method for preparing an essential oil that promotes skin regeneration, comprising the following steps:

[0064] Preparation of S1, polyaspartic acid;

[0065] 220g of L-aspartic acid was preheated in an oven at 100℃ for 5min, 40mL of 85% phosphoric acid was added, and the mixture was stirred evenly. Then it was placed in a vacuum dryer at 200℃ and 0.08MPa for 2h. After drying, it was pulverized to obtain crude polysuccinimide.

[0066] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0067] 20g of polysuccinimide and 110mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7.5 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0068] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0069] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 28g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 90℃ for 9h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0070] S3, Preparation of modified soybean lecithin;

[0071] 10 mL of n-hexane, 4.2 g of palmitic acid and 2 g of soybean lecithin were mixed, and then 0.4 g of lipase (Novozym435 enzyme) was added. The mixture was reacted at 55 °C for 50 h. After the reaction, the mixture was centrifuged and filtered to obtain a mixed solution. The n-hexane was removed by rotary evaporation under reduced pressure at 30 °C and 300 mbar. Then, 50 mL of cold acetone was added to precipitate the mixture. The precipitate was centrifuged and the addition of cold acetone was repeated. The process was repeated 5 times. The precipitate was then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0072] S4. Preparation of recombinant collagen flexible liposome suspension;

[0073] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.4 g of sodium deoxycholate and 0.7 g of recombinant collagen were dissolved in 35.49 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0074] S5. Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0075] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 90.91 mL of recombinant collagen flexible liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen flexible liposomes.

[0076] S6. Preparation of essential oils that promote skin regeneration;

[0077] Under stirring conditions, 55 parts rosehip oil, 7 parts lavender essential oil, and 9 parts rose essential oil were mixed, heated to 37°C, and stirred at 150 rpm for 35 minutes. While stirring, 2 parts stearyl alcohol were added to modify recombinant collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 5 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0078] Comparative Example 1: A method for preparing an essential oil that promotes skin regeneration. The specific steps are the same as in Example 1, except that in this comparative example, recombinant collagen is directly added to the essential oil without liposome encapsulation or stearyl alcohol modification. The specific steps are as follows:

[0079] Preparation of S1, polyaspartic acid;

[0080] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0081] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0082] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0083] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0084] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0085] S3. Preparation of essential oils that promote skin regeneration;

[0086] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part recombinant collagen was added, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0087] Comparative Example 2: A method for preparing an essential oil that promotes skin regeneration. The specific steps are the same as in Comparative Example 1, except that, since recombinant collagen is water-soluble and has poor compatibility with rosehip oil, an emulsifier is added to the essential oil in this comparative example to improve its stability. The specific steps are as follows:

[0088] Preparation of S1, polyaspartic acid;

[0089] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0090] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0091] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0092] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0093] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0094] S3. Preparation of essential oils that promote skin regeneration;

[0095] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed and heated to 40°C. The mixture was stirred at 150 rpm for 30 minutes. While stirring, 1 part recombinant collagen and 2 parts emulsifier soy lecithin were added, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and the mixture was stirred for 30 minutes before being cooled to room temperature to obtain an essential oil that promotes skin regeneration.

[0096] Comparative Example 3: A method for preparing an essential oil that promotes skin regeneration. The specific steps are the same as in Comparative Example 2, except that no modified polyaspartic acid penetration enhancer is added. The specific steps are as follows:

[0097] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part recombinant collagen and 2 parts emulsifier soy lecithin were added, and stirring was continued for 30 minutes. Then, the mixture was cooled to room temperature to obtain an essential oil that promotes skin regeneration.

[0098] Comparative Example 4: A method for preparing an essential oil that promotes skin regeneration, the specific steps of which are the same as in Example 1, except that sodium deoxycholate is not added during the preparation of recombinant collagen flexible liposomes. The specific steps are as follows:

[0099] Preparation of S1, polyaspartic acid;

[0100] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0101] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0102] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0103] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0104] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0105] S3, Preparation of modified soybean lecithin;

[0106] Mix 10 mL of n-hexane, 4 g of palmitic acid and 2 g of soybean lecithin, then add 0.4 g of lipase (Novozym435 enzyme) and react at 50 °C for 50 h. After the reaction, centrifuge and filter to obtain the mixture. Remove n-hexane by rotary evaporation under reduced pressure at 30 °C and 300 mbar, then add 50 mL of cold acetone to precipitate the mixture. Centrifuge the precipitate and repeat the process of adding cold acetone and centrifuging. Repeat this process 5 times. The precipitate obtained is then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0107] S4. Preparation of recombinant collagen flexible liposome suspension;

[0108] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.93 g of recombinant collagen was dissolved in 47.32 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen liposome suspension.

[0109] S5. Preparation of stearyl alcohol-modified recombinant collagen liposomes;

[0110] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 100 mL of recombinant collagen liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen liposomes.

[0111] S6. Preparation of essential oils that promote skin regeneration;

[0112] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part stearyl alcohol was added to modify recombinant collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0113] Comparative Example 5: A method for preparing an essential oil that promotes skin regeneration, the specific steps of which are the same as in Example 1, except that the recombinant collagen flexible liposomes are not modified with stearyl alcohol. The specific steps are as follows:

[0114] Preparation of S1, polyaspartic acid;

[0115] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0116] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0117] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0118] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0119] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0120] S3, Preparation of modified soybean lecithin;

[0121] Mix 10 mL of n-hexane, 4 g of palmitic acid and 2 g of soybean lecithin, then add 0.4 g of lipase (Novozym435 enzyme) and react at 50 °C for 50 h. After the reaction, centrifuge and filter to obtain the mixture. Remove n-hexane by rotary evaporation under reduced pressure at 30 °C and 300 mbar, then add 50 mL of cold acetone to precipitate the mixture. Centrifuge the precipitate and repeat the process of adding cold acetone and centrifuging. Repeat this process 5 times. The precipitate obtained is then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0122] S4. Preparation of recombinant collagen flexible liposome suspension;

[0123] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.4 g of sodium deoxycholate and 0.93 g of recombinant collagen were dissolved in 47.32 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0124] S5. Preparation of essential oils that promote skin regeneration;

[0125] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part recombinant collagen flexible liposome suspension was added, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0126] Comparative Example 6: A method for preparing an essential oil that promotes skin regeneration, the specific steps of which are the same as in Example 1, except that the polyaspartic acid is not modified with stearyl alcohol. The specific steps are as follows:

[0127] Preparation of S1, polyaspartic acid;

[0128] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0129] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0130] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0131] S2, Preparation of modified soybean lecithin;

[0132] Mix 10 mL of n-hexane, 4 g of palmitic acid and 2 g of soybean lecithin, then add 0.4 g of lipase (Novozym435 enzyme) and react at 50 °C for 50 h. After the reaction, centrifuge and filter to obtain the mixture. Remove n-hexane by rotary evaporation under reduced pressure at 30 °C and 300 mbar, then add 50 mL of cold acetone to precipitate the mixture. Centrifuge the precipitate and repeat the process of adding cold acetone and centrifuging. Repeat this process 5 times. The precipitate obtained is then vacuum dried at 40 °C and 0.08 MPa for 24 h to obtain modified soybean lecithin.

[0133] S3. Preparation of recombinant collagen flexible liposome suspension;

[0134] 2.8 g of modified soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.4 g of sodium deoxycholate and 0.93 g of recombinant collagen were dissolved in 47.32 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0135] S4. Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0136] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 100 mL of recombinant collagen flexible liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen flexible liposomes.

[0137] S5. Preparation of essential oils that promote skin regeneration;

[0138] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part stearyl alcohol was added to modify and reorganize collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 4 parts polyaspartic acid were added, and stirring was carried out for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0139] Comparative Example 7: A method for preparing an essential oil that promotes skin regeneration, the specific steps of which are the same as in Example 1, except that the soybean lecithin is not modified. The specific steps are as follows:

[0140] Preparation of S1, polyaspartic acid;

[0141] 200g of L-aspartic acid was preheated in an oven at 100℃ for 5 minutes. 40mL of 85% phosphoric acid was added and stirred evenly. The mixture was then vacuum dried at 200℃ and 0.08MPa for 2 hours. After drying, the mixture was pulverized to obtain crude polysuccinimide.

[0142] The crude polysuccinimide was dissolved in N,N-dimethylformamide and stirred in a 45°C water bath until completely dissolved. The solution was then slowly poured into cold water, and the precipitate was washed with distilled water until neutral. The precipitate was then dried in a 60°C oven to obtain polysuccinimide.

[0143] 20g of polysuccinimide and 100mL of 2.5mol / L sodium hydroxide solution were mixed and stirred at 60℃ for 24h. After the reaction was completed, the pH value was adjusted to 7 with hydrochloric acid. Then the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed. The precipitate was vacuum dried at 60℃ and 0.08MPa to obtain polyaspartic acid.

[0144] S2, Preparation of modified polyaspartic acid penetration enhancer;

[0145] 10g of polyaspartic acid was dissolved in 200mL of N,N-dimethylformamide and stirred at 60℃ until completely dissolved. 25g of stearyl alcohol and 1mL of 0.1mol / L concentrated sulfuric acid were added. The mixture was reacted at 80℃ for 12h under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered and dialyzed for 48h. The molecular weight cutoff was 1000Da. The solid was then vacuum dried at 40℃ and 0.08MPa to obtain the modified polyaspartic acid penetration enhancer.

[0146] S3. Preparation of recombinant collagen flexible liposome suspension;

[0147] 2.8 g of soybean lecithin was dissolved in 141.95 mL of anhydrous ethanol, and then 0.7 g of cholesterol was added to obtain the organic phase. 0.4 g of sodium deoxycholate and 0.93 g of recombinant collagen were dissolved in 47.32 mL of phosphate solution with pH 6.8 to obtain the aqueous phase. The organic and aqueous phases were mixed and sonicated at 250 W for 10 s. Then, the mixture was evaporated under reduced pressure in a water bath at 40 °C and 300 mbar to obtain a gel. The gel was hydrated in 50 mL of phosphate solution with pH 6.8 at 40 °C for 20 min and then immediately cooled. The mixture was filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension.

[0148] S4. Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes;

[0149] 0.1 g stearyl alcohol was dissolved in 1 L of anhydrous ethanol and stirred at 60 °C for 30 min to obtain a modified solution. The modified solution was slowly mixed with 100 mL of recombinant collagen flexible liposome suspension, incubated at 60 °C for 1 h, dialyzed for 48 h, the molecular weight cutoff was 1000 Da, and vacuum dried at 30 °C and 0.1 mbar for 12 h to obtain stearyl alcohol modified recombinant collagen flexible liposomes.

[0150] S5. Preparation of essential oils that promote skin regeneration;

[0151] Under stirring conditions, 50 parts rosehip oil, 6 parts lavender essential oil, and 8 parts rose essential oil were mixed, heated to 40°C, and stirred at 150 rpm for 30 minutes. While stirring, 1 part stearyl alcohol was added to modify recombinant collagen flexible liposomes, and stirring was continued for 30 minutes. Finally, 4 parts modified polyaspartic acid penetration enhancer were added, and stirring was continued for 30 minutes before cooling to room temperature to obtain an essential oil that promotes skin regeneration.

[0152] The transdermal experiments of the essential oils prepared in Examples 1-3 and Comparative Examples 1-7 were conducted as follows: Mice were euthanized by cervical dislocation, and the abdominal skin was peeled off. The abdominal hair was carefully removed, and then the subcutaneous fat and connective tissue were removed. The skin was then rinsed clean with fresh physiological saline to obtain the experimental ex vivo skin, which was stored for later use. Before the experiment, the integrity of the mouse skin was checked, ensuring there were no breaks. The surface water droplets were blotted dry with filter paper.

[0153] In vitro transdermal experiments were conducted using a vertical diffusion cell. Mouse skin was fixed in the diffusion cell (effective permeability area was 3.14 cm²). 2 Add essential oil sample to the supply tank, cover with plastic wrap, and secure with rubber bands; add 19 mL of preheated 32℃ physiological saline to the receiving tank to ensure close contact between the receiving solution and the skin, leaving no cavities. Then place the diffusion tank in a water bath, set the water bath temperature to (32±0.5)℃, and stir with a magnetic stirrer at 300 r / min. Samples were taken at 1h, 2h, 4h, 8h, 12h, and 24h, with 1 mL of sample taken each time and 1 mL of 32℃ physiological saline added simultaneously. The sample solution was filtered through a 0.22μm filter membrane, and 20μL of sample solution was added to a 96-well microplate. Then, 200μL of BCA solution was added, and the absorbance was measured at 562nm. The concentration of recombinant collagen was calculated by substituting the results into the linear equation of the standard curve. The results are shown in Table 1.

[0154] The linear equation of the standard curve is: absorbance = 0.45 × recombinant collagen concentration + 0.0072, r = 0.9932, and the method for determining the standard curve is existing technology.

[0155] Table 1. Transdermal test results of the essential oils prepared in Examples 1-3 and Comparative Examples 1-7

[0156]

[0157] As shown in Table 1, Comparative Examples 1-3 had the worst transdermal absorption effect. This is because neither Comparative Examples 1-2 had the recombinant collagen encapsulated with flexible liposomes, so even with the addition of penetration enhancers, the transdermal absorption effect could not be improved. In Comparative Example 3, no penetration enhancer was added on the basis of adding only recombinant collagen. Compared with Comparative Examples 1-2, the absorption effect was similar. This indicates that penetration enhancers only have a penetration-enhancing effect on essence oils with recombinant collagen flexible liposomes, and have almost no penetration-enhancing effect on essence oils with recombinant collagen.

[0158] In Comparative Example 4, only the recombinant collagen was coated with liposomes. The absorption effect was similar to that of Comparative Examples 1-2, indicating that sodium deoxycholate can deform the liposomes coated with recombinant collagen as they pass through the intercellular channels of skin cells, adapting them to the size of the intercellular channels and improving the absorption of recombinant collagen.

[0159] The transdermal absorption effects of the essential oils in Comparative Examples 5 and 6 were similar to those in Examples 1-3, indicating that the modification and alteration of stearyl alcohol have virtually no impact on the transdermal absorption effect.

[0160] The transdermal absorption effect of the essential oil in Comparative Example 7 was not as good as that in Examples 1-3. This is because the soybean lecithin was not modified, resulting in poor stability of the recombinant collagen flexible liposome suspension, which was easily oxidized, reducing the encapsulation rate of the recombinant collagen and thus worsening the transdermal absorption effect of the recombinant collagen.

[0161] The stability of the essential oils prepared in Examples 1-3 and Comparative Examples 1-7 was tested. All samples were placed in an environment of 50°C for 30 days and 45 days. The stability was determined by whether or not the samples separated into layers. The results are shown in Table 2.

[0162] Table 2. Stability results of the essential oils from Examples 1-3 and Comparative Examples 1-7

[0163]

[0164] As shown in Table 2, in Comparative Example 1, the lack of flexible liposome encapsulation of recombinant collagen led to poor compatibility between the recombinant collagen and rosehip oil, resulting in decreased stability of the essential oil. In Comparative Example 2, the addition of an emulsifier improved the compatibility between the recombinant collagen and rosehip oil. The essential oils in Comparative Examples 5 and 6 showed stratification after 45 days. This is because the polar groups in the flexible nanoliposomes are more active, causing their compatibility with rosehip oil to be affected by temperature and time.

[0165] The specific embodiments of the present invention described above do not constitute a limitation on the scope of protection of the present invention. Any other corresponding changes and modifications made in accordance with the technical concept of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A serum oil for promoting skin regeneration, characterized by, By weight, it includes the following components: 50-60 parts rosehip oil, 6-8 parts lavender essential oil, 8-10 parts rose essential oil, 4-6 parts modified polyaspartic acid penetration enhancer, and 1-3 parts stearyl alcohol modified recombinant collagen flexible liposomes. The modified polyaspartic acid penetration enhancer is prepared by modifying polyaspartic acid with stearyl alcohol as a modifier under acidic conditions. The specific steps are as follows: Polyaspartic acid is dissolved in N,N-dimethylformamide and stirred at 60°C until completely dissolved. Stearyl alcohol and 0.1 mol / L concentrated sulfuric acid are added to the solution. Under nitrogen protection, the reaction is carried out at 80~100°C for 6~12 hours. After the reaction is completed, the reaction solution is poured into cold ethanol to precipitate modified polyaspartic acid. The solid obtained by filtration is dialyzed and vacuum dried to obtain modified polyaspartic acid penetration enhancer. The method for preparing the stearyl alcohol-modified recombinant collagen flexible liposomes is as follows: Modified soybean lecithin was dissolved in anhydrous ethanol, and then cholesterol was added to obtain an organic phase. Sodium deoxycholate and recombinant collagen were dissolved in a phosphate solution with a pH of 6.8 to obtain an aqueous phase. The organic and aqueous phases were mixed and sonicated. Then, the mixture was evaporated under reduced pressure in a water bath at 40°C to obtain a gel. After hydration with phosphate solution at 40°C for 20 min, the mixture was immediately cooled and filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension. Stearyl alcohol was dissolved in anhydrous ethanol and stirred at 60°C for 30 min to obtain a modified solution. The modified solution was slowly added dropwise to a suspension of recombinant collagen flexible liposomes, incubated at 60°C for 1 h, dialyzed for 48 h, and then vacuum dried to obtain stearic acid modified recombinant collagen flexible liposomes. The modified soybean lecithin is prepared by mixing hexane, palmitic acid and soybean lecithin, then adding lipase, reacting at 50-60℃ for 50 hours, centrifuging and filtering after the reaction to obtain a mixture, removing organic solvent by rotary evaporation under reduced pressure, adding cold acetone, centrifuging, taking the precipitate, repeatedly adding cold acetone and centrifuging, repeating 5 times, and then vacuum drying the precipitate to obtain modified soybean lecithin.

2. The method of claim 1, wherein the preparation of the essence oil for promoting skin regeneration is characterized by, Includes the following steps: S1. Preparation of modified polyaspartic acid penetration enhancer; Polyaspartic acid was dissolved in N,N-dimethylformamide and stirred at 60°C until completely dissolved. Stearyl alcohol and 0.1 mol / L concentrated sulfuric acid were added to the solution. The mixture was reacted at 80-100°C for 6-12 hours under nitrogen protection. After the reaction was completed, the reaction solution was poured into cold ethanol to precipitate modified polyaspartic acid. The solid was filtered, dialyzed, and vacuum dried to obtain the modified polyaspartic acid penetration enhancer. S2, Preparation of stearyl alcohol-modified recombinant collagen flexible liposomes; Modified soybean lecithin was dissolved in anhydrous ethanol, and then cholesterol was added to obtain an organic phase. Sodium deoxycholate and recombinant collagen were dissolved in a phosphate solution with a pH of 6.8 to obtain an aqueous phase. The organic and aqueous phases were mixed and sonicated. Then, the mixture was evaporated under reduced pressure in a water bath at 40°C to obtain a gel. After hydration with phosphate solution at 40°C for 20 min, the mixture was immediately cooled and filtered through a 0.22 μm microporous membrane to obtain a recombinant collagen flexible liposome suspension. Stearyl alcohol was dissolved in anhydrous ethanol and stirred at 60°C for 30 min to obtain a modified solution. The modified solution was slowly added dropwise to a suspension of recombinant collagen flexible liposomes, incubated at 60°C for 1 h, dialyzed for 48 h, and then vacuum dried to obtain stearic acid modified recombinant collagen flexible liposomes. S3. Preparation of essential oils that promote skin regeneration; Under stirring conditions, rosehip oil, lavender essential oil, and rose essential oil are mixed and heated to 35-40℃. The mixture is stirred at 150 rpm for 30-40 minutes. While stirring, stearyl alcohol is added to modify recombinant collagen flexible liposomes, and stirring is continued for 30 minutes. Finally, modified polyaspartic acid penetration enhancer is added, and the mixture is stirred for 30 minutes before being cooled to room temperature to obtain an essential oil that promotes skin regeneration.

3. The method of claim 2, wherein the skin regeneration promoting essence oil is prepared by adding 0.1 to 10 parts by weight of the extract of claim 1 to 100 parts by weight of a base oil. In step S1, the preparation method of polyaspartic acid is as follows: L-aspartic acid is preheated in an oven at 100°C for 5 minutes, 85% by mass of phosphoric acid is added, the mixture is stirred evenly, and then it is vacuum dried at 200°C and 0.08MPa for 2 hours. After being taken out, it is pulverized to obtain crude polysuccinimide, which is then purified to obtain polysuccinimide. Polysuccinimide was mixed with 2.5 mol / L sodium hydroxide solution and stirred at 60 °C for 24 h. After the reaction was completed, the pH was adjusted to 7-8, and the solution was added dropwise to anhydrous ethanol. After standing and precipitation, the liquid was removed, and the precipitate was dried under vacuum to obtain polyaspartic acid.

4. The method for preparing the skin-regenerating essential oil according to claim 2, characterized in that, In step S2, the modified soybean lecithin is prepared by mixing hexane, palmitic acid and soybean lecithin, then adding lipase, reacting at 50-60℃ for 50 hours, centrifuging and filtering after the reaction to obtain a mixture, removing the organic solvent by rotary evaporation under reduced pressure, adding cold acetone, centrifuging, taking the precipitate, repeatedly adding cold acetone and centrifuging, repeating 5 times, and then vacuum drying the precipitate to obtain modified soybean lecithin.

5. The method for preparing the skin-regenerating essential oil according to claim 4, characterized in that, The ratio of soybean lecithin to n-hexane in the liquid-to-solid mixture is 0.2 g: 1 mL, and the mass ratio of soybean lecithin, palmitic acid, and lipase is 1:2 to 2.2:0.

2.

6. The method for preparing the skin-regenerating essential oil according to claim 2, characterized in that, In step S2, the mass concentration of modified soybean lecithin in the organic phase is 2.5%, the mass ratio of modified soybean lecithin, cholesterol and sodium deoxycholate is 28:7:4, the volume ratio of the organic phase to the aqueous phase is 3~5:1, and the mass ratio of recombinant collagen to modified soybean lecithin is 1:3~5.

7. The method for preparing the skin-regenerating essential oil according to claim 2, characterized in that, In step S2, the concentration of stearyl alcohol in the modification solution is 0.1 g / L, and the volume ratio of the modification solution to the recombinant collagen flexible liposome suspension is 10~12:

1.

8. The method for preparing the skin-regenerating essential oil according to claim 2, characterized in that, In step S1, the mass ratio of polyaspartic acid to stearyl alcohol is 1:2.5~3.

9. The method for preparing the skin-regenerating essential oil according to claim 3, characterized in that, The ratio of L-aspartic acid to phosphoric acid in the liquid solution is 5-6 g: 1 mL, and the ratio of polysuccinimide to sodium hydroxide solution in the liquid solution is 1 g: 5-6 mL.

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