Lindera reflexa hemsl essential oil liposome emulsifiable paste as well as preparation method and application thereof

Abstract

The invention relates to the technical field of emulsifiable paste preparation, in particular to lindera reflexa hemsl essential oil liposome emulsifiable paste and a preparation method and application thereof. The preparation method comprises the following steps: dissolving the lindera reflexa hemsl essential oil, lecithin and cholesterol in a solvent, and evaporating the solvent to form a reaction material; performing ultrasonic mixing on the reaction material and a pH regulator to obtain lindera reflexa essential oil liposome; heating and mixing the lindera reflexa hemsl essential oil liposome and a first emulsifier to form a water phase solution; heating and melting liquid paraffin, octadecanol and a second emulsifier to form an oil-phase mixture; and mixing the oil phase mixture with the water phase solution, and emulsifying to obtain the lindera reflexa hemsl essential oil liposome emulsifiable paste. The lindera reflexa hemsl essential oil and the preparation method thereof have the advantages that the problem of poor stability of the lindera reflexa hemsl essential oil is solved, the bioavailability and the release efficiency of the lindera reflexa hemsl essential oil are improved through the controlled release effect of the lindera reflexa hemsl essential oil liposome, meanwhile, the preparation method of the emulsifiable paste is simple and easy to implement and has the potential of industrial production, and a new way is opened up for deep processing of lindera reflexa

Description

Technical Field

[0001] The invention relates to the technical field of cream preparation, and in particular to a liposome cream containing bergamot essential oil and a preparation method and application thereof. Background Art

[0002] Sanchoke, also known as mountain ash and vice mountain ash, is the dried root of the lauraceae plant Sanchoke. It grows in valleys, hillside forests or bushes below an altitude of about 1,000 meters. It is distributed in provinces south of the Yangtze River, and its main production area is the Dabie Mountain area in Xinyang. Sanchoke has the effects of promoting qi and relieving pain, promoting blood circulation and reducing swelling, and strengthening the spleen and eliminating accumulation. It is mainly used to treat scabies, allergic dermatitis, stomachache, and bleeding from knife wounds. Studies have found that Sanchoke essential oil is one of its active ingredients, but Sanchoke essential oil has the disadvantages of poor stability, easy volatility, and easy oxidation and decomposition when exposed to light and heat, which seriously affects the efficacy of Sanchoke.

[0003] In the prior art, it is an effective method to make the ash essential oil into a β-cyclodextrin inclusion complex, which can improve the stability of the ash essential oil and reduce its volatility through inclusion. However, although this method improves the stability of the ash essential oil, the bioavailability and release efficiency of the inclusion complex are still limited by the properties of the inclusion material and its interaction with the ash essential oil. While the ash essential oil is directly mixed with an emulsifier and prepared into a cream form, the ash essential oil in the cream matrix is ​​still easy to evaporate quickly during preparation, storage and even use, which not only weakens the overall efficacy of the ash essential oil product, but also causes skin irritation due to the exposure of the ash essential oil, reducing the patient's comfort and treatment compliance. Summary of the invention

[0004] In view of the problems existing in the prior art, the present invention provides a kalanchoe essential oil liposome cream and a preparation method and application thereof. The present invention mixes kalanchoe essential oil with lecithin and cholesterol, adjusts the pH to obtain kalanchoe essential oil liposomes, and uses kalanchoe essential oil liposomes as raw materials and adopts a double emulsification system to prepare kalanchoe essential oil liposome cream. In the kalanchoe essential oil liposome cream obtained by the preparation method of the present invention, the lipid bilayer structure formed by lecithin and cholesterol overcomes the problem of poor stability of kalanchoe essential oil itself; the good biocompatibility of kalanchoe essential oil liposomes also improves the bioavailability of kalanchoe essential oil. At the same time, the preparation method of the kalanchoe essential oil liposome cream is simple and easy, and has the potential for industrial production, which opens up a new way for the deep processing of kalanchoe medicinal materials and the development and application of antibacterial agents.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] The first object of the present invention is to provide a method for preparing a liposome cream of yam essential oil, comprising the following steps:

[0007] S1. After dissolving the bergamot essential oil, lecithin and cholesterol in a solvent, a thin film dispersion method is used to evaporate the mixture in an eggplant-shaped bottle of a rotary evaporator to form a reaction material; wherein the dissolution temperature is 40°C.

[0008] S2. Ultrasonic mixing of the reaction material and the pH regulator to obtain the bergamot essential oil liposomes.

[0009] S3, heating and mixing the hawthorn essential oil liposomes and the first emulsifier to form an aqueous phase solution.

[0010] S4, heating and melting the liquid paraffin, stearyl alcohol and the second emulsifier to form an oil phase mixture.

[0011] S5. Mix the oil phase mixture with the aqueous phase solution and emulsify it to obtain the bergamot essential oil liposome cream.

[0012] Preferably, the volume of the ash essential oil, the mass ratio of lecithin and cholesterol is 1mL to 4mL: 1g to 5g: 1g. Among them, lecithin is a natural phospholipid with a hydrophilic head and a hydrophobic tail, which can spontaneously form a bilayer structure in an aqueous solution, the so-called lipid bilayer structure. The mass ratio of the volume of ash essential oil to lecithin directly affects the encapsulation rate and stability of ash essential oil liposomes. When the ratio of ash essential oil to lecithin is 2mL: 1g, the ash essential oil liposome encapsulation rate is the highest; when the amount of ash essential oil added is too much, it will exceed the carrying capacity of lecithin, and when the amount added is too little, the ash essential oil liposomes are unstable and prone to leakage, resulting in a reduced encapsulation rate.

[0013] The addition of cholesterol can adjust the fluidity and rigidity of the bilayer, enhance the stability of the lipid bilayer structure, and prevent it from breaking or leaking during preparation and storage. The mass ratio of lecithin to cholesterol determines the rigidity of the liposome membrane and the stability of the overall structure of the liposome. When the ratio of lecithin to cholesterol is 3g:1g, the liposome encapsulation rate of the liposome is the highest; when the amount of cholesterol increases, the bilayer of the liposome will exceed the capacity limit, causing the destruction of the bilayer of the liposome; and when the cholesterol is too little, the rigidity of the liposome membrane of the liposome is insufficient, which will also lead to the instability of the liposome membrane of the liposome.

[0014] Preferably, the pH regulator is a phosphate buffer, and the pH of the bergamot essential oil liposomes is adjusted to 7-7.2 by using the pH regulator to ensure the stability of the liposomes.

[0015] Preferably, the emulsification treatment is carried out under the conditions of: stirring at 75° C. to 80° C. for 2 to 3 minutes.

[0016] Preferably, the ratio of the oil phase mixture to the water phase solution is 1-2:3.

[0017] Preferably, the mass ratio of the hawthorn essential oil liposome to the first emulsifier is 10-15:1, and the mass ratio of the total mass of the liquid paraffin and octadecyl alcohol to the second emulsifier is 16-20:1; wherein the first emulsifier is Tween-80 and the second emulsifier is Span-80.

[0018] Preferably, in the emulsification treatment, the total mass of the first emulsifier and the second emulsifier accounts for 4% to 8% of the total mass of the mixture of the aqueous phase solution and the oil phase.

[0019] Preferably, the solvent is selected from anhydrous ethanol, ether, chloroform or dichloromethane, but anhydrous ethanol is commonly used in consideration of safety.

[0020] Preferably, the bergamot essential oil is prepared according to the following steps:

[0021] The agave calendula medicinal material is crushed, soaked in water and then distilled to obtain the agave calendula essential oil.

[0022] Preferably, the soaking time is 0 h to 2 h, and the distillation treatment time is 1 h to 4 h.

[0023] The second object of the present invention is to provide a liposome cream of yam essential oil prepared by the above preparation method.

[0024] The third object of the present invention is to provide the use of the above-mentioned yam essential oil liposome cream in the preparation of an antibacterial agent, wherein the antibacterial agent inhibits the activity of bacteria.

[0025] Preferably, the bacteria is Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli or Candida albicans.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. The present invention provides a method for preparing a kaempferia essential oil liposome cream. First, kaempferia essential oil, lecithin and cholesterol are dissolved in a solvent and evaporated to form a film, then a pH regulator is added to the film, and kaempferia essential oil liposomes are obtained by ultrasonic treatment; then, the kaempferia essential oil liposomes are heated and mixed with a first emulsifier to prepare an aqueous phase solution; at the same time, liquid paraffin, octadecyl alcohol and a second emulsifier are heated and melted to form an oil phase mixture; finally, the oil phase mixture is slowly added to the aqueous phase solution and emulsified to successfully prepare the kaempferia essential oil liposome cream. This method not only overcomes the problem of poor stability of kaempferia essential oil itself, but also improves its bioavailability and release efficiency through the controlled release effect of kaempferia essential oil liposomes, opening up a new way for the deep processing of kaempferia medicinal materials and the development and application of antibacterial agents.

[0028] Among them, lecithin and cholesterol are dissolved in a solvent, and a reaction material is formed by evaporating the solvent. Subsequently, a pH regulator is added to the reaction material, and ultrasonic treatment is used to promote the rearrangement of the phospholipid molecules of lecithin into a lipid bilayer structure. The kaempferia essential oil is encapsulated in the lipid bilayer structure to form kaempferia essential oil liposomes. The lipid bilayer structure not only provides a physical barrier for the kaempferia essential oil, preventing it from directly contacting the external environment and reducing the risk of oxidation and volatilization, but also protects the internally encapsulated kaempferia essential oil through its stable membrane structure.

[0029] As an artificially synthesized membrane structure, the bilayer membrane of the yam essential oil liposome is similar to the biological cell membrane in structure and composition. It is composed of two layers of parallel phospholipid molecules, the hydrophilic head of the phospholipid molecules faces the water phase, and the hydrophobic tails gather together on the inside to form a stable bilayer membrane structure. At the same time, the yam essential oil liposome is used as the oil phase and encapsulated in a larger water-in-oil emulsion to form a double emulsion structure. The double emulsion system further encapsulates the yam essential oil liposome in the inner phase of the water-in-oil emulsion. This double protection not only provides an additional protective layer for the yam essential oil liposome, enhances the overall stability, but also helps to control the release rate of the active ingredients. In addition, the cell membrane structure of the yam essential oil liposome and the amphiphilic properties of the emulsifier give the yam essential oil liposome cream excellent biocompatibility and permeability, so that the active ingredients of the yam essential oil can be more effectively absorbed and utilized by the skin, thereby exerting better health and therapeutic effects.

[0030] 2. The preparation method of the agave pine essential oil liposome cream of the present invention optimizes the preparation process of the agave pine essential oil liposome cream by investigating and screening the effects of soaking time, extraction time and amount of water added on the extraction rate of the agave pine essential oil, as well as the effect of the ratio between the volume of the agave essential oil, the mass of lecithin and the mass of cholesterol on the encapsulation rate of the agave essential oil liposome, thereby providing a theoretical basis for the resource utilization of local agave pine medicinal materials and the development of deep-processing products.

[0031] 3. The present invention provides a liposome cream of bergamot essential oil. The bergamot essential oil contains eucalyptol and camphor components, which can destroy the cell membrane and cell wall of bacteria, resulting in increased cell permeability and leakage of macromolecular substances, thereby inhibiting the growth and reproduction of bacteria and showing antibacterial activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a graph showing the effect of soaking time on the extraction rate of ash essential oil.

[0033] Figure 2 This is a graph showing the effect of extraction time on the extraction rate of ash essential oil.

[0034] Figure 3 This is a graph showing the effect of water addition on the extraction rate of kaempferia oxyphylla essential oil.

[0035] Figure 4 This is a UV detection scan of the bergamot essential oil in Example 3.

[0036] Figure 5 This is a graph showing the effect of the volume of the yam essential oil and the mass ratio of lecithin on the liposome encapsulation efficiency of the yam essential oil in Examples 1 to 5.

[0037] Figure 6 This is a graph showing the effect of the mass ratio of lecithin to cholesterol on the liposome encapsulation efficiency of the hawthorn essential oil in Examples 3 and 6 to 9.

[0038] Figure 7 This is a graph showing the effect of the volume of phosphate buffer solution on the liposome encapsulation efficiency of the hawthorn essential oil in Examples 3 and 10 to 13.

[0039] Figure 8 This is the appearance diagram of the liposomes of the hawthorn essential oil of Example 3.

[0040] Fig. 9 This is an optical microscope morphology of the hawthorn essential oil liposomes in Example 3.

[0041] Fig.10 This is the appearance picture of the finished product of the hawthorn essential oil liposome cream of Example 3.

[0042] Fig.11 The figures are the results of the antibacterial experiment of the hawthorn essential oil liposome cream in Example 3, wherein (a) is the result of the experimental group with the addition of hawthorn essential oil liposome cream, bacterial solution and corresponding culture medium, and (b) is the result of the positive control group with only bacterial solution and corresponding culture medium. DETAILED DESCRIPTION

[0043] The technical solution of the present invention will be clearly and completely described below in combination with the data in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0044] It should be noted that the professional terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the following experimental methods and detection methods of the present invention are conventional methods unless otherwise specified; the following reagents and raw materials are commercially available reagents and raw materials unless otherwise specified.

[0045] In the prior art, ashwagandha essential oil has attracted much attention due to its unique pharmacological effects, but its poor stability, volatility, and easy oxidative decomposition under light and high temperature conditions have always been technical bottlenecks that restrict its widespread application. Although researchers have tried to improve its stability by β-cyclodextrin inclusion, there are still deficiencies in bioavailability and release efficiency. At the same time, directly mixing ashwagandha essential oil with an emulsifier to prepare a cream will cause the ashwagandha essential oil in the cream to dissipate quickly, affecting the efficacy, and the irritating components in ashwagandha essential oil will cause skin discomfort and reduce patient compliance.

[0046] In view of the problems existing in the prior art, the present invention provides a method for preparing a kalanchoe essential oil liposome cream, comprising the following steps: dissolving kalanchoe essential oil, lecithin and cholesterol in a solvent, evaporating the solvent to form a reaction material; ultrasonically mixing the reaction material with a pH regulator to obtain kalanchoe essential oil liposomes; heating and mixing the kalanchoe essential oil liposomes with a first emulsifier to form an aqueous solution; heating and melting liquid paraffin, octadecyl alcohol and a second emulsifier to form an oil phase mixture; adding the oil phase mixture to the aqueous solution for emulsification treatment to obtain the kalanchoe essential oil liposome cream.

[0047] The present invention extracts kaempferia essential oil, prepares kaempferia essential oil liposomes, and uses kaempferia essential oil liposomes as raw materials to prepare kaempferia essential oil liposome cream. First, kaempferia essential oil is encapsulated in a lipid bilayer structure to form kaempferia essential oil liposomes, and then a double emulsification system is used to further encapsulate the kaempferia essential oil liposomes in the inner phase of an oil-in-water emulsion. This double protection not only greatly improves the stability of the kaempferia essential oil, but also prevents the oxidation and volatilization of the kaempferia essential oil liposomes during storage and use, and at the same time improves the transdermal absorption rate and bioavailability of the kaempferia essential oil.

[0048] The technical solution of the present invention is further explained by using embodiments and comparative examples, as shown below:

[0049] In order to investigate the effect of the ratio between koreana essential oil, lecithin and cholesterol on the liposome encapsulation efficiency of koreana essential oil, single factor experiments were performed with the mass ratio of koreana essential oil volume to lecithin, the mass ratio of lecithin to cholesterol and the volume of phosphate buffer as influencing factors, as shown in Examples 1 to 20:

[0050] Example 1

[0051] A method for preparing a liposome cream of yam essential oil comprises the following steps:

[0052] S1. Weigh 100 g of Rhizoma Cibotii medicinal materials, crush them, add 1000 mL of water to soak for 2 h, and then perform steam distillation extraction in a volatile oil extraction device for 5 h to obtain Rhizoma Cibotii essential oil.

[0053] S2. Place 0.45 mL of bergamot essential oil, 1.35 g of lecithin and 0.45 g of cholesterol in an eggplant-shaped bottle, add 10 mL of anhydrous ethanol, dissolve in a 40° C. water bath, perform rotary evaporation in a rotary evaporator and form a uniform film on the wall, i.e., the reaction material.

[0054] S3. Add 20 mL of phosphate buffer with a pH of 7.2 to the reaction material and perform ultrasonic treatment for 30 min to obtain the safflower essential oil liposomes.

[0055] S4. Add 1.5 mL of Tween-80 to the liposomes of the hawthorn essential oil, heat and mix, and form an aqueous phase solution.

[0056] S5. Heat 13.5 mL of liquid paraffin, 2.0 mL of octadecyl alcohol, and 0.5 mL of Span-80 at 70° C. to melt to form an oil phase mixture.

[0057] S6. Add the oil phase mixture into the aqueous phase solution for emulsification to obtain the bergamot essential oil liposome cream.

[0058] Example 2

[0059] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 1, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 1.5 mL: 3 g: 1 g, that is, in step S2, 0.45 mL of bergamot essential oil, 0.9 g of lecithin and 0.3 g of cholesterol are placed in an eggplant-shaped bottle.

[0060] Example 3

[0061] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 1, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 3 mL: 3 g: 1 g, that is, in step S2, 0.45 mL of bergamot essential oil, 0.45 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0062] Example 4

[0063] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 1, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 3 mL: 3 g: 1 g, that is, in step S2, 0.9 mL of bergamot essential oil, 0.45 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0064] Example 5

[0065] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 1, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 12 mL: 3 g: 1 g, that is, in step S2, 1.35 mL of bergamot essential oil, 0.45 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0066] Example 6

[0067] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 3, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 1 mL: 1 g: 1 g, that is, in step S2, 0.45 mL of bergamot essential oil, 0.45 g of lecithin and 0.45 g of cholesterol are placed in an eggplant-shaped bottle.

[0068] Example 7

[0069] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 3, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 2 mL: 2 g: 1 g, that is, in step S2, 0.3 mL of bergamot essential oil, 0.3 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0070] Example 8

[0071] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 3, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 4 mL: 4 g: 1 g, that is, in step S2, 0.6 mL of bergamot essential oil, 0.6 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0072] Example 9

[0073] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 3, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 5 mL: 5 g: 1 g, that is, in step S2, 0.75 mL of bergamot essential oil, 0.75 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0074] Example 10

[0075] A method for preparing a liposome cream containing bergamot essential oil is the same as the method in Example 3, except that in step S3, the volume of the phosphate buffer is 10 mL.

[0076] Embodiment 11

[0077] A method for preparing a safflower essential oil liposome cream is the same as the preparation method of Example 3, except that in step S3, the volume of the phosphate buffer is 30 mL.

[0078] Example 12

[0079] A method for preparing a liposome cream containing bergamot essential oil is the same as the method in Example 3, except that in step S3, the volume of the phosphate buffer is 40 mL.

[0080] Embodiment 13

[0081] A method for preparing a liposome cream containing bergamot essential oil is the same as the method in Example 3, except that in step S3, the volume of the phosphate buffer is 50 mL.

[0082] Embodiment 14

[0083] A method for preparing a safflower essential oil liposome cream is the same as the preparation method of Example 7, except that in step S3, the volume of the phosphate buffer is 10 mL.

[0084] Embodiment 15

[0085] A method for preparing a safflower essential oil liposome cream is the same as the preparation method of Example 8, except that in step S3, the volume of the phosphate buffer is 30 mL.

[0086] Example 16

[0087] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 10, except that in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 4 mL: 2 g: 1 g, that is, in step S2, 0.6 mL of bergamot essential oil, 0.3 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0088] Embodiment 17

[0089] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 11, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 8 mL:4 g:1 g, that is, in step S2, 1.2 mL of bergamot essential oil, 0.6 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0090] Embodiment 18

[0091] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 10, except that, in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 8 mL:4 g:1 g, that is, in step S2, 1.2 mL of bergamot essential oil, 0.6 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0092] Embodiment 19

[0093] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 11, except that in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 8 mL:2 g:1 g, that is, in step S2, 1.2 mL of bergamot essential oil, 0.3 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0094] Embodiment 20

[0095] A method for preparing a bergamot essential oil liposome cream is the same as the preparation method of Example 11, except that in step S2, the volume of the bergamot essential oil, the mass ratio of lecithin to cholesterol is 16 mL: 4 g: 1 g, that is, in step S2, 2.4 mL of bergamot essential oil, 0.6 g of lecithin and 0.15 g of cholesterol are placed in an eggplant-shaped bottle.

[0096] Comparative Example 1

[0097] A method for preparing a liposome cream of yam essential oil, which is prepared by a dry gel method, comprises the following steps:

[0098] S1. Weigh 100 g of Rhizoma Cibotii medicinal materials, crush them, add 1000 g of water and soak them for 2 h, then perform steam distillation extraction in a volatile oil extraction device for 5 h to obtain Rhizoma Cibotii essential oil.

[0099] S2. 0.45 mL of bergamot essential oil, 0.45 g of lecithin and 0.15 g of cholesterol were placed in an eggplant-shaped bottle, anhydrous ethanol was added to dissolve in a water bath at 40° C., and then rotary evaporated in a rotary evaporator to form a uniform film on the wall, i.e., the reaction material.

[0100] S3. Add 20 mL of phosphate buffer having a pH of 7.2 to the reaction material and perform ultrasonic treatment for 30 min to obtain bergamot essential oil liposomes.

[0101] S4. Weigh 5 g of gum arabic powder, add 28 mL of liquid paraffin to grind, then add 10 mL of bergamot essential oil liposome to grind to form water-in-oil colostrum, then add 40 mL of bergamot essential oil liposome to continue grinding to form an oil-in-water cream, i.e., bergamot essential oil liposome cream.

[0102] Comparative Example 2

[0103] A method for preparing a liposome cream of yam essential oil is prepared by a wet gel method, and the preparation steps are the same as those of comparative example 1, except that S4 comprises the following steps:

[0104] S4. Weigh 5 g of gum arabic powder, add 10 mL of bergamot essential oil liposomes and grind them, then add 28 mL of liquid paraffin and continue grinding to form water-in-oil colostrum, then add 40 mL of bergamot essential oil liposomes and continue grinding to form an oil-in-water cream, i.e., bergamot essential oil liposome cream.

[0105] Comparative Example 3

[0106] A method for preparing a liposome cream of yam essential oil is prepared by a wet gel method, and the preparation steps are the same as those of comparative example 1, except that S4 comprises the following steps:

[0107] S4. Weigh 50 mL of the bergamot essential oil liposomes, add 0.8 g of potassium carbonate to the weighed bergamot essential oil liposomes and mix well, then add 28 g of stearic acid and mix well to form an oil-in-water cream, i.e., the bergamot essential oil liposome cream.

[0108] In order to investigate the effects of soaking time, extraction time, and water addition on the extraction rate of kaempferia essential oil, single factor tests were conducted with soaking time, extraction time, and water addition, as shown below:

[0109] a. Soaking time: Weigh 5 portions of 100g of the Herba Aspergillus niger medicinal materials, extract for 4h, add 10 times the amount of water, soak for 0h, 0.5h, 1h, 1.5h, and 2h respectively, and calculate the extraction rate of Herba Aspergillus niger essential oil to investigate the effect of soaking time on the extraction rate of Herba Aspergillus niger essential oil.

[0110] b. Extraction time: Weigh 5 portions of 100g of the Rhizoma Cibotii medicinal material, soak for 1 hour, add 10 times the amount of water, and the extraction times are 2h, 3h, 4h, 5h, and 6h, respectively. Extraction is carried out in sequence, and the extraction rate of Rhizoma Cibotii essential oil is calculated to examine the effect of extraction time on the extraction rate of Rhizoma Cibotii essential oil.

[0111] c. Amount of water added: Weigh 5 portions of 100g of the Rhizoma Cibotii medicinal materials, soak for 1h, extract for 4h, add 6 times, 8 times, 10 times, 12 times, and 14 times of water respectively for extraction, and calculate the extraction rate of Rhizoma Cibotii essential oil to examine the effect of the amount of water added on the extraction rate of Rhizoma Cibotii essential oil.

[0112] On the basis of the single factor test, soaking time (A), extraction time (B), and water addition (C) were selected as influencing factors. According to the results of the single factor test, three levels were selected for each factor. The L9 (3 3) orthogonal table, as shown in Table 1, an orthogonal optimization test was carried out, and the extraction rate of the essential oil of the yam was used as the evaluation index. The results are shown in Table 2.

[0113] Table 1 Factor level table of orthogonal experimental design for extracting yam essential oil

[0114]

[0115]

[0116] Table 2 Orthogonal test results of extracting yam essential oil

[0117]

[0118] From the analysis of the results in Table 1-2, it can be concluded that the best combination for extracting ash essential oil is A3B3C2, that is, the soaking time is 2h, the extraction time is 5h, and the amount of water added is 10 times.

[0119] By comparing the range R, we know that the factor that has the greatest impact on the extraction rate of ash essential oil is the extraction time, followed by the amount of water added, and finally the soaking time. That is, the order of influencing factors is: extraction time> amount of water added> soaking time. Three groups of parallel experiments were carried out according to the optimal level combination A3B3C2 to obtain Table 3.

[0120] Table 3 Results of the validation test of the extraction of yam essential oil

[0121] Number of trials 1 2 3 Mean Extraction rate (%) 0.72 0.75 0.69 0.72

[0122] The average extraction rate of the essential oil of the yam is 0.72%, which is higher than any group in the orthogonal test group. Therefore, the experiment is considered feasible.

[0123] Figure 1 The effect of soaking time on the extraction rate of ash essential oil is shown in Figure 2. Figure 1 It was found that when the soaking time was 1.5h, the extraction rate of argyrocarpon essential oil was the highest, at 0.67%. When the soaking time was 1h and 2h, the extraction rate of argyrocarpon essential oil was higher. The extraction rate of argyrocarpon essential oil increased as the soaking time increased. After the soaking time was greater than 1.5h, the extraction rate of argyrocarpon essential oil reached its peak and no longer increased. It was found that the optimal soaking time was 1.5h.

[0124] Figure 2 The effect of extraction time on the extraction rate of Kaempferia oxyphylla essential oil is shown in Figure 2. Figure 2 The results show that when the extraction time is 4h, the extraction rate of agave ash essential oil is the highest, which is 0.66%. The extraction rate of agave ash essential oil increases with the increase of extraction time. When the extraction time is 4h, the extraction rate of agave ash essential oil reaches its peak, and the subsequent increase in extraction time has basically no effect on the extraction rate of agave ash essential oil. It can be concluded that the optimal extraction time for agave ash essential oil is 4h.

[0125] Figure 3 The effect of water addition on the extraction rate of Kaempferia oxyphylla essential oil is shown in Figure 2. Figure 3 It was concluded that the amount of water added in this experiment had little effect on the extraction rate of the kaempferia oxyphylla essential oil. When the amount of water added was 10 times the amount, the extraction rate of the kaempferia oxyphylla essential oil was the highest, at 0.66%. Therefore, the optimal amount of water added was 10 times the amount.

[0126] Figure 4 This is the ultraviolet detection scan of the yam essential oil of Example 3, Figure 4 It was concluded that the essential oil of Osmanthus fragrans had a maximum absorption peak near 296nm, so 296nm was selected as the detection wavelength for subsequent related measurements.

[0127] On the basis of the above-mentioned single factor test of the extraction of ash essential oil, the immersion time of 2h, the extraction time of 5h and the water addition amount of 10 times were taken as the best ash essential oil extraction conditions, and the ash essential oil liposome cream prepared in Examples 1 to 20 was obtained. On the basis of Examples 1 to 20, the mass ratio of the volume of ash essential oil to lecithin (A), the mass ratio of lecithin to cholesterol (B), and the volume of phosphate buffer solution (C) were selected as influencing factors. According to the results of the single factor test, three levels were selected for each factor, and L9 (3 3 ) orthogonal table, as shown in Table 4, an orthogonal optimization experiment was carried out, with the liposome encapsulation efficiency of the hawthorn essential oil as the evaluation index, and the results are shown in Table 5.

[0128] Table 4. Orthogonal experimental design factor level table for the preparation of liposomes of yam essential oil

[0129]

[0130]

[0131] Selection of UV detection wavelength for kaempferia essential oil: Place 0.02 mL of kaempferia essential oil in a 50 mL volumetric flask, add methanol solution, and dilute to the mark. Use methanol solution as a blank control and perform UV scanning at a wavelength of 200 nm to 400 nm. Determine the maximum absorption wavelength of kaempferia essential oil by scanning the spectrum.

[0132] Determination of the encapsulation efficiency of the shanju essential oil liposome: The encapsulation efficiency of the shanju essential oil liposome was determined by organic solvent extraction method. 1 mL of the shanju essential oil liposome was accurately measured, and 5 mL of methanol was added. The total absorbance of the free and encapsulated drugs was measured at a wavelength of 296 nm with methanol as the control, and recorded as A1; 2 mL of the shanju essential oil liposome was accurately measured and placed in a centrifuge tube, and petroleum ether was added for oscillation extraction. After standing and stratification, the lower layer of the suspension was taken, 1 mL of the suspension was accurately measured, 5 mL of methanol was added, and the absorbance was measured at a wavelength of 296 nm with methanol as the control, as the absorbance of the encapsulated shanju essential oil, recorded as A2. The encapsulation efficiency of the shanju essential oil liposome was calculated according to formula (1).

[0133] Encapsulation efficiency EE (%) = A2 / A1 × 100% .................... (1)

[0134] Table 5 Orthogonal test results for the preparation of liposomes of yam essential oil

[0135]

[0136]

[0137] From the results in Table 5, it can be seen that the optimal conditions for the preparation of liposomes of kaempferia essential oil are A1B2C2, that is, the ratio of kaempferia essential oil to lecithin is 1mL:1g, the ratio of lecithin to cholesterol is 3g:1g, and the volume of phosphate buffer solution is 20mL.

[0138] By comparing the range R, we know that the factor that has the greatest impact on the encapsulation rate of the liposomes of the kaempferia essential oil is the volume of the phosphate buffer solution, followed by the ratio of lecithin to cholesterol, and finally the ratio of the kaempferia essential oil to lecithin. That is, the order of the factors is: the volume of the phosphate buffer solution > the ratio of lecithin to cholesterol > the ratio of the kaempferia essential oil to lecithin. Table 6 is a table of repeated validation test results for the preparation of the liposomes of the kaempferia essential oil, as follows:

[0139] Table 6 Results of repeated validation test for preparation of liposomes of yam essential oil

[0140] Number of trials 1 2 3 Mean Encapsulation rate (%) 88.93 89.76 90.67 89.79

[0141] According to the optimal combination A1B2C2 of the above orthogonal results, three parallel experiments were carried out, and the average encapsulation rate of the liposomes of the hawthorn essential oil was 89.79%, which was higher than the encapsulation rate of any group in the orthogonal test. Therefore, this experimental combination, namely Example 3, is feasible.

[0142] Figure 5 The figure is the effect of the ratio of the ash essential oil and lecithin on the liposome encapsulation efficiency of the ash essential oil in Examples 1 to 5. Figure 5It was concluded that when the ratio of ashwagandha essential oil to lecithin was 2mL:1g, the liposome encapsulation rate of ashwagandha essential oil was the highest, at 86.27%. The liposome encapsulation rate of ashwagandha essential oil gradually increased and then decreased with the increase of the ratio of ashwagandha essential oil to lecithin. When too much ashwagandha essential oil was added, it would exceed the carrying capacity of lecithin; when too little was added, the liposome of ashwagandha essential oil was unstable and prone to leakage, resulting in a decrease in the encapsulation rate. Therefore, the optimal ratio of ashwagandha essential oil to lecithin was 2mL:1g.

[0143] Figure 6 The figure is the effect of the ratio of lecithin to cholesterol on the liposome encapsulation efficiency of the hawthorn essential oil in Examples 3 and 6 to 9. Figure 6 It was concluded that when the ratio of lecithin to cholesterol was 3mL:1g, the encapsulation rate of the liposomes of the mountain ash essential oil was the highest, which was 87.26%. The encapsulation rate of the liposomes of the mountain ash essential oil gradually increased and then decreased with the increase of the ratio of lecithin to cholesterol. Lecithin and cholesterol have a good synergistic effect. Cholesterol can stabilize the bilayer membrane structure of the liposome, but with the increase of the ratio of lecithin to cholesterol, the amount of cholesterol constituting the liposomes of the mountain ash essential oil decreases. In the liposomes of the mountain ash essential oil, cholesterol is embedded between the lecithin molecules, the hydroxyl end of cholesterol is combined with the head group of lecithin, and the other end is inserted into the hydrophobic alkyl chain of the bilayer. Cholesterol can increase the rigidity of the liposome membrane of the mountain ash essential oil and increase its stability, but excessive cholesterol will cause the bilayer to exceed the capacity limit, causing the destruction of the bilayer, causing the liposomes of the mountain ash essential oil to lose their stable structure and eventually lead to rupture. Too little cholesterol will cause the rigidity of the liposome membrane of the mountain ash essential oil to be insufficient, which will also lead to the instability of the liposome membrane of the mountain ash essential oil. Therefore, the optimal ratio of lecithin to cholesterol was 3 mL: 1 g.

[0144] Figure 7 The effect of the volume of phosphate buffer solution on the liposome encapsulation efficiency of the hawthorn essential oil in Examples 3 and 10 to 13 is shown in FIG. Figure 7 It was concluded that the liposome encapsulation rate of the kaempferia essential oil was the highest when the volume of the phosphate buffer solution was 10 mL, which was 89.72%. The liposome encapsulation rate of the kaempferia essential oil decreased with the increase of the volume of the phosphate buffer solution. The increase in the amount of phosphate buffer solution would cause the liposome of the kaempferia essential oil to rupture and the kaempferia essential oil to leak, thus affecting the encapsulation rate. Therefore, the optimal phosphate buffer solution volume selected was 10 mL.

[0145] Figure 8 This is the appearance diagram of the liposome of the yam essential oil of Example 3, Figure 8 The results showed that the hawthorn essential oil liposomes were uniform and fluid suspensions, meeting the appearance requirements of liposomes.

[0146] Fig. 9 This is an optical microscope morphology of the liposomes of the yam essential oil of Example 3, Fig. 9It was observed that when the agave liposomes were placed under an optical microscope, it could be clearly seen that the agave liposomes were spherical, with a lipid membrane wrapped on the surface, a complete structure, and relatively uniform size, indicating that the agave liposomes had stability and structural integrity.

[0147] The evaluation was conducted based on the appearance, properties and stability of the liposome cream containing bergamot essential oil. The test indicators are as follows:

[0148] a. Evaluation indicators of cream appearance properties: glossiness is to evaluate the visual experience during use, whiteness is to evaluate the color comfort during use, fineness is to evaluate the skin feel, and consistency and spreadability are to evaluate the drug application effect.

[0149] The creams prepared by each method were taken for visual observation and applied to the back of the hand.

[0150] Scoring rules for the five indicators: The comprehensive score is 100 points, each indicator is 20 points, and is divided into five levels according to the status of each indicator: very good (20 points), good (16 points), relatively good (12 points), appropriate (8 points), and poor (4 points), as shown in Table 7.

[0151] b. Stability investigation: The stability of the hawthorn essential oil liposome cream was investigated from three aspects: centrifugation test, heat resistance test, and cold resistance test.

[0152] Centrifugation test investigation: Take each group of hawthorn essential oil liposome cream and put it into a centrifuge tube with a scale to about 2 / 3 of the height, set the speed to 4000r / min, centrifuge for 30min, and observe whether there is oil separation, stratification, and particles.

[0153] Heat resistance test: Take each group of hawthorn essential oil liposome cream and place it in a sealed centrifuge tube, put it in an oven and keep it at a constant temperature (55°C), take it out after 6 hours, and observe whether there is discoloration or stratification after cooling to room temperature.

[0154] Cold resistance test: Take each group of hawthorn essential oil liposome cream and place it in a sealed centrifuge tube. Place it in a -15℃ refrigerator for 48 hours and then take it out. After it returns to room temperature, observe whether it becomes coarse, discolored, or delaminated.

[0155] Scoring rules for the three indicators: The total score is 30 points, with each indicator worth 10 points. They are divided into three levels according to the status: excellent (8 to 10 points), good (4 to 7 points), and poor (0 to 3 points), as shown in Table 8.

[0156] Table 7 Appearance and shape score table of Example 3 and Comparative Examples 1 to 3 of the yam essential oil liposome cream

[0157]

[0158] It can be seen from Table 7 that the appearance properties score of the hawthorn essential oil liposome cream prepared in Example 3 of the present invention is the highest, so the emulsification method used in the present invention is superior to the other three methods.

[0159] Table 8 Appearance and shape scores of creams prepared by different methods

[0160]

[0161] It can be seen from Table 8 that the stability score of the yam essential oil liposome cream prepared in Example 3 of the present invention is the highest. Therefore, the emulsification method adopted in the present invention is superior to the other three methods, and the yam essential oil liposome cream prepared by the emulsification method has the best appearance, properties and stability.

[0162] Table 9 Example 3 Preparation of the appearance and properties of the liposome cream of the mountain ash essential oil Repeated verification results table

[0163] Number of trials 1 2 3 Mean Score 79 83 84 82

[0164] Table 10 Example 3 Preparation of the stability of the liposome cream of the mountain ash essential oil Repeatability verification results

[0165] Number of trials 1 2 3 Mean Score 25 27 26 26

[0166] Fig.10 This is the finished product appearance diagram of the mountain ash essential oil liposome cream of embodiment 3, combined with Table 11, Table 12 and Fig.10 It was concluded that the appearance, properties and stability of the hawthorn essential oil liposome cream prepared in Example 3 were good, and the scores were higher than those of the other three methods, so the preparation process was feasible.

[0167] The dilution of the schisandra chinensis essential oil liposome cream prepared in Example 3 was used for antibacterial study:

[0168] Antibacterial observation:

[0169] Take 28 test tubes, add 1.0 mL of TSB medium to the bacterial experiment tube, add 1.0 mL of SDA medium to the fungal experiment tube, then dilute the mountain ash essential oil liposome cream sample by the two-fold dilution method, and then add 1.0 mL of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans liquid to the above test tubes respectively.

[0170] Take 1.0mL of Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli and inoculate them into 1.0mL of TSB medium respectively; take 1.0mL of Candida albicans liquid and inoculate it into 1.0mL of SDA medium as a positive control to observe whether bacteria and fungi grow normally.

[0171] Take two test tubes, add 2.0 mL of TSB medium and 2.0 mL of SDA medium respectively, and do not inoculate bacterial solution as blank control. All the above test tubes are cultured at 37°C for 24 hours to observe whether the liposome cream of yam essential oil has antibacterial properties.

[0172] Fig.11 The results of the antibacterial experiment of the liposome cream of the essential oil of the mountain ash in Example 3 are shown in Figure 3, wherein (a) is the experimental group with the addition of the liposome cream of the essential oil of the mountain ash, the bacterial solution and the corresponding culture medium, and (b) is the positive control group with only the bacterial solution and the corresponding culture medium added; Observation Fig.11 (a)(b) It was concluded that the experimental groups were clearer and more transparent than the positive control group, indicating that the liposome cream of the mountain ash essential oil has antibacterial properties.

[0173] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0174] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for preparing a liposome cream of yam essential oil, characterized in that: The following steps are involved: After dissolving the bergamot essential oil, lecithin and cholesterol in a solvent, the solvent is evaporated to form a reaction material; The reaction material and the pH regulator are ultrasonically mixed to obtain the yam essential oil liposomes; Heat and mix the bergamot essential oil liposomes and the first emulsifier to form an aqueous phase solution; Heat and melt the liquid paraffin, stearyl alcohol and the second emulsifier to form an oil phase mixture; The oil phase mixture is mixed with the aqueous phase solution and emulsified to obtain the bergamot essential oil liposome cream.

2. The method for preparing the liposome cream of the yam essential oil according to claim 1, wherein: The volume of the bergamot essential oil, the mass of lecithin and the mass ratio of cholesterol are 1mL-4mL: 1g-5g: 1g.

3. The method for preparing the liposome cream of the yam essential oil according to claim 1, wherein: The pH regulator is a phosphate buffer, and the pH of the kaempferia essential oil liposome is adjusted to 7-7.2 by using the pH regulator.

4. The method for preparing the safflower essential oil liposome cream according to claim 1, wherein: The emulsification treatment conditions are: stirring at 75°C to 80°C for 2min to 3min.

5. The method for preparing a liposome cream of yam essential oil according to claim 1, characterized in that: The mass ratio of the oil phase mixture to the water phase solution is 1-2:

3.

6. The method for preparing the liposome cream of the yam essential oil according to claim 1, characterized in that: The mass ratio of the hawthorn essential oil liposome to the first emulsifier is 10-15:

1.

7. The method for preparing the safflower essential oil liposome cream according to claim 1, characterized in that: The mass ratio of the total mass of the liquid paraffin and octadecyl alcohol to the mass ratio of the second emulsifier is 16 to 20:

1.

8. A liposome cream of bergamot essential oil prepared by the preparation method according to any one of claims 1 to 7.

9. An application of a liposome cream of yam essential oil in the preparation of an antibacterial agent, characterized in that: The antimicrobial agent inhibits the activity of bacteria.

10. The use of the yam essential oil liposome cream according to claim 9 in the preparation of an antibacterial agent, characterized in that: The bacteria are Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli or Candida albicans.

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