Lavender essential oil liposome emulsion as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of flavors and fragrances, and particularly relates to a lavender essential oil liposome emulsion as well as a preparation method and application thereof. According to the invention, sucrose / sodium chloride solid-phase particles and a wrapping material are combined together for the first time to prepare a no-load liposome, and the no-load liposome is coated with a drug lavender essential oil to prepare the lavender essential oil liposome emulsion. The sucrose / sodium chloride as solid-phase particles has excellent specific surface area, is green and environment-friendly, is rapid and mild in wrapping process, and is an excellent carrier. After the no-load liposome is prepared, the lavender essential oil is wrapped, so that the problems that the lavender essential oil is easy to volatilize, unstable, difficult to store, poor in absorption effect and the like are solved, the lavender essential oil can be prepared when being used, and the influence of storage and transportation processes on the lavender essential oil is avoided.

Description

Technical Field

[0001] The invention belongs to the technical field of flavors and fragrances, and particularly relates to a lavender essential oil liposome emulsion, a preparation method and application thereof. Background Art

[0002] Lavender essential oil is a plant essential oil with multiple physiological activities. It has antibacterial and anti-inflammatory effects, freckle removal and whitening, stress relief, blood pressure reduction, and regulation of skin oil secretion balance. Secondly, it has a certain improvement effect on insomnia. Linalool is one of the main components of lavender essential oil. As a naturally occurring terpenoid alcohol, it is mainly distributed in many flowers and spice plants. Linalool has a fresh floral fragrance and a slightly spicy smell. It has multiple physiological activities such as antibacterial, anti-inflammatory, anti-tumor, and antioxidant. It is an important component of many flavors, cosmetics, foods, and pesticides.

[0003] As a good transport system, liposomes are similar to biological membranes and have significant biocompatibility, biodegradability and low toxicity. Due to their flexible structure and practical functions, liposomes are increasingly widely used in the fields of medicine, food and cosmetics, and have very broad application prospects. According to the formulation and mode of action of the preparation, liposomes can be roughly divided into ordinary liposomes, environmentally sensitive liposomes, long-circulating liposomes, active targeting liposomes, multifunctional liposomes, etc. The commonly used preparation methods of liposomes include ethanol injection method, thin film dispersion method, ultrasonic dispersion method, melting method, reverse evaporation method, etc. However, the existing preparation process of liposomes also has certain defects, such as the preparation process is too complicated, the production cost is too high, a large amount of organic solvents are used, and it is difficult to achieve large-scale preparation, resulting in the inability to apply. Summary of the invention

[0004] In view of the problems that the existing liposome preparation process is cumbersome, the production cost is too high, the amount of organic solvent is large, and it is difficult to prepare on a large scale, the present invention combines sucrose / sodium chloride solid phase particles with a wrapping material for the first time, prepares empty liposomes, and coats the medicinal lavender essential oil to make a lavender essential oil liposome emulsion. Sucrose / sodium chloride has a huge surface area as a solid phase particle, is environmentally friendly, and the wrapping process is rapid and gentle, and is a kind of excellent carrier. After the empty liposome is prepared, the present invention wraps the lavender essential oil, which not only solves the problems that the lavender essential oil is volatile, unstable, difficult to preserve, and has a poor absorption effect, but also can be used and prepared, avoiding the impact of storage and transportation processes on the lavender essential oil.

[0005] To achieve the above object, the present invention provides a method for preparing a lavender essential oil liposome emulsion, comprising the following steps:

[0006] (a) adding a wrapping material into anhydrous ethanol to prepare a wrapping material solution;

[0007] (b) immersing the solid phase particles in a coating material solution, removing the anhydrous ethanol, and combining the solid phase particles with the coating material to obtain empty liposomes;

[0008] (c) mixing the empty liposomes with the lavender essential oil solution to obtain a lavender essential oil liposome emulsion.

[0009] The encapsulating material is one of lecithin and lecithin-cholesterol;

[0010] The mass ratio of lecithin to cholesterol is (4-8):1, preferably 6:1;

[0011] The solid phase particles are sucrose or sodium chloride.

[0012] The mechanism of action of the solid phase particles is to utilize the huge specific surface area of ​​the solid phase particles to allow the coating material solution to volatilize on the surface of the solid phase particles to form a film of the coating material with a huge specific surface area;

[0013] The particle size of the solid phase particles is 150-1250 μm.

[0014] Preferably, the particle size of the sucrose solid phase particles is 200 to 900 μm;

[0015] Preferably, the particle size of the sodium chloride solid phase particles is 300 to 1250 μm.

[0016] In step (b), the removal method of the anhydrous ethanol includes one of high-temperature water bath evaporation, standing at room temperature, and air drying; the high-temperature water bath evaporation includes normal pressure water bath evaporation and reduced pressure water bath evaporation.

[0017] Preferably, in step (c), the empty liposomes and the lavender essential oil solution are mixed in a manner comprising: vortexing, shaking, and ultrasound;

[0018] The mass concentration of the encapsulating material in the encapsulating material solution is 0.02363 to 0.2 g / mL, preferably 0.03544 to 0.2 g / mL.

[0019] The mass ratio of the solid phase particles to the encapsulating material is (1.39-6.5):1;

[0020] Preferably, the mass ratio of the sucrose solid phase particles to the encapsulating material is (1.79-6.5):1;

[0021] Preferably, the mass ratio of the sodium chloride solid phase particles to the encapsulating material is (1.56-6.25):1.

[0022] The mass ratio of the lavender essential oil to lecithin is (5.7-18.0):100;

[0023] The solvent of the lavender essential oil solution is water, and the volume fraction of the lavender essential oil solution is 0.025% to 0.1%;

[0024] Preferably, when the solid phase particles are sucrose, the mass ratio of the lavender essential oil to lecithin is (10-18):100; and the volume fraction of the lavender essential oil solution is 0.08%-0.1%.

[0025] Preferably, when the solid phase particles are sodium chloride, the volume fraction of the lavender essential oil solution is 0.025% to 0.05%.

[0026] The invention also provides lavender essential oil liposomes prepared by the preparation method.

[0027] The 24h permeability of the lavender essential oil liposome emulsion is not less than 23%;

[0028] The encapsulation efficiency of the lavender essential oil liposome is 69.8-100%;

[0029] Furthermore, the encapsulation efficiency of the lavender essential oil liposomes was measured by HPLC-ultrafiltration centrifugation method;

[0030] The ultrafiltration centrifugal speed is 6000-12000 rpm, and the centrifugal time is 10-20 min.

[0031] After granulation, the particle size of the lavender essential oil liposome is 78.8-124.5 nm, and the aggregation index (PDI) is 0.142-0.312;

[0032] The granulation method is one of the following: ultrasound, shearing, homogenization, and extrusion, preferably extrusion, and more preferably microfiltration extrusion.

[0033] The present invention also provides an application of the lavender essential oil liposome emulsion prepared by the above-mentioned preparation method in preparing skin care products with the functions of removing spots and whitening, antibacterial and anti-inflammatory, relieving pain and itching, and regulating skin oil secretion and balancing.

[0034] The dosage form of the lavender essential oil liposome emulsion is an emulsion.

[0035] The lavender essential oil liposome emulsion does not contain an organic solvent and does not need to be dried, thereby reducing the effect of drying on the stability of the liposome.

[0036] Beneficial Effects

[0037] 1. The present invention adopts sucrose / sodium chloride as solid phase particles for the first time, combines with a wrapping material to prepare empty liposomes, and then wraps lavender essential oil to prepare a lavender essential oil liposome emulsion. By utilizing the excellent specific surface area of ​​the solid phase particles, the wrapping material forms a thin and uniform film on the surface of the solid phase particles, thereby increasing its drug loading.

[0038] 2. Compared with the thin film dispersion method, in the preparation process, an organic solvent is required to dissolve the drug, and then the solvent is removed by rotary evaporation to form a thin film on the wall of the rotary evaporation bottle; the lavender essential oil of the present invention does not need to be dissolved in an organic solvent during the preparation process, which on the one hand avoids the influence of the organic solvent on the lavender essential oil, and on the other hand reduces the heating time caused by removing anhydrous ethanol, thereby greatly avoiding the influence of heat on the stability of the lavender essential oil; the solid phase particles provided by the present invention have excellent specific surface area compared with the rotary evaporation bottle, and the film thickness formed by the wrapping material on the surface of the solid phase particles is thin and uniform.

[0039] 3. Compared with the traditional rotary evaporation spray method, in the preparation process, the coating material solution is sprayed onto the surface of the solid particles to form a spot film with uneven thickness and discontinuousness; the present invention adopts an immersion film-forming method to immerse the solid particles in a saturated coating material solution to obtain a film with uniform thickness and continuity, thereby obtaining lavender essential oil liposomes with smaller and more uniform particle size.

[0040] 4. The present invention first prepares empty liposomes and then mixes them with a lavender essential oil solution to obtain a lavender essential oil liposome emulsion. During application, empty liposomes can be prepared first, and the lavender essential oil liposome emulsion can be prepared immediately without considering the impact of storage and transportation conditions on the lavender essential oil. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 HPLC spectra of the lavender essential oil liposome permeate and concentrate prepared in Examples 1-3;

[0042] Figure 2 HPLC spectra of the lavender essential oil liposome permeate and concentrate of Examples 5-7;

[0043] Figure 3 This is a particle size distribution diagram of the lavender essential oil liposomes prepared in Example 1 after granulation;

[0044] Figure 4 This is the particle size distribution diagram of the lavender essential oil liposomes prepared in Example 5 after granulation. DETAILED DESCRIPTION

[0045] The present invention will be further described in detail below through specific implementation examples, but it should not be understood that the present invention is limited to the following examples, and all technologies realized based on the above content of the present invention belong to the scope of the present invention.

[0046] The lecithin described in the examples was purchased from Beijing Aoboxing Biological Co., Ltd., cholesterol was purchased from Sinopharm Chemical Reagent Co., Ltd., lavender essential oil was provided by Shandong Freda Biological Co., Ltd., anhydrous ethanol was purchased from Tianjin Fuyu Fine Chemical Co., Ltd., sucrose and sodium chloride were conventional commercial products, and the artificial skin was Strat-MTM Membrane purchased from Merck Millipore Ltd.

[0047] Embodiment 1:

[0048] Weigh lecithin and add anhydrous ethanol solution, prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sucrose solid phase particles (average particle size of 550μm, particle size between 250-850μm) in an evaporating dish, heat in a 50°C water bath, then add to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.5:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add lavender essential oil solution with a volume fraction of 0.025% according to the mass ratio of lavender essential oil solution solid content to lecithin 6:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0049] Embodiment 2:

[0050] Weigh lecithin and add anhydrous ethanol solution, prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sucrose (average particle size of 550μm, particle size between 250-850μm) in an evaporating dish, heat in a 50°C water bath, then add to the lecithin solution at a solid phase particle to lecithin mass ratio of 6.5:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add a lavender essential oil solution with a volume fraction of 0.025% according to the mass ratio of lavender essential oil solution solid content to lecithin 18:100, shake for 10 minutes, and obtain a lavender essential oil liposome emulsion.

[0051] Embodiment 3:

[0052] Weigh lecithin and add anhydrous ethanol solution to prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sucrose (average particle size of 550μm, particle size between 250-850μm) in an evaporating dish, heat in a 50°C water bath, then add to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.5:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add lavender oil solution with a volume fraction of 0.1% according to the mass ratio of lavender essential oil solution solid content to lecithin 6:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0053] Embodiment 4:

[0054] The preparation conditions of this example are the same as those of Example 2, except that the lecithin in Example 2 is replaced by lecithin-cholesterol.

[0055] The mass ratio of lecithin to cholesterol is 6:1.

[0056] Embodiment 5:

[0057] Weigh lecithin and add anhydrous ethanol solution to prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sodium chloride (average particle size is 650μm, particle size is between 300 and 1000μm) in an evaporating dish, heat in a 50°C water bath, and then add it to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.39:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add lavender oil solution with a volume fraction of 0.025% according to the mass ratio of lavender essential oil solution solid content to lecithin 5.7:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0058] Embodiment 6:

[0059] Weigh lecithin and add anhydrous ethanol solution to prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sodium chloride (average particle size is 650μm, particle size is between 300 and 1000μm) in an evaporating dish, heat in a water bath at 50°C, then add sodium chloride solid phase particles to lecithin at a mass ratio of 6.25:1 to the lecithin solution, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add lavender oil solution with a volume fraction of 0.025% according to the mass ratio of lavender essential oil solution solid content to lecithin 17.4:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0060] Embodiment 7:

[0061] Weigh lecithin and add anhydrous ethanol solution to prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store under light-proof conditions. Weigh sodium chloride (average particle size of 650μm, particle size between 300 and 1000μm) in an evaporating dish, heat in a 50°C water bath, then add to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.39:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take empty liposomes, add lavender oil solution with a volume fraction of 0.1% according to the mass ratio of lavender essential oil solution solid content to lecithin 5.7:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0062] Comparative Example 1:

[0063] The preparation conditions of this comparative example are the same as those of Example 6, except that no sodium chloride solid phase particles are added.

[0064] The specific preparation method is as follows: weigh lecithin and add anhydrous ethanol solution to prepare a 0.2 g / mL lecithin solution, ultrasonicate for 10 minutes, and store in a dark environment. Place the lecithin solution in a rotary evaporator and evaporate at 50°C. Add 0.025% lavender essential oil aqueous solution according to the mass ratio of lavender essential oil solution solid content to lecithin 17.4:100, ultrasonicate for 10 minutes to obtain lavender essential oil liposome emulsion.

[0065] Comparative Example 2:

[0066] The preparation conditions of this comparative example are the same as those of Example 6, except that lavender essential oil is replaced by hydroxytyrosol.

[0067] Comparative Example 3:

[0068] Weigh lecithin and lavender essential oil, add them to an anhydrous ethanol solution, and perform ultrasonic treatment for 10 min to obtain a lavender essential oil organic solution, wherein the mass ratio of lavender essential oil to lecithin is 17.4:100, and the concentration of lecithin in the lavender essential oil organic solution is 0.2 g / mL.

[0069] The lavender essential oil organic solution was placed in an evaporating dish, and sodium chloride was added in a mass ratio of sodium chloride to lecithin of 6.25:1 under the condition of heating in a water bath at 50°C, stirring was performed while adding, and the solution was evaporated in a water bath, and then deionized water was added to obtain a lavender essential oil liposome emulsion (wherein the volume ratio of deionized water to lavender essential oil was 1000:0.25).

[0070] Comparative Example 4:

[0071] Weigh lecithin and add anhydrous ethanol solution to prepare 0.2g / mL lecithin solution, ultrasonicate for 10 minutes, and store in dark. Take sodium chloride and place it in a rotary evaporator, then spray the lecithin solution into the rotary evaporator at a mass ratio of sodium chloride to lecithin of 6.25:1, and the rotary evaporation temperature is 50°C to obtain empty liposomes. Take empty liposomes, add lavender essential oil aqueous solution with a volume fraction of 0.025% according to the mass ratio of lavender essential oil to lecithin of 17.4:100, shake for 10 minutes, and obtain lavender essential oil liposome emulsion.

[0072] Test example 1: Encapsulation efficiency test

[0073] The detection method of liposome encapsulation efficiency of lavender essential oil is as follows: take 0.1mL lavender essential oil emulsion in a centrifuge tube (CF), centrifuge at 8000rpm for 10min, take out the supernatant and add it to an ultrafiltration centrifuge tube (UF-CF, ultrafiltration membrane pore size is 30KD), then add 0.3mL pure water, centrifuge at 8000rpm for 10min, take the permeated liquid as the test solution for free essential oil detection (defined as permeated liquid). Continue to add 0.4mL anhydrous ethanol to the above CF, weigh, ultrasonically dissolve, add to the above UF-CF, ultrafiltration centrifuge at 8000rpm for 10min, take the permeated liquid to make up the lost weight with anhydrous ethanol, as the test solution for encapsulated essential oil detection (defined as concentrated solution).

[0074] HPLC conditions: bonded silica gel C 18 (4.6mm*250mm, 5μm), with acetonitrile (A)-0.2% phosphoric acid aqueous solution (B) as the mobile phase, gradient elution (0-30min, A% (35%~80%); 31-40min, A% 100%), flow rate 1.0mL·min, column temperature 30℃, detection wavelength: 203nm, injection volume: 10μL.

[0075] Here, encapsulation rate (100%) = amount of encapsulated essential oil / (amount of free essential oil+amount of encapsulated essential oil)×100%.

[0076] Table 1 Encapsulation efficiency of lavender essential oil liposomes (%)

[0077]

[0078]

[0079] As shown in Table 1, the encapsulation efficiency of the lavender essential oil liposomes prepared in each embodiment and comparative example is as follows:

[0080] Compared with Example 2, Example 1 uses a larger amount of lecithin, which may cause the lecithins to adhere to each other and fail to form a well-shaped sphere, resulting in a small drug holding capacity or a large thickness of the liposome shell, making it difficult for a small amount of drug to enter; when the amount of lecithin is small, a small number of liposomes may be formed, thereby affecting the encapsulation rate.

[0081] Compared with Example 1, the volume fraction of lavender essential oil in the lavender essential oil aqueous solution of Example 3 is increased, and its encapsulation efficiency is also significantly improved.

[0082] Compared with Example 2, Example 4 uses lecithin-cholesterol as the encapsulation material, and its encapsulation rate is increased to 100%.

[0083] According to Table 1, the encapsulation efficiency of Comparative Example 1 is much smaller than that of Example 6, because in the process of preparing lavender essential oil liposome by thin film dispersion method, the film thickness formed by the wrapping material on the wall of the rotary evaporation bottle is uneven, causing the encapsulation efficiency of lavender essential oil liposome to be low. This method has good effect on the encapsulation of hydrophobic active ingredients, and hydroxytyrosol is highly water-soluble, so the encapsulation efficiency of Comparative Example 2 is much smaller than that of Example 6. The encapsulation efficiency of Comparative Example 3 is much smaller than that of Example 6, and in the process of removing solvent from lavender oil and lecithin solution, due to the strong volatility of the component in lavender oil, the loss is large in the process of evaporation. The encapsulation efficiency of Comparative Example 4 is much smaller than that of Example 6, and the spraying film-forming can not make full use of the large advantage of the specific surface area of ​​solid particles, and the solvent evaporation speed is fast, and the spots of discontinuous thickness inhomogeneity are formed on the surface of solid particles, or directly form lecithin microspheres, which is unfavorable for the subsequent component encapsulation in lavender oil.

[0084] Figure 1 , Figure 2 The HPLC spectra of the permeate and concentrate of lavender essential oil liposome prepared in Examples 1-3 and 5-7 respectively show that both the permeate and the concentrate have peaks at the linalool chromatographic peak (chromatographic peak retention time is about 15 min), but the peak of the permeate is smaller and close to a straight line, while the concentrate has a strong peak, indicating that the encapsulated drug leaks little and has strong encapsulation.

[0085] Test Example 2: Determination of Particle Size

[0086] Take 0.1 mL of the lavender essential oil liposome emulsion prepared in Example 1 and Example 5, dilute it with water to 1 mL, and detect it three times by laser particle size analyzer Zetasizer Nano ZS90. The detection shows that the average particle size of the lavender essential oil liposome prepared in Example 1 is 122.7 nm, the average PDI is 0.388, and the average potential is -66.3 mV; the average particle size of the lavender essential oil liposome prepared in Example 5 is 164.6 nm, the average PDI is 0.345, and the average potential is -30.7 mV.

[0087] The lavender essential oil liposome emulsion prepared in Example 1 and Example 5 was filtered through a 0.22 μm microfiltration membrane, 0.1 mL of the filtrate was taken and diluted to 1 mL with water, and then tested three times by a laser particle size analyzer ZetasizerNano ZS90. Figure 3 , 4 The particle size distribution diagrams of the lavender essential oil liposomes prepared in Example 1 and Example 5 are shown respectively. From the diagrams, it can be seen that the average particle size after granulation in Example 1 is 78.8 nm, and the average PDI is 0.210; the average particle size after granulation in Example 5 is 101.58 nm, and the average PDI is 0.142.

[0088] Analysis shows that the particle size of the liposomes after microfiltration is small, about 100nm, and the dispersion coefficient is small, relatively stable, and evenly dispersed. The average potential of the liposomes is measured to be negative, the surface of the liposome particles is negatively charged, and the stability is good.

[0089] The particle size detection method of the lavender essential oil liposomes prepared in other embodiments and comparative examples before granulation is the same as that of Examples 1 and 5. The specific detection results are shown in Table 2.

[0090] Test Example 3: Transdermal Diffusion Experiment

[0091] Utilize the transdermal diffusion test apparatus, add the PEG400 solution with the mass concentration of 40% into the diffusion cell, take the lavender essential oil liposome emulsion and the lavender essential oil aqueous solution (volume fraction is 0.1%) prepared in each embodiment, each 0.3mL is evenly dispersed on the smooth surface of artificial skin, penetrated at room temperature, take out 0.2mL sample solution (lavender essential oil liposome emulsion permeation solution, lavender essential oil aqueous solution permeation solution) from the diffusion cell after an interval of 24h for liquid chromatography determination, and make up the volume with the PEG400 solution with the mass concentration of 40%, and calculate the permeability by high performance liquid chromatography.

[0092] Table 2 Transmission rate, average particle size, potential of lavender essential oil liposomes prepared in each embodiment and comparative example

[0093]

[0094]

[0095] It can be concluded from Table 2 that the particle size of the lavender essential oil liposomes prepared by this method after granulation is between 78.8 and 124.5 nm, the PDI is between 0.142 and 0.312, the dispersion coefficient is relatively small, relatively stable, and uniformly dispersed. The average potential of the liposome is between -26.3 and -66.3 mV, the surface of the liposome particles is negatively charged, and the stability is good. Compared with the lavender oil nanoliposomes prepared by using sucrose particles as carriers in Examples 1 to 4, the lavender oil nanoliposomes prepared by using sodium chloride as carriers in Examples 5 to 6 have smaller particle sizes, more ideal PDI, and the average potential value is also conducive to the stability of the lavender oil liposome emulsion. Compared with Comparative Examples 1, 3, and 4, Examples 1 to 7 involved in the present invention have smaller particle sizes, smaller dispersion coefficients, relatively stable, and uniform dispersion, and there is no significant difference in average potential and transmittance; Comparative Example 2 uses hydroxytyrosol for comparison, and it can be seen from Table 1 that its encapsulation rate is extremely low and the transdermal absorption effect is also poor.

[0096] Test Example 4: Acetylcholinesterase Inhibitory Activity

[0097] 1. Solution preparation

[0098] Phosphate buffered saline (PBS): Take 0.68 g of potassium dihydrogen phosphate, add 29.1 mL of 0.1 mol / L sodium hydroxide solution, and dilute with water to 100 mL.

[0099] Lavender essential oil sample solution: lavender essential oil was taken and dissolved in a 1 wt % Tween 20 aqueous solution to prepare a lavender essential oil sample solution with a volume fraction of 0.025%.

[0100] DTNB solution (0.01 M): Dissolve 3.9 mg DTNB in ​​1 mL DMSO and store at 4°C in the dark.

[0101] ATCI solution: 108.35 mg dissolved in 4 mL of water and stored at 4°C.

[0102] AChE solution: 3 mg dissolved in 3.5 mL PBS and stored at 4°C.

[0103] Huperzine A solution: 2 mg is dissolved in 1 mL of water and diluted to 0.01 mg / mL.

[0104] 2. Experimental Procedure

[0105] The experiment set up a blank reference group (BR), a blank control group (BC), a sample reference group (SR), a sample group (S), and a positive control group (C).

[0106] Group S: 5 μL of sample solution, 170 μL of phosphate buffer (0.1 mol / L, pH 7.0), 15 μL of AChE solution and 10 μL of DTNB solution were added to the wells in sequence, shaken thoroughly, and placed at 4°C for 20 minutes. After 20 minutes, 10 μL of ATCI solution was added dropwise and allowed to react in a 37°C water bath for 20 minutes. After the reaction, the absorbance A was measured in an ELISA reader. S .

[0107] The sample solution includes: a lavender essential oil sample solution; a permeation solution of the lavender essential oil liposome emulsion prepared in Examples 3 and 6 in Detection Example 3; and a permeation solution of the lavender essential oil aqueous solution in Detection Example 3.

[0108] SR group: 15 μL of PBS solution replaced 15 μL of AChE solution. Other steps were the same as those in S group. The absorbance A was measured. SR .

[0109] BR group: 175 μL of PBS solution replaced 5 μL of sample solution and 170 μL of phosphate buffer. Other steps were the same as the sample group. The absorbance A was measured. BR .

[0110] BC group: 20 μL of dimethyl sulfoxide (DMSO) and 140 μL of PBS solution replaced 5 μL of sample solution and 170 μL of phosphate buffer. Other steps were the same as the sample group. The absorbance A was measured. BC .

[0111] Group C: The sample solution was replaced with 5 μL of Huperzine A solution, and the other steps were the same as the sample group. The absorbance A was measured C .

[0112] The inhibition rate of lavender essential oil sample was calculated using the formula:

[0113] The inhibition rate of Huperzine A solution was calculated using the formula:

[0114] Table 3 Acetylcholinesterase inhibition rate

[0115]

[0116] It can be concluded from Table 3 that the lavender essential oil sample solution and its aqueous solution have obvious acetylcholinesterase inhibition, and two concentrations of huperzine A reference solution are used as positive controls, indicating the reliability of this method in determining acetylcholinesterase inhibition activity. Compared with the lavender essential oil liposome emulsion permeation solution prepared in Example 3 and the lavender essential oil aqueous solution permeation solution, the former detected significant acetylcholinesterase inhibition, while the latter did not detect it, indicating that the lavender oil prepared into liposomes significantly increased the transdermal ability of lavender; the comparison results of the lavender essential oil liposome emulsion permeation solution prepared in Example 6 and the lavender essential oil aqueous solution permeation solution also verified the above conclusion.

Claims

1. A method for preparing a lavender essential oil liposome emulsion, characterized in that: The steps include: (a) adding a wrapping material into anhydrous ethanol to prepare a wrapping material solution; (b) immersing the solid phase particles in a coating material solution, removing the anhydrous ethanol, and combining the solid phase particles with the coating material to obtain empty liposomes; (c) mixing the empty liposome with the lavender essential oil solution to obtain a lavender essential oil liposome emulsion; The solid phase particles are sucrose or sodium chloride.

2. The method for preparing lavender essential oil liposome emulsion according to claim 1, characterized in that, The packaging material is one of lecithin and lecithin-cholesterol.

3. The preparation method of lavender essential oil liposome emulsion according to claim 1, characterized in that, The particle size of the solid phase particles is 150-1250 μm. Preferably, the particle size of the sucrose solid phase particles is 200 to 900 μm; Preferably, the particle size of the sodium chloride solid phase particles is 300 to 1250 μm.

4. The method for preparing the lavender essential oil liposome emulsion according to claim 1, characterized in that: The mass concentration of the encapsulating material in the encapsulating material solution is 0.02363 to 0.2 g / mL, preferably 0.03544 to 0.2 g / mL.

5. The method for preparing the lavender essential oil liposome emulsion according to claim 1, characterized in that: The mass ratio of the solid phase particles to the encapsulating material is (1.39-6.5):1; Preferably, the mass ratio of the sucrose solid phase particles to the encapsulating material is (1.79-6.5):1; Preferably, the mass ratio of the sodium chloride solid phase particles to the encapsulating material is (1.56-6.25):

1.

6. The method for preparing the lavender essential oil liposome emulsion according to claim 2, characterized in that: The mass ratio of the lavender essential oil to lecithin is (5.7-18.0):100; Preferably, when the solid phase particles are sucrose, the mass ratio of the lavender essential oil to lecithin is (10-18):

100.

7. The method for preparing the lavender essential oil liposome emulsion according to claim 1, characterized in that: The solvent of the lavender essential oil solution is water, and the volume fraction of the lavender essential oil solution is 0.025% to 0.1%; Preferably, when the solid phase particles are sucrose, the volume fraction of the lavender essential oil solution is 0.08% to 0.1%; Preferably, when the solid phase particles are sodium chloride, the volume fraction of the lavender essential oil solution is 0.025% to 0.05%.

8. A lavender essential oil liposome emulsion prepared by the preparation method according to any one of claims 1 to 7, characterized in that: The 24h permeability of the lavender essential oil liposome emulsion is not less than 23%; The encapsulation rate of the lavender essential oil liposome emulsion is 69.8-100%; After granulation, the particle size of the lavender essential oil liposome is 78.8-124.5 nm, and the aggregation index is 0.142-0.

312.

9. Use of the lavender essential oil liposome emulsion prepared by the preparation method according to any one of claims 1 to 7 or the lavender essential oil liposome emulsion according to claim 8 in preparing skin care products for removing spots and whitening, antibacterial and anti-inflammatory, relieving pain and itching, and regulating skin oil secretion and balancing.

10. The use of the lavender essential oil liposome emulsion according to claim 9 in preparing skin care products for removing spots and whitening, antibacterial and anti-inflammatory, relieving pain and itching, and regulating skin oil secretion and balancing, characterized in that: The dosage form of the lavender essential oil liposome emulsion is an emulsion.

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