Acer truncatum oil liposome emulsion as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal preparations, and particularly relates to acer truncatum oil liposome emulsion as well as a preparation method and application thereof. The method comprises the following steps: (a) adding a wrapping material into absolute ethyl alcohol to prepare a wrapping material solution; (b) impregnating the solid-phase particles in a wrapping material solution, removing the absolute ethyl alcohol, and combining the solid-phase particles with a wrapping material to obtain a no-load liposome; and (c) mixing the no-load liposome with the acer truncatum oil-water dispersion liquid to obtain the acer truncatum oil liposome emulsion. According to the invention, sucrose / salt is used as solid-phase particles for the first time and is combined with a wrapping material to prepare a no-load liposome, and then acer truncatum oil is wrapped to prepare the acer truncatum oil liposome emulsion. By utilizing the excellent specific surface area of the solid-phase particles, the coating material forms a thin and uniform film on the surfaces of the solid-phase particles, so that the drug loading capacity of the solid-phase particles is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a maple oil liposome emulsion, a preparation method and application thereof. Background Art

[0002] Acer truncatum oil is a functional oil extracted from Acer truncatum seeds, mainly rich in unsaturated fatty acids such as oleic acid, linoleic acid, and neuraminic acid. Studies have shown that Acer truncatum oil has multiple pharmacological effects such as antibacterial, anti-tumor, anti-fatigue, antioxidant, regulating intestinal flora, improving immunity, and improving the body's inflammation level. Recent studies have shown that neuraminic acid can delay the aging of nerve cells, repair their damage, improve immunity, improve sleep quality, and lower blood lipids. The neuraminic acid content of Acer truncatum oil is as high as 5%. Acer truncatum oil is safe and non-toxic, has good antioxidant properties, meets the current requirements of the cosmetics industry for green, natural, safe and environmentally friendly, and has broad market application prospects in the cosmetics industry.

[0003] Unsaturated fatty acids such as linoleic acid and nervonic acid are the main chemical components of maple oil. They are easily oxidized and lose their biological activity during use and storage. Encapsulation technology can effectively improve their stability during use and storage. As a good transport system, liposome technology, similar to biological membranes, has significant biocompatibility, biodegradability and low toxicity. It has very broad application prospects in the fields of medicine, food and cosmetics. At present, the main methods for preparing liposomes include ethanol injection method, thin film dispersion method, ultrasonic dispersion method, melting method, reverse evaporation method and so on. However, the existing liposome preparation process 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, which makes it impossible to apply.

[0004] Chinese patent document CN114468299A discloses a dietary supplement rich in neuraminic acid that promotes brain function, which uses neuraminic acid-containing 65%-80% neuraminic acid extracted from neuraminic acid maple oil as the main raw material, supplemented with hawthorn kernel essential oil, emulsified with an emulsifier, encapsulated in chitosan-modified liposomes, and mixed with a sugar blocking solution to obtain a mixture. This patent uses neuraminic acid-containing liposomes modified with chitosan to encapsulate the emulsified neuraminic acid maple oil, which has the disadvantages of large amounts of solvents such as acetic acid and ethanol and complicated steps. Summary of the invention

[0005] The present invention aims at the problems of the existing liposome preparation process being cumbersome, the production cost being too high, the amount of organic solvent used being large, and the difficulty in large-scale preparation, and provides a liposome emulsion of maple oil, a preparation method thereof, and an application thereof. The present invention combines sucrose / salt solid phase particles with a packaging material for the first time to prepare empty liposomes, and further encapsulates the drug maple oil to prepare the maple oil liposome emulsion. Sucrose / salt as solid phase particles has a huge surface area, is green and environmentally friendly, and has a rapid and gentle encapsulation process, and is an excellent carrier. The present invention prepares empty liposomes and then encapsulates the maple oil, thereby solving the problems of low encapsulation rate, instability, difficulty in preservation, poor absorption effect, and low encapsulation rate of the maple oil in the prior art, and can be prepared on the spot, which is convenient for storage and transportation.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for preparing a maple oil liposome emulsion comprises the following steps:

[0008] (a) adding a wrapping material into anhydrous ethanol to prepare a wrapping material solution;

[0009] (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;

[0010] (c) mixing the empty liposomes with the Acer truncatum oil aqueous dispersion to obtain the Acer truncatum oil liposome emulsion.

[0011] Preferably, in step (a), the encapsulating material is one of lecithin and lecithin-cholesterol;

[0012] The mass ratio of lecithin to cholesterol is (4-8):1, preferably 6:1.

[0013] Preferably, in step (b), the solid phase particles are sucrose or salt, and the salt is sodium chloride. The mechanism of action of the solid phase particles is to utilize the huge specific surface area of ​​the solid phase particles to make the coating material solution volatilize on the surface of the solid phase particles to form a film of the coating material with a huge specific surface area;

[0014] The particle size of the solid phase particles is 150-1250 μm, and more preferably, the particle size of the sucrose solid phase particles is 300-900 μm; more preferably, it is 500-600 μm; more preferably, the particle size of the salt solid phase particles is 900-1250 μm, and more preferably, it is 600-700 μm.

[0015] In step (b), the removal method of the anhydrous ethanol includes one of high-temperature volatilization and room temperature standing volatilization; including water bath volatilization, which can be one of normal pressure, reduced pressure, and air drying.

[0016] The mass ratio of the solid phase particles to the encapsulating material is (1.56-12.5):1; the mass of the encapsulating material is calculated based on lecithin.

[0017] Preferably, the mass ratio of the sucrose solid phase particles to the encapsulating material is (1.56-6.25):1;

[0018] Preferably, the mass ratio of the salt solid phase particles to the encapsulating material is (1.56-12.5):1.

[0019] Preferably, in step (c), the empty liposomes and the Acer truncatum oil-water dispersion are mixed in a manner comprising: vortex oscillation, shaking, and ultrasound;

[0020] The mass concentration of the encapsulating material in the encapsulating material solution is 0.1 to 0.2 g / mL, preferably 0.2 g / mL;

[0021] The mass ratio of the maple oil to lecithin is (32.03-37.18):100;

[0022] The solvent of the Acer truncatum oil-water dispersion is water, and the volume fraction of the Acer truncatum oil-water dispersion is 0.04% to 0.08%.

[0023] The active ingredients of maple oil are unsaturated fatty acids such as linoleic acid and neuraminic acid.

[0024] The invention also provides a maple oil liposome emulsion prepared by the preparation method.

[0025] The 22-hour artificial skin retention rate of the maple oil liposome emulsion with sugar solid phase particles as carriers is not less than 15%, and the 22-hour skin retention rate of the maple oil liposome emulsion with salt solid phase particles as carriers is not less than 25%; the encapsulation rate of the maple oil liposome is 87.41-100%.

[0026] Further preferably, the method further comprises step (d) of a granulation method, wherein the liposome is subjected to one of the following treatments: ultrasound, shearing, homogenization, and extrusion, preferably extrusion, and more preferably microfiltration extrusion; after the granulation treatment, the particle size of the maple oil liposome is 151.5 to 156.5 nm, and the aggregation index (PDI) is 0.138 to 0.174. The emulsion potential is -26 to -37 mV.

[0027] The present invention also provides an application of the Acer truncatum oil liposome emulsion prepared by the preparation method in the preparation of anti-aging, antibacterial and anti-oxidation skin care products.

[0028] The Acer truncatum oil liposome emulsion does not contain an organic solvent, and there is no need to perform a desolventizing treatment on the liposomes loaded with the Acer truncatum oil, thereby reducing the influence of subsequent treatments on the stability of the liposomes.

[0029] Beneficial effects of the present invention:

[0030] 1. The present invention uses sucrose / salt as solid phase particles for the first time, combines with encapsulating materials to prepare empty liposomes, and then encapsulates Acer truncatum oil to prepare Acer truncatum oil liposome emulsion. By utilizing the excellent specific surface area of ​​the solid phase particles, the encapsulating materials form a thin and uniform film on the surface of the solid phase particles, thereby increasing the drug loading capacity.

[0031] 2. Compared with the thin film dispersion method, in the preparation process, an organic solvent is used to dissolve the film-forming material, and then the solvent is removed by rotary evaporation to form a thin film on the wall of the rotary evaporation bottle; the maple oil of the present invention does not need to be dissolved in an organic solvent during the preparation process. On the one hand, the influence of the organic solvent on the maple oil is avoided, and on the other hand, the heating time caused by the removal of the organic solvent is reduced, thereby greatly avoiding the influence of heat on the stability of the maple oil; the solid phase particles provided by the present invention have excellent specific surface area compared with the rotary evaporation bottle, and the film formed by the coating material on the surface of the solid phase particles is thin and uniform.

[0032] 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 phase particles to form a spot film with uneven thickness and discontinuousness; the present invention adopts an immersion film-forming method to immerse the solid phase particles in a high concentration of the coating material solution to obtain a film with uniform thickness and continuity, thereby obtaining Acer truncatum oil liposomes with smaller and more uniform particle size.

[0033] 4. The present invention first prepares empty liposomes and then mixes them with a maple oil aqueous dispersion to obtain a maple oil liposome emulsion. During application, empty liposomes can be prepared first, and the maple oil liposome emulsion can be prepared immediately without considering the impact of storage and transportation conditions on the maple oil. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 The HPLC spectra of the Acer truncatum oil liposome permeate and concentrate prepared in Examples 1-4;

[0035] Figure 2 The HPLC spectra of the permeate and concentrate of the Acer truncatum oil liposomes of Examples 5-8;

[0036] Figure 3 This is the particle size distribution diagram of the Acer truncatum oil liposomes prepared in Example 1;

[0037] Figure 4 This is the particle size distribution diagram of the Acer truncatum oil liposomes prepared in Example 5. DETAILED DESCRIPTION

[0038] 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.

[0039] The maple oil described in the examples was purchased from Ji'an Guoguang Spice Factory, lecithin was purchased from Beijing Aoboxing Biological Co., Ltd., cholesterol was purchased from Sinopharm Chemical Reagent Co., Ltd., anhydrous ethanol was purchased from Tianjin Fuyu Fine Chemical Co., Ltd., sucrose (containing more than 95% sucrose crystals) was purchased from Shandong Dongdu Food Co., Ltd., edible salt was purchased from Shandong Guangrao Minghua Salt Chemical Co., Ltd., and the transdermal diffusion test device was Strat-MTM Membrane (25mm Discs) purchased from Merck Millipore Ltd. Hydroxytyrosol was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0040] Embodiment 1:

[0041] Preparation of Acer truncatum oil liposome emulsion using sucrose as solid phase particles:

[0042] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh sucrose (average particle size of 550μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.56:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.04% of the Acer truncatum oil aqueous dispersion (equivalent concentration of 0.32mg / ml) at a mass ratio of Acer truncatum oil to lecithin of 32.03:100, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0043] Embodiment 2:

[0044] Preparation of Acer truncatum oil liposome emulsion using sucrose as solid phase particles:

[0045] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh sucrose (average particle size of 550μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a solid phase particle to lecithin mass ratio of 6.25:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.04% of the Acer truncatum oil water dispersion at a mass ratio of 37.17:100 to the Acer truncatum oil, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0046] Embodiment 3:

[0047] Preparation of Acer truncatum oil liposome emulsion using sucrose as solid phase particles:

[0048] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh sucrose (average particle size of 550μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a solid phase particle to lecithin mass ratio of 1.56:1, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.08% of the Acer truncatum oil water dispersion at a mass ratio of 32.03:100 to the Acer truncatum oil, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0049] Embodiment 4:

[0050] The preparation conditions of this embodiment are the same as those of embodiment 2, except that cholesterol is added to lecithin, and the mass ratio of lecithin to cholesterol is 6:1.

[0051] Embodiment 5:

[0052] Preparation of Acer truncatum oil liposome emulsion using NaCl as solid phase particles:

[0053] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh salt (average particle size is 650μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a mass ratio of 1.56:1 of salt solid phase particles to lecithin, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.04% of the Acer truncatum oil water dispersion at a mass ratio of 32.03:100 of Acer truncatum oil to lecithin, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0054] Embodiment 6:

[0055] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh salt (average particle size is 650μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a mass ratio of 12.5:1 of salt solid phase particles to lecithin, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.04% of the Acer truncatum oil water dispersion at a mass ratio of 34.62:100 of Acer truncatum oil to lecithin, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0056] Embodiment 7:

[0057] Preparation of Acer truncatum oil liposome emulsion using NaCl as solid phase particles:

[0058] Weigh lecithin and add it to anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / ml lecithin solution, and store it in dark conditions. Weigh salt (average particle size is 650μm) in an evaporating dish, heat it in a water bath at 50°C, then add it to the lecithin solution at a mass ratio of 1.56:1 of salt solid phase particles to lecithin, stir while adding, evaporate in a water bath to obtain empty liposomes. Take the empty liposomes, add 0.08% of the Acer truncatum oil water dispersion at a mass ratio of 32.03:100 of Acer truncatum oil to lecithin, shake for 10 minutes, and obtain the Acer truncatum oil liposome emulsion.

[0059] Preparation of Acer truncatum oil liposome emulsion using NaCl as solid phase particles:

[0060] Embodiment 8:

[0061] The preparation conditions of this embodiment are the same as those of embodiment 6, except that cholesterol is added to lecithin, and the mass ratio of lecithin to cholesterol is 6:1.

[0062] Comparative Example 1:

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

[0064] Comparative Example 2:

[0065] The preparation conditions of this comparative example are the same as those of Example 6, except that the Acer truncatum oil is replaced by hydroxytyrosol.

[0066] Comparative Example 3:

[0067] Weigh lecithin and maple oil, add them to anhydrous ethanol solution, and perform ultrasonic treatment for 10 minutes to obtain a maple oil organic solution, wherein the mass ratio of maple oil to lecithin is 34.62:100, and the concentration of lecithin in the maple oil organic solution is 0.2 g / ml.

[0068] The maple oil organic solution was placed in an evaporating dish, and edible salt was added in a mass ratio of edible salt to lecithin of 12.5:1 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. Subsequently, deionized water was added to obtain a maple oil liposome emulsion (wherein the volume ratio of deionized water to maple oil was 100:0.04).

[0069] Comparative Example 4:

[0070] Weigh lecithin and add anhydrous ethanol solution, ultrasonicate for 10 minutes, prepare 0.2g / mL lecithin solution, and store in dark. Take edible salt and place it in a rotary evaporator, then spray the lecithin solution into the rotary evaporator at a mass ratio of edible salt to lecithin of 12.5:1, and the rotary evaporation temperature is 50°C to obtain empty liposomes. Take empty liposomes, add 0.04% of the volume fraction of Acer truncatum oil and lecithin in a mass ratio of 34.62:100, shake for 10 minutes, and obtain Acer truncatum oil liposome emulsion.

[0071] Test example 1: Encapsulation efficiency test

[0072] Take 0.1mL of the liposome emulsion prepared in each example 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 of pure water, centrifuge at 8000rpm for 10min, take the permeate as the test solution for free component detection (the present invention refers to the permeate). Continue to add 0.4mL of anhydrous ethanol to the above CF, weigh, ultrasonically dissolve, add to the above UF-CF, ultrafiltration centrifuge at 8000rpm for 10min, take the permeate to make up the lost weight with anhydrous ethanol, as the test solution for the detection of the encapsulated element component (the present invention refers to the concentrated solution).

[0073] HPLC conditions: bonded silica gel C 18 (4.6 mm*250 mm, 5 μm), acetonitrile (A)-0.1% phosphoric acid aqueous solution (B) as mobile phase, gradient elution (0-30 min, A% (35%-80%); 31-40 min, A% 100%), flow rate 1.0 ml min -1 , column temperature 30℃, detection wavelength: 203nm, injection volume: 10μL.

[0074] Among them, encapsulation rate (100%) = amount of encapsulated components / (amount of free components + amount of encapsulated components) × 100%. That is, encapsulation rate (100%) = peak area of ​​concentrated solution / (peak area of ​​permeate + peak area of ​​concentrated solution) × 100%.

[0075] Table 1 Encapsulation efficiency of maple oil liposomes (%) (encapsulation material is based on lecithin)

[0076]

[0077]

[0078] As shown in Table 1, the encapsulation rate of the Acer truncatum oil liposomes prepared in each embodiment and comparative example, it can be seen from Table 1 that the mass ratio of solid phase particles and encapsulation materials, the dosage ratio of Acer truncatum oil and encapsulation materials, and the concentration of Acer truncatum oil all have an important influence on the encapsulation rate. Lecithin is the raw material for forming the liposome shell. Within a certain dosage range of lecithin, the dosage of lecithin is small, the number of liposomes formed is small, and the embedding rate is poor; the dosage of lecithin is too much, causing the lecithin to adhere to each other, unable to form a spherical shape with good morphology, small drug capacity or large thickness of the liposome shell, causing a small amount of drug to be difficult to enter, and the embedding rate is small, while within the appropriate dosage range, the embedding rate is high. When the empty liposomes prepared by using sucrose solid phase particles to embed Acer truncatum oil, the concentration of Acer truncatum oil in the aqueous dispersion has little effect on the embedding rate. The present invention has significant advantages over the prior art, can obtain a higher embedding rate within a wider parameter range, and has a simple preparation process, a small amount of organic solvent, and a short preparation time, and can be prepared on a large scale.

[0079] Figure 1 , Figure 2 The HPLC spectra of the permeate and concentrate of the Acer truncatum oil liposome prepared in Examples 1-4 and 5-8 respectively show that both the permeate and the concentrate have peaks at the linoleic acid 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.

[0080] Test Example 2: Determination of Particle Size

[0081] 0.1 mL of the Acer truncatum oil liposome emulsion prepared in Example 5 was taken and diluted to 1 mL with water, and then detected by a laser particle size analyzer Zetasizer Nano ZS90. The average particle size of the Acer truncatum oil liposome prepared in Example 5 was 238.1 nm, the average PDI was 0.333, and the average potential was -28.4 mV;

[0082] The prepared Acer truncatum oil liposome emulsion in Example 5 was filtered through a 0.22 μm microfiltration membrane, and the average particle size of the Acer truncatum oil liposome was 153.7 nm, the average PDI was 0.155, and the average potential was -31.4 mV, indicating that microfiltration extrusion can effectively control the particle size of the Acer truncatum oil liposome.

[0083] The average particle size of the maple oil liposomes in Example 1 before microfiltration and extrusion was 484.5 nm, the average PDI was 0.513, and the average potential was -74 mV; the average particle size after microfiltration and extrusion was 154.1 nm, the average PDI was 0.159, and the average potential was -36 mV, which also verified the effect of microfiltration and extrusion in effectively controlling the particle size of the maple oil liposomes.

[0084] Table 2 Average particle size and potential of Acer truncatum oil liposomes prepared in each embodiment and comparative example

[0085] Average particle size / nm PDI Potential (mV) Example 1 154.1 0.159 -36 Example 2 154.2 0.165 -34.2 Example 3 153.3 0.140 -26.1 Example 4 152.3 0.160 -34.0 Example 5 153.7 0.155 -31.4 Example 6 156.5 0.164 -36.7 Example 7 154.4 0.174 -36.1 Example 8 151.5 0.138 -35.2 Comparative Example 1 186.3 0.247 -33.8 Comparative Example 2 110.6 0.267 -43.9 Comparative Example 3 160.9 0.225 -37.4 Comparative Example 4 176.7 0.366 -33.1

[0086] It can be concluded from Table 2 that the particle size of the Acer truncatum oil nanoliposomes prepared by this method is between 151.5 and 156.5 nm, the PDI is between 0.138 and 0.174, the liposome particle size and dispersion coefficient are both small, the dispersion is uniform, the average potential of the liposome is between -26.1 and -36.7 mV, the surface of the liposome particles is negatively charged, and the stability is good. Compared with Comparative Examples 1, 3, and 4, Examples 1 to 8 of the present invention have smaller particle sizes, smaller dispersion coefficients, are more stable, and are evenly dispersed, and there is no significant difference in average potential and transmittance; Comparative Example 2 is a hydroxytyrosol liposome prepared by the same method. It can be seen from Table 1 that its encapsulation rate is extremely low, indicating that fat-soluble components such as linoleic acid are easier to encapsulate. In summary, the method for preparing nanoliposomes proposed in the present invention is suitable for the preparation of Acer truncatum oil liposome emulsions.

[0087] Test Example 3: Transdermal Diffusion Experiment

[0088] Using a transdermal diffusion test apparatus, a PEG400 solution with a mass concentration of 40% was added to the diffusion cell, and 0.4 mL of the Acer truncatum oil liposome emulsion prepared in Example 3 and Example 7 and a 0.08% Acer truncatum oil aqueous dispersion were uniformly dispersed on the smooth surface of the artificial skin and permeated at room temperature. After 22 hours, the permeated liquid was taken out from the diffusion cell, extracted with an equal volume of ethyl acetate, and the solvent was evaporated in a water bath. 1 ml of 0.5 mol / L potassium hydroxide methanol solution was added, saponified in a 60°C water bath for 15 min, cooled, 1 ml of boron trifluoride methanol solution was added, esterified in a 60°C water bath for 5 min, cooled, 1 ml of n-hexane was accurately added, shaken, 1 ml of saturated sodium chloride solution was added, shaken well, and the upper layer was taken after stratification as the permeability test solution.

[0089] Retention rate test solution, take the artificial skin treated with 22h, soak it in 2.5ml anhydrous ethanol for 2h, evaporate the solvent in a water bath, and treat it in the same way as above.

[0090] Gas chromatography was carried out in accordance with the general gas chromatography method (general rule 0521) of the 2020 edition of the Chinese Pharmacopoeia, using a capillary column (HP-WAX, 3 m) with polyethylene glycol as the stationary liquid as the chromatographic column, with an initial temperature of 200 °C, and then heated to 220 °C at a rate of 2 °C per minute and maintained for 10 minutes. The injection port temperature was 250 °C, and the detector temperature was 250 °C.

[0091] Test example 4: DPPH free radical scavenging activity assay

[0092] 1. Solution preparation:

[0093] DPPH-ethanol solution: weigh 1 mg of DPPH, add 22.2 ml of anhydrous ethanol, and dissolve by ultrasonication.

[0094] Sample solution: Take an appropriate amount of Acer truncatum essential oil, weigh it accurately, and dissolve it in DMSO to make a volume concentration of 2%,

[0095] 1.25% and 0.625% Acer truncatum essential oil sample solutions.

[0096] Permeability test solution concentrate: evaporate the permeability test solution to dryness, add 0.1 ml DMSO to dissolve, and obtain the concentrate.

[0097] Retention rate test sample solution concentrate: evaporate the retention rate test sample solution to dryness, add 0.1 ml DMSO to dissolve, and obtain the concentrate.

[0098] Vitamin C solution: Take an appropriate amount of vitamin C, weigh it accurately, and dissolve it in 60% ethanol to prepare a vitamin C sample solution with a concentration of 0.004 mg / ml.

[0099] 2. Experimental steps:

[0100] The experiment set up a test group, a control group, and a blank group.

[0101] Determination group: Take 40 μl of the test solution into a 96-well plate, add 60 μl of DPPH free radical solution, shake well, and stand at room temperature (25°C) in the dark for 10 min. Measure its absorbance, which is recorded as A1;

[0102] Control group: According to the above method, the DPPH free radical solution was replaced by an equal volume of 60% ethanol, and its absorbance was measured and recorded as A2;

[0103] Blank group: Pipette 40 μL of 60% ethanol into a 96-well plate, add 60 μL of DPPH free radical solution, shake well, and stand at room temperature away from light for 10 min. Measure the absorbance and record it as A0.

[0104] Add the solutions to the wells in order, keep them at room temperature and away from light for 10 min, measure the absorbance at 517 nm in an ELISA reader, and calculate the clearance rate according to the following formula:

[0105]

[0106] Table 3 DPPH free radical scavenging rate of maple oil

[0107]

[0108]

[0109] The results in Table 3 show that the maple oil has a certain DPPH free radical scavenging effect. The 22h permeability of the maple oil liposome emulsion using sucrose solid phase particles as carriers through the test solution is 0 in Example 3, but its 22h skin retention rate is detected to be 15%, indicating that the maple oil liposome emulsion using salt solid phase particles as carriers can penetrate into artificial skin, but it is extremely difficult to penetrate artificial skin. The 22h permeability of the maple oil liposome emulsion using salt solid phase particles as carriers through the test solution is 25%, and its 22h skin retention rate is not detected, indicating that the maple oil liposome emulsion using sugar solid phase particles as carriers has a certain artificial skin permeability, and the DPPH free radical scavenging effect also confirms that it has a certain permeability. The maple oil aqueous dispersion is detected to penetrate the test solution and the skin retention test solution, indicating that the maple oil can significantly improve its artificial skin retention or permeability by forming liposomes, which is beneficial to the absorption and utilization of the effective components of the maple oil.

Claims

1. A method for preparing a maple 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 liposomes with the Acer truncatum oil aqueous dispersion to obtain the Acer truncatum oil liposome emulsion.

2. The preparation method according to claim 1, characterized in that: In step (a), the encapsulating material is one of lecithin and lecithin-cholesterol; the mass ratio of lecithin to cholesterol is (4-8):1, preferably 6:

1.

3. The preparation method according to claim 1, characterized in that: In step (b), the solid phase particles are sucrose or salt; and the particle size of the solid phase particles is 150-1250 μm.

4. The preparation method according to claim 3, characterized in that: The particle size of the sucrose solid phase particles is 300-900 μm, preferably 500-600 μm; the particle size of the salt solid phase particles is 900-1250 μm, preferably 600-700 μm.

5. The preparation method according to claim 1, characterized in that The mass ratio of the solid phase particles to the encapsulating material is (1.56-12.5):1; preferably, the mass ratio of the sucrose solid phase particles to the encapsulating material is (1.56-6.25):1; preferably, the mass ratio of the salt solid phase particles to the encapsulating material is (1.56-12.5):

1.

6. The preparation method according to claim 1, characterized in that: The mass concentration of the encapsulating material in the encapsulating material solution is 0.1-0.2 g / mL; the mass ratio of the Acer truncatum oil to lecithin is (32.03-37.18):100; The solvent of the Acer truncatum oil-water dispersion is water, and the volume fraction of the Acer truncatum oil-water dispersion is 0.04% to 0.08%.

7. The preparation method according to claim 1, characterized in that: The method further comprises a step (d) of a granulation method, wherein the granulation method comprises subjecting the liposomes to one of the following treatments: ultrasound, shearing, homogenization, and extrusion, preferably extrusion, and more preferably microfiltration extrusion.

8. The Acer truncatum oil liposome emulsion prepared by the method according to any one of claims 1 to 7.

9. The emulsion according to claim 8, characterized in that The encapsulation rate of the Acer truncatum oil liposome is 87.41-100%. Preferably, after the whole particle treatment, the particle size of the Acer truncatum oil liposome emulsion is 151.5 to 156.5 nm, the aggregation index (PDI) is 0.138 to 0.174, and the emulsion potential is -26 to -37 mV.

10. Use of the Acer truncatum oil liposome emulsion according to claim 8 in the preparation of anti-aging, antibacterial or antioxidant skin care products.

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