Buddha fragrance essential oil liposome, Buddha fragrance essential oil liposome gel, and preparation method and application

Abstract

The present invention discloses buddha fragrance essential oil liposomes, buddha fragrance essential oil liposome gel, and preparation method and application, belonging to the field of pharmaceutical technology. The raw materials of the buddha fragrance essential oil liposomes of the present invention include buddha fragrance essential oil, lecithin and cholesterol. The preparation method of the buddha fragrance essential oil liposomes comprises: dissolving buddha fragrance essential oil, lecithin and cholesterol in a solvent, performing vacuum rotary evaporation to form a thin film of lipid on the wall of the container; adding a buffer solution and continuing rotary evaporation to make the thin film fall off, ultrasonically hydrating; and then high-pressure homogenization to obtain the product. The raw materials of the buddha fragrance essential oil liposome gel of the present invention include: a gel matrix, a moisturizing agent, triethanolamine and buddha fragrance essential oil liposomes. The preparation method of the buddha fragrance essential oil liposome gel comprises: taking the gel matrix and adding the buddha fragrance essential oil liposomes to swell, homogenizing, adding propylene glycol and triethanolamine, adding water to a specified amount, and stirring evenly to obtain the product. The present invention has a precise and safe therapeutic effect on acne; improves the stability and water solubility of essential oils and delays the release time of essential oils.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines, and in particular relates to foeniculum odoratum essential oil liposome, foeniculum odoratum essential oil liposome gel, and a preparation method and application thereof. Background Art

[0002] Acne is one of the most common skin diseases worldwide. Research estimates suggest it will become the eighth most prevalent disease globally, affecting 9.4% of the global population. Epidemiological studies show that acne is most common among adolescents, with boys being the primary sufferers. Acne manifests clinically as a rash consisting of comedones, pustules, and deep suppurative lesions. Severe cases can cause disfigurement and scarring, severely diminishing patients' self-esteem and impacting their mental health.

[0003] Studies have shown that the onset of acne is closely related to the proliferation of epidermal bacteria, which leads to skin inflammation. Clinically, oral antibiotics are commonly used to treat moderate to severe acne caused by P. acnes. Although antibiotics have certain efficacy in treating acne, long-term use can easily lead to drug resistance, making the treatment effect no longer significant, and produce adverse reactions such as higher rates of upper respiratory tract infections. Currently, the problem of bacterial resistance has become a serious problem that hinders the clinical treatment of acne. Another commonly used drug, retinoids, has good efficacy, but it has serious adverse reactions, which hinders the clinical use of such drugs. Based on the above background, the search for topical anti-acne drugs with precise efficacy and few side effects has become a worldwide concern. The development of anti-acne drugs with good effects and less likelihood of developing resistant strains has become very urgent and necessary.

[0004] Plant essential oils (EOs) are aromatic, oily liquids obtained through steam distillation of plants and are green, natural antibacterial materials. Therefore, providing an acne medication using plant essential oils as raw materials has become an urgent problem to be solved by those skilled in the art. Summary of the Invention

[0005] One of the purposes of the present invention is to provide a liposome of chrysanthemum essential oil, which has good antibacterial effect, can significantly inhibit acne disease-related strains, improve the stability and water solubility of essential oil, and delay the release time of essential oil.

[0006] The second object of the present invention is to provide a method for preparing the Buddha's fragrance essential oil liposome.

[0007] The third object of the present invention is to provide a liposome gel of ginseng essential oil prepared with ginseng essential oil liposome as raw material.

[0008] A fourth object of the present invention is to provide a method for preparing the Buddha's fragrance essential oil liposome gel.

[0009] A fifth object of the present invention is to provide a use of the liposomes of the ginseng essential oil in the preparation of external-use medicines for preventing and treating acne.

[0010] The sixth object of the present invention is to provide a use of the liposome gel of the Buddha's fragrance essential oil in the preparation of an external-use medicine for preventing and treating acne.

[0011] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0012] The present invention provides a buddha fragrance essential oil liposome. The raw materials of the buddha fragrance essential oil liposome include buddha fragrance essential oil, lecithin and cholesterol. The buddha fragrance essential oil is composed of bergamot essential oil and clove essential oil in a mass ratio of 1 to 3:1, the mass ratio of lecithin to cholesterol is 4 to 6:1, and the total mass ratio of buddha fragrance essential oil, lecithin and cholesterol is 1 to 2.5:7.

[0013] In some embodiments of the present invention, the mass ratio of bergamot essential oil to clove essential oil is 1:1; or / and the mass ratio of lecithin to cholesterol is 5-6:1; or / and the total mass ratio of bergamot essential oil to lecithin and cholesterol is 2:7.

[0014] The present invention provides a method for preparing a liposome of Buddha's fragrance essential oil, which comprises the following steps:

[0015] S1. After dissolving the Buddha's essential oil, lecithin and cholesterol in a solvent, vacuum rotary evaporation was performed to form a thin film of lipids on the wall;

[0016] S2. Add buffer solution, continue rotary evaporation to remove the film, and ultrasonically hydrate;

[0017] S3. The ultrasonically hydrated material is homogenized by a high-pressure homogenizer to obtain.

[0018] In some embodiments of the present invention, in S1, the rotary evaporation under reduced pressure is carried out at 30-50°C, preferably at 35°C;

[0019] In said S2, the pH value of said buffer solution is 7.0-8.0, preferably 7.5-8.0;

[0020] In said S2, after adding the buffer solution, rotary evaporation is continued at 40-60°C, preferably 45°C;

[0021] In S2, the ultrasonic hydration time is 10-40 min, preferably 20-30 min;

[0022] In S3, the material after ultrasonic hydration is circulated and homogenized in a high-pressure homogenizer at 60-100 MPa for 90-150 seconds; preferably, the homogenization pressure is 80 MPa and the homogenization time is 120 seconds.

[0023] The invention provides a liposome gel of buddhist fragrance essential oil, the raw materials of which include: a gel matrix, a moisturizing agent, triethanolamine and the liposome of buddhist fragrance essential oil according to claim 1 or 2.

[0024] In some embodiments of the present invention, the gel matrix comprises carbomer, sodium carboxymethyl cellulose, sodium alginate, and xanthan gum, preferably carbomer;

[0025] The moisturizing agent includes at least one of glycerin and propylene glycol, preferably propylene glycol.

[0026] In some embodiments of the present invention, the gel matrix accounts for 1.0-1.5% of the total mass of the Buddha's fragrance essential oil liposome gel, preferably 1.25%;

[0027] The propylene glycol accounts for 7.5-15% of the total mass of the Buddha's fragrance oil liposome gel, preferably 12.5%;

[0028] Triethanolamine accounts for 0.5-1% of the total mass of the Buddha's fragrance oil liposome gel, preferably 0.5%;

[0029] The Buddha's fragrance essential oil liposomes account for 60-75% of the total mass of the Buddha's fragrance essential oil liposome gel.

[0030] The preparation method of the Buddha fragrance essential oil liposome gel provided by the present invention comprises the following steps:

[0031] Take the gel matrix, add the Buddha fragrance essential oil liposome to swell, homogenize with a high-speed dispersing homogenizer, add propylene glycol, stir evenly, then add triethanolamine, add water to the specified amount, stir evenly, and obtain the Buddha fragrance essential oil liposome gel.

[0032] The invention provides an application of the Buddha's fragrance essential oil liposome in preparing an external-use medicine for preventing and treating acne.

[0033] The invention provides an application of the Buddha fragrance essential oil liposome gel in preparing an external-use medicine for preventing and treating acne.

[0034] Compared with the prior art, the present invention has the following beneficial effects:

[0035] The present invention creatively uses the Buddha's fragrance essential oil as raw material to prepare an external medicine for preventing and treating acne, which has a definite therapeutic effect. In addition, the natural plant essential oil is safe and has no drug resistance, which provides another option for the development of acne medicines.

[0036] The present invention prepares essential oil into liposomes and then into a gel, thereby improving the stability and water solubility of the essential oil and delaying the release time of the essential oil, so as to prepare an essential oil product with good safety, strong stability and easier skin absorption. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1This is the appearance of Buddha fragrance essential oil liposome suspension.

[0038] Figure 2 This is a transmission electron micrograph of liposomes of Buddha's fragrance essential oil.

[0039] Figure 3 This is the particle size distribution diagram of Buddha fragrance essential oil liposomes.

[0040] Figure 4 This is the Zeta potential diagram of Buddha essential oil liposomes.

[0041] Figure 5 The figure is a chromatogram for determination of organic residues; A is a chromatogram of chloroform solution, B is a chromatogram of liposome solution of jojoba essential oil, 1 is the O2 chromatographic peak, and 2 is the chloroform chromatographic peak.

[0042] Figure 6 DSC graphs of (A) Buddha's fragrance essential oil and (B) Buddha's fragrance essential oil liposome freeze-dried powder.

[0043] Figure 7 This is the FT-IR image of Buddha fragrance essential oil.

[0044] Figure 8 This is the FT-IR image of lyophilized powder of Buddha's fragrance essential oil liposome.

[0045] Figure 9 This is the Qt curve of eugenol.

[0046] Figure 10 This is the appearance and properties of Buddha fragrance essential oil liposome gel.

[0047] Figure 11 This is the particle size distribution diagram of Buddha essential oil liposome gel.

[0048] Figure 12 This is the Qn-t curve of eugenol.

[0049] Figure 13 Figure 2 shows the skin retention of D-limonene and eugenol. DETAILED DESCRIPTION

[0050] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0051] The method for determining the encapsulation efficiency in the embodiment of the present invention is:

[0052] 4 mL of the liposome suspension of the essential oil of the Buddha was accurately drawn and centrifuged at 40,000 r / min for 30 min (to separate the liposomes and the free volatile oil). After removing the supernatant, an equal volume of chromatographic methanol was added, and the mixture was ultrasonically treated in an ice-water bath for 1 h, and then centrifuged at 10,000 r / min for 20 min at 4°C. The supernatant was filtered through a 0.22 μm microporous filter membrane, and the filtrate was taken. The concentration of encapsulated D-limonene C1 and the concentration of eugenol C2 in the essential oil liposome were determined by gas chromatography. Another 4 mL of the essential oil liposome suspension was accurately drawn and demulsified with an equal volume of methanol, ultrasonicated for 30 min, and centrifuged at 10,000 r / min for 20 min. The concentration of D-limonene C in the essential oil liposome suspension was determined by the same method. 1总 , the concentration of eugenol C 2总 The encapsulation efficiency calculation formula is:

[0053] EE D-柠檬烯 =C1 / C 1总 ×100%,EE 丁香酚 =C2 / C 2总 ×100%

[0054] The chromatographic conditions of the gas chromatography method were as follows: Agilent DB-5MS capillary column (30 m × 0.25 mm × 0.25 μm); heating program: initial temperature 50°C, hold for 2 min, heat to 150°C at 15°C / min, hold for 3 min, then heat to 250°C at 25°C / min, hold for 2 min; injection port temperature 250°C; carrier gas was He, with a flow rate of 1.0 ml / min; injection mode was pulse split injection, split ratio 10:1, pulse pressure 120 kPa, time 0.5 min, and injection volume 1 μL.

[0055] Example 1

[0056] This embodiment discloses a method for preparing the Buddha's fragrance essential oil liposome of the present invention, specifically:

[0057] 240 mg of egg yolk lecithin, 40 mg of cholesterol and 80 mg of ginseng essential oil were dissolved in an appropriate amount of chloroform, shaken and mixed in a round-bottom flask, and rotary evaporated under reduced pressure at 35°C to form a thin film of lipid on the wall. 20 mL of PBS buffer solution, pH 8.0, was added with glass beads, and rotary evaporated at 45°C for 20 min to remove the thin film. The lipid was then hydrated by water bath ultrasound for 20 min, and then homogenized in a high-pressure homogenizer at 80 MPa for 120 s to obtain a suspension of ginseng essential oil liposomes.

[0058] Example 2

[0059] This embodiment discloses a screening experiment for the types and proportions of essential oils of the present invention.

[0060] 1. Screening of essential oil types

[0061] Using the inhibition zone assay, eight plant essential oils were tested for their ability to inhibit acne: bergamot, clove, turmeric, Chuanxiong, eucalyptus, lavender, fructus aurantii, and mugwort. Essential oils with strong antibacterial properties were selected for compounding to explore the combined antibacterial effects, aiming to achieve synergistic and additive effects with the essential oils and enhance their antibacterial effects.

[0062] The specific method is:

[0063] Use a punch to press the filter paper into discs with a diameter of 6 mm, divide them into clean and dry test tubes, sterilize them in a high-pressure steam cooker at 121°C for 15 minutes, and place them in a clean bench for later use.

[0064] Place the cooled and solidified solid culture medium plate on a clean bench, mark the area on the back of the culture plate and number it. Pipette 100 μL of bacterial suspension onto the plate and spread it slowly and evenly over the entire culture medium plate with a coating stick. The concentration of bacterial suspension is OD value 0.8 (about 1.5×10 8 After 2-3 minutes, use sterile tweezers to gently place the filter paper onto the culture medium at the marked site. Gently press the filter paper to ensure good contact without puncturing the culture medium surface. Then, vertically drip 5 μL of each of the eight plant essential oils onto the filter paper. One filter paper serves as a blank control (normal saline). After 3 minutes, invert the plate and incubate in an incubator for 24 hours (aerobically incubate Staphylococcus epidermidis at 37°C; anaerobic incubate Propionibacterium acnes in an anaerobic bag at 37°C). After 24 hours, measure the diameter of the inhibition zone on the filter paper using the cross-hatch method.

[0065] The results are shown in Table 1:

[0066] Table 1 Diameter of inhibition zone of plant essential oils

[0067]

[0068] Note: *p<0.05, **p<0.01 compared with blank control group.

[0069] Verification experiment:

[0070] Table 2 Diameters of inhibition zones of plant essential oils

[0071]

[0072] Note: *p<0.05, **p<0.01 compared with blank control group.

[0073] The antibacterial effects of plant essential oils on Propionibacterium acnes and Staphylococcus epidermidis were determined based on the diameter of the inhibition zone. The results showed that the eight plant essential oils exhibited varying degrees of antibacterial activity against the test bacteria, with clove essential oil showing the greatest effect, followed by bergamot essential oil. The inhibition zone diameters for all oils ranged from 10mm to 15mm, indicating moderate sensitivity. The remaining plant essential oils were all below 10mm, indicating low sensitivity. Clove and bergamot essential oils were selected from the eight plant essential oils for subsequent experiments.

[0074] 2. Screening of compound essential oil ratios

[0075] Bergamot and clove essential oils, both of which have the best antibacterial properties, were selected and mixed in varying proportions for antibacterial testing. The antibacterial effects of the compound essential oils and individual essential oils were compared by measuring the size of the inhibition zone. Erythromycin was used as the positive control, and saline was used as the blank control. The specific methods were the same as those for the antibacterial test under "Essential Oil Selection." The results of the antibacterial efficacy test are shown in Table 3, and the results of the validation test for the optimal formulation are shown in Table 4.

[0076] Table 3 Diameters of inhibition zones of different proportions of bergamot and clove essential oils

[0077]

[0078]

[0079] Note: *p<0.05, **p<0.01 compared with blank control group.

[0080] Verification experiment:

[0081] Table 4 Diameters of inhibition zones for the ratio of bergamot and clove essential oils (1:3) (n=3)

[0082]

[0083] Note: *p<0.05, **p<0.01 compared with blank control group.

[0084] The above results show that the antibacterial effect of the bergamot and clove essential oil ratio of (1:3) is the best. According to the measurement results of the inhibition zone, the antibacterial effect of the mixture of bergamot and clove essential oils is greater than that of the single essential oils of bergamot and clove.

[0085] 3. Determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of Plant Essential Oils

[0086] Use the MIC continuous test tube dilution method to prepare 12 10mL test tubes, add 3.875mL of culture medium to test tube No. 1, then add 2mL of culture medium to test tubes No. 2-12 respectively, add 0.125mL of essential oil to test tube No. 1, mix repeatedly with a pipette, take 2mL and add to tube No. 2, dilute to tube No. 10 in sequence, aspirate 2mL from tube No. 10 and discard. Tube No. 11 is a blank control tube for sterile growth of culture medium, and tube No. 12 is a positive control tube for bacterial growth. Except for tube No. 11, add 100μL of bacterial suspension to each tube and culture in a 37℃ constant temperature shaker for 24-48h. The OD value of the bacterial suspension concentration is 0.8 (about 1.5×10 8 cfμ / mL).

[0087] The final concentrations of the bergamot essential oil in test tubes 1 to 10 were 53.125, 26.562, 13.281, 6.640, 3.320, 1.660, 0.83, 0.415, 0.207, and 0.103 mg / mL;

[0088] Final concentrations of clove essential oil: 60.0, 30.0, 15.0, 7.50, 3.75, 1.875, 0.937, 0.468, 0.234, 0.117 mg / mL;

[0089] Concentrations of Buddha fragrance essential oil: 59.375, 29.687, 14.843, 7.421, 3.710, 1.855, 0.927, 0.463, 0.231, 0.115 mg / mL.

[0090] After 24 hours of incubation, take 100 μL of the suspension from each test tube and add it to the corresponding culture medium plate of bacteria. Use a spreader to slowly and evenly spread it, invert and incubate at 37°C. Return the remaining suspension to a 37°C constant temperature shaker for incubation. Incubate the culture medium plates for 18-24 hours. Remove the test and blank culture plates and compare whether there is bacterial growth. The minimum drug concentration in the absence of visible bacteria is the minimum inhibitory concentration (MIC).

[0091] The remaining culture suspension was taken out after 24 hours, and 100 μL of the suspension was taken from the test tube of each concentration and added to the corresponding solid culture medium plate. It was evenly spread with a spreader and inverted to culture at 37°C for 18-24 hours. The growth of bacteria in the culture plates of the test group and the blank group was compared, and the complete sterile growth was defined as the minimum bactericidal concentration (MBC).

[0092] Evaluation of the antibacterial effect of the combined effects of buddha fragrance essential oil using the FIC index calculation formula

[80] :

[0093]

[0094] If the FIC index is ≤0.5, it is a synergistic effect; if 0.5<FIC index ≤1, it is an additive effect; if 1<FIC index ≤2, it is an irrelevant effect; and if the FIC index is >2, it is an antagonistic effect.

[0095] The results are shown in the following table:

[0096] Table 5 Antibacterial effect of essential oils on Propionibacterium acnes (n=3)

[0097]

[0098] Table 6 Antibacterial effect of essential oils on Staphylococcus epidermidis (n=3)

[0099]

[0100] The results showed that the MIC and MBC of compound buddha's fragrance essential oil against Propionibacterium acnes and Staphylococcus epidermidis were both 3.710 mg / mL; while the MIC and MBC of clove essential oil against Propionibacterium acnes were both 7.50 mg / mL, and the MIC and MBC against Staphylococcus epidermidis were 3.750 mg / mL and 7.50 mg / mL; the MIC of bergamot essential oil against Propionibacterium acnes was 6.640 mg / mL and the MBC was 13.280 mg / mL, and the MIC and MBC against Staphylococcus epidermidis were both 13.280 mg / mL. The antibacterial effect of plant essential oils against Propionibacterium acnes and Staphylococcus epidermidis was compound buddha's fragrance essential oil > clove essential oil > bergamot essential oil.

[0101] Through the FIC index effect evaluation, the FIC values ​​of the compound Buddha fragrance essential oil against Propionibacterium acnes and Staphylococcus epidermidis were both 0.5<FIC index≤1, indicating that the combined antibacterial effect of the compound Buddha fragrance essential oil on Propionibacterium acnes and Staphylococcus epidermidis is additive.

[0102] Example 3

[0103] This example investigates the process conditions of the liposomes of the Buddha's fragrance essential oil.

[0104] 1. Single Factor Investigation of Liposome Preparation

[0105] Taking the encapsulation efficiency as the evaluation index, the thin film dispersion method was used to investigate the effects of high-pressure homogenizer homogenization pressure, homogenization time, lecithin to cholesterol mass ratio, essential oil addition amount, hydration medium pH, and ultrasonic time on liposomes.

[0106] 1.1 Homogenization pressure

[0107] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. The homogenization pressure was set to 40 MPa, 60 MPa, 80 MPa, and 100 MPa, respectively. The results are shown in Table 7.

[0108] Table 7 Effect of homogenization pressure on liposomes of buddhist essential oil

[0109] Mean pressure / MPa D-limonene encapsulation rate / % Eugenol encapsulation rate / % 40 25.02 33.81 60 35.43 48.04 80 42.38 52.98 100 43.56 53.06

[0110] The results showed that as the homogenization pressure increased, the encapsulation efficiency first increased. The liposome encapsulation efficiency values ​​of liposomes homogenized at 80MPa and 100MPa did not change significantly, and the pressure of the high-pressure homogenizer was unstable when homogenizing at 100MPa.

[0111] 1.2 Homogenization time

[0112] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. The homogenization time was 60 s, 90 s, 120 s, 150 s, 180 s, and 210 s, respectively, and the encapsulation efficiency was measured. The results are shown in the following table.

[0113] Table 8 Effect of homogenization time on liposomes of Buddha's fragrance essential oil

[0114] Homogenization time / s D-limonene encapsulation rate / % Eugenol encapsulation rate / % 60 35.49 38.08 90 44.25 44.50 120 45.56 54.37 150 43.18 43.88 180 36.31 40.68

[0115] The results showed that with the increase of homogenization time, the liposome encapsulation efficiency first increased and then decreased. The liposome encapsulation efficiency of 90s, 120s, and 150s homogenization did not change significantly, but the liposomes were more uniform and clear at 120s and were not easy to stratify.

[0116] 1.3 Lecithin / Cholesterol Ratio

[0117] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. The mass ratios of lecithin: cholesterol were 2:1, 3:1, 4:1, 5:1, and 6:1, respectively, and the encapsulation efficiency was measured. The results are shown in the following table.

[0118] Table 9 Effect of lecithin-cholesterol mass ratio on liposomes of Buddha's fragrance essential oil

[0119] Lecithin to cholesterol mass ratio D-limonene encapsulation rate / % Eugenol encapsulation rate / % 2﹕1 22.91 31.79 3﹕1 28.79 34.59 4﹕1 42.23 43.87 5﹕1 45.58 64.45 6﹕1 44.64 58.27

[0120] The results showed that as the ratio of lecithin to cholesterol increased, the liposome encapsulation efficiency first increased and then decreased. When the ratio was 5:1 and 6:1, the liposome encapsulation efficiency did not change significantly.

[0121] 1.4 Amount of essential oil added

[0122] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. The addition amounts of essential oil were 20, 40, 60, 80, 100, and 120 mg, and the encapsulation efficiency was measured. The results are shown in the following table.

[0123] Table 10 Effect of essential oil addition on liposomes of buddha fragrance essential oil

[0124] Essential oil addition amount / mg D-limonene encapsulation rate / % Eugenol encapsulation rate / % 20 21.67 30.14 40 37.46 45.96 60 44.80 51.28 80 53.82 69.00 100 45.03 55.83 120 28.87 32.58

[0125] The results showed that with the increase of the amount of essential oil added, the encapsulation efficiency of essential oil liposomes first increased and then decreased.

[0126] 1.5 water pH

[0127] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. The pH values ​​of PBS hydration medium were examined at 5.5, 6, 6.5, 7, 7.5, and 8, and the encapsulation efficiency was measured. The results are shown in the following table.

[0128] Table 11 Effect of hydration medium pH on liposomes of buddha fragrance essential oil

[0129] Water pH D-limonene encapsulation rate / % Eugenol encapsulation rate / % 5.5 19.29 30.75 6 26.78 37.82 6.5 29.07 39.80 7 50.32 60.70 7.5 53.71 65.95 8 52.74 63.31

[0130] The results showed that with the increase of the pH value of the hydration medium, the liposome encapsulation efficiency first increased and then decreased. The liposome encapsulation efficiency did not change significantly when pH = 7.5 and pH = 8.0, so the pH value of the hydration medium was selected as 7.5.

[0131] 1.6 Ultrasound time

[0132] Liposomes were prepared according to the method of Example 1, with other conditions kept unchanged. Ultrasonication time was examined at 10, 20, 30, 40, 50, and 60 min, and the encapsulation efficiency was measured. The results are shown in the following table.

[0133] Table 12 Effect of ultrasonic time on liposomes of Buddha's fragrance essential oil

[0134]

[0135]

[0136] The results showed that the encapsulation efficiency of essential oil liposomes decreased with the increase of ultrasonic time. If the ultrasonic time was less than 10 min, the film would stick to the round-bottom flask and could not be completely removed.

[0137] 2. Liposome Preparation Process Optimization

[0138] 2.1 Orthogonal experimental design

[0139] In order to optimize the formulation of liposomes of ginseng essential oil, four factors were screened out based on single-factor experiments, including the ratio of phospholipids to cholesterol (A), the amount of essential oil added (B), the hydration medium (C), and the ultrasonic time (D). Three different levels were selected for each factor (see Tables 13-14). The encapsulation efficiency was used as the optimization index, and the L9 (34) orthogonal experimental method was used to optimize the preparation process of liposomes of ginseng essential oil.

[0140] Nine groups of experiments were conducted according to the orthogonal design table. The results of the orthogonal experiments are shown in Table 13, and the results of the variance analysis are shown in Table 14.

[0141] Table 13 Factor level table

[0142]

[0143] Table 14 Orthogonal test results

[0144]

[0145]

[0146] Table 15 ANOVA results

[0147]

[0148] Note: F 0.05 (2, 2) = 19, F 0.01 (2, 2) = 99, * indicates a significant difference

[0149] The results showed that the range (R) for D-limonene and eugenol was RA > RB > RC > RD. The order of influence of each factor on the encapsulation efficiency of essential oil liposomes was A > B > C > D, that is, lecithin to cholesterol ratio > essential oil addition amount > hydration medium pH > ultrasonication time. Analysis of variance showed that the lecithin to cholesterol ratio had a significant effect on the encapsulation efficiency of essential oil liposomes, and the difference was statistically significant (P < 0.05). The optimal preparation process for essential oil liposomes was A3B2C3D2, namely, a lecithin to cholesterol ratio of 6:1, essential oil addition amount of 80 mg, hydration medium pH = 8.0, and ultrasonication time of 20 min.

[0150] 2.2 Verification test

[0151] Dissolve 240 mg of egg yolk lecithin, 40 mg of cholesterol and 80 mg of ginseng essential oil in an appropriate amount of chloroform, shake and mix in a round-bottom flask, and rotary evaporate under reduced pressure at 35°C to form a thin film of lipid on the wall. Add 20 mL of PBS buffer solution, pH 8.0, add glass beads, and rotary evaporate at 45°C for 20 minutes to remove the thin film. Then, ultrasonicate in a water bath for 20 minutes to hydrate the lipid, and then homogenize in a high-pressure homogenizer at 80 MPa for 120 seconds to obtain the product.

[0152] Table 16 Buddha essential oil liposome validation test (n=3)

[0153] Serial number D-limonene encapsulation rate / % Eugenol encapsulation rate / % 1 64.72 84.63 2 63.85 84.35 3 64.09 83.93 average value 64.22 84.30 RSD / % 0.70 0.41

[0154] The results showed that three batches of eugenol essential oil liposomes were prepared according to the optimal orthogonal design scheme, with the D-limonene encapsulation efficiency of 64.22% and RSD value of 0.70%; the eugenol encapsulation efficiency of 84.30 and RSD value of 0.41%.

[0155] Test Example 1

[0156] This test example investigated the physicochemical properties of the Buddha's fragrance essential oil liposomes prepared according to the method of Example 1.

[0157] 1. Appearance

[0158] As attached Figure 1 As shown, the Buddha fragrance essential oil liposomes of the present invention are a light yellow suspension with good uniform fluidity and no particles visible to the naked eye.

[0159] 2. Determination of the Morphology of Foxiang Liposomes

[0160] The morphology of the liposomes of the essential oil of the Buddha's fragrance was observed using a transmission electron microscope (TEM). Take an appropriate amount of the liposomes of the essential oil of the Buddha's fragrance, dilute it to an appropriate concentration, drop a small amount on a carbon copper grid, let it stand for 1 minute, then dry it with a filter paper, then drop 1% phosphotungstic acid solution on the copper grid for negative staining for 1 minute, evaporate it naturally, and observe the morphology of the liposomes. The transmission electron microscope image of the liposomes of the essential oil of the Buddha's fragrance is shown in the attached figure. Figure 2 As shown: The liposomes are spherical, with complete structure and are non-sticky.

[0161] 3. Determination of particle size, PDI and Zeta potential

[0162] As attached Figure 3 To the attached Figure 4 As shown, the average particle size of the Buddha's fragrance essential oil liposomes was 178±2.30 nm, the PDI was 0.18±0.02, and the Zeta potential was -32±0.30 mV.

[0163] 4. Organic residue test of Buddha fragrance essential oil liposome

[0164] According to the general chapter 0861 Residual Solvent Determination Method (Method 2) of the "Chinese Pharmacopoeia" (2020 edition, Volume IV), the capillary column headspace injection programmed temperature method was used to detect the residual amount of chloroform in the liposomes of ginseng essential oil.

[0165] Chromatographic conditions: Agilent DB-5MS capillary column (30 m × 0.25 mm × 0.25 μm), detector temperature 150°C, temperature program: start temperature 40°C, hold for 3 min, increase to 150°C at a heating rate of 10°C per minute, hold for 5 min; increase to 250°C at a heating rate of 30°C per minute, hold for 3 min, split ratio 10:1, hydrogen flame ionization detector (FID).

[0166] Take 0.5 mL of chloroform solution and liposome solution of ginseng essential oil respectively, place them in a 10 mL volumetric flask, dilute to the mark with N,N-dimethylformamide (DMF), mix well, and put them into a headspace injection bottle. Detect according to the above chromatographic conditions. The chromatogram is shown in the attached figure. Figure 5 shown.

[0167] 5. Differential Scanning Calorimetry

[0168] Accurately weigh the euphorbia essential oil and euphorbia essential oil liposome freeze-dried powder and perform differential scanning calorimetry analysis. Related parameters: initial temperature 30℃, end temperature 400℃, 30℃·min -1 Heating, filling gas is N2, control: empty aluminum crucible. The results are shown in the attached Figure 6 shown.

[0169] Depend on Figure 6 It can be seen that the lyophilized powder of buddha fragrance essential oil and buddha fragrance essential oil liposomes have an exothermic peak at 250°C and 300°C, respectively. The shift of the exothermic peak indicates that the buddha fragrance essential oil is encapsulated in the liposomes.

[0170] 6Infrared spectrum test

[0171] Take appropriate amount of buddhist essential oil and buddhist essential oil liposome freeze-dried powder samples respectively, use KBr as dispersant to press them into tablets, and observe the results at wave number 4000-400cm -1 Scan within the range. The results are shown in the attached Figure 7 and attached Figure 8 shown.

[0172] From the infrared spectrum of Buddha fragrance oil, at (1514±1cm -1 ) is the C=C stretching vibration absorption peak of eugenol molecule, at (1640±1cm -1 ) is the C=C stretching vibration absorption peak of the limonene molecule, but this characteristic peak is not found in the freeze-dried powder of the Buddha's fragrance essential oil liposome, which proves that the essential oil is encapsulated into the liposome.

[0173] 7 In vitro release assay test

[0174] The determination was carried out according to the third method (small cup method) in the general dissolution and release determination method of the "Chinese Pharmacopoeia" (2020 edition, Part IV): the release medium was 100 mL of degassed normal saline. 2 mL of the liposome suspension of the euphorbia pilosa essential oil and the euphorbia pilosa essential oil solution were respectively aspirated and placed in a treated dialysis bag (molecular weight 7000Da cutoff), a glass bead was placed in it, and both ends were clamped with a seal. 100 mL of degassed normal saline was added to the beaker, the temperature was set to 38±1°C, the speed was 50 r / min, and 1 mL of the medium solution was aspirated at 1, 2, 4, 6, 8, 10, and 12 h (1 mL of the same temperature medium was supplemented at the same time), filtered with a 0.22 μm microporous filter membrane, and the filtrate was taken. The content was determined by gas chromatography. The chromatographic conditions were the same as those for the encapsulation rate determination. Calculate the cumulative release percentage Q of the sample at different time points and plot it against time t, see. Figure 9 .

[0175] The calculation formula is as follows:

[0176]

[0177] Among them, C t is the concentration at the sampling point, V o is the volume of the release medium - 100 mL in this test, V is the sampling volume, C t-1 is the mass concentration of the previous sampling point, M o is the total amount of eugenol in the system.

[0178] From the drug release curve, it can be seen that the 24-hour cumulative release rate of eugenol in the eugenol essential oil solution is 70.09%, but there is no drug release between 12 and 24 hours, indicating that the drug release is basically completed within 12 hours; while the 24-hour cumulative release rate of eugenol in the eugenol essential oil liposome solution is 63.18%, and the drug is still slowly released between 12 and 24 hours, indicating that the liposome has a sustained-release effect; the cumulative release rate of eugenol in the eugenol essential oil liposome solution is less than 40% within the first hour, and there is no burst release effect, which meets the in vitro release test evaluation requirements for sustained-release, controlled-release and delayed-release preparations in the 2020 edition of the "Chinese Pharmacopoeia".

[0179] Equation fitting of the release curve:

[0180] The average release data of eugenol in eugenol liposome solution and eugenol in eugenol liposome solution at each time point were processed and fitted according to the first-order kinetic model and Higuchi model respectively to obtain the kinetic equation, release model fitting equation and correlation coefficient (R 2 ), the results showed that the in vitro release curves were best fitted with the first-order kinetic model, indicating that the essential oil liposome solution had a certain sustained-release effect.

[0181] Table 17 Equation fitting of eugenol release curve

[0182]

[0183]

[0184] 8 Effect of storage time on liposome stability

[0185] The prepared Buddha's fragrance essential oil liposomes were stored in a refrigerator at 4°C away from light, and samples were taken out at 0, 1, 2, 5, and 10 days to observe the appearance of the liposomes.

[0186] After short-term storage, the appearance of the essential oil liposomes did not change significantly, and no stratification or large-scale precipitation occurred, indicating that the liposomes had good stability.

[0187] Example 4

[0188] This embodiment discloses a method for preparing liposome gel of buddhist essential oil.

[0189] Take 0.375g of carbomer and add 20g of Buddha's fragrance essential oil liposome to swell overnight, use a high-speed dispersion homogenizer at 3000r / min, homogenize for 2min, add 3.75g of propylene glycol, stir on a magnetic stirrer for 10min, then add 0.125g of triethanolamine and 0.1g of methylparaben, add water to 30g, and stir to obtain Buddha's fragrance essential oil liposome gel. The Buddha's fragrance essential oil liposome of the present embodiment is prepared by the method of Example 1.

[0190] Example 5

[0191] This embodiment discloses an experiment to screen and optimize the process conditions of Buddha's essence liposome gel.

[0192] 1. Selection of gel matrix

[0193] A gel based on carbomer, sodium carboxymethylcellulose, sodium alginate, and xanthan gum was tested by swelling with purified water to create a 1 wt.% gel. The gel matrix was observed for appearance, viscosity, spreadability, uniformity, and high and low temperature and centrifugation tests to determine the optimal gel matrix. The results are shown in Table 18.

[0194] Table 18 Comparison of different gel matrices

[0195] Traits Carbomer CMC-NA Sodium alginate Xanthan gum Dosage 1％ 1％ 1％ 1％ Appearance Transparent, semi-solid Slightly yellow, fluid Transparent, fluid Light yellow, slightly thin viscosity suitable Rare Rare Rare Spreadability Easy to apply Not easy to apply Not easy to apply Not easy to apply Uniformity exquisite More delicate More delicate More delicate High temperature test No stratification No stratification No stratification No stratification Low temperature test No stratification No stratification No stratification No stratification Centrifugation test No stratification No stratification No stratification No stratification

[0196] The results showed that carbomer as the gel matrix had the best performance in all the parameters tested.

[0197] 2. Single factor experiment to screen the preparation formula of liposome gel

[0198] The appearance, viscosity, pH value, water loss rate and stability test score of the gel were used as intuitive indicators to examine the formation of liposome gel. The comprehensive scoring criteria are shown in Table 19.

[0199] Table 19 Comprehensive scoring criteria

[0200]

[0201] The viscosity evaluation method is as follows: take an appropriate amount of Buddha's fragrance essential oil liposome gel and place it in a 10 mL centrifuge tube, set the temperature to 20°C, and use a rotary viscometer to measure the viscosity of the gel.

[0202] The pH value determination method is as follows: take 2 g of liposome gel of ginseng essential oil, add 40 mL of water, heat in a 50°C water bath, stir until dissolved, and determine the pH value using a pH meter.

[0203] The water loss rate evaluation method is as follows: weigh an appropriate amount of liposome gel of ginseng essential oil and place it in a dry culture dish, recorded as M (the total mass of the blank culture dish and the liposome gel of ginseng essential oil), put it in an oven to dry at 55°C, take it out for 3 hours, let it cool, weigh it, and record it as M1 (the total mass of the weighing bottle and the liposome gel of ginseng essential oil at this time), M0 is the mass of the blank culture dish, and calculate the water loss rate (S) of each sample. The water loss rate calculation formula is:

[0204]

[0205] Stability test evaluation method

[0206] Centrifugation test: Weigh an appropriate amount of liposome gel of ginseng essential oil into a test tube, centrifuge at 4000r / min for 30min, and check whether the gel has stratification or agglomeration.

[0207] Low temperature test: Weigh an appropriate amount of liposome gel of ginseng essential oil into a test tube, place it in a refrigerator (-20℃) for 24 hours, and after returning to room temperature, check whether the gel has stratification or agglomeration.

[0208] High temperature test: Weigh an appropriate amount of Buddha's fragrance essential oil liposome gel into a test tube, place it in an oven (40℃) for 24 hours, and after returning to room temperature, check whether the gel has stratification or agglomeration.

[0209] 2.1 Selection of Carbomer Concentration

[0210] According to the method of Example 4, liposome gel of Buddha fragrance essential oil was prepared, and the carbomer concentration was 0.75%,

[0211] Liposome gels at 1.0%, 1.25%, 1.5%, 1.75%, and 2.0% were evaluated and scored. The results are shown in the table below.

[0212] Table 20 Effect of different mass concentrations of Carbomer 940 on the preparation of gel

[0213]

[0214] The results showed that when the concentration of Carbomer 940 was 1.25%, the comprehensive score was the highest, the appearance was better, and the viscosity was the best.

[0215] 2.2 Selection of moisturizer

[0216] According to the method of Example 4, liposome gels containing no moisturizer, 10% glycerol, 10% (glycerol: propylene glycol 1:1), and 10% propylene glycol were prepared and evaluated. The results are shown in the table below.

[0217] Table 21 Effects of different moisturizers on the preparation of gel

[0218]

[0219] The results showed that when the moisturizer was 1% propylene glycol, the comprehensive score was the highest, the appearance was better, and the water loss rate was lower than the other three groups.

[0220] 2.3 Selection of moisturizer dosage

[0221] According to the method of Example 4, liposome gels with mass concentrations of 7.5%, 10%, 12.5%, 15% and 17.5% were prepared respectively, and evaluated and scored. The results are shown in Table 22.

[0222] Table 22 Effect of moisturizer dosage on gel preparation

[0223]

[0224] The results showed that when the mass concentration of moisturizer propylene glycol was 12.5%, the comprehensive score was the highest and the appearance was better. When the amount of propylene glycol was larger, the paste became sticky.

[0225] 2.4 Investigation of triethanolamine dosage

[0226] According to the method of Example 4, 0.05 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g and 0.3 g of liposome gel were added respectively, and the results were evaluated and scored. The results are shown in Table 23.

[0227] Table 23 Effect of triethanolamine dosage on gel preparation

[0228]

[0229] The results showed that when the dosage of triethanolamine was 0.15g, the comprehensive score was the highest and the appearance was better. When the dosage of triethanolamine was larger, the paste became semi-solid and sticky.

[0230] 3. Optimization design of the preparation process of Buddha fragrance essential oil liposome gel

[0231] 3.1 Orthogonal optimization design

[0232] According to the results of single factor investigation, the carbomer dosage (A), propylene glycol dosage (B), and triethanolamine dosage (C) were used as investigation factors, and 3 levels were selected for each factor. The gel's appearance, viscosity, stability (centrifugation, low temperature, high temperature), pH value, water loss rate, and spreadability were used for comprehensive scoring. The gel comprehensive score (Y 综合评分 ) was used as the evaluation index, and orthogonal experimental design was used to determine the optimal prescription.

[0233] Table 24 Orthogonal Experiment Factor Levels

[0234]

[0235] Table 25 Orthogonal test results

[0236]

[0237] Table 26 Variance analysis results

[0238] Sources of variance Sum of Squares of Deviations degrees of freedom F ratio F critical value P A 8.81 2.00 24.33 19.00 P<0.05* B 1.20 2.00 3.32 19.00 C 2.54 2.00 7.01 19.00 D(error) 0.36 2.00

[0239] Note: F 0.05 (2, 2) = 19, F 0.01 (2, 2) = 99, * indicates a significant difference

[0240] From the value of the range R, we can know that: R A >R C >R B The influence of various factors on the preparation process of essential oil liposome gel was in the order of A > C > B, i.e., carbomer concentration > triethanolamine dosage > propylene glycol concentration. Analysis of variance showed that carbomer had a significant impact on the gel preparation process, with the difference being statistically significant (P < 0.05), while triethanolamine dosage and propylene glycol concentration had little effect. The optimal preparation process for essential oil liposome gel was A2B2C2, i.e., carbomer concentration of 1.25%, propylene glycol concentration of 12.5%, and triethanolamine dosage of 0.125g.

[0241] 3.2 Prescription Verification

[0242] 0.375 g of carbomer was added to 20 g of liposomes of ginseng essential oil and swollen overnight. The mixture was homogenized for 2 min using a high-speed dispersing homogenizer at 3000 r / min. 3.75 g of propylene glycol was added and stirred on a magnetic stirrer for 10 min. 0.125 g of triethanolamine and 0.1 g of methylparaben were then added, and the mixture was made up to 30 g with water. The mixture was stirred evenly to obtain a liposome gel. Three batches of liposome gels of ginseng essential oil were prepared and evaluated for comprehensive scores (Table 27).

[0243] Table 27 Prescription verification experiment

[0244] batch number <![CDATA[Y 综合评分 ]]> 20201202 18.4 20201203 18.6 20201204 18.8 average value 18.6 RSD / % 1.07

[0245] The best process test results are Y 综合评分 The values ​​were 18.4, 18.6, and 18.8, respectively, with an RSD of 1.07%, showing no significant difference. This indicated that the preparation process of the liposome gel of ginseng essential oil was stable and had good reproducibility.

[0246] Test Example 2

[0247] This test example investigates the physical and chemical properties of the liposome gel of Buddha's fragrance essential oil. This test uses the liposome gel of Buddha's fragrance essential oil batch number 20201202.

[0248] 1. Characteristics

[0249] The Buddha fragrance essential oil liposome gel of the present invention is milky white, translucent, has an aromatic odor, is uniform, fine and glossy, and is in a semisolid state, and has a cooling effect when applied to the skin.

[0250] 2. Inspection

[0251] 2.1 pH determination: The pH value of the liposome gel of Buddha's fragrance essential oil was measured to be 5.80±0.25, which is in line with the pH value of 5.5-6.5 specified in the 2020 edition of the Pharmacopoeia.

[0252] 2.2 Viscosity determination: The viscosity of the liposome gel of buddha fragrance essential oil was measured to be 32.5±2.0 Pa·s, which is in line with the viscosity requirements of the 2020 edition of the Pharmacopoeia (30-49 Pa·s).

[0253] 2.3 Particle size determination: Take an appropriate amount of Buddha's fragrance essential oil liposome gel and dilute it with pure water. Use a quartz cell to measure the particle size. The average particle size distribution of the gel was measured to be 182±3.50nm, which is in line with the requirement of the 2020 edition of the Chinese Pharmacopoeia that it should be less than 180μm. The results are shown in the attached Figure 11 shown.

[0254] 2.4 Stability test

[0255] Centrifugation test: Weigh an appropriate amount of liposome gel of ginseng essential oil into a test tube and centrifuge at 4000r / min for 30min. The results showed that the appearance and properties of the liposome gel of ginseng essential oil did not change, and no stratification occurred, which complies with the requirement of the 2020 edition of the Pharmacopoeia that the gel should not sink or agglomerate.

[0256] Low temperature test: Weigh an appropriate amount of liposome gel of ginseng essential oil into a test tube and place it in a refrigerator (-20℃) for 24 hours. After returning to room temperature, the appearance and properties of the liposome gel of ginseng essential oil did not change, and no stratification occurred, which complies with the requirement of the 2020 edition of the Pharmacopoeia that the gel should not sink or agglomerate.

[0257] High temperature test: Weigh an appropriate amount of liposome gel of ginseng essential oil into a test tube and place it in an oven (40°C) for 24 hours. After returning to room temperature, the appearance and properties of the liposome gel of ginseng essential oil did not change, and no stratification occurred, which complies with the requirement of the 2020 edition of the Pharmacopoeia that the gel should not sink or agglomerate.

[0258] Test Example 3

[0259] This test example investigated the stability of the liposome gel of ginseng essential oil.

[0260] 1. Strong light test

[0261] The liposome gel of the Buddha's fragrance essential oil was placed in a transparent sample bottle, opened and placed under 25°C and 4500±500LX light conditions. Samples were taken on the 5th, 10th and 30th day respectively. The results are shown in the following table:

[0262] Table 28 Strong light stability test results

[0263]

[0264] The results showed that during the strong light stability test, the gel's appearance became slightly yellow and its content decreased, indicating that it should be kept away from light during storage. Therefore, this product is intended to be stored in light-proof packaging materials.

[0265] 2. High temperature test

[0266] The liposome gel of ginseng essential oil was placed in a sample bottle and protected from light with tin foil. The bottle was opened and placed at 40°C. Samples were taken on the 5th, 10th and 30th day respectively. The results are shown in Table 29.

[0267] Table 29 High temperature gel stability test results

[0268]

[0269] During the high-temperature stability test, the gel's appearance became slightly yellow and its content decreased, indicating that high-temperature environments should be avoided during storage to improve the stability of the preparation.

[0270] 3. Room temperature test

[0271] The liposome gel of the Buddha's fragrance essential oil was placed in a sample bottle and protected from light with tin foil. The bottle was opened and placed at room temperature. Samples were taken on the 5th, 10th and 30th day respectively. The results are shown in Table 30.

[0272] Table 30 Room temperature stability test results

[0273]

[0274] During the room temperature stability test, the appearance, properties and content of the gel remained basically unchanged, indicating that the sample was stable at room temperature.

[0275] 4. Low temperature test

[0276] The liposome gel of ginseng essential oil was placed in a sample bottle and protected from light with tin foil. The bottle was opened and placed at 4°C. Samples were taken on the 5th, 10th and 30th day respectively. The results are shown in Table 31.

[0277] Table 31 Low temperature stability test results

[0278]

[0279] During the low-temperature stability test, the appearance, properties and content of the gel remained basically unchanged, indicating that the sample was stable at low temperatures.

[0280] 5 Accelerated testing

[0281] The liposome gel of ginseng essential oil was placed in a brown sample bottle and placed under the conditions of 30±2°C and 65±5% relative humidity. Samples were taken at 1, 2, 3 and 6 months respectively. The results are shown in Table 32.

[0282] Table 32 Accelerated test results

[0283]

[0284] After the Buddha fragrance essential oil liposome gel was placed at a temperature of 30±2℃ and a relative humidity of 65%±5% for 3 months, the sample was stable in appearance, properties, content, etc., and met the relevant regulations on the stability of the gel.

[0285] Test Example 4

[0286] This test example discloses an in vitro transdermal absorption test of the present invention's liposome gel of the Buddha's fragrance essential oil. The Buddha's fragrance essential oil liposome gel used in this test example was prepared according to the method of Example 4. The Buddha's fragrance essential oil gel used in this example was prepared according to the method of Example 4, except that the Buddha's fragrance essential oil liposomes were replaced with Buddha's fragrance essential oil.

[0287] 1. Preparation of Ex vivo Skin

[0288] The mice were killed, part of the hair on their backs was shaved with a shaver, and all the hair on their backs was removed with a depilatory cream. The skin on the back of the mice was peeled off, washed with physiological saline, and the subcutaneous fat layer and connective tissue were wiped off. After absorbing the moisture with filter paper, the skin was wrapped with tin foil and stored at -20°C for later use.

[0289] 2 In vitro permeation test

[0290] In vitro drug transdermal test was conducted using a Franz diffusion cell. The water bath temperature was set at 32°C and the stirring speed was 300 r / min. The treated skin was fixed between the supply cell and the receiving cell with the stratum corneum facing upwards. 1 g each of 1 g of fujian essential oil liposome gel and 1 g of fujian essential oil gel were added to the supply cell. The transdermal penetration area was 2.2 cm. 2 ; 30% ethanol-normal saline solution with bubbles removed was injected into the receiving cell as the receiving medium, so that the skin was in complete contact with the receiving cell solution. The volume of the receiving cell was 6.5 mL. 1 mL of the medium solution was removed at 4, 6, 8, 12, 16, 20, and 24 hours, and 1 mL of the receiving medium at the same temperature was immediately added to the receiving cell. The removed medium solution was filtered through a filter membrane and the content was determined by gas chromatography. The chromatographic conditions were the same as those for the encapsulation efficiency determination. tThe transdermal absorption rate curve was fitted with a model to obtain the transdermal kinetic equation, and the slope was the transdermal rate constant (J, μg·h -1 cm -2 ).

[0291] Cumulative transdermal drug dose Q t Calculation formula:

[0292] Q t =(V 总 C t +∑C t-1 V 取 ) / A

[0293] Where: Q t is the cumulative permeation at time t, V 总 is the volume of the receiving cell, which is 6.5 mL in this study, C t The concentration of this sample is determined, V 取 is the sampling volume each time, A is the diffusion and penetration area, which is 2.2 cm in this study. 2 .

[0294] Model fitting of the Qn-t cumulative transdermal amount revealed that the transdermal equation for the liposome gel of eugenol was Q = 54.126t - 7.8413, r = 0.9935, and the transdermal rate J = 54.126. The transdermal equation for the eugenol in the liposome gel of eugenol was Q = 86.77t - 34.996, r = 0.9972, and the transdermal rate J = 86.77. These results indicate that the transdermal rate of eugenol in the liposome gel of eugenol is lower than that of eugenol in the liposome gel of eugenol, indicating that the liposome gel of eugenol exhibits sustained release.

[0295] 3. Skin retention test of liposome gel of Buddha essential oil

[0296] Take the Buddha's fragrance essential oil liposome gel and the Buddha's fragrance essential oil gel respectively, and conduct transdermal experiments. Take the skin after transdermal test administration at 4, 8, 12, 16, 20, and 24 hours, wash off the residual substances on the skin surface, dry the water, cut the skin into pieces and place it in a centrifuge tube, first treat it with a vortex oscillator for 2 minutes, then add a certain amount of methanol and sonicate for 60 minutes, centrifuge at 10000r / min for 15 minutes, take the supernatant, and filter it with a 0.22μm microporous membrane. The content was determined by gas chromatography. The chromatographic conditions were the same as those for the encapsulation efficiency determination. The results are shown in Figure 2. Figure 13 .

[0297] Drug skin retention Q s Calculation formula:

[0298] Q S =C S / A

[0299] Where: Cs is the drug concentration in the skin sample liquid measured at the nth time point, and A is the penetration area.

[0300] After 24 hours of transdermal permeation, the skin retention of eugenol in the liposome gel group and the gel group was 126.16 μg / cm 2 、104.65μg / cm 2 The D-limonene content in the Buddha's fragrance essential oil liposome gel group and the Buddha's fragrance essential oil gel group was 63.91 μg / cm 2 , 44.95μg / cm 2 After 24 hours of transdermal permeation, the skin retention of eugenol and D-limonene in the liposome gel group was higher than that in the liposome gel group, producing a sustained-release effect in the form of a reservoir.

[0301] Finally, it should be noted that the above embodiments are merely preferred embodiments of the present invention and are intended to illustrate the technical solutions of the present invention, rather than limiting them, and certainly not limiting the patent scope of the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some or all of the technical features therein may be replaced by equivalents. These modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention. In other words, any changes or refinements made to the main design concept and spirit of the present invention that have no substantive significance, provided that the technical problems they solve are still consistent with those of the present invention, should be included in the protection scope of the present invention. In addition, the direct or indirect application of the technical solutions of the present invention to other related technical fields should also be included in the patent protection scope of the present invention.

Claims

1. A method for preparing a liposome gel of Buddha's fragrance essential oil, characterized in that: The following steps are involved: Take the gel matrix, add the Buddha's fragrance essential oil liposome to swell, homogenize with a high-speed dispersing homogenizer, add propylene glycol, stir evenly, then add triethanolamine, add water to the specified amount, stir evenly, and obtain the Buddha's fragrance essential oil liposome gel; The gel matrix accounts for 1.0-1.5% of the total mass of the Buddha's fragrance oil liposome gel, propylene glycol accounts for 7.5-15% of the total mass of the Buddha's fragrance oil liposome gel; triethanolamine accounts for 0.5-1% of the total mass of the Buddha's fragrance oil liposome gel, and the Buddha's fragrance oil liposome accounts for 60-75% of the total mass of the Buddha's fragrance oil liposome gel; The raw materials of the buddha fragrance essential oil liposome are buddha fragrance essential oil, lecithin and cholesterol; the buddha fragrance essential oil is composed of bergamot essential oil and clove essential oil in a mass ratio of 1:3, the mass ratio of lecithin to cholesterol is 4-6:1; the total mass ratio of buddha fragrance essential oil to lecithin and cholesterol is 1-2.5:7; The preparation method of Buddha fragrance essential oil liposome comprises the following steps: S1. After dissolving the Buddha's essential oil, lecithin and cholesterol in a solvent, vacuum rotary evaporation was performed to form a thin film of lipids on the wall; S2. Add buffer solution, continue rotary evaporation to remove the film, and ultrasonically hydrate; S3. The ultrasonically hydrated material is homogenized by a high-pressure homogenizer to obtain.

2. The method for preparing the liposome gel of Buddha's fragrance essential oil according to claim 1, wherein: In said S1, rotary evaporation under reduced pressure at 30-50°C; In said S2, the pH value of said buffer solution is 7.0-8.0; In said S2, after adding the buffer solution, rotary evaporation is continued at 40-60°C; In S2, the ultrasonic hydration time is 10-40 min; In S3, the ultrasonically hydrated material is homogenized in a high-pressure homogenizer at 60-100 MPa for 90-150 seconds.

3. The method for preparing the liposome gel of Buddha's fragrance essential oil according to claim 1, wherein: In the above-mentioned S1, the mixture was evaporated under reduced pressure and rotary evaporated at 35°C.

4. The method for preparing the liposome gel of Buddha's fragrance essential oil according to claim 1, wherein: In S2, the pH value of the buffer solution is 7.5-8.

0.

5. The method for preparing the liposome gel of Buddha's fragrance essential oil according to claim 1, wherein: In the S2, after adding the buffer solution, rotary evaporation is continued at 45°C.

6. The method for preparing the liposome gel of Buddha's fragrance essential oil according to claim 1, wherein: In S2, the ultrasonic hydration time is 20-30 minutes.

7. The method for preparing the Buddha's fragrance essential oil liposome gel according to claim 1, wherein: In S3, the ultrasonically hydrated material is homogenized in a high-pressure homogenizer at 80 MPa for 120 seconds.

8. The method for preparing the Buddha's fragrance essential oil liposome gel according to claim 1, wherein The gel matrix comprises carbomer, sodium carboxymethyl cellulose, sodium alginate and xanthan gum.

9. The Buddha's fragrance essential oil liposome gel according to claim 1, characterized in that The gel matrix is ​​carbomer.

10. The Buddha fragrance essential oil liposome gel according to claim 1, characterized in that The gel matrix accounts for 1.25% of the total mass of the Buddha's fragrance essential oil liposome gel.

11. The Buddha's fragrance essential oil liposome gel according to claim 8, characterized in that The propylene glycol accounts for 12.5% ​​of the total mass of the Buddha's fragrance oil liposome gel.

12. The Buddha fragrance essential oil liposome gel according to claim 1, characterized in that Triethanolamine accounts for 0.5% of the total mass of the Buddha's fragrance essential oil liposome gel.

13. Use of the Buddha's fragrance essential oil liposome gel according to any one of claims 1 to 12 in preparing an external-use medicament for preventing and treating acne.

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