Rhodiola rosea nano-emulsion and preparation method thereof

Abstract

The invention provides a rhodiola rosea nano-emulsion. The rhodiola rosea nano-emulsion comprises a rhodiola rosea extract, an interfacial agent, a co-interfacial agent and oil. The surfactant comprises diethylene glycol monoethyl ester or polysorbate 80, the co-surfactant comprises polysorbate 80, sorbitan oleate 80, or caprylic / capric polyethylene glycol glyceride, and the oil comprises medium chain triglyceride or soybean oil, and coats the rhodiola rosea extract in an oil-in-water manner. The invention also provides a method for preparing the rhodiola rosea nano-emulsion, so as to provide the rhodiola rosea nano-emulsion suitable for drug delivery.

Description

Technical Field

[0001] The present disclosure relates to a nanoemulsion and a preparation method thereof, and in particular to a Rhodiola rosea nanoemulsion and a preparation method thereof. Background Art

[0002] Nanoemulsion is one of the drug delivery systems in dosage form, which is used to deliver active ingredients to target sites in our body. Nanoemulsion consists of an oil phase, water and a surfactant (co-surfactant). The droplet size of nanoemulsion varies in the nanometer range, ranging from 10 nanometers (nm) to 1000nm. Nanoemulsions with a particle size below 500nm have clear emulsification and good stability. Therefore, the Brownian motion of small droplets is sufficient to overcome their low gravity characteristics, resulting in good physical stability, easy penetration and properties similar to transparent solutions. Nanoemulsions can generally be divided into two types: water-in-oil (W / O) and oil-in-water (O / W).

[0003] Rhodiola rosea (RR), also known as the "golden root", is traditionally considered a plant powerful enough to fight viral infections, relieve depression, reduce weight, increase energy and even improve sexual function. Rhodiola rosea has a wide range of pharmacological activities, including neuroprotection, anti-inflammatory, anti-aging, cardioprotection, anti-cancer and antioxidant. Rhodiola rosea contains a variety of secondary metabolites, including flavonoids, phenylethanol, phenylpropanoids, lignans, phenolic acids, essential oils, polysaccharides, and also contains cyanogenic glucosides.

[0004] However, there is no drug delivery formulation for Rhodiola rosea extracts available at present. Therefore, the prior art needs to be improved. Summary of the invention

[0005] Some embodiments of the present disclosure provide a Rhodiola rosea nanoemulsion, comprising a Rhodiola rosea extract, a surfactant, a co-surfactant, and oil. The surfactant comprises diethylene glycol monoethyl ester (diethylene glycolmonoethyl ester, ) or polysorbate 80 (polysorbate 80, Tween 80), the co-surfactant includes polysorbate 80, sorbitan oleate 80 (sorbitan oleate 80, Span80), or caprylic capric macrogolglycerides (caprylic capric macrogolglycerides, ), the oil is medium-chain triglycerides (Labrafac TM) or soybean oil, and the Rhodiola rosea extract is coated in an oil-in-water (O / W) manner. In one embodiment, the surfactant is diethylene glycol monoethyl ester, the co-surfactant is sorbitan oleate 80, and the oil is soybean oil.

[0006] In some embodiments, the Rhodiola rosea extract comprises an alcoholic Rhodiola rosea root extract.

[0007] In some embodiments, the Rhodiola rosea extract comprises 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranoside (TMU060615B) and 1,2,3,6-tetra-O-galloyl-4-Op-hydroxybenzoyl-β-D-glucopyranoside.

[0008] In some embodiments, based on 100% of the total weight of the Rhodiola rosea nanoemulsion, the weight percentage of the surfactant is 10% to 30%; and the weight percentage of the co-surfactant is 10% to 30%.

[0009] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; the co-surfactant is polysorbate 80, and its weight percentage is 10% to 30%; and the oil is medium chain triglyceride, and its weight percentage is 1% to 5%.

[0010] In some embodiments, the weight percentage of the surfactant is 10%, and the weight percentage of the co-surfactant is 16.63%.

[0011] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is polysorbate 80, and its weight percentage is 10% to 30%; the co-surfactant is sorbitan oleate 80, and its weight percentage is 10% to 30%; and the oil is soybean oil, and its weight percentage is 1% to 5%. In one embodiment, the combination of 10% by weight of diethylene glycol monoethyl ester as the surfactant, 10% by weight of sorbitan oleate 80 as the co-surfactant, and 3% by weight of soybean oil as the oil is excluded.

[0012] In some embodiments, the weight percentage of the surfactant is 10%, and the weight percentage of the co-surfactant is 29.87%.

[0013] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; the co-surfactant is caprylic acid capric acid macrogol glyceride, and its weight percentage is 10% to 30%; and the oil is medium chain triglyceride, and its weight percentage is 1% to 5%.

[0014] In some embodiments, the weight percentage of the surfactant is 28.41%, and the weight percentage of the co-surfactant is 16.63%.

[0015] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; the co-surfactant is polysorbate 80, and its weight percentage is 10% to 30%; and the oil is soybean oil, and its weight percentage is 1% to 5%.

[0016] In some embodiments, the weight percentage of the surfactant is 29.61%, and the weight percentage of the co-surfactant is 30%.

[0017] In some embodiments, the size of the Rhodiola rosea nanoemulsion is 10 nm to 500 nm, the polydispersity index (PDI) is 0.173 to 0.667, and the zeta potential is -96.7 mV to -7.39 mV.

[0018] Another embodiment of the present disclosure provides a method for preparing a Rhodiola rosea nanoemulsion, comprising: providing a Rhodiola rosea extract; mixing the Rhodiola rosea extract with oil to obtain an oil phase component; mixing a surfactant and a co-surfactant to obtain an aqueous phase component; adding the oil phase component to the aqueous phase component to form an oil-in-water mixture until emulsified to obtain a Rhodiola rosea nanoemulsion, wherein the surfactant comprises diethylene glycol monoethyl ester or polysorbate 80, wherein the co-surfactant comprises polysorbate 80, sorbitan oleate 80, or caprylic capric macrogol glyceride, wherein the oil comprises medium-chain triglycerides and soybean oil, wherein the combination of the surfactant being diethylene glycol monoethyl ester, the co-surfactant being sorbitan oleate 80, and the oil being soybean oil is excluded.

[0019] In some embodiments, the step of adding the oil phase ingredients to the water phase ingredients comprises dropping the oil phase ingredients into the water phase ingredients.

[0020] In some embodiments, in the step of adding the oil phase component to the water phase component, the emulsification method is ultrasonic vibration to emulsify.

[0021] In some embodiments, based on 100% of the total weight of the Rhodiola rosea nanoemulsion, the weight percentage of the surfactant is 10% to 30%; and the weight percentage of the co-surfactant is 10% to 30%. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The various aspects of the present disclosure will be most easily understood when the following detailed description is read in conjunction with the accompanying drawings. It should be noted that various features may not be drawn to scale in accordance with industry standard operating procedures. In fact, the dimensions of various features may be arbitrarily increased or decreased for clarity of discussion. In order to make the above and other objects, features, advantages and embodiments of the present disclosure more apparent, the accompanying drawings are described as follows:

[0023] Figures 1A to 1G Contour plots and three-dimensional plots of one embodiment of the present disclosure are shown, showing the effects of the independent variables on the size of the F-group spheres (Y1), PDI (Y2), and Zedar potential (Y3). Figure 1A , Figure 1B , Figure 1C Contour plots of surfactant and co-surfactant concentrations versus sphere size, PDI and Zed potential. Figure 1D For expectation. Figure 1E , Figure 1F , Figure 1G The effects of surfactant and co-surfactant concentrations on the sphere size, PDI and Zed potential in the three-dimensional graph of group F.

[0024] Figures 2A to 2G Contour plots and three-dimensional plots of one embodiment of the present disclosure are shown, showing the effects of the independent variables on the G-group ball size (Y1), PDI (Y2), and Zedar potential (Y3). Figure 2A , Figure 2B , Figure 2C Contour plots of surfactant and co-surfactant concentrations versus sphere size, PDI and Zed potential. Figure 2D For expectation. Figure 2E , Figure 2F , Figure 2G The effects of surfactant and co-surfactant concentrations on the sphere size, PDI and Zed potential in the three-dimensional graph of group G.

[0025] Figures 3A to 3G Contour plots and 3D plots are shown of one embodiment of the present disclosure, showing the effects of the independent variables on the size of the H-group spheres (Y1), PDI (Y2), and Zedar potential (Y3). Figure 3A , Figure 3B , Figure 3C Contour plots of surfactant and co-surfactant concentrations versus sphere size, PDI and Zed potential. Figure 3D For expectation. Figure 3E , Figure 3F, Figure 3G The effects of surfactant and co-surfactant concentrations on the sphere size, PDI and Zed potential in the three-dimensional graph of group H.

[0026] FIG. 4A to FIG. 4G Contour plots and three-dimensional plots of one embodiment of the present disclosure are shown, showing the effects of the independent variables on the J-group ball size (Y1), PDI (Y2), and Zedar potential (Y3). Figure 4A , Figure 4B , Figure 4C Contour plots of surfactant and co-surfactant concentrations versus sphere size, PDI and Zed potential. Figure 4D For expectation. Figure 4E , Figure 4F , Figure 4G The effects of surfactant and co-surfactant concentrations on the sphere size, PDI and Zed potential in the three-dimensional graph of group J.

[0027] FIG. 5A to FIG. 5D An embodiment of the present disclosure is shown in FIG. Figure 5A )、Group G( Figure 5B )、Group H( Figure 5C ) and Group J ( Figure 5D ) pH line graph at 25°C, 40°C and 4°C on day 1, day 30, day 60 and day 90.

[0028] FIG. 6A to FIG. 6D FIG. 1 is a diagram showing an optimized F group (Opt F: Fig. 6A )、Opt G Group( Figure 6B )、Opt H Group( Figure 6C ) and Opt J Group ( Fig.6D ) Transmission electron microscope (TEM) photograph at a scale of 500 nm. DETAILED DESCRIPTION

[0029] In order to make the description of the present disclosure more detailed and complete, the following is an illustrative description of the implementation and specific embodiments of the present disclosure, but this is not the only form of implementing or using the specific embodiments of the present disclosure. The various embodiments disclosed below can be combined or replaced with each other in beneficial situations, and other embodiments can be added to one embodiment without further recording or explanation. In the following description, many specific details will be described in detail so that the reader can fully understand the following embodiments. However, the embodiments of the present disclosure can also be practiced without such specific details.

[0030] In this document, unless the context specifically limits the article, "a", "an" and "the" may refer to a single or multiple. It will be further understood that "comprising", "including", "having" and similar words used in this document indicate the features, regions, integers, steps, operations, elements and / or components described therein, but do not exclude the described or additional one or more other features, regions, integers, steps, operations, elements, components, and / or groups thereof.

[0031] As used herein, a "co-surfactant" is a chemical moiety that, when used in combination with a surfactant, further reduces the surface tension of a liquid.

[0032] As used herein, "self-assembly method" means that in self-assembly emulsification, an organic phase consisting of oil, surfactant and co-surfactant is simply added to an aqueous phase and gently stirred. The surfactant present in the organic phase has a high affinity for the continuous phase. Therefore, when mixing the two phases (organic phase or dispersed phase and aqueous phase or continuous phase), turbulence (turbulence) will be generated, and the surfactant will quickly diffuse into the aqueous phase and form a covering / film around the dispersed oil droplets by reducing the interfacial tension, thereby causing the spontaneous formation of a nanoemulsion system. The co-surfactant further helps to cause turbulence, reduce the interfacial tension between the two immiscible phases, and alleviate the formation of dispersions by staying in unoccupied places around the dispersed oil droplets that are not protected by surfactant molecules. The screening of surfactants for self-assembly emulsification is based on hydrophilic-lipophilic balance (HLB), which is the strength and size of the hydrophilic and lipophilic parts of the surfactant molecule.

[0033] As used herein, "Rhodiola rosea (RR)" refers to about 140 compounds present in the underground part of Rhodiola rosea. The root of Rhodiola rosea contains phenolic substances, rosin, rosin, rosin, organic acids, terpenoids, phenolic acids and their derivatives, flavonoids, anthraquinones, alkaloids, tyrosol and salidroside.

[0034] In some embodiments, the Rhodiola rosea nanoemulsion comprises a Rhodiola rosea extract, a surfactant, a co-surfactant, and oil.

[0035] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the co-surfactant is polysorbate The ester 80 has a weight percentage of 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the oil is a medium chain triglyceride, and its weight percentage is 1% to 5%, for example, 2%, 3%, 4%, or any value between any two of these values.

[0036] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is polysorbate 80, and its weight percentage is 10% to 30%, such as 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the co-surfactant is sorbitan Oleate 80, its weight percentage is 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the oil is soybean oil, its weight percentage is 1% to 5%, for example, 2%, 3%, 4%, or any value between any two of these values.

[0037] In some embodiments, the total weight of the Rhodiola rosea nanoemulsion is 100%, and the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the co-surfactant is caprylic acid capric acid polyethylene The glycol glyceride has a weight percentage of 10% to 30%, for example 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the oil is a medium chain triglyceride, and its weight percentage is 1% to 5%, for example 2%, 3%, 4%, or any value between any two of these values.

[0038] In some embodiments, based on the total weight of the Rhodiola rosea nanoemulsion as 100%, the surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the co-surfactant is polysorbate. The alcohol ester 80 has a weight percentage of 10% to 30%, for example, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, or any value between any two of these values; the oil is soybean oil, and its weight percentage is 1% to 5%, for example, 2%, 3%, 4%, or any value between any two of these values.

[0039] Several embodiments and experimental examples are listed below to further illustrate the Rhodiola rosea nanoemulsion disclosed in the present invention. However, they are only for illustrative purposes and are not intended to limit the present invention. The protection scope of the present invention shall be determined by the following claims.

[0040] Although a series of operations or steps are used below to illustrate the methods disclosed herein, the order in which these operations or steps are shown should not be construed as a limitation of the present disclosure. For example, certain operations or steps may be performed in different orders and / or simultaneously with other steps. In addition, it is not necessary to perform all of the operations, steps, and / or features illustrated to implement the embodiments of the present disclosure. In addition, each operation or step described herein may include a number of sub-steps or actions.

[0041] For the sake of clarity, features and elements that are known in the art and are not necessary for understanding the described principles may be omitted.

[0042] Preparation Example

[0043] Surfactants, co-surfactants and oil phase are key parts in preparing nanoemulsion formulations. The selection of surfactants and co-surfactants is based on the emulsifying ability in the formation of nanoemulsions. The preparation of emulsifying ability adopts the spontaneous method. Surfactants and co-surfactants are selected based on the difference in hydrophilic-lipophilic balance (HLB) combination. HP, Tween 80, Span 80 and As a surfactant and co-surfactant. HLB values ​​in the range of (3-6) are more likely to form water-in-oil (w / o) nanoemulsions, while those in the range of (8-18) are more likely to form oil-in-water (o / w) nanoemulsions. For the use of oil phases with different carbon chain lengths (C8-C18) to prepare nanoemulsions by self-assembly emulsification, the selected oil is Labrafac TM and soybean oil as the oil phase.

[0044] After the roots of Rhodiola rosea are dried in the shade, they are crushed with a grinder. The crushed roots of Rhodiola rosea are soaked and extracted with 95% alcohol at a weight-to-volume ratio of 1:20 (w / v) for 1 week to obtain an extract. The extract is concentrated by a vacuum concentrator to obtain a dry Rhodiola rosea crude extract. The above alcohol extraction steps are repeated three times, and the concentrates obtained three times are combined to obtain an alcohol extract of the roots of Rhodiola rosea.

[0045] The root of Rhodiola rosea (RR) was extracted with alcohol, and the evidence specimen (voucherspecimen, number TMU060615) was identified and preserved in the School of Pharmacy of Taipei Medical University, Taipei. The alcohol extract of the root of Rhodiola rosea contains at least 1,2,3,4,6-penta-O-galloyl pyranoglucoside and 1,2,3,6-tetra-O-galloyl-4-Op-hydroxybenzoyl-β-D-pyranoglucoside. The nanoemulsions of this preparation example are all prepared by the low-energy nanoemulsion preparation method, and the nanoemulsions are prepared by the following steps. (1) The alcohol extract of the root of Rhodiola rosea is mixed with oil (oil or essential oil) to obtain an oil phase component. (2) The surfactant and the co-surfactant are mixed and mixed at a speed of 500 rpm for 15 minutes to obtain an aqueous phase component. (3) dropping the oil phase component into the water phase component, stirring at 1000 rpm for 60 minutes at 25° C. to form an oil-in-water (o / w) mixture, then mixing with ultrasonic vibration at 25° C. for 15 minutes and then standing for 15 minutes until emulsification, to obtain a Rhodiola rosea nanoemulsion. The Rhodiola rosea nanoemulsion is stored in an environment not higher than 25° C.

[0046] Example 1 Oil phase selection test of Rhodiola rosea nanoemulsion

[0047] Please refer to Table 1 below. The test formulas are divided into Group F, Group Fx and Group Fy. The difference between the test formulas of each group lies in the choice of oil phase. Group F is The Fx group is corn oil and the Fy group is sunflower oil. After the preparation is completed, the results show that the experimental formula F group is clear and the particle size is 19.13nm. As for the Fx group and the Fy group, they are both turbid and the particle size is greater than 1000nm. In other words, when sunflower oil or corn oil is used as the oil phase formula of the Rhodiola rosea nanoemulsion, the effect of clarification and nanoparticles cannot be obtained.

[0048] Table 1

[0049]

[0050]

[0051] Example 2 Design of Rhodiola rosea nanoemulsion

[0052] The formulations were experimented with response surface method (RSM) to test the effect of the percentage of surfactant and co-surfactant and the two oils with the same percentage. As independent variables, the combination of surfactant (10% to 30%), co-surfactant (10% to 30%) and oil was divided into four groups: Group F ( Tween 80, Labrafac TM )、Group G (Tween 80, Span 80, soybean oil), Group H ( Labrafac TM ) and Group J ( Tween 80, soybean oil). A three-factor central composite design (CCD) was used to evaluate the effects of surfactants and co-surfactants on three dependent variables (or response variables), such as particle size, polydispersity index (PDI), and zeta potential, where particle size and polydispersity index were measured by dynamic light scattering (DLS) and zeta potential was measured by zetasizer. Software (Stat-Ease, Inc., Minneapolis, USA) was used for experimental design and data analysis. A total of 13 experimental conditions were generated for each group of preparations (as shown in Table 2, Table 3, Table 4, and Table 5), and the results were statistically analyzed and used to calculate the nanoemulsion with the smallest particle size, the lowest PDI and the best zeta potential to examine the characteristics of the nanoemulsion. The analysis was performed at 25°C with a backscattering angle of 173°.

[0053] Table 2. Independent variables, dependent variables and parameters

[0054]

[0055] * Rhodiola rosea root alcohol extract 1w / w%, alcohol 10w / w% (emulsifier), sodium ethylenediaminetetraacetate (Na.EDTA) 0.2w / w% (buffer), sodium metabisulfite (Na.Metabisulfit) 0.3w / w% (preservative), and the rest is water to make up to 100w / w% (the total weight of the Rhodiola rosea nanoemulsion is 100%).

[0056] The influence of independent variable factors is determined by Design The software uses three-dimensional response surface and contour plots ( Figures 1A to 4G) was examined for the strain terms (3 strain terms). Quadratic effect was suitable for all parameters as there was a good influence or effect both individually and in combination. The analysis of variance (ANOVA) of the individual strains was managed by Design-Expert software and then the results showed that the model fit all the data sets. The independent variable factors: surfactant concentration (X1) and co-surfactant concentration (X2) were evaluated at three levels (low -1, medium 0 and high +1) to formulate Rhodiola rosea nanoemulsion. The surfactant concentration (X1) was used in different concentrations such as 10% w / w, 20% w / w and 30% w / w. The co-surfactant concentration had three different concentrations of 10% w / w, 20% w / w and 30% w / w. A total of 13 formulations were prepared for each group with 4 different groups.

[0057] Table 3. Independent variable factors

[0058]

[0059] *-1 = minimum, 0 = medium, 1 = maximum

[0060] Table 4. Four groups of 13 experiments were performed in different groups

[0061]

[0062]

[0063] Table 5. Response surface methodology for four groups of 13 experiments

[0064]

[0065] *Y1 = particle size (nm), Y2 = PDI, Y3 = ZD potential (mV)

[0066] The particle size distribution of the prepared nanoemulsions is shown in Table 5, including group F particle size (13.41 to 39.88 nm), PDI (0.183 to 0.441), Zed potential (-10.2 to -58.6 mV), group G size (96.66 to 233.70 nm), PDI (0.053 to 0.667), Zed potential (-27.3 to -96.7 mV), group H size (19.80 to 85.29 nm), PDI (0.173 to 0.504), Zed potential (-16.6 to -33.0 mV), and group I size (219.13 to 267.63 nm), PDI (0.224 to 0.376), Zed potential (-7.39 to -32.33 mV).

[0067] The particle size of the nanoemulsion is proportional to the oil phase (dispersed phase in the o / w emulsion). The increase in particle size is attributed to the competition of the oil particles for the limited emulsifier retained in the chamber. The PDI value varies between 0.0 and 1.0, and the closer the value is to 0, the more uniform the particle distribution. The PDI value of the obtained Rhodiola rosea nanoemulsion is <1, which shows that the droplet size distribution of all groups of formulations is uniform. For the zeta potential results, the higher the zeta potential value, the more stable the nanoemulsion system formed, which will prevent the prepared Rhodiola rosea nanoemulsion from flocculation. These results show that the emulsion system formed has good stability and will not form flocculation. The zeta potential value greater than ± 30mV indicates that the colloidal system has medium stability, the emulsion system formed does not flocculate or form aggregates, and has high stability (see Table 5 for complete results).

[0068] Enter the above results into Design Software Figures 1A to 4G , showing the contour and 3D plots of the effect of the percentage of surfactant and co-surfactant combination on three strain terms (e.g., ball size, PDI, and Zed potential) and desirability. The red area indicates the highest value for each strain term (ball size, PDI, and Zed potential), followed by yellow, green, and blue, with blue indicating the lowest value for each strain term. The software calculates the best formula for the choice of Figure 2D , Figure 3D , Figure 4D and Figure 5D As can be seen in the figure, based on the expectancy values ​​of each group, the F group is 0.722, the G group is 0.817, the H group is 0.766, and the J group is 0.987. The closer the expectancy value is to 1, the more the proposed solution formula can achieve the ideal optimal solution. The closer the value is to 1, the closer the predicted value is to the result between the predicted value of the best formula and the experimental result value.

[0069] Example 3 pH value test

[0070] The pH values ​​of the blank and drug-loaded nanoemulsions were measured at 25°C for the same group as in Example 2. The measurements were performed at three different temperatures, 4°C, 25°C, and 40°C, for 90 days, with three replicates, and the data are shown as mean ± SD. The pH values ​​were measured on the 1st day, 30th day, 60th day, and 90th day. The results are shown in Table 1. FIG. 5A to FIG. 5D As shown, Group F (6.19 to 6.96), Group G (6.21 to 6.56), Group H (5.21 to 5.98) and Group J (5.19 to 5.98). The pH range for topical or dermal administration is required to be 4-8. If the pH value of the preparation is lower, it will irritate the skin, and if the pH value is higher, it will make the skin dry.

[0071] Example 4 Centrifugal Test

[0072] The same group as in Example 2 was centrifuged at 5000 rpm for 30 minutes and observed for phase separation, creaming and cracking. The nanoemulsion should have maximum stability and no phase separation (emulsification and cracking). The successful formulation was tested for other thermodynamic stability and was tested in triplicate.

[0073] Table 6. Thermodynamic stability test of Rhodiola rosea nanoemulsion

[0074]

[0075] *ND: Not Detected

[0076] The results are shown in Table 6. There was no phase separation in groups F, G, H and J, and all Rhodiola rosea nanoemulsions were found to be stable. Therefore, the physical properties of the nanoemulsions did not change after the centrifugation test, indicating that the nanoemulsions have long-term stability.

[0077] Example 5 Freeze-thaw cycle test

[0078] The same groups as in Example 2 were subjected to freeze-thaw cycles for 6 cycles (12 days) of Rhodiola rosea nanoemulsions in an oscillating incubator at 40°C and 25°C, and it was observed that there was no phase separation in the nanoemulsions of groups G, H and J. After the freeze-thaw test, the prepared nanoemulsions were subjected to freeze-thaw cycles to determine the thermodynamic stability.

[0079] Table 7. Freeze-thaw cycle test results

[0080]

[0081] ND: Not Detected, D: Detected, S: Stable, *This formula is not stable

[0082] As shown in Table 7 above, after freezing and thawing cycles, nanoemulsions F1-F13, G2-G13, H1-H13, and J1-J13 will not emulsify and phase separate, which confirms that the nanoemulsion is thermodynamically stable. As for group G, formulation G1 is stable in cycles 1 to 4, and is separated and unstable in cycles 5 to 6.

[0083] Example 6 Optimization of Rhodiola rosea nanoemulsion

[0084] Input the independent variables and dependent variables of the four groups in Example 2 into Design After the software is run, the four optimized formulas are output. If the optimization criteria produce the smallest droplet size, the narrowest PDI and the value of the barrier potential as strain items, multiple strain items will be considered for optimization. Based on the statistical analysis of the predicted solutions conducted by the software for the combination ratio of surfactants and co-surfactants with higher expected points, the following four groups of solutions are given. Optimized Formula Group F (Opt F): The percentage of surfactants and co-surfactants (10% 16.63% tween 80), the corresponding strain terms of size, PDI and Zed potential are 16.08nm, 0.301 and -47.69mV respectively. Optimized formula G group (Opt G): the percentage of surfactant and co-surfactant (10% tween 80, 29.87% span 80), the corresponding strain terms of size, PDI and Zed potential are 136.97, 0.200 and -42.629mV respectively. Optimized formula H group (Opt H): the percentage of surfactant and co-surfactant (28.41% 16.63% ), the corresponding strain values ​​of size, PDI and ZD potential are 26.64nm, 0.366 and -24.608mV respectively. The last group is the optimized formula J group (Opt J), the percentage of surfactant and co-surfactant (29.608% 30% tween 80), the corresponding strain values ​​of size, PDI and ZD potential are 210.34nm, 0.366 and -31.341mV respectively. Then, the four groups of optimization formulas suggested by the software were subjected to specific experiments, and the experimental results are shown in Table 8.

[0085] Table 8. Results of verifying the optimization formula

[0086]

[0087] The results are shown in Table 8, Difference % = (experimental value - predicted value) / experimental value x 100%, the difference is within 10%, which is within the acceptable range. Therefore, subsequent experiments were conducted on the four optimized formulas.

[0088] Example 7 Stability Study of Optimized Rhodiola Rosea Nanoemulsion

[0089] According to the four groups of formulations optimized in Example 6, stability tests or stability studies were performed to determine the stability of the API under various environmental conditions (such as temperature, light and humidity). The impact of external factors on the quality of the API, physical preparation or preparation product was evaluated in the stability test. The stability test of the optimized Rhodiola rosea nanoemulsion can be carried out at different times and temperatures.

[0090] Table 9. Stability study of optimized Rhodiola rosea nanoemulsion at different temperatures on the 1st, 30th, 60th and 90th days

[0091]

[0092] In this study, visual observation and physical tests such as pH, centrifugation test and viscosity test were examined. The changes in pH of each optimized formulation were examined at different temperatures (25°C, 40°C and 4°C) on day 1, day 30, day 60 and day 90. The pH values ​​of each optimized formulation were all within the range of 5.13-6.39, which met the requirements of topical drug delivery systems (Table 9).

[0093] Also the organoleptic conditions such as smell, color and texture were checked after the centrifugation test and no phase separation after 3000 rpm (30 minutes).Next was the viscosity stability test to check the 90th day and at the end of the 90th day (Table 10).

[0094] Table 10. Stability test of optimized Rhodiola rosea nanoemulsion using a rheometer for viscosity testing at 25°C

[0095]

[0096] The results showed that no changes in smell, color, texture were detected, and there was no phase separation and precipitation after the centrifugation test (not shown), indicating that the optimized Rhodiola rosea nanoemulsion had long-term stability. The viscosity test of each sample was checked before and after centrifugation to compare the results (Table 10). The stability indexes were Opt F 67.18%, Opt G 88.62%, Opt H 87.20, and Opt J 89.27%, respectively.

[0097] Example 8 Morphological Study

[0098] Transmission electron microscopy (TEM) images of the optimized Rhodiola rosea nanoemulsion were taken to observe the droplet diameter and surface morphology at a scale of 500 nm. FIG. 6A to FIG. 6D As shown, the results of the micrographs show spherical oil globules with a dense core surrounded by a darker color like a corona. The morphology of the Rhodiola rosea nanoemulsion droplets is clearly visible in the micrographs, the globules are spherical and evenly distributed, and their sizes are very close to the results obtained from the Zeta Potential Analyzer. In addition, no signs of globules or droplet coalescence were observed, thus proving the physical stability of the Rhodiola rosea nanoemulsion.

[0099] Although the present disclosure has been disclosed in the above embodiments, it is not intended to limit the present disclosure. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present disclosure. Therefore, the protection scope of the present disclosure shall be determined by the definition of the attached claims.

Claims

1. A Rhodiola rosea nanoemulsion, characterized in that: Include: Rhodiola rosea extract; Surfactants, including diethylene glycol monoethyl ester or polysorbate 80; a co-surfactant comprising polysorbate 80, sorbitan oleate 80, or caprylic / capric macrogol glycerides; and The oil is medium chain triglyceride or soybean oil, and is used to coat the Rhodiola rosea extract in an oil-in-water (O / W) manner.

2. The Rhodiola rosea nanoemulsion according to claim 1, wherein the Rhodiola rosea extract comprises an alcohol extract of Rhodiola rosea roots.

3. The Rhodiola rosea nanoemulsion according to claim 1, wherein the Rhodiola rosea extract comprises 1,2,3,4,6-penta-O-galloyl glucopyranoside and 1,2,3,6-tetra-O-galloyl-4-Op-hydroxybenzoyl-β-D-glucopyranoside.

4. The Rhodiola rosea nanoemulsion according to claim 1, wherein based on the total weight of the Rhodiola rosea nanoemulsion as 100%, The weight percentage of the surfactant is 10% to 30%; and The weight percentage of the co-surfactant is 10% to 30%.

5. The Rhodiola rosea nanoemulsion according to claim 1, wherein based on the total weight of the Rhodiola rosea nanoemulsion as 100%, The surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; The co-surfactant is polysorbate 80, and its weight percentage is 10% to 30%; and The oil is medium chain triglycerides, and its weight percentage is 1% to 5%. 6 . The Rhodiola rosea nanoemulsion according to claim 5 , wherein the weight percentage of the surfactant is 10%, and the weight percentage of the co-surfactant is 16.63%.

7. The Rhodiola rosea nanoemulsion according to claim 1, wherein based on the total weight of the Rhodiola rosea nanoemulsion as 100%, The surfactant is polysorbate 80, and its weight percentage is 10% to 30%; The co-surfactant is sorbitan oleate 80, and its weight percentage is 10% to 30%; and The oil is soybean oil, and its weight percentage is 1% to 5%. 8 . The Rhodiola rosea nanoemulsion according to claim 7 , wherein the weight percentage of the surfactant is 10%, and the weight percentage of the co-surfactant is 29.87%.

9. The Rhodiola rosea nanoemulsion according to claim 1, wherein based on the total weight of the Rhodiola rosea nanoemulsion as 100%, The surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; The co-surfactant is caprylic acid capric acid macrogol glyceride, and its weight percentage is 10% to 30%; and The oil is medium chain triglycerides, and its weight percentage is 1% to 5%. 10 . The Rhodiola rosea nanoemulsion according to claim 9 , wherein the weight percentage of the surfactant is 28.41%, and the weight percentage of the co-surfactant is 16.63%.

11. The Rhodiola rosea nanoemulsion according to claim 1, wherein based on the total weight of the Rhodiola rosea nanoemulsion as 100%, The surfactant is diethylene glycol monoethyl ester, and its weight percentage is 10% to 30%; The co-surfactant is polysorbate 80, and its weight percentage is 10% to 30%; and The oil is soybean oil, and its weight percentage is 1% to 5%. 12 . The Rhodiola rosea nanoemulsion according to claim 11 , wherein the weight percentage of the surfactant is 29.61%, and the weight percentage of the co-surfactant is 30%.

13. The Rhodiola rosea nanoemulsion according to claim 1, wherein the size of the Rhodiola rosea nanoemulsion is 10 nm to 500 nm, the polydispersity index is 0.173 to 0.667, and the zeta potential is -96.7 milliFord to -7.39 milliFord.

14. A method for preparing a Rhodiola rosea nanoemulsion, characterized in that: Include: Provides Rhodiola rosea extract; mixing the Rhodiola rosea extract with oil to obtain an oil phase component; mixing a surfactant and a co-surfactant to obtain a water phase component; The oil phase component is added to the water phase component to form an oil-in-water emulsion until emulsification, thereby obtaining the Rhodiola rosea nanoemulsion. Wherein, the surfactant comprises diethylene glycol monoethyl ester or polysorbate 80, Wherein, the co-surfactant comprises polysorbate 80, sorbitan oleate 80, or caprylic / capric macrogol glyceride. The oil contains medium chain triglycerides and soybean oil. The combination where the surfactant is diethylene glycol monoethyl ester, the co-surfactant is sorbitan oleate 80, and the oil is soybean oil is excluded.

15. The method according to claim 14, wherein the step of adding the oil phase component to the water phase component comprises dropping the oil phase component into the water phase component.

16. The method according to claim 14, wherein the step of adding the oil phase component to the water phase component is performed by emulsification by ultrasonic vibration.

17. The method according to claim 14, wherein the total weight of the Rhodiola rosea nanoemulsion is 100%, The weight percentage of the surfactant is 10% to 30%; and The weight percentage of the co-surfactant is 10% to 30%.

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