Stevioside-traditional Chinese medicine monomer self-assembly system and application thereof

Abstract

The invention discloses a stevioside-traditional Chinese medicine monomer self-assembly system and application thereof.The self-assembly system comprises stevioside, traditional Chinese medicine monomers and water, and the traditional Chinese medicine monomers are one or more of flavonoid substances, alkaloid substances, anthraquinone substances and organic acid substances; the flavonoid substance comprises one or more of quercetin, baicalein and glabridin, the alkaloid substance comprises one or more of glaucine, sophocarpidine and oxymatrine, the anthraquinone substance comprises one or more of resveratrol, oxyresveratrol and alpha-mangostin, and the anthraquinone substance comprises one or more of mangostin. The organic acid substance comprises one or more of ferulic acid, chlorogenic acid and gallic acid, the concentration of stevioside in water is greater than or equal to 100 mg / mL, and the concentration of the traditional Chinese medicine monomer in water is greater than or equal to 0.3 mg / mL. The self-assembly system prepared by the invention not only can increase the water solubility of the traditional Chinese medicine monomer, but also can be prepared into liquid crystal cream, and compared with common cream, under the condition of the same dosage, the liquid crystal cream is relatively high in skin retention rate and has a certain sustained and controlled release effect.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly to a stevioside - traditional Chinese medicine monomer self - assembly system and its application. Background Art

[0002] In cosmetics and personal care products, common active ingredients include antioxidants, whitening ingredients, anti - inflammatory ingredients, moisturizing ingredients, antibacterial ingredients, anti - wrinkle and firming ingredients, etc. However, most of the above - mentioned active ingredients are poorly soluble in water or have a solubility in water that is not high enough. This results in poor compatibility when these ingredients are formulated with other ingredients in the formula, may be unstable in the formula, and are prone to precipitation or aggregation, leading to a decrease in their bioavailability. In addition, the skin is a natural barrier, and ingredients that are poorly soluble in water may not be easily absorbed by the skin, thus reducing their efficacy.

[0003] To address the problem that the above - mentioned substances are poorly soluble in water or have insufficient water solubility, emulsification technology, nanotechnology, or special carrier systems are usually used to increase the solubility and stability of these active ingredients in an aqueous environment. This can not only improve the efficacy of the product but also ensure their uniform distribution in the formula. However, the preparation processes of the above - mentioned methods are relatively complex, and various raw materials need to be added to ensure the stability and safety of the formula system. Summary of the Invention

[0004] The object of the present invention is to provide a stevioside - traditional Chinese medicine monomer self - assembly system and its application to solve the problems of the relatively complex preparation method for improving the water solubility of traditional Chinese medicine monomers and their poor stability.

[0005] To achieve the above object, in the first aspect of the present invention, a stevioside - traditional Chinese medicine monomer self - assembly system is provided. The self - assembly system includes stevioside, traditional Chinese medicine monomers, and water.

[0006] Preferably, the traditional Chinese medicine monomers are one or more of flavonoids, alkaloids, anthraquinones, and organic acids.

[0007] Preferably, the flavonoids include one or more of quercetin, baicalein, and glabridin; the alkaloids include one or more of glauvin, matrine, and oxymatrine; the anthraquinones include one or more of resveratrol, oxyresveratrol, and α - mangostin; the organic acids include one or more of ferulic acid, chlorogenic acid, and gallic acid.

[0008] Preferably, the concentration of stevioside in water is ≥100 mg / mL, and the concentration of traditional Chinese medicine monomers in water is ≥0.3 mg / mL.

[0009] More preferably,

[0010] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of quercetin in water ≥ 463.42 μg / mL;

[0011] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of baicalein in water ≥ 536.43 μg / mL;

[0012] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of glabridin in water ≥ 5713.09 μg / mL;

[0013] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of glaucarubine in water ≥ 443.2 μg / mL;

[0014] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of matrine in water ≥ 2354.24 μg / mL;

[0015] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of oxymatrine in water ≥ 3212.42 μg / mL;

[0016] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of resveratrol in water ≥ 2180.24 μg / mL;

[0017] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of oxyresveratrol in water ≥ 6724.31 μg / mL;

[0018] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of α - mangostin in water ≥ 339.60 μg / mL;

[0019] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of ferulic acid in water ≥ 8023.40 μg / mL;

[0020] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of chlorogenic acid in water ≥ 5366.12 μg / mL;

[0021] When the concentration of stevioside in water ≥ 100 mg / mL and the concentration of gallic acid in water ≥ 16147.716 μg / mL.

[0022] Preferably, the concentration of stevioside in water is 100 mg / mL, and the concentration of the traditional Chinese medicine monomer in water is 0.3 - 17 mg / mL.

[0023] Preferably, the solubility of the traditional Chinese medicine monomer in the mixture of stevioside and water is 1 - 7500 times that of the traditional Chinese medicine monomer in pure water.

[0024] In view of the problem that the active ingredients used in cosmetics in the prior art are difficult to dissolve in water or have insufficient water solubility, the present invention specifically proposes a method with a simple preparation process and capable of achieving the water solubility of active ingredients. The Chinese medicine monomers are mixed with stevioside. During the mixing process, there is a synergistic effect between stevioside and these Chinese medicine monomers, which changes the physical properties of the Chinese medicine monomers and further improves their solubility.

[0025] Preferably, the self-assembly system further includes a surfactant.

[0026] Preferably, the surfactant is one or more of liquid surfactants and solid surfactants.

[0027] Preferably, the liquid surfactant is one or more of Tween 80, Tween 60, and Tween 20, and the solid surfactant is cetostearyl alcohol.

[0028] Preferably, the addition amount of the surfactant is 0.1-20% of the total mass of stevioside, Chinese medicine monomers, and water.

[0029] The present invention further studies and finds that although the above self-assembly system of stevioside, Chinese medicine monomers, and water can improve the water solubility of Chinese medicine monomers, during the long-term storage process of this self-assembly system, due to the instability of the system, solid substances precipitate. To solve this problem, the present invention further adds a surfactant to the above self-assembly system. After being placed at room temperature for up to 30 days, the solution can still remain clear, and the self-assembly system has good stability. By adding an appropriate amount of surfactant, the aqueous solution of stevioside and Chinese medicine monomers can be made more stable. This is because the surfactant can reduce the interfacial tension, form micelles, generate electrostatic repulsion, provide steric hindrance, adsorb and improve the dispersibility, etc., through the combined action of various mechanisms. The present invention improves the water solubility of glabridin through the synergistic effect of stevioside and surfactant and maintains its stability.

[0030] The second aspect of the present invention provides a preparation method of a stevioside-Chinese medicine monomer self-assembly system, including the following steps:

[0031] Add Chinese medicine monomers and stevioside to water, stir evenly until dissolution equilibrium is reached to obtain a mixed solution, add a surfactant to the mixed solution for mixing, and vortex and ultrasonicate at 40°C for 10-60 min until the solution is clear and transparent to obtain a self-assembly system.

[0032] In the present invention, the mixing method of Chinese medicine monomers, stevioside, and water includes, but is not limited to, mixing Chinese medicine monomers and stevioside and then adding them to water for dissolution, or first adding stevioside to water, mixing evenly, and then adding Chinese medicine monomers and continuing to mix and stir until a uniform mixed solution is obtained.

[0033] The third aspect of the present invention provides an application of a stevioside - traditional Chinese medicine monomer self - assembly system, specifically the application of the stevioside - traditional Chinese medicine monomer self - assembly system in the preparation of cosmetics.

[0034] More preferably, it is the application of the stevioside - traditional Chinese medicine monomer self - assembly system in the preparation of a liquid crystal cream.

[0035] The stevioside - traditional Chinese medicine monomer self - assembly system prepared by the present invention can be directly used in the preparation process of cosmetics, and the cosmetics include but are not limited to aqueous, paste - like, and milk - like forms. However, the research of the present invention finds that when the stevioside - traditional Chinese medicine monomer self - assembly system is applied to different cosmetics, the state of the cosmetics will affect the skin absorption effect of the self - assembly system to a certain extent. When the self - assembly system is applied to the preparation of a liquid crystal cream, the transdermal absorption effect of the active ingredients of the traditional Chinese medicine monomer is better.

[0036] Therefore, by adopting the above - mentioned stevioside - traditional Chinese medicine monomer self - assembly system and its application, the present invention has the following beneficial effects:

[0037] (1) By directly mixing stevioside, traditional Chinese medicine monomers, and water, the present invention can achieve the technical effect of improving traditional Chinese medicine monomers, and the preparation process of the present invention is simple and feasible.

[0038] (2) By further adding a surfactant to the system of stevioside, traditional Chinese medicine monomers, and water, the surfactant and stevioside act synergistically to further improve the stability of the self - assembly system.

[0039] (3) By using the stevioside - traditional Chinese medicine monomer self - assembly system to prepare a liquid crystal cream, compared with ordinary creams, under the same dosage, the liquid crystal cream has a higher skin retention rate and has a certain sustained - release effect.

[0040] The technical solution of the present invention will be further described in detail below through the accompanying drawings and examples. Description of the Drawings

[0041] Figure 1 It is the appearance diagram of the stability test of the Tween 80 - stevioside - glabridin self - assembly system;

[0042] Figure 2 It is the appearance diagram of the stability test of the Tween 60 - stevioside - glabridin self - assembly system;

[0043] Figure 3 It is the appearance diagram of the stability test of the Tween 20 - stevioside - glabridin self - assembly system;

[0044] Figure 4 It is the appearance diagram of the stability test of the cetostearyl alcohol - stevioside - glabridin self - assembly system;

[0045] Figure 5Appearance diagram of the stability test of hydrogenated soy phosphatidylcholine - stevioside - glabridin self - assembly system;

[0046] Figure 6 Appearance diagram of the stability test of the STE control group;

[0047] Figure 7 Appearance of the cream;

[0048] Figure 8 Liquid crystal structure of blank liquid crystal cream and liquid crystal cream containing drugs;

[0049] Figure 9 Particle size diagrams of 4 kinds of creams (100×);

[0050] Figure 10 Radar diagram of the sensory evaluation of 4 kinds of cream samples;

[0051] Figure 11 Results of specificity investigation;

[0052] Figure 12 Standard curve of GLA;

[0053] Figure 13 Retention rate of GLA in the skin of the liquid crystal cream group and the ordinary cream group;

[0054] Figure 13 Retention rate of GLA in fat of the liquid crystal cream group and the ordinary cream group;

[0055] Figure 13 Appearance diagram of the stability test of the Tween 80 - stevioside - glaucinol self - assembly system;

[0056] Figure 14 Appearance diagram of the stability test of the Tween 60 - stevioside - glaucinol self - assembly system;

[0057] Figure 14 Appearance diagram of the stability test of the Tween 20 - stevioside - glaucinol self - assembly system;

[0058] Figure 18 Appearance diagram of the stability test of the cetearyl alcohol - stevioside - glaucinol self - assembly system;

[0059] Figure 19 Appearance diagram of the stability test of the hydrogenated soy phosphatidylcholine - stevioside - glaucinol self - assembly system;

[0060] Figure 20 Appearance diagram of the stability test of the Tween 80 - stevioside - resveratrol self - assembly system;

[0061] Figure 21Appearance diagram for the stability test of the Tween 60 - stevioside - resveratrol self - assembly system;

[0062] Figure 22 Appearance diagram for the stability test of the Tween 20 - stevioside - resveratrol self - assembly system;

[0063] Figure 23 Appearance diagram for the stability test of the cetearyl alcohol - stevioside - resveratrol self - assembly system;

[0064] Figure 24 Appearance diagram for the stability test of the hydrogenated soy phosphatidylcholine - stevioside - resveratrol self - assembly system;

[0065] Figure 25 Appearance diagram for the stability test of the Tween 80 - stevioside - ferulic acid self - assembly system;

[0066] Figure 26 Appearance diagram for the stability test of the Tween 60 - stevioside - ferulic acid self - assembly system;

[0067] Figure 27 Appearance diagram for the stability test of the Tween 20 - stevioside - ferulic acid self - assembly system;

[0068] Figure 28 Appearance diagram for the stability test of the cetearyl alcohol - stevioside - ferulic acid self - assembly system;

[0069] Figure 29 Appearance diagram for the stability test of the hydrogenated soy phosphatidylcholine - stevioside - ferulic acid self - assembly system;

[0070] Figure 30 Retention rate of quercetin in the skin for the liquid crystal cream group and the ordinary cream group;

[0071] Figure 31 Retention rate of quercetin in the fat for the liquid crystal cream group and the ordinary cream group;

[0072] Figure 32 Retention rate of glaucine in the skin for the liquid crystal cream group and the ordinary cream group;

[0073] Figure 33 Retention rate of glaucine in the fat for the liquid crystal cream group and the ordinary cream group;

[0074] Figure 34 Retention rate of oxymatrine in the skin for the liquid crystal cream group and the ordinary cream group;

[0075] Figure 35 Retention rate of oxymatrine in the fat for the liquid crystal cream group and the ordinary cream group;

[0076] Figure 36is the retention rate of resveratrol in the skin of the liquid crystal cream group and the ordinary cream group;

[0077] Figure 37 is the retention rate of resveratrol in the fat of the liquid crystal cream group and the ordinary cream group;

[0078] Figure 38 is the retention rate of α-mangostin in the skin of the liquid crystal cream group and the ordinary cream group;

[0079] Figure 39 is the retention rate of α-mangostin in the fat of the liquid crystal cream group and the ordinary cream group;

[0080] Figure 40 is the retention rate of ferulic acid in the skin of the liquid crystal cream group and the ordinary cream group;

[0081] Figure 41 is the retention rate of ferulic acid in the fat of the liquid crystal cream group and the ordinary cream group;

[0082] Figure 42 is the retention rate of gallic acid in the skin of the liquid crystal cream group and the ordinary cream group;

[0083] Figure 43 is the retention rate of gallic acid in the fat of the liquid crystal cream group and the ordinary cream group. Detailed implementation mode

[0084] The present invention will be further described below. It should be noted that this embodiment is based on the present technical solution and gives detailed implementation methods and specific operation processes, but the present invention is not limited to this embodiment.

[0085] Example 1

[0086] A preparation method of a stevioside-quercetin self-assembly system includes the following steps:

[0087] Take 5 mg of quercetin powder, accurately weigh it, add it to a beaker, and then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a stevioside-quercetin self-assembly system.

[0088] Example 2

[0089] A preparation method of a stevioside-scutellarin self-assembly system includes the following steps:

[0090] Take 6 mg of scutellarin powder, accurately weigh it, add it to a beaker, and then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a stevioside-scutellarin self-assembly system.

[0091] Example 3

[0092] A preparation method of a stevioside-glabridin self-assembly system, comprising the following steps:

[0093] Take 60.0 mg of glabridin (GLA) powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-glabridin self-assembly system.

[0094] Example 4

[0095] A preparation method of a stevioside-glaucine self-assembly system, comprising the following steps:

[0096] Take 5 mg of glaucine powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-glaucine self-assembly system.

[0097] Example 5

[0098] A preparation method of a stevioside-matrine self-assembly system, comprising the following steps:

[0099] Take 30.0 mg of matrine powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-matrine self-assembly system.

[0100] Example 6

[0101] A preparation method of a stevioside-oxymatrine self-assembly system, comprising the following steps:

[0102] Take 40.0 mg of oxymatrine powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-oxymatrine self-assembly system.

[0103] Example 7

[0104] A preparation method of a stevioside-resveratrol self-assembly system, comprising the following steps:

[0105] Take 30.0 mg of resveratrol powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-resveratrol self-assembly system.

[0106] Example 8

[0107] A preparation method of a stevioside-veratryl alcohol self-assembly system, comprising the following steps:

[0108] Take 70.0 mg of veratryl alcohol powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-veratryl alcohol self-assembly system.

[0109] Example 9

[0110] A preparation method of a stevioside-α-mangostin self-assembly system, comprising the following steps:

[0111] Take 4.0 mg of α-mangostin powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-α-mangostin self-assembly system.

[0112] Example 10

[0113] A preparation method of a stevioside-ferulic acid self-assembly system, comprising the following steps:

[0114] Take 90.0 mg of ferulic acid powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-ferulic acid self-assembly system.

[0115] Example 11

[0116] A preparation method of a stevioside-chlorogenic acid self-assembly system, comprising the following steps:

[0117] Take 60.0 mg of chlorogenic acid powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-chlorogenic acid self-assembly system.

[0118] Example 12

[0119] A preparation method of a stevioside-gallic acid self-assembly system, comprising the following steps:

[0120] Take 170.0 mg of gallic acid powder, accurately weigh it, add it to a beaker, then add it to 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, and place it in a magnetic stirrer to stir until dissolution equilibrium is reached to obtain a stevioside-gallic acid self-assembly system.

[0121] Test Example 1

[0122] The water solubilities of flavonoids, alkaloids, anthraquinones, and organic acid compounds in the stevioside - traditional Chinese medicine monomer self - assembly systems of Examples 1 to 12 were detected. The detection method was: high - performance liquid chromatography, which is a detection method already disclosed in the prior art. The detection results are shown in Table 1.

[0123] The chromatographic conditions were as follows:

[0124] Flavonoids:

[0125] ① Quercetin

[0126] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100mm×2.1mm, 1.7μm); Mobile phase: methanol - water (60∶40); Detection wavelength: 254nm; Flow rate: 0.2mL / min; Column temperature: 30℃; Injection volume: 2μL.

[0127] ② Baicalein

[0128] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100mm×2.1mm, 1.7μm); Mobile phase: methanol - water (60∶40); Detection wavelength: 275nm; Flow rate: 0.2mL / min; Column temperature: 30℃; Injection volume: 2μL.

[0129] ③ Glabridin

[0130] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100mm×2.1mm, 1.7μm); Mobile phase: acetonitrile - water (60∶40); Detection wavelength: 280nm; Flow rate: 0.2mL / min; Column temperature: 30℃; Injection volume: 2μL.

[0131] Alkaloids:

[0132] ① Glaucine

[0133] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100mm×2.1mm, 1.7μm); Mobile phase: methanol - triethylamine water (30∶70); Detection wavelength: 235nm; Flow rate: 0.2mL / min; Column temperature: 30℃; Injection volume: 2μL.

[0134] ② Matrine

[0135] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - triethylamine water (35∶65); Detection wavelength: 220 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0136] ③ Oxymatrine

[0137] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - triethylamine water (35∶65); Detection wavelength: 206 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0138] Anthraquinones:

[0139] ① Resveratrol

[0140] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - water (60∶40); Detection wavelength: 306 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0141] ② Oxidized resveratrol

[0142] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: acetonitrile - water (30∶70); Detection wavelength: 329 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0143] ③ α - Mangostin

[0144] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - water (80∶20); Detection wavelength: 317 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0145] Organic acids:

[0146] CD Chlorogenic acid

[0147] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - water (65∶35); Detection wavelength: 225 nm; Flow rate: 0.2 mI / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0148] ②Gallic acid

[0149] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - water (65∶35); Detection wavelength: 273 nm; Flow rate: 0.2 mL / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0150] ③Ferulic acid

[0151] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100 mm × 2.1 mm, 1.7 μm); Mobile phase: methanol - water (60∶40); Detection wavelength: 325 nm; Flow rate: 0.2 mI / min; Column temperature: 30 °C; Injection volume: 2 μL.

[0152] Table 1 Detection results of water solubility of traditional Chinese medicine monomers

[0153]

[0154] It can be seen from Table 1 that flavonoids and anthraquinones have low solubility in water. Stevioside can significantly improve the solubility of flavonoids and anthraquinones in water. For alkaloids and organic acids with relatively high solubility in water, stevioside can also further improve the solubility of alkaloids and organic acids in water to a certain extent.

[0155] Example 13

[0156] A preparation method of a stevioside - glabridin self - assembly system, comprising the following steps:

[0157] Take 60.0 mg of glabridin powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / m1, stir and mix evenly, place it in a magnetic stirrer and stir until dissolution equilibrium is reached to obtain a mixed solution. Weigh different surfactants according to Table 2 respectively. The addition amount of the surfactant is the mass percentage of the mixed solution. Place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, place it in a 10.0 mL centrifuge tube for mixing, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, then the stevioside - glabridin self - assembly system is obtained.

[0158] Table 2 Formulation table of stevioside - glabridin self - assembly system under different surfactants

[0159]

[0160]

[0161] Experimental Example 2

[0162] Study on the effect of different surfactants on the stability of the stevioside - glabridin self - assembly system. Experimental process: The stevioside - glabridin self - assembly systems prepared with different surfactants were placed in a 4°C refrigerator. The stevioside - glabridin self - assembly system (STE) in Example 1 was used as the control group. Samples were taken out at 0, 1, 2, 4, 6, 8, 10....30 days to observe, record the experimental phenomena, and take pictures.

[0163] From Figures 1 - 6 the experimental results, it can be seen that the effects of different liquid surfactants on the performance of the stevioside - glabridin self - assembly system are different. The stevioside - glabridin solution without adding liquid surfactant began to precipitate at 3 days.

[0164] (1) Tween 80

[0165] When the addition amount of Tween 80 was 1%, 5%, the stevioside - glabridin solution precipitated at 3 days, with the same stability as the stevioside - glabridin solution without adding Tween 80. When the addition amount of Tween 80 was 10%, the stevioside - glabridin solution precipitated at 10 days, which was more stable than the stevioside - glabridin solution without adding Tween 80. When the addition amount of Tween 80 was 20%, the stevioside - glabridin solution still did not precipitate at 30 days, which was very stable compared to the stevioside - glabridin solution without adding Tween 80. It shows that high - concentration Tween 80 can greatly increase the stability of the stevioside - glabridin self - assembly system.

[0166] (2) Tween 60

[0167] When the addition amount of Tween 60 was 1%, 5%, the stevioside - glabridin solution precipitated at 3 days, with the same stability as the stevioside - glabridin solution without adding Tween 60. When the addition amount of Tween 60 was 10%, the stevioside - glabridin solution precipitated at 8 days, which was more stable than the stevioside - glabridin solution without adding Tween 60. When the addition amount of Tween 60 was 20%, the stevioside - glabridin solution still did not precipitate at 30 days, which was very stable compared to the stevioside - glabridin solution without adding Tween 60. It shows that high - concentration Tween 60 can greatly increase the stability of the stevioside - glabridin self - assembly system.

[0168] (3) Tween 20

[0169] When the addition amount of Tween 20 is 1% and 5%, the stevioside - glabridin solution precipitates at 4 days, and it is more stable than the stevioside - glabridin solution without adding Tween 20. When the addition amount of Tween 20 is 10%, the stevioside - glabridin solution precipitates at 9 days, and it is more stable than the stevioside - glabridin solution without adding Tween 20. When the addition amount of Tween 20 is 20%, the stevioside - glabridin solution still does not precipitate at 30 days, and it is very stable compared with the stevioside - glabridin solution without adding Tween 20. It shows that high - concentration Tween 20 can greatly increase the stability of the stevioside - glabridin self - assembly system.

[0170] From Figure 2 it can be seen that the effects of different solid surfactants on the performance of the stevioside - glabridin self - assembly system are different. The stevioside - glabridin solution without adding liquid surfactant starts to precipitate at 3 days.

[0171] (4) Cetearyl alcohol

[0172] When the addition amount of cetearyl alcohol is 1%, the stevioside - glabridin solution precipitates at 5 days, and it is more stable than the stevioside - glabridin solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 5%, the stevioside - glabridin solution precipitates at 8 days, and it is more stable than the stevioside - glabridin solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 10%, the stevioside - glabridin solution precipitates at 9 days, and it is more stable than the stevioside - glabridin solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 20%, the stevioside - glabridin solution precipitates at 20 days, and it is more stable than the stevioside - glabridin solution without adding cetearyl alcohol. It shows that high - concentration cetearyl alcohol can preferably increase the stability of the stevioside - glabridin self - assembly system.

[0173] (5) Hydrogenated soy phosphatidylcholine

[0174] When the addition amount of hydrogenated soy phosphatidylcholine is 5%, the stevioside - glabridin solution precipitates at 1 day, and it is less stable than the stevioside - glabridin solution without adding hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 10%, the stevioside - glabridin solution precipitates at 2 days, and it is less stable than the stevioside - glabridin solution without adding hydrogenated soy phosphatidylcholine. When the addition amounts of hydrogenated soy phosphatidylcholine are 1% and 20%, the stevioside - glabridin solution precipitates at 3 days, and it has the same stability as the stevioside - glabridin solution without adding hydrogenated soy phosphatidylcholine. It shows that hydrogenated soy phosphatidylcholine cannot increase the stability of the stevioside - glabridin self - assembly system.

[0175] Example 14

[0176] The difference between this example and Example 13 is that the stevioside and glabridin added in Example 5 are respectively replaced with the stevioside and other traditional Chinese medicine monomer components (quercetin, baicalein, glaucerine, matrine, oxymatrine, resveratrol, oxyresveratrol, α-mangostin, ferulic acid, chlorogenic acid and gallic acid) in Examples 1-2 and Examples 4-12. Different types and contents of surfactants are also added, and finally a stevioside-traditional Chinese medicine monomer self-assembly system with different components is obtained. The component composition is shown in Table 3.

[0177] Test Example 3

[0178] The stability of each stevioside-traditional Chinese medicine monomer self-assembly system in Example 14 was tested. The test method was the same as that in Test Example 2, and the test results are shown in Table 3.

[0179] Table 3 Stability test results of stevioside-traditional Chinese medicine monomers under different surfactants

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193] It can be seen from Table 3 that the stabilizing effects of different surfactants are different, and among them, Tween 20, Tween 60 and Tween 80 have better stabilizing effects.

[0194] For the stevioside-glaucine self-assembly system, Table 4 shows the formulation table of the stevioside-glaucine self-assembly system under different surfactants. As can be seen from Figures 15 - 19 the experimental results in it, the effects of different liquid surfactants on the performance of the stevioside-glaucine self-assembly system are different. The stevioside-glaucine solution without adding liquid surfactant starts to precipitate after 2 days.

[0195] Table 4 Formulation table of stevioside-glaucine self-assembly system under different surfactants

[0196]

[0197]

[0198] (1) Tween 80

[0199] When the addition amount of Tween 80 is 1%, the stevioside-glaucine solution starts to precipitate at 4 days, which is more stable than the stevioside-glaucine solution without adding Tween 80. When the addition amount of Tween 80 is 5%, the stevioside-glaucine solution starts to precipitate at 5 days, which is more stable than the stevioside-glaucine solution without adding Tween 80. When the addition amount of Tween 80 is 10%, the stevioside-glaucine solution starts to precipitate at 8 days, which is more stable than the stevioside-glaucine solution without adding Tween 80. When the addition amount of Tween 80 is 20%, the stevioside-glaucine solution still does not precipitate at 30 days, which is very stable compared to the stevioside-glaucine solution without adding Tween 80. This shows that a high concentration of Tween 80 can greatly increase the stability of the stevioside-glaucine self-assembly system.

[0200] (2) Tween 60

[0201] When the addition amount of Tween 60 is 1%, the stevioside-glaucine solution starts to precipitate at 4 days, which is more stable than the stevioside-glaucine solution without adding Tween 60. When the addition amount of Tween 60 is 5%, the stevioside-glaucine solution starts to precipitate at 5 days, which is more stable than the stevioside-glaucine solution without adding Tween 60. When the addition amount of Tween 60 is 10%, the stevioside-glaucine solution starts to precipitate at 7 days, which is more stable than the stevioside-glaucine solution without adding Tween 60. When the addition amount of Tween 60 is 20%, the stevioside-glaucine solution still does not precipitate at 30 days, which is very stable compared to the stevioside-glaucine solution without adding Tween 60. This shows that a high concentration of Tween 60 can greatly increase the stability of the stevioside-glaucine self-assembly system.

[0202] (3) Tween 20

[0203] When the addition amount of Tween 20 is 1%, precipitation occurs in the stevioside-glaucine solution at 4 days, which is more stable than the stevioside-glaucine solution without adding Tween 20. When the addition amount of Tween 20 is 5%, precipitation occurs in the stevioside-glaucine solution at 6 days, which is less stable than the stevioside-glaucine solution without adding Tween 20. When the addition amount of Tween 20 is 10%, precipitation occurs in the stevioside-glaucine solution at 9 days, which is more stable than the stevioside-glaucine solution without adding Tween 20. When the addition amount of Tween 20 is 20%, precipitation still does not occur in the stevioside-glaucine solution at 30 days, which is very stable compared to the stevioside-glaucine solution without adding Tween 20. It shows that high-concentration Tween 20 can greatly increase the stability of the stevioside-glaucine self-assembly system.

[0204] (4) Cetearyl alcohol

[0205] When the addition amount of cetearyl alcohol is 1%, precipitation occurs in the stevioside-glaucine solution at 5 days, which is more stable than the stevioside-glaucine solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 5%, precipitation occurs in the stevioside-glaucine solution at 8 days, which is more stable than the stevioside-glaucine solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 10%, precipitation occurs in the stevioside-glaucine solution at 9 days, which is more stable than the stevioside-glaucine solution without adding cetearyl alcohol. When the addition amount of cetearyl alcohol is 20%, precipitation occurs in the stevioside-glaucine solution at 20 days, which is more stable than the stevioside-glaucine solution without adding cetearyl alcohol. It shows that high-concentration cetearyl alcohol can preferably increase the stability of the stevioside-glaucine self-assembly system.

[0206] (5) Hydrogenated soy phosphatidylcholine

[0207] When the addition amount of hydrogenated soy phosphatidylcholine is 5%, precipitation occurs in the stevioside-glaucine solution at 1 day, which is less stable than the stevioside-glaucine solution without adding hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 10%, precipitation occurs in the stevioside-glaucine solution at 2 days, which is less stable than the stevioside-glaucine solution without adding hydrogenated soy phosphatidylcholine. When the addition amounts of hydrogenated soy phosphatidylcholine are 1% and 20%, precipitation occurs in the stevioside-glaucine solution at 3 days, and the stability is the same as that of the stevioside-glaucine solution without adding hydrogenated soy phosphatidylcholine. It shows that hydrogenated soy phosphatidylcholine cannot increase the stability of the stevioside-glaucine self-assembly system.

[0208] For the stevioside-resveratrol self-assembly system, Table 5 shows the formulation table of the stevioside-resveratrol self-assembly system under different surfactants. From Figures 20 - 24 the experimental results in it, it can be seen that different liquid surfactants have different effects on the performance of the stevioside-resveratrol self-assembly system. The stevioside-resveratrol solution without adding liquid surfactant begins to precipitate after 3 days.

[0209] Table 5 Formulation table of stevioside-resveratrol self-assembly system under different surfactants

[0210]

[0211]

[0212] (1) Tween 80

[0213] When the addition amount of Tween 80 is 1%, the stevioside-resveratrol solution precipitates after 3 days, and its stability is the same as that of the stevioside-resveratrol solution without adding Tween 80. When the addition amount of Tween 80 is 5%, the stevioside-resveratrol solution precipitates after 3 days, and its stability is the same as that of the stevioside-resveratrol solution without adding Tween 80. When the addition amount of Tween 80 is 10%, the stevioside-resveratrol solution precipitates after 10 days, and it is more stable than the stevioside-resveratrol solution without adding Tween 80. When the addition amount of Tween 80 is 20%, the stevioside-resveratrol solution still does not precipitate after 30 days, and it is very stable compared with the stevioside-resveratrol solution without adding Tween 80. It shows that high-concentration Tween 80 can greatly increase the stability of the stevioside-resveratrol self-assembly system.

[0214] (2) Tween 60

[0215] When the addition amount of Tween 60 is 1%, the stevioside-resveratrol solution precipitates after 3 days, and its stability is the same as that of the stevioside-resveratrol solution without adding Tween 60. When the addition amount of Tween 60 is 5%, the stevioside-resveratrol solution precipitates after 5 days, and it is more stable than the stevioside-resveratrol solution without adding Tween 60. When the addition amount of Tween 60 is 10%, the stevioside-resveratrol solution precipitates after 9 days, and it is more stable than the stevioside-resveratrol solution without adding Tween 60. When the addition amount of Tween 60 is 20%, the stevioside-resveratrol solution still does not precipitate after 30 days, and it is very stable compared with the stevioside-resveratrol solution without adding Tween 60. It shows that high-concentration Tween 60 can greatly increase the stability of the stevioside-resveratrol self-assembly system.

[0216] (3) Tween 20

[0217] When the addition amount of Tween 20 is 1%, the stevioside-resveratrol solution precipitates at 4 days, which is more stable than the stevioside-resveratrol solution without Tween 20. When the addition amount of Tween 20 is 5%, the stevioside-resveratrol solution precipitates at 4 days, which is more stable than the stevioside-resveratrol solution without Tween 20. When the addition amount of Tween 20 is 10%, the stevioside-resveratrol solution precipitates at 10 days, which is more stable than the stevioside-resveratrol solution without Tween 20. When the addition amount of Tween 20 is 20%, the stevioside-resveratrol solution still does not precipitate at 30 days, which is very stable compared to the stevioside-resveratrol solution without Tween 20. This shows that high-concentration Tween 20 can greatly increase the stability of the stevioside-resveratrol self-assembly system.

[0218] (4) Cetearyl alcohol

[0219] When the addition amount of cetearyl alcohol is 1%, the stevioside-resveratrol solution precipitates at 5 days, which is more stable than the stevioside-resveratrol solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 5%, the stevioside-resveratrol solution precipitates at 8 days, which is more stable than the stevioside-resveratrol solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 10%, the stevioside-resveratrol solution precipitates at 9 days, which is more stable than the stevioside-resveratrol solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 20%, the stevioside-resveratrol solution precipitates at 22 days, which is more stable than the stevioside-resveratrol solution without cetearyl alcohol. This shows that high-concentration cetearyl alcohol can preferably increase the stability of the stevioside-resveratrol self-assembly system.

[0220] (5) Hydrogenated soy phosphatidylcholine

[0221] When the addition amount of hydrogenated soy phosphatidylcholine is 1%, the stevioside-resveratrol solution precipitates at 3 days, and its stability is the same as that of the stevioside-resveratrol solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 5%, the stevioside-resveratrol solution precipitates at 1 day, and it is less stable than the stevioside-resveratrol solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 10%, the stevioside-resveratrol solution precipitates at 1 day, and it is less stable than the stevioside-resveratrol solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 20%, the stevioside-resveratrol solution precipitates at 3 days, and its stability is the same as that of the stevioside-resveratrol solution without the addition of hydrogenated soy phosphatidylcholine. This shows that hydrogenated soy phosphatidylcholine cannot increase the stability of the stevioside-resveratrol self-assembly system.

[0222] For the stevioside-ferulic acid self-assembly system, Table 6 shows the formulation table of the stevioside-ferulic acid self-assembly system under different surfactants. From Figures 25 - 29 the experimental results in it, it can be seen that different liquid surfactants have different effects on the performance of the stevioside-ferulic acid self-assembly system. The stevioside-ferulic acid solution without the addition of liquid surfactant starts to precipitate at 4 days.

[0223] Table 6 Formulation table of the stevioside-ferulic acid self-assembly system under different surfactants

[0224]

[0225] (1) Tween 80

[0226] When the addition amount of Tween 80 is 1%, the stevioside-ferulic acid solution precipitates at 3 days, and its stability is the same as that of the stevioside-ferulic acid solution without the addition of Tween 80. When the addition amount of Tween 80 is 5%, the stevioside-ferulic acid solution precipitates at 5 days, and it is more stable than the stevioside-ferulic acid solution without the addition of Tween 80. When the addition amount of Tween 80 is 10%, the stevioside-ferulic acid solution precipitates at 9 days, and it is more stable than the stevioside-ferulic acid solution without the addition of Tween 80. When the addition amount of Tween 80 is 20%, the stevioside-ferulic acid solution still does not precipitate at 30 days, and it is very stable compared to the stevioside-ferulic acid solution without the addition of Tween 80. This shows that high-concentration Tween 80 can greatly increase the stability of the stevioside-ferulic acid self-assembly system.

[0227] (2) Tween 60

[0228] When the addition amount of Tween 60 is 1%, the stevioside-ferulic acid solution precipitates at 2 days, which is more unstable than the stevioside-ferulic acid solution without Tween 60. When the addition amount of Tween 60 is 5%, the stevioside-ferulic acid solution precipitates at 3 days, and has the same stability as the stevioside-ferulic acid solution without Tween 60. When the addition amount of Tween 60 is 10%, the stevioside-ferulic acid solution precipitates at 5 days, which is more stable than the stevioside-ferulic acid solution without Tween 60. When the addition amount of Tween 60 is 20%, the stevioside-ferulic acid solution still does not precipitate at 30 days, which is very stable compared with the stevioside-ferulic acid solution without Tween 60. It shows that high concentration of Tween 60 can greatly increase the stability of the stevioside-ferulic acid self-assembly system.

[0229] (3) Tween 20

[0230] When the addition amount of Tween 20 is 1%, the stevioside-ferulic acid solution precipitates at 2 days, which is more unstable than the stevioside-ferulic acid solution without Tween 20. When the addition amount of Tween 20 is 5%, the stevioside-ferulic acid solution precipitates at 3 days, and has the same stability as the stevioside-ferulic acid solution without Tween 20. When the addition amount of Tween 20 is 10%, the stevioside-ferulic acid solution precipitates at 7 days, which is more stable than the stevioside-ferulic acid solution without Tween 20. When the addition amount of Tween 20 is 20%, the stevioside-ferulic acid solution still does not precipitate at 30 days, which is very stable compared with the stevioside-ferulic acid solution without Tween 20. It shows that high concentration of Tween 20 can greatly increase the stability of the stevioside-ferulic acid self-assembly system.

[0231] (4) Cetearyl alcohol

[0232] When the addition amount of cetearyl alcohol is 1%, the stevioside-ferulic acid solution precipitates at 5 days, which is more stable than the stevioside-ferulic acid solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 5%, the stevioside-ferulic acid solution precipitates at 7 days, which is more stable than the stevioside-ferulic acid solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 10%, the stevioside-ferulic acid solution precipitates at 8 days, which is more stable than the stevioside-ferulic acid solution without cetearyl alcohol. When the addition amount of cetearyl alcohol is 20%, the stevioside-ferulic acid solution precipitates at 20 days, which is more stable than the stevioside-ferulic acid solution without cetearyl alcohol. It shows that high concentration of cetearyl alcohol can preferably increase the stability of the stevioside-ferulic acid self-assembly system.

[0233] (5) Hydrogenated soy phosphatidylcholine

[0234] When the addition amount of hydrogenated soy phosphatidylcholine is 1%, precipitation occurs in the stevioside-ferulic acid solution at 3 days, and the stability is the same as that of the stevioside-ferulic acid solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 5%, precipitation occurs in the stevioside-ferulic acid solution at 2 days, and it is less stable than the stevioside-ferulic acid solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 10%, precipitation occurs in the stevioside-ferulic acid solution at 4 days, and it is more stable than the stevioside-ferulic acid solution without the addition of hydrogenated soy phosphatidylcholine. When the addition amount of hydrogenated soy phosphatidylcholine is 20%, precipitation occurs in the stevioside-ferulic acid solution at 2 days, and it is less stable than the stevioside-ferulic acid solution without the addition of hydrogenated soy phosphatidylcholine. This shows that hydrogenated soy phosphatidylcholine cannot increase the stability of the stevioside-ferulic acid self-assembly system.

[0235] Example 15

[0236] A preparation method of a stevioside-glabridin self-assembly system includes the following steps:

[0237] Take 60.0 mg of glabridin powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, then the stevioside-glabridin self-assembly system is obtained.

[0238] A preparation method of a liquid crystal cream includes the following steps:

[0239] (1) Place the obtained stevioside-glabridin self-assembly system or the stevioside-glabridin self-assembly system prepared in Example 1 in a -80 °C refrigerator overnight, take it out and freeze-dry it, and grind the obtained solid into powder to obtain the solubilized powder.

[0240] (2) Weigh each raw material of phase A according to Table 7, heat it in a water bath at 72 °C, stir and dissolve it, and keep the temperature at 72 °C;

[0241] Disperse carbomer into the mixture of glycerol and butanediol, stir evenly with a glass rod, then add other raw materials of phase B, heat it in a water bath at 75 °C, stir and dissolve it, and keep the temperature at 75 °C;

[0242] After the phases A and B are completely dissolved, pour phase A into phase B, maintain the temperature at 75 °C, stir evenly, homogenize and emulsify at 7000 r / min for 3 min, add the raw materials of phase C, stir evenly, homogenize and emulsify at 4000 r / min for 4 min, and cool down to 45 °C;

[0243] Add the raw materials of phase D and the solubilizer powder, stir evenly, and cool to room temperature to obtain the medicated liquid crystal cream.

[0244] The medicated liquid crystal cream is divided into the stevioside - glabridin liquid crystal cream prepared in Example 1 and the Tween 20 - stevioside - glabridin liquid crystal cream prepared in Example 13.

[0245] Table 7 Self - made basic formula table of liquid crystal cream

[0246]

[0247]

[0248] Comparative Example 1

[0249] The difference from Example 15 is that the solubilizer powder is not added, and a blank liquid crystal cream is prepared.

[0250] Comparative Example 2

[0251] The difference from Example 15 is that the solubilizer powder is added to the preparation process of the ordinary cream to obtain a medicated ordinary cream.

[0252] The preparation process of the ordinary cream is as follows:

[0253] (1) Place the stevioside - glabridin self - assembly system obtained in Example 15 above or the stevioside - glabridin self - assembly system prepared in Example 1 in an - 80 °C refrigerator overnight, take it out and freeze - dry, and grind the obtained solid into powder to obtain the solubilizer powder.

[0254] (2) Weigh each raw material of phase A according to Table 8, heat it in a water bath at 72 °C, stir to dissolve, and maintain the temperature at 72 °C;

[0255] Disperse carbomer into the mixture of glycerol and butanediol, stir evenly with a glass rod, then add other raw materials of phase B, heat it in a water bath at 75 °C, stir to dissolve, and maintain the temperature at 75 °C;

[0256] After the phases A and B are completely dissolved, pour phase A into phase B, maintain the temperature at 75 °C, stir evenly, homogenize and emulsify at 7000 r / min for 3 min, add the raw materials of phase C, stir evenly, homogenize and emulsify at 4000 r / min for 4 min, and cool down to 45 °C;

[0257] Add the raw material of Phase D and solubilizer powder, stir evenly, and cool to room temperature to obtain a medicated ordinary cream. The medicated ordinary cream is divided into the stevioside-glabridin ordinary cream prepared in Example 1 and the Tween 20-stevioside-glabridin ordinary cream prepared in Example 7.

[0258] Table 8 Basic formula table of self-made ordinary cream

[0259]

[0260]

[0261] Comparative Example 3

[0262] The difference from Comparative Example 2 is that solubilizer powder was not added during the preparation of the ordinary cream to obtain a blank ordinary cream.

[0263] Test Example 4

[0264] Characterize the cosmetics prepared in Example 15 and Comparative Examples 1-3.

[0265] (1) Appearance

[0266] As Figure 7 、 Figure 8 and Figure 9 shown, take an appropriate amount of blank liquid crystal cream, medicated liquid crystal cream (2 kinds), blank ordinary cream and medicated ordinary cream (2 kinds), and observe at room temperature without direct sunlight. It is observed that the blank liquid crystal cream and the blank ordinary cream are white and delicate creams; the medicated liquid crystal cream and the medicated ordinary cream are slightly yellow and delicate creams. The colors and textures of the four creams are uniform. Observe the particle size and crystal precipitation of the creams under an optical microscope. It is observed under a polarized light microscope that the liquid crystal structures of the blank liquid crystal cream and the medicated liquid crystal cream are complete. At the same time, it can be seen under an optical microscope that the solubilizer powder can be dissolved in the blank liquid crystal cream and the blank ordinary cream. The particle size of the liquid crystal cream is generally significantly smaller than that of the ordinary cream and is more uniform.

[0267] (2) Sensory evaluation of cream

[0268] The sensory evaluation group consists of 10 members, aged 19 to 23 years. Before the test, professionals explained the test content to make the results highly reliable. The evaluators need to clean the inner forearm area and dry it with a clean tissue before the test. The same sampling amount is required for each test. When testing, 6 cream samples (blank liquid crystal cream, blank ordinary cream, two kinds of medicated liquid crystal cream and two kinds of medicated ordinary cream) are compared. Referring to the ASTM sensory evaluation vocabulary and standards for cream products, 5 evaluation vocabulary are listed in this sensory evaluation test, and the specific contents are the pick-up property before use, the spreadability during use, the moisturizing feeling, the absorption speed and the greasiness. The specific descriptions are shown in Table 9. After calculating the average score of the scoring results, the results are statistically asFigure 10 As can be seen from the figure, the liquid crystal cream sample has a low stickiness and poor pick-up property; it is superior to the ordinary cream sample in terms of spreadability, moisturizing feeling and absorption speed.

[0269] Table 9 Sensory Evaluation Indexes and Definitions

[0270] Index Name Definition Description Scoring Range Provocativeness Ease of Dipping the Paste 1 (Weak) - 5 (Strong) Spreadability Ease of Spreading the Paste 1 (Weak) - 5 (Strong) Moisturizing Feeling Degree of Moisturization Perceivable by the Skin 1 (Weak) - 5 (Strong) Absorption Rate Absorption Amount after a Fixed Time of Applying the Paste 1 (Weak) - 5 (Strong) Stickiness Degree of Adhesion between the Finger and the Skin 1 (Weak) - 5 (Strong)

[0271] (3) Establishment of Ultra Performance Liquid Chromatography Detection Method

[0272] Chromatographic Conditions

[0273] Chromatographic column: Waters ACQUITY UPLC BEH C18 (100mm×2.1mm, 1.7μm); Mobile phase: acetonitrile - water (60∶40); Detection wavelength: 280nm; Flow rate: 0.2mL / min; Column temperature: 30℃; Injection volume: 2μL.

[0274] Preparation of GLA Reference Substance Solution

[0275] Take about 10.00mg of GLA reference substance, accurately weigh it, place it in a 10mL volumetric flask, and make up the volume with methanol to prepare a reference substance stock solution with a mass concentration of 1mg / mL.

[0276] Preparation of Test Samples

[0277] Accurately weigh 1.00g of liquid crystal cream, add 5mL of methanol solution, ultrasonically demulsify for 1h, take the supernatant, filter it through a 0.22μm filter membrane, and dilute it with the mobile phase by an appropriate multiple for liquid phase analysis.

[0278] (4) Specificity Test

[0279] Respectively take blank skin, blank fat, medicated skin and medicated fat samples, and perform liquid phase analysis according to the above chromatographic conditions to obtain the chromatograms of blank skin, blank fat, medicated skin, medicated fat and GLA reference substance. Figure 11 。From Figure 11 it can be seen that characteristic peaks of glabridin appear in the chromatograms of medicated skin and medicated fat, indicating that the active ingredients in the self-assembly system are absorbed by the skin and retained in the skin.

[0280] (5) GLA High Performance Liquid Chromatography Test

[0281] Investigation of Linear Relationship

[0282] Dilute the GLA reference stock solution to obtain a series of GLA solutions with different mass concentrations. Filter through a 0.22 μm microporous membrane and perform liquid chromatography analysis under the above chromatographic conditions. Using the mass concentration of GLA reference (μg / mL) as the abscissa (X) and the peak area as the ordinate (Y) for linear fitting, the regression equation is Y = 16.157X - 12.529, with a correlation coefficient f = 0.9998. In the concentration range of 2.25 - 260 μg / mL for GLA, there is a good linear relationship between the peak area and the concentration. The standard curve is shown in Figure 12 .

[0283] Precision investigation

[0284] Take an appropriate concentration of GLA reference solution and inject it continuously 6 times under the above chromatographic conditions. The injection volume is 2 μL each time. Record the peak area. The RSD of the measured peak area is 0.320%, indicating good precision of the instrument.

[0285] Stability investigation

[0286] Take the complex sample solution at different storage times, namely 0, 2, 4, 8, 12, and 24 h. Inject and measure according to the above chromatographic conditions. The injection volume is 2 μL each time. Record the peak area value and calculate the RSD. After investigation, the GLA test solution remains stable within 24 h, with an RSD of 1.399%, indicating good stability of the sample.

[0287] Repeatability investigation

[0288] Prepare 6 parallel samples of the test solution, inject and measure. The injection volume is 2 μL each time. Record the peak area value and calculate the RSD. After investigation, the RSD of the repeatability test result of GLA is 1.094%, indicating good repeatability of this method.

[0289] Spiked recovery investigation

[0290] Take 9 samples of the test solution with known concentration, add GLA reference according to 80%, 100%, and 120% of the GLA content in the sample within the linear range, vortex and mix well. Take the supernatant, filter through a 0.22 μm microporous membrane, inject and measure. Record the peak area value and calculate the spiked recovery rate. After investigation, the average spiked recovery rate of the 9 samples is 99.44%, with an RSD of 1.94%, indicating good accuracy of this method.

[0291] (6) In vivo percutaneous penetration test

[0292] Experimental grouping and treatment

[0293] Grouping: 144 mice weighing 18 - 22 g were taken and divided into 2 major groups (liquid crystal cream group, ordinary cream group). Each major group was further divided into 6 groups (n = 6) according to time points (0.5, 1, 2, 4, 6, 8 h), with 6 mice in each group, totaling 24 groups.

[0294] Pretreatment: The mice were fixed with their backs facing up, and the hair on their backs was shaved off with an electric shaver and then washed with warm water. After 24 h, the drug was applied, with the application area being 2 cm × 2 cm and the application amount being 0.1 g. The four limbs of the mice were fixed with 3M tape to prevent the mice from licking.

[0295] The unabsorbed drug on the backs of the mice was wiped off with a cotton swab moistened with physiological saline, and the mice were sacrificed by dislocation. The skin at the drug administration site was cut off with surgical scissors, and the subcutaneous fat was scraped off with a razor and placed in a 2 mL centrifuge tube for weighing and standby.

[0296] Skin sample treatment method: Weigh the skin, add a fixed amount of methanol, cut it into pieces, homogenize at -50 °C, vortex for 2 min, seal and ultrasonically treat for 30 min, then centrifuge at 4 °C and 12000 r / min for 5 min. Quantitatively take the upper layer liquid and dry it with nitrogen at 37 °C. The residue was redissolved with 0.4 mL of ethyl acetate, centrifuged at 8000 r / min for 5 min, quantitatively take the supernatant, and dry it with nitrogen at 37 °C. The residue was redissolved with a fixed amount of methanol, vortexed for 2 min, and filtered through a 0.22 μm filter membrane for standby.

[0297] Fat sample treatment method: Weigh the fat, add a fixed amount of methanol, cut it into pieces, homogenize at -50 °C, vortex for 2 min, seal and ultrasonically treat for 30 min, centrifuge at 4 °C and 12000 r / min for 5 min, then quantitatively take the upper layer liquid and dry it with nitrogen at 37 °C. The residue was redissolved with 0.4 mL of ethyl acetate, centrifuged at 8000 r / min for 5 min, quantitatively take the supernatant, and dry it with nitrogen at 37 °C. The residue was redissolved with a fixed amount of methanol, vortexed for 2 min, and the supernatant was taken and filtered through a 0.22 μm filter membrane for standby.

[0298] The skin / fat retention rate W of the drug s / % was calculated according to the following formula:

[0299]

[0300] C / (μg·mL -1 ) The concentration of the drug in the skin sample; V 1 / mL: The volume of methanol added to treat the skin / fat sample; V 2 / μL: The volume of the supernatant taken after centrifugation after homogenization; V 3 / μL: The volume of ethyl acetate added after the first nitrogen blowing, i.e., 400 μL; V 4 / μL: The volume of the supernatant taken after centrifugation after redissolving with ethyl acetate, i.e., 300 μL; V 5 / μL: The volume of methanol added during reconstitution after the second nitrogen blow, which is 500 μL; Q a / μg: The mass of GLA contained in the Tween 20 - stevioside - glabridin liquid crystal cream, stevioside - glabridin liquid crystal cream, stevioside - glabridin ordinary cream, and Tween 20 - stevioside - glabridin ordinary cream applied to each mouse.

[0301] From Figure 13 It can be seen that under the same dosage, the skin retention rates of GLA at 0.5 h, 1 h, 2 h, 4 h, 6 h, and 8 h (P < 0.001) in the stevioside - glabridin liquid crystal cream group are 1.67, 0.79, 1.73, 1.47, 1.17, and 1.15 times those in the stevioside - glabridin ordinary cream group (P < 0.001), respectively. In summary, the liquid crystal cream can significantly increase the retention amount of GLA in the skin of mice.

[0302] At the same time, from Figure 13 It can be seen that the maximum skin retention rate of the stevioside - glabridin liquid crystal cream group appears at 2 h, which is 10.918 ± 1.465%; the maximum skin retention rate of the stevioside - glabridin ordinary cream group appears at 1 h, which is 8.737 ± 1.605%. The time to reach the peak and the retention time of GLA in the skin of the liquid crystal cream group are longer than those in the ordinary cream group, indicating that the liquid crystal cream has a certain sustained - release and controlled - release effect.

[0303] Moreover, from Figure 13 It can be seen that the skin retention rates of GLA at 0.5 h, 1 h, 2 h, 4 h, and 6 h (P < 0.001) in the Tween 20 - stevioside - glabridin liquid crystal cream group are higher than those in the stevioside - glabridin liquid crystal cream group, and the skin retention rates of GLA at 1 h, 2 h, 4 h, 6 h, and 8 h (P < 0.001) in the Tween 20 - stevioside - glabridin ordinary cream group are higher than those in the stevioside - glabridin liquid crystal cream group. It can be seen that Tween 20 has a promoting effect on the skin retention of GLA.

[0304] From Figure 14It can be seen that under the same dosage, the fat retention rate of stevioside - glabridin ordinary cream group at 1 h for GLA was 4.717 ± 1.350%, which was 1.87 times that of stevioside - glabridin liquid crystal cream group at 1 h for GLA (2.525 ± 0.850%). The fat retention rate of stevioside - glabridin ordinary cream group at 2 h for GLA was 3.423 ± 1.053%, which was 1.37 times that of stevioside - glabridin liquid crystal cream group at 2 h for GLA (2.504 ± 1.542%). Similarly, it can be obtained in turn that the fat retention rate of GLA in stevioside - glabridin ordinary cream group (P < 0.001) at 4 h, 6 h and 8 h was 1.20, 1.35 and 1.19 times that of stevioside - glabridin liquid crystal cream group (P < 0.05). In summary, the liquid crystal cream can make GLA stay more in the skin rather than in the fat, showing certain skin targeting property.

[0305] And it can be seen from Figure 14 that the skin retention rates of GLA at 0.5 h, 1 h, 2 h, 4 h, 6 h and 8 h in the Tween 20 - stevioside - glabridin liquid crystal cream group (P < 0.001) were basically the same as those in the stevioside - glabridin liquid crystal cream group, and the skin retention rates of GLA at 0.5 h, 1 h, 2 h, 4 h, 6 h and 8 h in the Tween 20 - stevioside - glabridin ordinary cream group (P < 0.001) were basically the same as those in the stevioside - glabridin liquid crystal cream group. Thus, it can be known that Tween 20 can effectively promote the retention of GLA in the skin without affecting the penetration of GLA through the skin into the blood.

[0306] Example 16

[0307] The difference between this example and Example 15 is: different types of traditional Chinese medicine monomers. The specific preparation method of this example is as follows:

[0308] A preparation method of a stevioside - quercetin self - assembly system, comprising the following steps:

[0309] Take 5 mg of quercetin powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix evenly, place it in a magnetic stirrer and stir until dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, place it in a 10.0 mL centrifuge tube for mixing, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, then the stevioside - quercetin self - assembly system is obtained.

[0310] A preparation method of a stevioside - glaucinone self - assembly system, comprising the following steps:

[0311] Take 5 mg of glauvin powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-glauvin self-assembly system.

[0312] A preparation method of a stevioside-oxymatrine self-assembly system, comprising the following steps:

[0313] Take 40 mg of oxymatrine powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-oxymatrine self-assembly system.

[0314] A preparation method of a stevioside-resveratrol self-assembly system, comprising the following steps:

[0315] Take 30.0 mg of resveratrol powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / ml, stir and mix well, place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-resveratrol self-assembly system.

[0316] A preparation method of a stevioside-α-mangostin self-assembly system, comprising the following steps:

[0317] Take 4 mg of α-mangostin powder, accurately weigh it, add it to a beaker, add 10.0 mI of stevioside with a concentration of 100.0 mg / ml, stir and mix well, place it in a magnetic stirrer and stir until the dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 ml of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-α-mangostin self-assembly system.

[0318] A preparation method of a stevioside-ferulic acid self-assembly system, comprising the following steps:

[0319] Take 90.0 mg of ferulic acid powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / mL, stir and mix evenly, place it in a magnetic stirrer and stir until dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 mL of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-ferulic acid self-assembly system.

[0320] A preparation method of a stevioside-gallic acid self-assembly system, comprising the following steps:

[0321] Take 170.0 mg of gallic acid powder, accurately weigh it, add it to a beaker, add 10.0 mL of stevioside with a concentration of 100.0 mg / mL, stir and mix evenly, place it in a magnetic stirrer and stir until dissolution equilibrium is reached to obtain a mixed solution. Weigh 20% Tween 80, place it in a plastic centrifuge tube, add 6.0 mL of the mixed solution, mix it in a 10.0 mL centrifuge tube, vortex, and ultrasonicate at 40 °C for 30 min until the solution is clear and transparent, thus obtaining the stevioside-gallic acid self-assembly system.

[0322] Respectively prepare different types of medicated liquid crystal creams from the above seven self-assembly systems according to the preparation method of the liquid crystal cream in Example 15.

[0323] Test Example 5

[0324] Test the transdermal absorption effect of the medicated liquid crystal cream in Example 16 according to the process of the in vivo percutaneous penetration test in Test Example 4. Specific detection details not mentioned in this test example are carried out according to the methods disclosed in the prior art. The skin retention results and fat retention results of the medicated liquid crystal cream are shown in Figures 30 - 43 . It can be seen from the figure that the liquid crystal cream can make more traditional Chinese medicine monomers stay in the skin rather than in the fat, having a certain skin targeting property.

[0325] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that they can still modify or equivalently replace the technical solutions of the present invention, and these modifications or equivalent replacements cannot make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A stevioside-traditional Chinese medicine monomer self-assembly system, characterized in that: The self-assembly system includes stevioside, traditional Chinese medicine monomers and water.

2. A stevioside-traditional Chinese medicine monomer self-assembly system according to claim 1, characterized in that: The monomers of traditional Chinese medicine are one or more of flavonoids, alkaloids, anthraquinones and organic acids.

3. A stevioside-traditional Chinese medicine monomer self-assembly system according to claim 2, characterized in that: The flavonoids include one or more of quercetin, baicalein, and glabridin; the alkaloids include one or more of papaverine, matrine, and oxymatrine; the anthraquinones include one or more of resveratrol, oxyresveratrol, and α-mangostin; and the organic acids include one or more of ferulic acid, chlorogenic acid, and gallic acid.

4. The stevioside-traditional Chinese medicine monomer self-assembly system according to claim 1, characterized in that: The concentration of stevioside in water is ≥100 mg / mL, and the concentration of Chinese medicine monomer in water is ≥0.3 mg / mL.

5. The stevioside-traditional Chinese medicine monomer self-assembly system according to claim 1, characterized in that: Surfactants are also included in the self-assembly system.

6. A stevioside-traditional Chinese medicine monomer self-assembly system according to claim 5, characterized in that: The surfactant is one or more of a liquid surfactant and a solid surfactant.

7. A stevioside-traditional Chinese medicine monomer self-assembly system according to claim 6, characterized in that: The liquid surfactant is one or more of Tween 80, Tween 60 and Tween 20, and the solid surfactant is cetearyl alcohol.

8. The stevioside-traditional Chinese medicine monomer self-assembly system according to claim 5, characterized in that: The added amount of surfactant is 0.1-20% of the total mass of stevioside, Chinese medicine monomer and water.

9. The method for preparing a stevioside-traditional Chinese medicine monomer self-assembly system according to claims 5 to 8, characterized in that: The following steps are involved: The Chinese medicine monomer and stevioside are added to water, stirred evenly until the solution is in equilibrium, and a mixed solution is obtained. A surfactant is added to the mixed solution for mixing. The mixed solution is vortexed and ultrasonicated for 10 to 60 minutes at 40° C. until the solution is clear and transparent, thereby obtaining a self-assembly system.

10. An application of a stevioside-traditional Chinese medicine monomer self-assembly system, characterized in that: Use of the stevioside-traditional Chinese medicine monomer self-assembly system according to any one of claims 1 to 8 in the preparation of cosmetics.

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