CN113413404B - Traditional Chinese medicine preparation with anti-inflammation and acne-removing effects and preparation method and application thereof - Google Patents

Traditional Chinese medicine preparation with anti-inflammation and acne-removing effects and preparation method and application thereof Download PDF

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CN113413404B
CN113413404B CN202110805109.2A CN202110805109A CN113413404B CN 113413404 B CN113413404 B CN 113413404B CN 202110805109 A CN202110805109 A CN 202110805109A CN 113413404 B CN113413404 B CN 113413404B
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essential oil
chrysanthemum
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宿树兰
段金廒
刘夏进
严辉
魏丹丹
钱大玮
郭盛
孙晓东
丁凤清
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Nanjing University of Chinese Medicine
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Abstract

The invention discloses a traditional Chinese medicine preparation with anti-inflammation and acne-removing effects and a preparation method thereof, wherein the traditional Chinese medicine preparation comprises the following components: 0.05-5 parts of chrysanthemum stem leaf essential oil, 0.05-5 parts of mint essential oil, 0.1-5 parts of oil phase auxiliary material, 0.01-5 parts of baicalin extract, 1-5 parts of surfactant, 0.1-5 parts of cosurfactant and the balance of chrysanthemum stem leaf water extract. The preparation method comprises the following steps: preparing chrysanthemum stem leaf essential oil by adopting a steam distillation method, and adding an oil phase auxiliary material to prepare an oil phase; adding cosurfactant, surfactant and baicalin extracted from Chinese medicinal materials, stirring, and slowly adding water phase into oil phase. According to the invention, the anti-inflammation and acne-removing microemulsion and gel are prepared by matching the essential oil of the chrysanthemum morifolium and the non-medicinal parts thereof with the traditional Chinese medicine extract, so that the stability of active ingredients is effectively protected, the absorption of the skin to the medicine is promoted, and the anti-inflammation and acne-removing activity of the microemulsion and gel prepared by matching the essential oil with the baicalin is enhanced.

Description

Traditional Chinese medicine preparation with anti-inflammation and acne-removing effects and preparation method and application thereof
Technical Field
The invention belongs to the field of medicinal preparations, and particularly relates to a Chinese medicinal preparation with anti-inflammatory and acne-removing effects and a preparation method thereof.
Background
Acne is a chronic skin disease with a corresponding inflammatory response; in recent years, acne removing consumers in China are growing at 400-600 ten thousand people per year, and the common inflammatory disease troubles 80% of teenagers and adults in China.
Although acne is not an infectious disease, its causative factors are complex and diverse, with bacterial infection being one of the important causative factors, with propionibacterium acnes (p.acnes) and staphylococcus aureus (s.aureus) present at various stages of their development and progression.
Acne seriously affects the appearance and mind of people and has certain negative influence on the work and life of people, so that the method and the product for diminishing inflammation and removing acnes are one of the hot spots of research. According to related researches, hormones or antibiotics are often used for treating acne, but side effects, drug resistance and the like are caused by the hormones or the antibiotics; therefore, the development of natural green traditional Chinese medicine external preparations is very necessary.
The microemulsion is a good entrapment technology, can solve the problems of volatile oil instability and the like, and can increase the solubility of insoluble drugs, so that the drugs can achieve the permeation promoting effect through skin appendages. The water gel has no greasy feeling, is easy to spread, and does not interfere with normal functions of skin. The microemulsion and the gel are combined, so that the bioavailability of volatile components and indissolvable punishment components is greatly increased.
The Chrysanthemum is a dry head-shaped inflorescence of Chrysanthemum (Chrysanthemum morifolium Ramat.) of the Compositae, is a traditional Chinese medicinal material in China, has various varieties, and is mainly used for treating wind-heat type common cold, light headedness, red eyes, swelling and pain, sores and carbuncle pyogenic infections and other symptoms clinically; according to survey, high-quality Hangzhou white chrysanthemum dry head-shaped inflorescences produced in Zhejiang Tongxiang city every year are about 7000t, while the non-medicinal parts such as Hangzhou white chrysanthemum stems and the like produced along with the high-quality Hangzhou white chrysanthemum dry head-shaped inflorescences reach 2.5 ten thousand tons, and the yield of the non-medicinal parts is 3.5 times of that of the medicinal parts; in the process of culturing and harvesting the inflorescence of the chrysanthemum, the non-medicinal parts of the chrysanthemum are not reasonably applied and are treated as production waste and are burnt or discarded; only a few chrysanthemum stems and leaves are used as the feed for livestock, which causes great waste in resource utilization.
Research shows that the active components contained in the non-medicinal parts of the chrysanthemum and the pharmacological action thereof are very similar to the inflorescence, and have a plurality of biological activities of remarkable bacteriostasis, anti-inflammation, free radical removal and the like. The volatile oil components in the stems and leaves of Chrysanthemum mainly include monoterpene, sesquiterpene and its oxygen-containing derivatives. Has antibacterial, anti-infectious, antiinflammatory, analgesic, antiaging, skin and mucosa healing promoting, and immunity enhancing effects.
Clinically, scutellaria baicalensis is a common traditional Chinese medicine for treating skin inflammatory diseases such as acne. Baicalin (C) 21 H 18 O 11 ) Is a flavonoid compound extracted from scutellaria root, is the main active component of scutellaria and has obvious anti-inflammatory activity. Baicalin has a wide antibacterial range, has a strong effect on staphylococcus aureus and pseudomonas aeruginosa, can absorb ultraviolet rays, remove oxygen radicals and inhibit the generation of melanin, and therefore can be used for medicines and cosmetics, and is a good functional cosmetic raw material.
The project adopts chrysanthemum stem leaf essential oil and traditional Chinese medicine baicalin for compatibility, the anti-inflammatory activity of the chrysanthemum stem leaf essential oil and the traditional Chinese medicine baicalin are evaluated from an in vitro cell experiment, the bacteriostatic action of the chrysanthemum stem leaf essential oil and the baicalin is evaluated from a bacterial level, and the microemulsion gel for diminishing inflammation and removing acnes is prepared.
Disclosure of Invention
The invention aims to provide an essential oil and Chinese patent medicine compound traditional Chinese medicine preparation with anti-inflammatory and acne-removing effects and a preparation method thereof, the traditional Chinese medicine preparation provided by the invention has good solubilizing capability on fat-soluble components and water-soluble components, so that effective components such as chrysanthemum stem and leaf essential oil, water extract, mint essential oil baicalin and the like are utilized to the maximum extent, and meanwhile, the traditional Chinese medicine preparation has the advantages of good stability, high moisture retention, high permeability and the like; and improves the cell inflammation on the basis of no stimulation to the skin, and also has the effect of inhibiting the growth of the pox-causing bacteria.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides an anti-inflammation acne-removing traditional Chinese medicine preparation which comprises the following components: 0.05-5 parts of chrysanthemum stem leaf essential oil, 0.05-5 parts of mint essential oil, 0.1-5 parts of oil phase auxiliary material, 0.01-5 parts of baicalin extract, 1-5 parts of surfactant, 0.1-5 parts of cosurfactant, 0.1-5 parts of antioxidant and the balance of chrysanthemum stem leaf water extract.
A preparation method of a traditional Chinese medicine preparation with anti-inflammation and acne-removing effects comprises the following steps:
(1) Extracting chrysanthemum stem and leaf essential oil by adopting a steam distillation method to obtain chrysanthemum stem and leaf essential oil and chrysanthemum stem and leaf water extract, dehydrating the essential oil by using anhydrous sodium sulfate, and storing in a dark place;
(2) Mixing the chrysanthemum stem and leaf essential oil extracted in the step (1) with the mint essential oil, and adding an oil-phase auxiliary material to prepare an oil phase; taking the water extract of the chrysanthemum stems and leaves prepared in the step (2), and adding water phase auxiliary materials to prepare a water phase; adding mixed surfactant and baicalin, stirring under magnetic stirring at normal temperature, and dropwise adding water phase into oil phase while stirring to obtain clear and transparent microemulsion;
(3) And (3) preparing the microemulsion prepared in the step (2) into gel.
As a preferred scheme, the preparation method of the microemulsion gel for diminishing inflammation and removing acne is required, and the optimal process for extracting the chrysanthemum stem leaf essential oil by the water vapor in the step (1) is that the grinding degree is 100-120 meshes, the soaking time is 6h, and the material-liquid ratio is 1:8, distilling for 2 times, 6h each time, and 12h.
As a preferred scheme, the preparation method of the traditional Chinese medicine preparation with anti-inflammatory and acne-removing effects comprises the steps of diluting the microemulsion by a proper multiple by using the water phase prepared in the step (2), and adding 1-5% of gel matrix, 5-20% of mixed humectant, 1-5% of antioxidant, 5-10% of solubilizer and 0.05-0.1% of preservative by weight of the total gel; the total weight of the gel is 100g, the rest is water phase, and the clear transparent gel is obtained after even stirring at 37 ℃.
As a preferred scheme, in the preparation method of the traditional Chinese medicine preparation with anti-inflammatory and anti-acne effects, the oil phase auxiliary materials in the microemulsion comprise one or more of peppermint oil, oleic acid, linoleic acid, ethyl oleate, wheat germ oil, castor oil, peanut oil, ethyl linoleate and triglycerides with medium fatty chain length, and preferably ethyl oleate; the water phase auxiliary materials in the microemulsion comprise one or more of aloe extract, sorbitol, propylene glycol, parabens and vitamin C.
As a preferable scheme, the preparation method of the traditional Chinese medicine preparation with the anti-inflammatory and anti-acne effects is required, wherein the mixed surfactant in the microemulsion comprises a surfactant and a cosurfactant in a mass ratio of 3; wherein the surfactant comprises one or more of poloxamer, soybean phospholipid, tween 80, tween 60, span 80 and peregal; preferably tween 80. Wherein the cosurfactant comprises one or more of anhydrous alcohol, polyethylene glycol 400, propylene glycol, isopropanol, n-butanol and ethylene glycol, preferably glycerol.
According to the preferable scheme, in the preparation method of the traditional Chinese medicine preparation with the effects of diminishing inflammation and removing acnes, the weight of the oil phase accounts for 2-12% of the total weight of the microemulsion carrier, the weight of the surfactant accounts for 15-25% of the total weight of the microemulsion carrier, the weight of the cosurfactant accounts for 2-12% of the total weight of the microemulsion carrier, the weight of the antioxidant accounts for 1-10% (w/w) of the total weight of the microemulsion carrier, and the weight of the water phase accounts for 50-80% of the total weight of the microemulsion carrier.
Preferably, the weight of the oil phase accounts for 4.87% (w/w) of the total weight of the microemulsion carrier, the weight of the surfactant accounts for 21.82% (w/w) of the total weight of the microemulsion carrier, the weight of the cosurfactant accounts for 7.26% (w/w) of the total weight of the microemulsion carrier, the weight of the antioxidant accounts for 3.1% (w/w) of the total weight of the microemulsion carrier, and the weight of the water phase accounts for 58.00% (w/w) of the total weight of the microemulsion carrier.
The optimal technical scheme is that 100g of microemulsion contains 0.25g of chrysanthemum stem and leaf essential oil, 0.25g of mint essential oil, 4.37g of ethyl oleate, 8021.82g of tween, 7.26g of glycerol, 0.10g of baicalin, 3.00g of antioxidant vitamin C, 0.10g of antioxidant sodium metabisulfite and 58.00g of water phase (crude drug amount is 20.45 mg/mL).
As a preferred scheme, the preparation method of the traditional Chinese medicine preparation with the effects of diminishing inflammation and removing acne is characterized in that the aqueous gel matrix in the gel comprises one or more of carboxymethyl cellulose derivatives, alginate, gelatin, pectin, starch and derivatives thereof, and polyacrylic acids (such as carbomer, polyacrylic acid and the like);
the water phase adjuvants comprise one or more of ethanol, glycerol, aloe extract, sorbitol, propylene glycol, nipagin, microemulsion with proper dilution, and vitamin C.
Preferably, the weight of the gel matrix accounts for 1-10% of the total weight of the gel carrier, the weight of the microemulsion in the gel accounts for 15-30% of the total weight of the gel carrier, the weight of the moisture retention agent in the gel accounts for 15-30% of the total weight of the gel carrier, the weight of the antioxidant agent in the gel accounts for 1-10% of the total weight of the gel carrier, the weight of the cosolvent in the gel accounts for 5-10% of the total weight of the gel carrier, the weight of the preservative in the gel accounts for 0.05-0.1% of the total weight of the gel carrier, and the balance is the chrysanthemum stem leaf water extract (30-60%).
Preferably, the weight of the gel matrix accounts for 3% (w/w) of the total weight of the gel carrier, the weight of the microemulsion in the gel accounts for 20% (w/w) of the total weight of the gel carrier, the weight of the moisture preserving agent in the gel accounts for 10% (w/w) of the total weight of the gel carrier, the weight of the antioxidant in the gel accounts for 3.1% (w/w) of the total weight of the gel carrier, the weight of the cosolvent in the gel accounts for 10% (w/w) of the total weight of the gel carrier, the weight of the preservative in the gel accounts for 0.1% (w/w) of the total weight of the gel carrier, and the weight of the chrysanthemum stem and leaf water extract accounts for 53.8% (w/w).
As the most preferable scheme, in the preparation method of the traditional Chinese medicine preparation with the effects of diminishing inflammation and removing acnes, 20g of microemulsion, 9403g of gel matrix carbomer, 3g of humectant glycerol, 7g of humectant aloe barbadensis, 3g of antioxidant vitamin C, 0.1g of antioxidant sodium metabisulfite, 10g of cosolvent propylene glycol, 0.1g of preservative ethylparaben and 53.8g of aqueous phase with the crude drug amount of 20.45mg/ml are contained in 100g of gel.
The humectant accounts for 5-10% of the total amount of the carrier, and comprises one or more of hyaluronic acids, allantoin, aloe extract, glycerol, sorbitol and propylene glycol.
The antioxidant added in the invention accounts for 3.1-3.2% of the total amount of the carrier, and comprises one or more of vitamin C, sulfite, pyrosulfite and bisulfite.
The method utilizes resources of the abandoned or burnt chrysanthemum stem leaves originally, adopts an optimal extraction process to extract the chrysanthemum stem leaf essential oil, and then carries out a large number of in vitro cell experiments and bacterial experiments to determine the anti-inflammation and acne-removing activity of the chrysanthemum stem leaf essential oil; and then the composition is compatible with the baicalin extracted from the traditional Chinese medicine, so that the irritation of the baicalin and essential oil to the skin is greatly reduced, and the natural green anti-inflammation acne-removing micro-emulsion and gel are prepared.
Drawings
FIG. 1 is a graph of the effect of chrysanthemum essential oil on the survival rate of THP-1
FIG. 2 shows the effect of baicalin on the survival rate of THP-1
FIG. 3 shows the effect of the combination of chrysanthemum essential oil and baicalin on the induction of THP-1 inflammation by P.acnes
FIG. 4 is a plot of the area of the microemulsion regions for different co-surfactants
FIG. 5 is a plot of the area of the microemulsion region for different surfactant screens
FIG. 6 is a micro-emulsion area diagram of surfactants and co-surfactants with different mass ratios
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1 investigation of extraction Process of Chrysanthemum stem and leaf essential oil
1. Optimization of chrysanthemum stem leaf essential oil extraction process
The invention selects main factors of pulverization degree (A), water adding amount (B), distillation time (C) and soaking time (D) which influence a steam distillation method, firstly screens out three factors through a single-factor experiment, selects three superior factors according to a single-factor experiment result (the result is shown in a table 1), designs an L9 (34) orthogonal table, performs experiments according to the table, and inspects the optimal extraction process of the chrysanthemum stem leaf volatile oil.
TABLE 1 Quadrature experiments three-factor four-level
Figure BDA0003166207040000041
2. The optimal extraction rate of the chrysanthemum stem and leaf essential oil is determined by adopting an orthogonal experiment (table 2), the change of the chrysanthemum stem and leaf essential oil components under different conditions is observed by a gas chromatograph-mass spectrometer, and the finally determined optimal process is a process 7, wherein the yield of the chrysanthemum stem and leaf essential oil extracted by selecting 80-120 meshes of chrysanthemum stem and leaf, adding 8 times of water addition amount, distilling for 12h and soaking for 6h is 0.50%.
TABLE 2 results of orthogonal experiments
Figure BDA0003166207040000051
3. Result verification
To further examine the stability of the above extraction process, the extraction was repeated 3 times according to the selected optimal process conditions, and the results are shown in table 3.
TABLE 3 verification of the results of the volatile oil extraction Process
Experiment number Volatile oil mass (mg) Yield of volatile oil (%)
1 490 0.49
2 497 0.50
3 518 0.52
The verification result is basically consistent with the orthogonal experiment result, so that the extraction process has good stability and can be used for extracting volatile components.
Example 2 anti-inflammatory action
1. Survival rate research of chrysanthemum essential oil (chrysanthemum essential oil and chrysanthemum stem leaf essential oil) and baicalin on human mononuclear acute cell (THP-1)
The influence of chrysanthemum essential oil and baicalin on THP-1 proliferation is detected by adopting a cck8 method. Taking THP-1 in exponential growth phase at 9X 10 4 Inoculating at density of one/mL into 96-well plate, adding different mass concentrations (6.25, 12.5,25, 50, 100mg/L) of flos Chrysanthemi essential oil and baicalin, and culturing for 36h. Then, the drug and the supernatant were centrifuged, 10% cck8 was added to each well, and the mixture was left for 3 hours to detect absorbance at 450 nm.
The result shows that the chrysanthemum essential oil has no inhibition effect on the growth of cells when the concentration of the chrysanthemum essential oil is within 100mg/mL, and also has the effect of promoting the growth of the cells when the concentration of the chrysanthemum essential oil is 50mg/L, so the administration concentration of the chrysanthemum essential oil is selected to be 50mg/L; baicalin has obvious inhibition effect on cells at 25mg/mL, and the administration concentration of the baicalin is selected to be 12.5mg/L. The maximum safe use concentrations of the chrysanthemum essential oil and the baicalin on the cells are respectively 100mg/mL and 12.5mg/mL preliminarily obtained from the experimental results. Then, the concentration of the chrysanthemum stem and leaf essential oil is expanded to carry out an experiment, and the maximum safe use concentration of the chrysanthemum stem and leaf essential oil on cells is determined to be 500mg/mL (shown in the attached figure 1 and the attached figure 2).
2. Effect of chrysanthemum essential oil and baicalin alone or in combination on Propionibacterium acnes (P.acnes) induced THP-1 inflammation
Propionibacterium acnes (p.acnes) was inoculated into liquid medium and cultured anaerobically at 37 ℃ to OD600=1.0 (logarithmic growth phase) to obtain a bacterial suspension. Centrifuging the bacterial suspension at 4 deg.C for 15min at 5000g to obtain culture supernatant and bacterial precipitate. Washing the precipitate with PBS for 3 times, suspending in PBS, placing in 80 deg.C water bath for 30min to achieve heat inactivation, i.e. Propionibacterium acnes inactivated matter, and storing at 4 deg.C for use.
THP-1 was inoculated into 96-well plates and the cell concentration was adjusted to 9X 10 4 And culturing for 24h, adding fresh culture medium containing the medicine again, culturing for 4h, and then adding the propionibacterium acnes inactivated solution, and culturing for 8h. Centrifuge at 3000 rpm for 20min and carefully collect the supernatant in EP without ribozyme. The level of IL-1. Beta. In the cell supernatants was detected using an ELISA kit.
The results show that both 50mg/L of chrysanthemum essential oil and 12.5mg/L of baicalin can reduce the concentration of an intracellular inflammatory factor IL-1 beta caused by P.acnes heat inactivation liquid, and the result of relieving the cell inflammation by combining (25 mg/L) chrysanthemum stem and leaf essential oil and (6.25 mg/L) baicalin is superior to the effect of relieving the cell inflammation by combining the chrysanthemum stem and leaf essential oil and the baicalin (shown in figure 3).
According to the results of the anti-inflammation experiments, the chrysanthemum essential oil, the chrysanthemum stem leaf essential oil and the baicalin have the effect of relieving the THP-1 inflammation induced by P.acnes under the concentration of safety and no stimulation to cells;
example 3 bacteriostatic action
1. Preparation of test article
Preparing chrysanthemum essential oil into 1000mg/mL essential oil mother liquor by using absolute ethyl alcohol as a solvent; DMSO is adopted as a cosolvent to dissolve baicalin, the concentration of a mother solution is 100mg/mL, and the concentration of the DMSO is ensured to be lower than 1%.
Extracting volatile oil (80-120 mesh flos Chrysanthemi or flos Chrysanthemi stem and leaf essential oil obtained by distilling with 8 times of water for 12 hr and soaking for 6 hr) with water phase, wherein the crude drug concentration is 125 mg/mL;
preparation of positive drug penicillin G sodium and erythromycin lactobionate: weighing a certain amount of positive drug, adding a certain volume of pure water, and mixing uniformly to prepare a penicillin G sodium solution with the concentration of 2.5 mu G/mL and an erythromycin lactobionate solution with the concentration of 1.22 mg/mL.
2. Studies on inhibitory Activity against Staphylococcus aureus and Propionibacterium acnes
S.aureus was cultured overnight in an incubator (12h, 37 ℃,5% CO2), P.acnes was cultured in an anaerobic oven (48h, 37 ℃,10% CO2,10% H2,80% N2), and the final concentration of each strain was adjusted to 3X 104CFU/mL. Preparing serial doubled dilutions of the test sample in a 96-well plate, sequentially adding 100 μ L of bacterial liquid, and culturing in an incubator; adding 30 mu L (0.01%) of 2,3, 5-triphenyltetrazolium chloride (TTC) solution into the bacteria solution cultured for 12h and 24h respectively, and observing whether red precipitate is generated after 2 h; if no red precipitate is generated, the growth activity of the bacteria is inhibited; the concentration of the essential oil at this time is MIC; MIC is defined as the lowest concentration of essential oil that is bacteriostatic, at which the microorganism does not show visible growth. The experiment was set with a blank control (only solvent was added, no other species was identified), a negative control (only solvent and bacterial solution were added, no inhibition of the solvent on the bacteria was identified) and a positive control (positive drug was added, the drug that had an inhibition on the species tested was identified). Penicillin and erythromycin lactobionate were used as positive antibiotics for s. All inhibition assays were performed in triplicate.
The minimum inhibitory concentration of each sample to S.aureus is shown in Table 4, and the chrysanthemum stem leaf essential oil and the water extract have an inhibitory effect on S.aureus; but the inhibiting effect of the chrysanthemum stem and leaf essential oil and the chrysanthemum stem and leaf water extract on S.aureus is slightly lower than that of the chrysanthemum essential oil and the chrysanthemum stem and leaf water extract. In addition, baicalin has strong inhibiting effect on s.
Table 4 inhibition results on staphylococcus aureus (s. Aureus)
Figure BDA0003166207040000071
Remarking: "-" indicates no bacterial growth and "+" indicates bacterial growth
The minimum inhibitory concentrations of the various samples on p.acnes are shown in table 5, the minimum inhibitory concentrations of the chrysanthemum stem and leaf essential oil and the chrysanthemum essential oil on p.acnes are both 25mg/ml, and the minimum inhibitory concentration of the chrysanthemum and chrysanthemum stem and leaf aqueous extract on p.acnes is 12.5mg/ml.
Table 5 inhibition results of propionibacterium acnes (p.acnes)
Figure BDA0003166207040000072
Remarking: "-" indicates no bacterial growth, "+" indicates bacterial growth
EXAMPLE 4 preparation of microemulsion gel formulations
1. Preparation of microemulsion
1.1 drawing of pseudo-ternary phase diagram
Precisely weighing a surfactant and a cosurfactant which are in a fixed mass ratio of 1. A mixed surfactant with Km =1 and an oil phase auxiliary material according to the mass ratio of 1. And recording the quality of the added water phase when the microemulsion turns from turbid to clear for the second time, and determining the area for forming the O/W microemulsion.
1.2 oil phase determination
The chrysanthemum essential oil anti-inflammation experiment can obtain that the chrysanthemum stem leaf essential oil contained in the microemulsion or gel preparation has anti-inflammation effect as long as the content of the chrysanthemum stem leaf essential oil is more than 50mg/L; and then, the chrysanthemum stem leaf essential oil, the coarse penetration-promoting mint essential oil and the oil-phase auxiliary material are mixed according to four proportions, and finally the chrysanthemum stem leaf essential oil is determined: mint essential oil: ethyl oleate =1.
1.3 determination of the aqueous phase
As can be seen from example 3, the minimum inhibitory concentration of the aqueous extract of the chrysanthemum stem and leaf on P.acnes is 12.5mg/mL, i.e., the crude drug amount of the final aqueous phase should be 20.45mg/mL.
1.4 screening of cosurfactants
The invention fixes Km =1, and the oil phase is chrysanthemum stem leaf essential oil: mint essential oil: ethyl oleate =1, and the emulsifier is tween 80, and the size of the area forming a microemulsion when absolute ethanol, glycerol, and polyethylene glycol-400 are used as cosurfactants, respectively, is examined, wherein the absolute ethanol and tween 80 do not form a microemulsion in combination, and the three-phase diagrams of the other two groups are shown in fig. 4 below.
As is apparent from fig. 4, when other conditions were maintained, the area of the microemulsion region formed by using peg-400 as a co-surfactant was larger, and therefore, peg-400 was selected as a co-surfactant.
1.5 screening of surfactants
The invention fixes Km =1, and the oil phase is chrysanthemum stem leaf essential oil: mint essential oil: ethyl oleate =1, the co-emulsifier is polyethylene glycol-400, and the size of the area of the microemulsion formed when poloxamer 188 and tween 80 are respectively used as the co-surfactants is examined, and the result is shown in fig. 5 below.
Because poloxamer 188 and auxiliary emulsifier, oil phase and oil phase auxiliary material can not form microemulsion in the experimental range, tween 80 is selected as the surfactant.
1.6 screening of optimal Km values (surfactant/co-surfactant)
Selecting a Km value of 1,2;1, 3.
As can be seen from fig. 6, km =3 is selected because the area of the microemulsion region is largest when Km = 3.
1.7 determination of baicalin drug-loading
As determined by an anti-inflammation example and a bacteriostatic experiment, the microemulsion or gel preparation has the anti-inflammation effect as long as the concentration of baicalin in the microemulsion or gel preparation is more than 12.5 mg/L; when the baicalin content is 7.5mg/mL, the composition has antibacterial effect when being combined with 25mg/mL chrysanthemum stem and leaf essential oil.
According to the content result and double investigation of baicalin solubility (baicalin content is 0.1%,0.15%,0.2% and 0.25% of the total microemulsion), the clarification of the microemulsion is optimal when the content of the screened baicalin is greater than the content of an anti-inflammation experiment and the content of the baicalin is 0.1% of the total microemulsion, so that the content of the baicalin is determined to be 0.10g.
1.7 determination of optimal prescription
The formulation was determined by selecting points in the microemulsion region at and near the center of gravity (oil: surfactant 1,4, 1.
Table 6 investigation results of prescription particle size
Oil-mixed surfactant ratio Particle size (nm) Pdl
1:3 59.37±1.18 0.46±0.01
1:4 44.38±0.38 0.29±0.03
1:5 25.74±0.30 0.58±0.01
1:6 14.08±0.08 0.18±0.01
1:7 15.55±0.15 0.36±0.03
1:8 15.01±0.68 0.31±0.04
According to the investigation result, when the ratio of the oil-mixed surfactant is 1.
Through the examination of the above experiment, the final proportion of the microemulsion formula is determined to be that each 100g of microemulsion contains 0.25g of chrysanthemum stem and leaf essential oil, 0.25g of mint essential oil, 4.37g of ethyl oleate, 8021.82g of tween, 7.26g of glycerol, 0.10g of baicalin, 3g of antioxidant vitamin C, 0.1g of antioxidant sodium metabisulfite and 58.00g of water phase (the crude drug amount is 20.45 mg/mL).
1.8 best prescription Process validation
The microemulsion was prepared according to the above formulation, and the particle size, polydispersity index (PDI) and Zeta potential were measured, and the results are shown in table 7.
TABLE 7 measurement results of particle diameter, polydispersity index (PDI) and Zeta potential
Batches of Particle size (nm) Polydisperse coefficient (PDI) Zeta potential (mV)
1 14.59±0.55 0.20±0.04 -5.72±0.49
2 14.60±0.29 0.19±0.02 -5.03±0.23
3 14.40±0.15 0.21±0.12 -5.70±0.53
2. Preparation of gel
2.1 screening of gel matrix dosage
The gel matrix adopts carbomer-940 which is easy to swell and has better spreadability; firstly, fixing the dilution multiple (10 times) of the microemulsion, the dosage of the humectant (3% glycerol and 7% aloe juice) and the pH value (pH = 6.5), and investigating the influence of 1%,2%,3%,4% and 5% of carbomer-940 on the appearance, the viscosity and the like of the microemulsion gel, wherein the result is shown in table 8, which indicates that the optimal dosage of carbomer-940 is 3%.
TABLE 8 examination of the amount of carbomer-940
The amount of carbomer-940 is Microemulsion gel property
1% The fluidity is too strong and the gel state is not obvious
2% Has low viscosity and is not easy to adhere to skin
3% Good viscosity and spreadability
4% Has high viscosity and is not easy to be smeared
5% The gel is not easy to swell and is difficult to stir
2.2 investigating the dosage and the type of the humectant, wherein the dosage, the pH value (pH = 6.5) and the microemulsion dilution multiple of the fixed gel matrix are investigated, and the influence of glycerin and aloe which are respectively used as the humectant on the appearance, the viscosity and the like of the microemulsion gel is investigated.
The result shows that the glycerin has good moisturizing effect, and is too sticky when being applied on the skin, so that the use feeling is poor; aloe vera juice, however, is fresh and non-tacky when applied to the skin, but has a short moisturizing time, so glycerin was chosen for mixing with aloe vera juice as a humectant (aloe vera juice: glycerin =3
2.3 examination of the amount of microemulsion to be added
In order to ensure that the microemulsion gel has the effects of diminishing inflammation and removing acne, the contents of chrysanthemum stem and leaf essential oil, baicalin and water extract in the final gel need to be ensured, 20g of microemulsion diluted by 10 times is added according to screening, the contents of the chrysanthemum stem and leaf essential oil and the baicalin in the gel are 3 times of the minimum anti-inflammation content, and the crude drug content in the gel is 12.5mg/mL.
2.4 optimal formulation of microemulsion gelling agent
Through the examination of the above experiment, the final proportion of the microemulsion gel formula is determined to be that each 100g of the microemulsion gel contains 20g of microemulsion which is diluted by 10 times, carbomer-9403 g, 3g of glycerol, 10g of propylene glycol, 7g of aloe juice, 3g of vitamin C, 0.1g of sodium metabisulfite, 0.1g of ethylparaben and 53.8g of water phase (the crude drug amount is 20.45 mg/mL).
In the invention, the chrysanthemum stem and leaf essential oil and baicalin in the anti-inflammatory acne-removing preparation have the main anti-inflammatory effect, and the chrysanthemum stem and leaf water extract has the acne-removing effect; in the later stage, according to the requirement, the contents of the chrysanthemum stem and leaf essential oil and the baicalin are increased, so that the anti-inflammation and acne-removing effects can be achieved, the appearance of the acne-removing microemulsion is improved, and the convenience of industrial production is also improved.
The invention also compares the chrysanthemum essential oil and the chrysanthemum water extract with the chrysanthemum stem leaf essential oil and the chrysanthemum stem leaf water extract for research, and the two essential oils and the two water extracts have the activities of diminishing inflammation and removing acne and have the curative effect when being matched with baicalin. Both essential oils and aqueous extracts are within the scope of this patent.

Claims (3)

1. A preparation method of a traditional Chinese medicine preparation with anti-inflammation and acne-removing effects is characterized by comprising the following steps:
(1) Extracting chrysanthemum stem and leaf essential oil by adopting a steam distillation method to obtain chrysanthemum stem and leaf essential oil and chrysanthemum stem and leaf water extract, dehydrating the essential oil by using anhydrous sodium sulfate, and storing in a dark place;
(2) Mixing the chrysanthemum stem leaf essential oil extracted in the step (1) with the mint essential oil, and adding an oil phase auxiliary material ethyl oleate to prepare an oil phase; adding mixed surfactant composed of tween 80 and glycerol and baicalin into the oil phase; taking the water extract of the chrysanthemum stems and leaves prepared in the step (1), and adding water-phase auxiliary materials to prepare a water phase; stirring uniformly at normal temperature under magnetic stirring, and dropwise adding a water phase into the oil phase while stirring to obtain a clear and transparent microemulsion; the water phase auxiliary materials comprise one or more of aloe extract, sorbitol, propylene glycol, parabens and vitamin C;
the microemulsion is prepared from the following components: 0.25g of chrysanthemum stem and leaf essential oil, 0.25g of mint essential oil, 4.37g of ethyl oleate, 21.82g of tween 80, 7.26g of glycerol, 0.10g of baicalin and 58g of water phase with the crude drug amount of 20.45mg/ml of chrysanthemum stem and leaf;
(3) And (3) preparing the microemulsion prepared in the step (2) into gel.
2. The preparation method of the traditional Chinese medicine preparation with the effects of diminishing inflammation and removing acnes as claimed in claim 1, wherein each 100g of the gel prepared in step (3) contains 20g of microemulsion, 3g of gel matrix carbomer 940, 3g of humectant glycerol, 7g of humectant aloe vera, 3g of antioxidant vitamin C, 0.1g of antioxidant sodium metabisulfite, 10g of cosolvent propylene glycol, 0.1g of preservative ethylparaben, and 53.8g of aqueous phase with the crude drug amount of the chrysanthemum stem and leaf prepared in step (2) being 20.45mg/ml.
3. Use of the Chinese medicinal preparation prepared according to claim 1 for the preparation of a medicament for inhibiting staphylococcus aureus and propionibacterium acnes.
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