Composite antibacterial agent based on rose water and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical preparations and preparation and application thereof, and particularly relates to a composite antibacterial agent based on rose water and application thereof. The compound antibacterial agent based on the rose water is prepared from the following components in parts by weight: 54.1-78.9 parts of rose water, 0.2-4.0 parts of folium cortex eucommiae extract and 20.9-43.3 parts of latex matrix, the compound antibacterial agent based on the rose water has excellent antibacterial activity on staphylococcus aureus, escherichia coli and propionibacterium acnes, and the compound antibacterial agent based on the rose water has excellent antibacterial activity on staphylococcus aureus, escherichia coli and propionibacterium acnes through scientific compatibility of the latex matrix. Therefore, the preparation has the advantages of stability, easiness in smearing and natural rose fragrance.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a rose water-based composite antibacterial agent and application thereof. Background Art

[0002] Acne is an extremely common skin disease that primarily occurs in areas with well-developed sebaceous glands, including the face, back, and trunk. Propionibacterium acnes (P. acnes) is an anaerobic pathogen that plays a key role in the pathogenesis of acne. P. acnes breaks down triglycerides in sebum, releasing free fatty acids and propionic acid, which stimulate abnormal keratinization and inflammation of hair follicles. Staphylococcus aureus can cause secondary infection, leading to suppurative acne. If patients scratch their acne lesions, causing rupture, or improperly squeeze them with instruments, they can introduce Escherichia coli from the environment, causing secondary infection (manifested as suppurative inflammation and exudation). P. acnes is the target of antimicrobial therapy for acne vulgaris. Acne strains from various geographic locations have reportedly developed acquired resistance to erythromycin, clindamycin, and tetracyclines. Currently, acne treatments primarily rely on topical isotretinoin and antibiotics, as well as oral antibiotics, retinoids, and corticosteroids. However, increasing resistance has reduced clinical efficacy and is inconsistent with the "safe, natural, and green" product development strategy.

[0003] Eucommia leaves (Folium Eucommiae) are the leaves of the Eucommia bark, a rare and endemic medicinal herb in my country. Compared to Eucommia bark, Eucommia leaves are more abundant and readily available, with a significant annual harvest. In 2017, Eucommia leaves were officially designated as a medicinal and edible resource. Eucommia leaves possess important pharmacological properties, including antioxidant, antibacterial, lipid-lowering, and anti-inflammatory activities. However, there are few reports on the use of Eucommia leaves in the treatment and prevention of acne.

[0004] Chinese patent publication number CN108420938A discloses a Chinese herbal decoction for treating acne, which is made from the following Chinese herbal ingredients in parts by weight: 30 parts of dandelion, 10 parts of raw earth, 5 parts of sophora flavescens, 3 parts of talc, 15 parts of red vine, 12 parts of isatis indigotica, 25 parts of cnidium monnieri, 20 parts of loquat leaves, 10 parts of oldenlandia diffusa, 25 parts of mulberry bark, and 12 parts of houttuynia cordata. 15 parts of honeysuckle, 15 parts of glossy privet fruit, 20 parts of litchi seed, 20 parts of aloe, 15 parts of elsholtzia, 10 parts of chicken's gizzard lining, 30 parts of herba agrimonii, 15 parts of cicada shells, 20 parts of schisandra chinensis, 30 parts of kochia scoparia, 9 parts of purslane, 5 parts of rose, 20 parts of alfalfa, 30 parts of eucommia, 15 parts of antler frost, 12 parts of salvia miltiorrhiza, 20 parts of ligusticum root, 15 parts of longan, 6 parts of raw rhubarb, and 25 parts of liquorice. The above herbs, after syndrome differentiation and compatibility, can effectively play the effects of clearing away heat and detoxifying, promoting blood circulation and dispersing blood stasis, relieving exterior symptoms and expelling wind, dispersing stagnation and reducing swelling, and can also reduce sebum secretion and reduce the formation of scars, thereby achieving the purpose of curing acne. However, this plan is based on Yujisan in "The Authentic Surgery·Freckles" and Wuxiangsan in "Qianjin Yaofang·Seven Aperture Diseases·Facial Medicine", combined with clinical experience. Therefore, the raw materials are complex, the preparation is rough, and it has a bitter Chinese medicine taste. The user experience is not good enough, and it cannot effectively realize the resource utilization of rose water. Summary of the Invention

[0005] In view of the deficiencies of the prior art, the present invention provides a composite antibacterial agent based on rose water and application thereof.

[0006] This is achieved specifically through the following technical solutions:

[0007] 1. A composite antibacterial agent based on rose water, comprising, by weight, rose water: eucommia leaf extract: latex matrix = 54.1-78.9: 0.2-4.0: 20.9-43.3.

[0008] Furthermore, the formula of 10 grams of the rose water-based composite antibacterial agent is: 5.35-5.41 g of rose water, 0.32 g of eucommia leaf extract, and 4.27-4.33 g of latex matrix.

[0009] The rose water is obtained from roses of the floribunda variety picked in May and is the remaining aqueous phase after the rose essential oil is separated when the rose essential oil is produced by steam distillation.

[0010] The eucommia leaf extract is a 60% ethanol eucommia leaf extract, wherein the chlorogenic acid content is ≥315 mg / g.

[0011] The eucommia leaf extract is prepared by drying the eucommia leaves at 55° C. for 24 hours, crushing and sieving, crushing, and passing through a 40-mesh sieve. 100 g of the eucommia leaf powder is placed in a 3000 mL round-bottom flask, and reflux extraction with ethanol is performed. The extracts are combined, concentrated by vacuum rotary evaporation at 55° C., and vacuum dried at 55° C. for 12 hours to obtain the eucommia leaf extract (solid). The eucommia leaf extract obtained after crushing is a brown-green powder that is easy to store.

[0012] The process conditions of the reflux extraction are: ethanol volume concentration of 60%, liquid-solid ratio of 16 mL:1 g, extraction times of 3 times, each time of 55 minutes.

[0013] Furthermore, in the rose water-based composite antibacterial agent, the concentration of the Eucommia ulmoides leaf extract is 320 mg / mL, and the antibacterial rate against Propionibacterium acnes is 100%.

[0014] Furthermore, the minimum inhibitory concentration of the Eucommia ulmoides leaf extract in the rose water-based composite antibacterial agent against Staphylococcus aureus and Escherichia coli is 20 mg / mL.

[0015] The emulsion matrix consists of an oil phase matrix and an aqueous phase matrix, wherein the oil phase matrix consists of glyceryl monostearate, cetearyl alcohol, liquid paraffin, dimethicone, and propyl parahydroxybenzoate; and the aqueous phase matrix consists of methyl parahydroxybenzoate, glycerin, carbomer, and triethanolamine.

[0016] In the rose water-based composite antibacterial agent, when the added amount of the eucommia leaf extract in every 10g of the latex is ≥320mg, the mass proportion of glyceryl monostearate in the latex matrix is ​​≥6.4%.

[0017] The formula of 10 grams of the rose water-based composite antibacterial agent is: 0.64-0.7g of glyceryl monostearate, 0.7g of cetearyl alcohol, 0.06g of dimethicone, 2.4g of liquid paraffin, 0.025g of propyl paraben, 0.025g of methyl paraben, 0.3g of glycerin, 0.05g of carbomer 934, 0.07g of triethanolamine, 0.32g of eucommia leaf extract, and 5.35-5.41g of rose water.

[0018] 2. Application of a rose water-based composite antibacterial agent in cosmetics for improving acne.

[0019] Beneficial effects:

[0020] The rose water-based composite antibacterial agent of the present invention has excellent antibacterial activity against Staphylococcus aureus, Escherichia coli and Propionibacterium acnes.

[0021] The invention uses non-toxic and safe ethanol with a volume fraction of 60% to extract the eucommia leaves, the yield of the eucommia leaf extract is 45.71%, and the content of chlorogenic acid in the eucommia leaf extract is 315 mg / g.

[0022] The invention uses a scientific latex matrix to make the rose water-based composite antibacterial agent stable, easy to spread and have the natural fragrance of roses and the herbal aroma of eucommia leaves.

[0023] Experimental Example 1 Antibacterial Test of Single Components of Rose Water and Eucommia Leaf Extract

[0024] 1. Experimental instruments and reagents

[0025] Experimental materials: Eucommia ulmoides leaves were collected from Zunyi City, Guizhou Province; rose water came from the manufacturers of Floribunda roses in Zunyi and double roses in Bijie.

[0026] Table 1 Main reagents for the experiment

[0027]

[0028]

[0029] Table 2 Main experimental instruments

[0030]

[0031] Table 3 Antibacterial test strains

[0032]

[0033] 2. Experimental Methods

[0034] 2.1 Rose Water Analysis and Quality Evaluation

[0035] 1) Gas chromatography and mass spectrometry conditions

[0036] Gas chromatography conditions: J&W CD-WAX column (60 mm × 250 μm × 2.5 μm); injection volume: 1 μL, injection temperature: 250°C, split ratio: 5:1; carrier gas: helium (99.999%); flow rate: 1.2 mL / min; heating program: initial temperature 100°C, hold for 1 min; increase to 160°C at 10°C / min, hold for 2 min; increase to 200°C at 2°C / min, then increase to 230°C at 20°C / min, hold for 1 min.

[0037] Mass spectrometry conditions: interface temperature 260°C, ion source temperature 230°C, quadrupole temperature 150°C; ionization mode EI+, energy: 70 eV; detector voltage: 2047 V; scan mode: full scan; mass range: 20–400; comparison library: NIST 17.L.

[0038] 2) Pretreatment method

[0039] Take 5 mL of rose water sample and quickly inject 2 mL of a mixed solution of extractant and dispersant (the extractant is 0.6 mL of dichloromethane and the dispersant is 1.4 mL of tetrahydrofuran) with a syringe. After ultrasonication for 5 minutes, centrifuge at a centrifugal speed of 3000 r / min and a centrifugal temperature of 20°C for 5 minutes. The lower organic phase is dried with anhydrous sodium sulfate and filtered with a 0.45 μm microporous filter membrane for GC-MS analysis.

[0040] 3) Content of nine volatile substances in rose water produced in Zunyi and Bijie, Guizhou Province in May 2022

[0041] Rose water produced in May 2022 in Bijie and Zunyi, Guizhou, which have good sensory evaluation, was selected for analysis. The contents of nine volatile substances in rose water produced in May 2022 in Bijie and Zunyi, Guizhou (μg / mL) are as follows:

[0042] The contents of linalool, citral, α-terpineol, citronellol, phenylethyl acetate, geraniol, phenylethyl alcohol, methyl eugenol, and eugenol in rose water from Bijie area are: 3.11, 5.34, 3.45, 45.68, 0.35, 3.11, 551.5, 39.31, and 46.81 respectively. The contents of linalool, citral, α-terpineol, citronellol, phenylethyl acetate, geraniol, phenylethyl alcohol, methyl eugenol, and eugenol in rose water from Zunyi area are: 39.94, 27.44, 4.44, 147.24, 0.82, 14.92, 490.99, 24.43, and 69.86 respectively.

[0043] Floribunda rose water produced in Zunyi has relatively high levels of nine aroma components, ranging from 0.82 to 490.99 μg / mL. Double-petal rose water produced in Bijie, with the exception of a higher level of phenylethanol, has lower levels of the other eight aroma components than that found in Zunyi, ranging from 0.35 to 551.53 μg / mL. Floribunda rose water produced in Zunyi is generally of good quality.

[0044] 4) Sensory evaluation of rose water produced from roses harvested in different months

[0045] A sensory evaluation of rose water produced in May, June, and July in 2022 and 2023 by the Fenghua rose product manufacturer in Zunyi found that rose water harvested and produced in May outperformed rose water harvested and produced in June and July. Considering product safety and aroma requirements, the following tests used rose water harvested and produced in May by the Fenghua rose product manufacturer in Zunyi, Guizhou.

[0046] 2.2 Study on the antibacterial effect of rose water

[0047] Rose water was diluted with sterile water to five different concentrations: 20%, 40%, 60%, 80%, and 100%. Staphylococcus aureus (SA) and Escherichia coli (E. coli) were tested, with DMSO as a negative control and levofloxacin standard as a positive control (test concentrations: SA: 25 μg / mL, E. coli: 6 μg / mL). The antibacterial effect of rose water was evaluated using the disc diffusion method, using the total inhibition zone size as the evaluation metric. Sterile filter paper discs were placed in the different dilutions of rose water and soaked for 30 minutes to allow the discs to fully absorb the rose water. After draining the soaked filter paper discs under sterile conditions, they were affixed to the center of a bacterial plate, one disc per plate. Each treatment with rose water concentration was repeated three times. The discs were then placed in a constant-temperature incubator at 37°C for 24 hours. The size of the inhibition zone was then measured and recorded. The results are as follows:

[0048] Table 4 Antibacterial effect of rose water

[0049]

[0050] 2.3 Preparation of Eucommia ulmoides leaf extract

[0051] Air-dry the Eucommia leaves in a cool, dry place, and oven-dry at 55°C for 24 hours. Pulverize and pass through a 40-mesh sieve. Accurately weigh 100g of Eucommia leaf powder into a 3000mL round-bottom flask. Add 60% ethanol at a liquid-to-solid ratio of 16mL:1g. Reflux and extract three times for 55 minutes each. Combine the extracts, rotary evaporation at 55°C, and vacuum dry for 12 hours to obtain the Eucommia extract. The dry extract was weighed and the calculated yield was 45.71%.

[0052] 2.4 Determination of chlorogenic acid content in Eucommia ulmoides leaf extract

[0053] 1) Establishment of chlorogenic acid standard curve

[0054] Accurately weigh 5 mg of chlorogenic acid reference substance, dilute to 25 mL volumetric flask with 60% ethanol to obtain 0.2 mg / mL standard stock solution. Pipette 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.4, and 0.5 mL of standard stock solution respectively. Prepare a series of chlorogenic acid standard working solutions. Measure its absorption peak at a wavelength of 330 nm. With absorbance A as the ordinate and chlorogenic acid content C as the abscissa, a standard curve is obtained. The standard curve equation is R2=0.999, and the regression equation is y=45.886x-0.0035. See Figure 1 .

[0055] 2) Determination of chlorogenic acid content in Eucommia ulmoides leaf extract

[0056] Accurately weigh 5 g of Eucommia leaf extract powder and dilute to 100 mL with 60% ethanol to obtain the Eucommia leaf extract mother solution. Take 0.02 mL of the Eucommia leaf extract mother solution and dilute to 10 mL with 60% ethanol to obtain the Eucommia leaf extract working solution. Measure the absorbance of the working solution at a wavelength of 330 nm as described above. Substituting the absorbance into the standard curve equation, the chlorogenic acid content in the Eucommia leaf extract is calculated to be 0.0315 mg / mL. This translates to 1575 mg of chlorogenic acid in 5 g of Eucommia leaf extract powder, or 315 mg / g.

[0057] 2.5 Antibacterial test of Eucommia ulmoides leaf extract

[0058] 1) Antibacterial experiment of Eucommia ulmoides leaf extract against SA and E.col

[0059] SA and E.coli were used as test strains, DMSO as negative control, and levofloxacin standard as positive control (test concentrations: SA: 25 μg / mL, E.coli: 6 μg / mL). The antibacterial test was carried out using the paper diffusion method, and the total inhibition zone size was used as the evaluation index to evaluate the antibacterial effect of Eucommia ulmoides leaf extract. Sterile filter paper was placed in Eucommia ulmoides extracts of different dilution concentrations and soaked for 30 minutes to allow the paper to fully absorb the extract. After draining the soaked filter paper under sterile conditions, it was pasted on a bacterial plate. Each concentration of Eucommia ulmoides leaf extract was treated in parallel three times. The culture dish with the filter paper was placed in a constant temperature incubator. After constant temperature incubation at 37°C for 24 hours, the size of the inhibition zone was measured and recorded. The results are shown in Table 5. See Table 5 for details. Figure 2 and Figure 3 .

[0060] Table 5 Antibacterial effect of Eucommia ulmoides extract

[0061]

[0062] 2) Minimum inhibitory concentration (MIC) of Eucommia ulmoides leaf extract against SA and E.

[0063] The lowest dilution concentration at which the culture medium does not become turbid with naked eyes is the MIC of Eucommia ulmoides leaf extract to the strain. 8CFU / mL was set aside, and the drug concentration was reduced by continuous two-fold dilution to determine the minimum drug concentration that could inhibit microbial growth. 5mL of liquid culture medium was added to each test tube, and samples of Eucommia leaf extract with a set concentration gradient were added in sequence. The E. coli antibacterial experiment was set at a concentration range of 2.5, 5, 10, 20, 40, 80, and 160mg / mL; the SA antibacterial experiment was set at a concentration range of 0.625, 1.25, 2.5, 5, 10, 20, and 40mg / mL (DMSO or culture medium dilution). 100μL of a concentration of 10 was added to each test tube containing different Eucommia leaf extract concentrations. 8 CFU / mL bacterial suspension, place the bacterial suspension in a 37℃ constant temperature incubator and culture for 24 to 72 hours, then take out the test tube and observe the growth of microorganisms, such as Figure 4 It can be seen that when the concentration of Eucommia ulmoides leaf extract is 2.5, 5, and 10 mg / mL, obvious bacterial film is produced on the surface of the culture medium containing SA ( Figure 4 -A), indicating that at this concentration, bacterial growth cannot be inhibited. When the concentration of Eucommia ulmoides leaf extract was 0.625, 1.25, 2.5, and 10 mg / mL, the surface of the culture medium containing E. coli produced obvious bacterial film ( Figure 4 -B), indicating that at this concentration, bacterial growth was not inhibited. The minimum inhibitory concentration (MIC) of Eucommia ulmoides leaf extract against E. coli was finally determined to be 20 mg / mL, and the MIC against SA was 20 mg / mL.

[0064] 2) Antibacterial test of Eucommia ulmoides leaf extract against P. acnes

[0065] Propionibacterium acnes (P. acnes) was used as the test strain, sterile culture medium was used as the negative control, and clindamycin was used as the positive control (test concentration: 20 μg / mL). Three parallel experiments were performed for each group of samples.

[0066] (1) Material pretreatment: UV sterilization for 2 h.

[0067] (2) Streak the test strain on a plate culture medium and culture it anaerobically (10% CO2, 10% H2, 80% N2) at 37°C for about 5-6 days until obvious bacterial plaques are visible.

[0068] (3) Pick an appropriate amount of mature colonies and culture them anaerobically in 5 ml of BHI fetal bovine serum liquid culture medium (10% CO2, 10% H2, 80% N2) at 37°C for 6-12 h, adjust the OD600 to 0.6, and adjust the pH to 0.6 with normal saline.

[0069] (4) Take 5 PA bottles, inoculate 5 ml of the test bacterial solution (i.e., add 100 μl of bacterial solution with an OD600 value of 0.6 to 100 ml of culture medium) at a ratio of 0.1%, add three concentrations of the extract (80 mg / ml; 160 mg / ml, 320 mg / ml), clindamycin gel (10 g / tube: containing 0.1 g of clindamycin hydrochloride) with a drug concentration of 4 mg / ml and a blank control, and place in a 37°C incubator for anaerobically cultured for 24 h.

[0070] (5) Preparation of BHI fetal bovine serum solid culture medium (referred to as plate culture medium): Weigh 28.1 g of BHI culture medium powder and 15 g of agar, dissolve in 1000 ml of distilled water with heating and stirring, and dispense into Erlenmeyer flasks. Autoclave at 121°C for 15 min, cool to about 50°C, add fetal bovine serum at a ratio of 10%, 20 ml of LB per plate, mix well, and pour into culture dishes to cool and solidify.

[0071] (6) Gradient dilution: Take several centrifuge tubes, mark them from 1 to n, add 900ul of physiological saline to each tube, take 100ul of the original solution and add it to tube 1 to make a 10-fold dilution. After mixing, take 100ul from tube 1 and add it to tube 2. Dilute to tube n in sequence, then take 100ul of liquid from the appropriate dilution gradient tubes and spread it on a BHI plate. After anaerobically incubating at 37°C for 48h, take photos and count.

[0072] (7) Select one appropriate dilution gradient for each group to calculate the inhibition rate.

[0073]

[0074] Table 6 Experimental results of the inhibition rate of P. acnes

[0075]

[0076]

[0077] 2.6 Experimental Conclusion

[0078] In this experiment, the antibacterial activity of rose water and Eucommia leaf extract against SA and E. coli was determined using the disc diffusion method. The inhibition rate of Eucommia leaf extract against P. acnes was determined using the plate count method. Rose water showed no significant antibacterial activity against SA and E. coli at dilution concentrations of 20%, 40%, 60%, 80%, and 100%. The results indicate that rose water has no antibacterial effect against SA and E. coli, primarily for its moisturizing, whitening, and fragrance properties. Eucommia leaf extract exhibited inhibitory activity against both SA and E. coli at concentrations of 20, 40, and 80 mg / mL, with the MIC of Eucommia leaf extract against both SA and E. coli being 20 mg / mL. At a concentration of 320 mg / mL, the Eucommia leaf extract exhibited 100% inhibition against P. acnes.

[0079] Experimental Example 2 Dosage Form Screening

[0080] Considering that the MIC of Eucommia leaf extract against SA and E. coli is 20 mg / mL, and the inhibitory concentration for 100% inhibition against P. acnes is 320 mg / mL, the present invention necessitates achieving a high drug loading capacity of Eucommia leaf extract in order to translate these experimental results into a product, thereby obtaining a composite Eucommia antibacterial agent utilizing rose water. We explored various systems, including carbomer gel, carbomer / HPMC gel, carbomer / chitosan gel, and latex, with the following details:

[0081] 1. Rose and Eucommia Complex Carbomer Gel:

[0082] Table 7 Rose Eucommia composite carbomer gel formula (g)

[0083]

[0084]

[0085] The preparation method of the rose and eucommia composite carbomer gel is as follows: Carbomer 934 is added to rose water, stirred to disperse it, and allowed to stand overnight to hydrate to form a carbomer sol; polyethylene glycol 400 (PEG 400), propylene glycol, and an appropriate amount of methylparaben and propylparaben solution dissolved in rose water are added to the carbomer sol, and after thorough mixing, triethanolamine is added to adjust the pH to 5.8 to form a gel, and then the remaining amount of rose water is added, followed by eucommia leaf extract, and stirred evenly to form a gel containing rose water and eucommia leaf extract. The amount of eucommia leaf extract that can be added to this gel system is only 0.8 mg, with a total amount of 10 g. Test Example 1 Figure 10 The third sample from the left.

[0086] 2. Rose Eucommia Complex Carbomer / HPMC Gel:

[0087] Table 8 Rose Eucommia composite carbomer / HPMC gel (g)

[0088]

[0089] The preparation method of the rose and eucommia composite carbomer / HPMC gel comprises accurately weighing the required amounts of the gel materials, carbomer 934 and HPMC (K4M), and uniformly dispersing them in rose water (the amount added is 60% of the total rose water volume) with moderate stirring. The mixture is then soaked overnight to form a carbomer sol and an HPMC sol. Polyethylene glycol 4000, propylene glycol, methylparaben dissolved in an appropriate amount of rose water, and the remaining rose water are then added to the carbomer and HPMC mixed sol, or to the HPMC sol alone. The mixture is stirred evenly, and the pH is adjusted to 5.8 with triethanolamine. Finally, the eucommia leaf extract is added and stirred evenly. The prepared preparation is placed in a suitable container and labeled accordingly. Note: pH adjustment with triethanolamine is not necessary if carbomer is not used. Test Examples 5 and 6 both exhibited homogeneous systems upon initial preparation. However, after one week, the system in Test Example 6 became thinner, whereas Test Example 5 exhibited good stability. However, when we increased the amount of Eucommia leaf extract to 100 mg in Test Example 5, the system became unstable, thinned, and produced water. This indicates that the amount of Eucommia leaf extract that can be added to the HPMC gel system is 80 mg, with a total amount of 10 g. Figure 10 The fourth sample from the left.

[0090] 3. Rose Eucommia Complex Carbomer / Chitosan Gel:

[0091] Table 9 Rose Eucommia composite carbomer / chitosan gel (g)

[0092]

[0093] Preparation method of rose and eucommia composite carbomer / chitosan gel:

[0094] Test Example 7: According to the formula in Table 9, take an appropriate amount of rose water, evenly sprinkle the formulated amount of carbomer 940 powder, let it stand, and naturally swell for 24 hours to obtain a 1% carbomer sol; take an appropriate amount of rose water and evenly sprinkle the formulated amount of carboxymethyl chitosan, and naturally swell for 24 hours to obtain a 1% carboxylated chitosan sol; slowly add the carboxylated chitosan sol to the carbomer sol under stirring, and then add methyl parahydroxybenzoate dissolved in an appropriate amount of rose water and the remaining amount of rose water, mix well, and adjust the pH to about 5.8 with triethanolamine to obtain a 1% chitosan / carbomer composite hydrogel. Finally, add the eucommia leaf extract to the gel, stir until the system is uniform, and put the prepared preparation into a suitable container and mark it accordingly. The amount of eucommia leaf extract that can be added to this gel system is 40 mg. See Test Example 7. Figure 10The fifth sample from the left. Increasing the extract content further results in instability in the system.

[0095] 4. Rose and Eucommia composite latex

[0096] Table 10 Rose Eucommia composite latex (g)

[0097]

[0098]

[0099] Preparation method of rose and eucommia composite latex:

[0100] 1) Gel phase preparation: Carbomer 934 was dispersed in an appropriate amount of rose water using a magnetic stirrer to obtain a uniform sol. The pH was adjusted to approximately 5.8 with triethanolamine to prepare the gel phase. An ultrasonic water bath was used to remove air bubbles for 1 h.

[0101] 2) Preparation of aqueous phase: Dissolve methyl parahydroxybenzoate in an appropriate amount of rose water and mix well with glycerin to obtain an aqueous phase;

[0102] 3) Preparation of oil phase: Glyceryl monostearate, cetearyl alcohol, liquid paraffin, dimethicone, and propyl paraben are uniformly mixed to obtain an oil phase;

[0103] 4) Use a constant temperature water bath to heat the oil phase and the water phase to 65°C respectively. Under magnetic stirring (250-1000 rpm), slowly add the water phase to the oil phase to form an emulsion. Then, add the gel phase under continuous stirring to form a latex system. Under stirring, add the Eucommia leaf extract to the latex system and continue stirring for 5 minutes until the system is uniform; the total amount is 10g. This process is always carried out under magnetic stirring. The prepared latex is placed in a suitable container and labeled accordingly. The samples of Test Example 8 and Test Example 9 are shown in Figure 10. Figure 10 First and second samples from the left.

[0104] 5) Preparation of Eucommia ulmoides leaf extract: Eucommia ulmoides leaves were air-dried in a cool place, oven-dried at 55°C for 24 h, crushed, and passed through a 40-mesh sieve. 100 g of Eucommia ulmoides leaf powder was weighed into a 3000 mL round-bottom flask, and 60% ethanol was added at a liquid-to-solid ratio of 16:1 (mL / g). Reflux extraction was performed three times (55 min each time). The combined extracts were concentrated by rotary evaporation at 55°C and vacuum-dried at 55°C for 12 h to obtain a solid block, which was then crushed to obtain a powdered Eucommia ulmoides leaf extract.

[0105] 6) Rose water preparation: Rose water is obtained from roses of the floriferous rose variety harvested in May, and the remaining aqueous phase is separated after the rose essential oil is produced by steam distillation;

[0106] In the above preparation process, a blank latex was set, that is, no Eucommia leaf extract was added, and rose water was used instead of Eucommia leaf extract. The blank latex obtained was a white paste. Figure 11 First sample from the left.

[0107] The gels and emulsions prepared in Test Examples 1 to 9 were tested or evaluated as follows:

[0108] 1. Antibacterial test on Staphylococcus aureus and Escherichia coli

[0109] 1. Preparation before the test

[0110] Preparation of bacterial suspension (slant strain): Two types of bacteria were inoculated into test tubes containing LB medium, cultured on a shaking platform for 12 hours, and activated continuously for 3 days before use.

[0111] Preparation of culture medium: Weigh 33 g of culture medium, add 1000 mL of deionized water, adjust the pH to between 7.0 and 7.2, and sterilize at 121°C for 20 min; this is the culture medium required for the experiment.

[0112] 2. Strain recovery:

[0113] Remove Staphylococcus aureus and Escherichia coli from -20°C and thaw in a 37°C water bath. Pipette 200 μL each of the thawed Staphylococcus aureus and E. coli into autoclaved LB medium, label the tubes, and incubate in a shaker at 37°C (180 rpm) for 12-16 hours.

[0114] 3. Preparation of antibacterial culture medium

[0115] 200 μL of revived SA and E. coli strains were each pipetted and evenly spread onto a pre-prepared plate. Sterile filter paper was immersed in each preparation for 30 minutes. Under sterile conditions, the soaked filter paper was drained and applied to a prepared bacterial plate. Each sample was treated three times in parallel and then incubated in a constant temperature incubator at 37°C for one day. A blank preparation was used as a negative control, and levofloxacin was used as a positive control. The size of the inhibition zone was measured and recorded.

[0116] 2. Formula Evaluation

[0117] 1. Performance and sensory evaluation

[0118] The spreadability was evaluated by oiliness, spreadability, ductility, and absorbability; the sensory properties were evaluated by observing water dilution characteristics, uniformity, odor, and color.

[0119] 2. pH

[0120] The pH of the formulations was measured using a digital pH meter by dissolving 1 g of the formulation in 100 mL of distilled water and allowing to stand for two hours. The pH of each formulation was measured in triplicate and the average was calculated.

[0121] 3. Stability evaluation

[0122] This experiment was conducted after the optimal formulation was obtained. The preparation was stored in a sealed glass container with a lid at room temperature or -5°C for 1 month and at 45°C for 12 hours, and the system changes were checked regularly.

[0123] 4. The stability of the above test examples 1 to 9 is as follows:

[0124] Table 11 Gel stability of test examples 1-4

[0125]

[0126] The present invention investigated the effects of different amounts of Eucommia leaf extract added on the formation of a rose Eucommia composite carbomer gel. As shown in Table 11, the carbomer gel system alone had poor amalgamation with Eucommia leaf extract. The maximum amount of Eucommia leaf extract added per 100 g of the formula was only 8 mg. As the dosage increased, the system was severely damaged, such as thinning and water precipitation. This indicates that the carbomer gel system alone had poor agglomeration with Eucommia leaf extract, and the minimum inhibitory concentration (MIC) of Eucommia leaf extract against E. coli was 20 mg / mL, and the MIC against SA was 20 mg / mL) of Eucommia leaf extract could not be achieved.

[0127] Performance and sensory evaluation of Test Examples 5 and 6 indicate that the rose and eucommia composite gels prepared in both Test Examples 5 and 6 were able to achieve a drug loading of 80 mg of eucommia leaf extract, meeting the required loading capacity. However, drug sedimentation occurred in Test Example 6 after standing. The formulation in Test Example 5 demonstrated good stability, and therefore was determined to be representative of the rose and eucommia composite HPMC gel formulation.

[0128] The 1% chitosan / carbomer composite hydrogel prepared in Experimental Example 7 is a translucent solid with an appropriate consistency, a light rose fragrance and a herbal green fragrance. It has a cooling sensation after application and good stability after placement. However, the drug loading capacity of the eucommia leaf extract in this system is 40 mg, which cannot meet the requirements of a higher encapsulation capacity.

[0129] In the latex system, as in Test Example 8, when the Eucommia leaf extract content was increased to 80 mg, the system exhibited good stability, a non-greasy feel upon application, good ductility, no sedimentation after prolonged storage, and a natural rose fragrance. Test Example 9 increased the Eucommia leaf extract loading to 100 mg. The system also exhibited good stability, a non-greasy feel upon application, good ductility, no sedimentation after prolonged storage, and a natural rose fragrance.

[0130] According to the above encapsulation and stability tests, there are four types of composite antibacterial agents that can encapsulate Eucommia ulmoides leaf extract, namely, rose Eucommia ulmoides composite carbomer gel (0.8 mg Eucommia ulmoides extract / 10 g), rose Eucommia ulmoides composite HPMC gel (80 mg Eucommia ulmoides extract / 10 g), rose Eucommia ulmoides composite carbomer / chitosan gel (40 mg Eucommia ulmoides extract / 10 g), and rose Eucommia ulmoides composite latex (20-100 mg Eucommia ulmoides extract / 10 g) (see Tables 10 and 12). Obviously, the latex system has the largest encapsulation capacity for Eucommia ulmoides leaf extract.

[0131] In the antibacterial experiment of rose eucommia composite latex, the latex containing 20 mg (chlorogenic acid content 0.63 mg / mL) or 40 mg (chlorogenic acid content 1.26 mg / mL) of eucommia leaf extract had no antibacterial effect on SA and E. coli. When the addition amount was 80 mg (chlorogenic acid content 2.52 mg / mL), the diameter of the inhibition zone produced on SA was 15 mm, and the diameter of the inhibition zone produced on E. coli was 8 mm ( Figure 13 This indicates that the only rose eucommia compound antibacterial agents that can resist SA and E. coli are the HPMC gel system (80 mg eucommia extract / 10 g) and the latex system (80-100 mg eucommia extract / 10 g).

[0132] In experiments conducted against P. acnes, neither 80mg of Eucommia leaf extract nor the latex showed activity against P. acnes. Therefore, increasing the dosage of Eucommia leaf extract, the results showed that the 160mg / ml Eucommia leaf extract group exhibited activity against P. acnes, with an inhibition rate of 31.29%. The 320mg / ml Eucommia leaf extract group achieved a 100% inhibition rate against P. acnes.

[0133] Therefore, it is necessary to explore the latex formulation with a higher eucommia leaf extract loading (320 mg / 10 g).

[0134] Experimental Example 3 Screening of high-load latex formulations

[0135] Table 12 Formulations of Samples 1 to 8

[0136]

[0137] Note: The above recipe is for 10 servings.

[0138] Emulsion samples obtained from formulations F1-F8 were tested and evaluated according to the aforementioned preparation, testing, and evaluation methods. When the eucommia leaf extract dosage was 20 mg, the rose eucommia composite emulsion prepared using formulation F1 in Table 12 exhibited a light licorice-colored, stable emulsion system. The formulation had a uniform paste, good ductility, and a delicate skin feel. It was non-greasy and non-slip after application, rapidly absorbed through the skin, and exhibited excellent film-forming properties. It exhibited a herbal, green fragrance with a subtle rose aroma and was stable at room temperature, refrigerated at -5°C, and at 45°C.

[0139] When the dosage of Eucommia ulmoides leaf extract was 40 mg, using the formula F2 in Table 12, but increasing the extract dosage, the prepared rose Eucommia ulmoides composite emulsion exhibited a lighter licorice-colored stable emulsion system, slightly darker than the 20 mg rose Eucommia ulmoides composite emulsion. The preparation had a uniform paste, good ductility, and a delicate skin feel. It was non-greasy and non-slip after application, was rapidly absorbed by the hand skin, and had good film-forming properties. It had a herbal green fragrance and a light rose aroma and was stable at room temperature, refrigerated at -5°C, and at 45°C.

[0140] When the amount of Eucommia ulmoides leaf extract was 80 mg, the formula of F2 in Table 12 was used. However, when the amount of extract was increased, the latex system was unstable. After being placed for 1 hour, the Eucommia ulmoides leaf extract in the latex settled and the system was clearly layered. Therefore, the 40 mg rose Eucommia ulmoides composite latex formula was not suitable for the preparation of the 80 mg rose Eucommia ulmoides composite latex system. Therefore, the formula was adjusted to increase the amount of the oil phase matrix and the gel matrix components in the system, such as the F3 formula in Table 12. Under 65 ° C magnetic stirring conditions, the oil phase and the aqueous phase were heated and mixed uniformly. The aqueous phase was then slowly added to the oil phase. After forming an emulsion, the gel phase was added and magnetic stirring was performed until a white latex was formed. 80 mg of Eucommia ulmoides leaf extract was added and magnetic stirring was continued under non-heating conditions to obtain 80 mg of rose Eucommia ulmoides composite latex. The adjusted formula of the 80mg rose and eucommia complex emulsion system is stable, with a uniform paste, good ductility when applied to the hands, and a delicate skin feel. It is non-greasy and absorbs quickly through the skin, forming a good film. It has a herbal green and rose aroma and is stable at room temperature, refrigerated at -5°C, and at 45°C.

[0141] When the dosage of Eucommia leaf extract is 100 mg, an 80 mg formulation is sufficient to meet the addition requirements of 100 mg of rose and eucommia compound emulsion. See formula F4. The prepared rose and eucommia compound emulsion containing 100 mg of Eucommia leaf extract is stable, has a uniform paste, spreads well on the hands, and has a delicate skin feel. It is non-greasy and non-slip, absorbs quickly through the skin, and forms a good film. It has a herbal green and rose aroma and is stable at room temperature, refrigerated at -5°C, and at 45°C.

[0142] When a 160mg rose eucommia emulsion system was prepared with reference to the 100mg formula F4, it was found that the emulsion system of this formula was unstable. The oil and water phases were mixed under magnetic heating and stirring at 65°C to form an emulsion, and then the gel phase was added to form a stable emulsion system. The magnetic stirring was continued, and 160mg eucommia leaf extract was added to the emulsion system. The stable emulsion system gradually broke, the eucommia leaf extract settled, and the two phases separated. This shows that the formula is not suitable for the encapsulation of 160mg eucommia leaf extract. Therefore, the amounts of cetearyl alcohol, liquid paraffin, dimethicone, and monostearic glycerol in the oil phase components of this formula were screened in a gradient dosage. The results showed that the addition amount of cetearyl alcohol and monostearic glycerol had a greater influence on the stability of the emulsion, especially the amount of monostearic glycerol was crucial, while increasing the amount of liquid paraffin and dimethicone did not improve the stability of the emulsion system. The specific experimental results are as follows:

[0143] 1) Exploring the formula of rose and eucommia compound latex (160mg)

[0144] (1) Cetearyl Alcohol

[0145] The amount of cetearyl alcohol used was increased and set to 0.8g, 1.0g, and 1.2g.

[0146] When the dosage was 0.8 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and obvious sedimentation still occurred, indicating that the dosage could not meet the encapsulation requirements.

[0147] When the dosage is 1.0g and the dosage of other formulas remains unchanged, the above operation is repeated. The latex system is stable, olive-colored, and the paste has a uniform color. It has poor ductility when applied on the hands, feels rough and greasy on the skin, rubs into mud severely after application, cannot be absorbed by the skin, and has poor film formation.

[0148] When the dosage is 1.2g and the dosage of other formulas remains unchanged, the above operation is repeated. The latex system is stable, olive-colored, and the paste has a uniform color. It has poor ductility when applied on the hands, feels rough and greasy on the skin, rubs into mud severely after application, cannot be absorbed by the skin, and has poor film formation.

[0149] Note that in this formula, the amount of cetearyl alcohol added has a significant impact on the stability of the emulsion. Excessive cetearyl alcohol will cause the emulsion paste to be rough, poorly spread on the skin surface, and increase the greasy feeling, making it difficult to absorb. Therefore, the amount of cetearyl alcohol used is still 0.7g.

[0150] (2) Liquid paraffin

[0151] The amount of liquid paraffin increased and was set to 2.6g, 2.8g, and 3.0g

[0152] When the dosage was 2.6 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and the crude extract of Eucommia ulmoides leaves still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0153] When the dosage was 2.8 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and the crude extract of Eucommia ulmoides leaves still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0154] When the dosage was 3.0 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and the crude extract of Eucommia ulmoides leaves still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0155] This indicates that increasing the amount of liquid paraffin in this formulation did not improve the stability of the emulsion system. The Eucommia extract still precipitated, and the overall stability of the system was not enhanced. Therefore, the amount of liquid paraffin remained at 2.4 g.

[0156] (3) Dimethicone

[0157] The dosage of dimethicone was increased and set to 0.08g, 0.10g, and 0.12g

[0158] When the dosage was 0.08 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and the Eucommia extract still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0159] When the dosage was 0.10 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was unstable and the Eucommia leaf extract still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0160] When the dosage was 0.12 g and the dosage of other formulas remained unchanged, the above operation was repeated. The latex system was stable, but the Eucommia extract still showed obvious sedimentation, indicating that the dosage could not meet the encapsulation requirements.

[0161] This indicates that increasing the amount of dimethicone in this formulation has no effect on the stability of the latex system. Therefore, the amount of dimethicone used remains at 0.06 g.

[0162] (4) Glyceryl monostearate

[0163] The amount of glyceryl monostearate was increased and set to 0.25g, 0.3g, and 0.35g.

[0164] When the amount of glyceryl monostearate was 0.25 g and the amounts of other ingredients remained unchanged, the above operation was repeated, but the latex system was still unstable and obvious sedimentation occurred;

[0165] When the amount of glycerol monostearate was 0.3 g and the amounts of other ingredients remained unchanged, the above operation was repeated. The sedimentation of the latex system was improved, but demulsification still existed and the latex texture was thinner.

[0166] When the monostearate dosage was 0.35g and the other formula dosages remained unchanged, the above steps were repeated. The emulsion system was stable, dark olive-colored, and capable of encapsulating 160mg of Eucommia ulmoides extract. The preparation had a uniform paste consistency, spread well on the hands, and had a delicate feel. It was non-greasy and non-slip, absorbed quickly through the skin, and had good film-forming properties. It had a strong herbal and rose aroma and was stable at room temperature, refrigerated at -5°C, and at 45°C.

[0167] Since 0.35g of glyceryl monostearate can achieve system stability, we tried reducing the amount of glyceryl monostearate to 0.32g, keeping the other formula amounts unchanged, and repeating the above steps to examine the amount of glyceryl monostearate. The emulsion system with 0.32g of glyceryl monostearate was stable, had a dark olive color, and could encapsulate 160mg of Eucommia ulmoides leaf extract. The preparation had a uniform paste, spread well on the hands, had a delicate skin feel, was non-greasy, absorbed quickly through the skin, and had good film-forming properties. It had a strong natural rose aroma and was stable at room temperature, -5°C, and refrigerated at 45°C.

[0168] 2) Exploration of the formula of rose and eucommia composite emulsion (320mg)

[0169] When a 320mg rose eucommia emulsion system was prepared using the same formulation as 160mg, F5, it was found that this formulation was unable to encapsulate 320mg of eucommia extract. After the oil and water phases were mixed under magnetic heating and stirring at 65°C to form an emulsion, the gel phase was added to form a stable emulsion system. Magnetic stirring was continued, and 320mg of eucommia leaf extract was added to the emulsion system. The stable emulsion system gradually demulsified, the eucommia leaf extract settled, and the two phases separated. This indicates that this formulation is unsuitable for encapsulating 320mg of eucommia leaf extract. The dosage of glyceryl monostearate, a component of the formulation, was adjusted accordingly.

[0170] The dosage of glyceryl monostearate was increased to 0.64g, 0.68g, and 0.70g for the experiment:

[0171] When the dosage is 0.64 g and the dosage of other formulas remains unchanged, the above operation is repeated, and the latex system is stable, dark olive in color, and can encapsulate 320 mg of Eucommia ulmoides leaf extract; it has good ductility when applied to the hands, the paste is uniform, the skin feel is delicate, it is quickly absorbed by the skin of the hands, is not greasy, and has good film-forming properties on the skin surface after application.

[0172] When the dosage is 0.68 g and the dosage of other formulas remains unchanged, the above operation is repeated, and the latex system is stable, dark olive in color, and can encapsulate 320 mg of Eucommia ulmoides extract; it has good ductility when applied to the hands, the paste is uniform, the skin feel is delicate, it is quickly absorbed by the skin of the hands, is not greasy, and has good film-forming properties on the skin surface after application.

[0173] When the dosage is 0.70 g and the dosage of other formulas remains unchanged, the above operation is repeated, and the latex system is stable, dark olive in color, and can encapsulate 320 mg of Eucommia ulmoides extract; it has good ductility when applied to the hands, the paste is uniform, the skin feel is delicate, it is quickly absorbed by the skin of the hands, is not greasy, and has good film-forming properties on the skin surface after application.

[0174] It is shown that in the formula of 320mg rose eucommia emulsion, the amount of glyceryl monostearate added is crucial to the stability of the emulsion. After appropriately increasing the amount of glyceryl monostearate, the stability of the emulsion system is significantly enhanced, which can meet the encapsulation requirements of 320mg eucommia extract. The formula has a strong herbal green fragrance and rose fragrance, which is in line with the sensory experience of herbal emulsions. It has good ductility when applied on the hands, the paste is uniform, has good ductility, and a delicate skin feel. It is quickly absorbed by the skin of the hands and is not greasy. It has good film-forming properties on the skin surface after application. It has a strong natural rose aroma and is stable at room temperature, -5℃ refrigeration, and 45℃. The optimal formula F6 was determined, and the amount of glyceryl monostearate was 0.64g, see Figure 12 shown.

[0175] Table 13 Evaluation of the formula of rose and eucommia compound antibacterial agent

[0176]

[0177] Note: 0 (unusable), + (medium), ++ (good), +++ (best).

[0178] The preparation of Test Example 8 is olive green, with a uniform texture and a good rose herbal fragrance. It has good ductility when applied on the hands and is absorbed quickly. It does not feel greasy after application, can form a film on the skin surface, and has a good dilution effect in water. The preparation of Test Example 9 is dark olive, with a uniform texture and a light rose and herbal green fragrance. It has good ductility when applied on the hands and is absorbed quickly. It does not feel greasy after application, has a good film-forming effect, and has a good dilution effect in water. The rose eucommia latex sample of Test Example 3 is tea brown, with a rose herbal fragrance, and good viscosity, uniformity, spreadability, and spreadability. The system of Test Example 5 is black-green, with good moldability after application, but the odor is slightly worse than that of Test Example 8, Test Example 9, and Test Example 1. Test Example 7 is brown-yellow, with good spreadability and ductility, can meet the use requirements, and has good stability after placement. Figure 10 of 5 samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0179] Figure 1 : Standard curve of chlorogenic acid at 330 nm;

[0180] Figure 2 : Bacteriostatic result of Eucommia ulmoides Oliv. leaf extract on S.A; from left to right, the concentration of Eucommia ulmoides Oliv. leaf extract is 5 mg / mL, 10 mg / mL, 20 mg / mL, 40 mg / mL, 80 mg / mL, respectively;

[0181] Figure 3 : Bacteriostatic result of Eucommia ulmoides Oliv. leaf extract on E. coli; from left to right, the concentration of Eucommia ulmoides Oliv. leaf extract is 5 mg / mL, 10 mg / mL, 20 mg / mL, 40 mg / mL, 80 mg / mL, respectively;

[0182] Figure 4 : Minimum bacteriostatic concentration of Eucommia ulmoides Oliv. leaf extract on S.A and E. coli; (A) is the growth of S.A; (B) is the growth of E. coli;

[0183] Figure 5 : Bacteriostatic result of blank control sample on P. acnes, the left figure is the sample colony picture before SCAN1200 analysis, and the right figure is the sample colony picture analyzed by SCAN1200;

[0184] Figure 6 : Bacteriostatic result of experimental sample 1 on P. acnes, the left figure is the sample colony picture before SCAN1200 analysis, and the right figure is the sample colony picture analyzed by SCAN1200;

[0185] Figure 7 : Bacteriostatic result of experimental sample 2 on P. acnes, the left figure is the sample colony picture before SCAN1200 analysis, and the right figure is the sample colony picture analyzed by SCAN1200;

[0186] Figure 8 : Bacteriostatic result of experimental sample 3 on P. acnes, the left figure is the sample colony picture before SCAN1200 analysis, and the right figure is the sample colony picture analyzed by SCAN1200;

[0187] Figure 9 : Bacteriostatic result of positive control sample on P. acnes, the left figure is the sample colony picture before SCAN1200 analysis, and the right figure is the sample colony picture analyzed by SCAN1200;

[0188] Figure 10: Appearance of rose and eucommia compound antibacterial agent, from left to right: rose and eucommia compound emulsion (80 mg), rose and eucommia compound emulsion (100 mg), rose and eucommia compound carbomer gel (0.8 mg), rose and eucommia compound HPMC gel (80 mg), rose and eucommia compound carbomer / chitosan gel (40 mg);

[0189] Figure 11 : Emulsion samples, from left to right: blank emulsion, rose and eucommia composite emulsion F3, F4, F5, F6.

[0190] Figure 12 : Photos of the color and spreadability of the rose and eucommia composite latex prepared in Example 1;

[0191] Figure 13 : The antibacterial activity of rose and eucommia composite latex against SA and E.coli in Experimental Example 2;

[0192] Figure 14 : Experimental photos of the antibacterial rate of Eucommia ulmoides leaf extract against P. acnes at different concentrations extracted according to the example method; A-80 mg / mL, B-160 mg / mL, C-320 mg / mL. DETAILED DESCRIPTION

[0193] The specific embodiments of the present invention are further described in detail below, but the present invention is not limited to these embodiments. Any improvement or replacement based on the basic spirit of the present embodiment still falls within the scope of protection required by the claims of the present invention.

[0194] Examples 1-3

[0195] Table 14 Formulation of Examples 1-3

[0196]

[0197] Note: The above formula is for 10 grams.

[0198] Preparation method:

[0199] 1) Gel phase preparation: Carbomer 934 was dispersed in an appropriate amount of rose water using a magnetic stirrer to obtain a uniform sol. The pH was adjusted to approximately 5.8 with triethanolamine to prepare the gel phase. An ultrasonic water bath was used to remove air bubbles for 1 h.

[0200] 2) Preparation of aqueous phase: Dissolve methyl parahydroxybenzoate in an appropriate amount of rose water and mix well with glycerin to obtain an aqueous phase;

[0201] 3) Preparation of oil phase: Glyceryl monostearate, cetearyl alcohol, liquid paraffin, dimethicone, and propyl paraben are uniformly mixed to obtain an oil phase;

[0202] 4) Preparation of Eucommia Leaf Extract: The Eucommia leaves were air-dried in a cool place, dried in an oven at 55°C for 24 h, crushed, and passed through a 40-mesh sieve. 100 g of Eucommia leaf powder was accurately weighed and placed in a 3000 mL round-bottom flask. 60% ethanol was added at a liquid-to-solid ratio of 16:1 (mL / g). Extraction was performed three times (55 min each time). The combined extracts were rotary evaporated at 55°C under reduced pressure and dried in vacuo to obtain the Eucommia extract as a solid.

[0203] 5) Rose water preparation: Rose water is obtained from roses of the floriferous rose variety picked in May, and the remaining aqueous phase is separated after the rose essential oil is produced by distillation;

[0204] 6) Preparation of latex:

[0205] Heat the oil and water phases to 65°C in a constant-temperature water bath. Slowly add the water phase to the oil phase under magnetic stirring (250-1000 rpm) to form an emulsion. Add the gel phase with continuous stirring to form a stable latex. Continue adding the Eucommia leaf extract to the latex under magnetic stirring (250 rpm) and stir evenly. A total of 10 g of the extract is added. This process is performed under magnetic stirring. Place the prepared latex into suitable containers and label accordingly.

[0206] The emulsion system of Example 1-3 was stable, dark olive in color, and capable of encapsulating 320 mg of Eucommia leaf extract. It spread well on the hands, applied evenly, had a smooth feel, was rapidly absorbed by the skin, and was non-greasy. It formed a good film on the skin surface after application and had a natural rose fragrance. It was stable at room temperature, refrigerated at -5°C, and at 45°C. The optimal formula was ultimately determined and is shown in Table 15.

[0207] Table 15 Rose Eucommia latex formulation (320mg formulation)

[0208]

[0209] Based on the previous experiments, the inventors explored an optimal process, comprising the following steps: according to the formula in Table 15: the oil phase material and the water phase material were weighed separately, and then the oil phase and the water phase (methylparaben, glycerol, carbomer 934 and rose water) were heated to 65 ° C. in a constant temperature water bath, and after magnetic stirring (250 rpm) mixing, the water phase was slowly added to the oil phase, and magnetic stirring (1000 rpm) was continued to form an emulsion. After continuous stirring (250 rpm), the eucommia leaf extract was added, and continued to stir evenly (250 rpm). The pH was adjusted to about 5.8 with triethanolamine to obtain a rose eucommia composite emulsion. Figure 12 The prepared formulation is filled into suitable containers and labeled accordingly. Figure 11 Right one.

[0210] Compared with the process of preparing the gel phase, water phase and oil phase in steps and then mixing them, the optimized process has no significant difference in results. However, this process is also easy to operate and has fewer steps. It is not only suitable for operations in the laboratory research stage, but also for production practice. It can greatly improve production efficiency and reduce process energy consumption.

[0211] At the same time, the antibacterial test was carried out on the Eucommia ulmoides leaf extract in this example, and the results are shown in Table 16:

[0212] Table 16 Antibacterial effect of Eucommia ulmoides leaf extract

[0213]

[0214] Note: Highly sensitive (>15mm); moderately sensitive (10-15mm); lowly sensitive (7-9mm); insensitive: \.

[0215] Through qualitative experiments, it can be seen that the Eucommia leaf extract at concentrations of 20, 40, 60, and 80 mg / mL has obvious inhibition zones against SA and E.colin, but no inhibition zone is produced against P.acnes. Usually, an obvious inhibition zone will be produced only when the inhibition rate is above 95%. Therefore, the concentration of Eucommia leaf extract was further increased, and the plate count method was used to examine its inhibition rate against P.acnes. When the concentration of Eucommia leaf extract was 80 mg / mL, the inhibition rate against P.acnes was -30.94%, when the concentration of Eucommia leaf extract was 160 mg / mL, the inhibition rate against P.acnes was 31.29%, and when the concentration of Eucommia leaf extract was 320 mg / mL, the inhibition rate against P.acnes was 100%. The results are shown in Tables 17 and Figure 14 As shown;

[0216] Table 17 Experimental results of the antibacterial rate of Eucommia ulmoides leaf extract against P. acnes

[0217]

[0218] Typical application example 1

[0219] A female volunteer, Ms. Peng, who often suffered from facial acne, applied the rose and eucommia composite emulsion (320 mg / 10 g) of Example 1 to one acne. The acne began to shrink after the second day of application and healed after five days.

Claims

1. A composite antibacterial agent based on rose water, characterized in that, The composition includes, by weight ratio: rose water: eucommia leaf extract: latex matrix = 54.1-78.9: 0.2-4.0: 20.9-43.

3.

2. The rose water-based composite antibacterial agent according to claim 1, wherein The formula of 10 grams of the rose water-based composite antibacterial agent is: 5.35-5.41 g of rose water, 0.32 g of eucommia leaf extract, and 4.27-4.33 g of latex matrix.

3. The rose water-based composite antibacterial agent according to claim 1, wherein The rose water is obtained from roses of the floribunda variety picked in May and is the remaining aqueous phase after the rose essential oil is separated when the rose essential oil is produced by steam distillation.

4. The rose water-based composite antibacterial agent according to claim 1, wherein The eucommia leaf extract is a 60% ethanol eucommia leaf extract, wherein the chlorogenic acid content is ≥315 mg / g.

5. A composite antibacterial agent based on rose water according to claim 1 or 4, characterized in that: The concentration of the Eucommia ulmoides leaf extract is 320 mg / mL, and the antibacterial rate to Propionibacterium acnes is 100%.

6. A composite antibacterial agent based on rose water according to claim 1 or 4, characterized in that, The minimum inhibitory concentration of the eucommia leaf extract to Staphylococcus aureus and Escherichia coli is 20 mg / mL.

7. The rose water-based composite antibacterial agent according to claim 1, wherein: The emulsion matrix consists of an oil phase and a water matrix phase, wherein the oil phase matrix consists of glyceryl monostearate, cetearyl alcohol, liquid paraffin, dimethicone, and propyl parahydroxybenzoate; and the water phase matrix consists of methyl parahydroxybenzoate, glycerin, carbomer, and triethanolamine.

8. The rose water-based composite antibacterial agent according to claim 7, wherein: In the rose water-based composite antibacterial agent, when the added amount of eucommia leaf extract in every 10g of the preparation is ≥320mg, the mass proportion of glyceryl monostearate in the emulsion is ≥6.4%.

9. The rose water-based composite antibacterial agent according to claim 7, wherein: The formula of 10 grams of the rose water-based composite antibacterial agent is: 0.64-0.7g of glyceryl monostearate, 0.7g of cetearyl alcohol, 0.06g of dimethicone, 2.4g of liquid paraffin, 0.025g of propyl paraben, 0.025g of methyl paraben, 0.3g of glycerin, 0.05g of carbomer 934, 0.07g of triethanolamine, 0.32g of eucommia leaf extract, and 5.35-5.41g of rose water.

10. Use of the rose water-based composite antibacterial agent according to any one of claims 1 to 9 in cosmetics for improving acne.

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