Application of fragrant round essential oil in preparation of skin pathogenic bacterium bacteriostatic drug
By extracting scent essential oil from scented leaves and fruits and applying it to inhibit skin pathogenic bacteria, the problem of drug resistance caused by antibiotic drugs is solved, and broad-spectrum antibacterial activity and effective antibacterial effect are achieved.
Patent Information
- Application Number
- CN202411852999.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-16
- Publication Date
- 2025-03-14
AI Technical Summary
The frequent use of existing antibiotics has led to resistance to skin pathogenic bacteria, and the effect of treating skin diseases is not ideal.
Extract the essential oil of fragrant round leaves and fragrant round fruits and is used in the preparation of antibacterial drugs used to inhibit skin pathogenic bacteria.
Xiangyuan essential oil exhibits broad spectrum and significant antibacterial activity at lower concentrations, which can effectively inhibit a variety of bacteria and fungi that cause skin diseases, avoid their drug resistance, and have broad application prospects.
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Figure CN119607072A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of application of angelica sinensis essential oil, and in particular to application of angelica sinensis essential oil in preparing bacteriostatic drugs for skin pathogenic bacteria. Background Art
[0002] There is a close and complex relationship between the skin and microorganisms, which is essential for maintaining human health. The skin is not only the largest organ of the human body, but also the first line of defense against the external environment. The balance of interactions between the human body and microorganisms also plays a key role in the coordination of skin homeostasis. When microorganisms gain growth advantage, it is extremely easy to cause skin diseases, such as dermatitis, psoriasis, acne, etc., which have an impact on people's lives. In order to treat skin diseases, people often use fast-acting antibiotics and other drugs, but with the frequent and large-scale use of antibiotics, skin pathogens develop drug resistance, and the treatment effect is not ideal. Plant essential oils have pharmacological activities such as antioxidant, anti-inflammatory, anti-allergic, antibacterial, and anti-tumor. Their antibacterial properties are not easy to lead to drug resistance. Therefore, their application in the preparation of skin pathogens has become a new way to deal with drug-resistant bacteria.
[0003] Citron comes from Citrus medica L. or Citrus wilsonii Tanaka, a plant of the Rutaceae family. It has been included in the "Chinese Pharmacopoeia" of all dynasties. Its fruit, leaves, and roots can be used as medicine, and have the effects of soothing the liver and regulating qi, relieving fullness and resolving phlegm. In ancient times, citron was the mainstream variety, but now commercial citrons come from two sources: citron and citron, and citron has the largest output and is also widely used. In addition, citron is one of the oldest known citrus fruits and a traditional Chinese medicine and food variety. It first appeared in the "List of Varieties that are Both Food and Medicine (First Batch)" released by my country in 1987. Citron (CITRUS MEDICA LIMONUM) flowers / leaves / stems / fruits / fruit oils / extracts have been included in the "Catalogue of Used Cosmetic Raw Materials (2021 Edition)" and can be used as high-quality raw materials for medicines, foods, spices, flavors, and cosmetics. It has market potential for developing big health products such as "medicine, food, and cosmetics".
[0004] Citron has a strong aroma and was often used as a spice in ancient times. The Tang Dynasty's "Zhenglei Bencao" called it "fragrant". In the West, citron can also be used to treat indigestion, antibacterial and antiseptic, and repel mosquitoes. According to modern research, citron contains a variety of ingredients such as volatile oils, flavonoids, coumarins, alkaloids and terpenes, and has antioxidant, anti-inflammatory, anti-allergic, antibacterial, and anti-tumor pharmacological activities. It was found from the literature that whether from the genetics of citron's germplasm resources or chemical composition and pharmacological activity, more research is focused on another source of citron, Citrus medica L., and the relevant research on citron needs to be further developed. At present, the development and utilization of fragrant round fruits are mainly based on their fruits, such as slicing and drying fragrant round fruits to make medicinal materials, or using their fruits to prepare beverages, preserves, canned foods, etc., while few people have developed and utilized the aromatic oil in the leaves of fragrant round fruits. The lush branches and leaves of fragrant round trees and the large number of fragrant round leaves left behind after annual pruning are thrown away as waste. If the effective ingredients contained in the leaves of fragrant round trees can be reasonably developed and utilized, it will not only promote the extension and development of the fragrant round tree industry chain, but also increase the economic added value of the fragrant round tree. Summary of the invention
[0005] In view of this, the present invention proposes the use of osmanthus essential oil in the preparation of antibacterial drugs for inhibiting skin pathogens.
[0006] The technical solution of the present invention is achieved in this way:
[0007] Application of hollyhock essential oil in the preparation of antibacterial drugs for inhibiting skin pathogens.
[0008] Furthermore, the Cyperus rotundus essential oil is Cyperus rotundus leaf essential oil and / or Cyperus rotundus fruit essential oil.
[0009] Furthermore, the extraction method of the fragrant round leaf essential oil is: take fresh fragrant round leaves and place them in a ventilated place to dry in the shade for 3-4 days, crush them into 40-60 mesh to obtain fragrant round leaf powder, and then mix the fragrant round leaf powder and deionized water according to a solid-liquid ratio of g / ml of 1:10-12, and then place them in an essential oil extraction device and extract them for 2-4 hours at a distillation temperature of 200-250°C to obtain the fragrant round leaf essential oil.
[0010] Furthermore, the extraction method of the fragrant round fruit essential oil is: take fresh fragrant round fruit slices and place them in a ventilated place to dry in the shade for 3-4 days, crush them into 40-60 mesh to obtain fragrant round fruit powder, and then mix the fragrant round fruit powder and deionized water according to a solid-liquid ratio of g / ml of 1:10-12, and then place them in essential oil extraction equipment and extract them at a distillation temperature of 200-250°C for 2-4 hours to obtain the fragrant round fruit essential oil.
[0011] Furthermore, the skin pathogens include bacteria and / or fungi.
[0012] Furthermore, the safflower essential oil is used in the preparation of antibacterial drugs for bacterial and / or fungal skin pathogens.
[0013] Furthermore, the bacteria are one or more of Staphylococcus aureus, Bacillus subtilis, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa.
[0014] Furthermore, the fungus is one or more of Candida albicans, Candida tropicalis, Malassezia furfur, and Trichophyton rubrum.
[0015] Compared with the prior art, the present invention has the following beneficial effects:
[0016] The invention extracts the essential oil of the scutellaria leaf and the essential oil of the scutellaria fruit from the scutellaria leaf and the scutellaria fruit, respectively, and applies the essential oils to the antibacterial process of Staphylococcus aureus, Bacillus subtilis, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Candida tropicalis, Malassezia furfur, Trichophyton rubrum and other bacteria and fungi that cause skin diseases. The essential oil of the scutellaria leaf and the essential oil of the scutellaria rubrum show broad-spectrum and significant antibacterial activity at relatively low concentrations, have broad application prospects in preparing antibacterial drugs for skin pathogens, can also prevent the bacteria from developing drug resistance, and have excellent effect. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 It is a physical picture of the fragrant round leaf and fragrant round fruit of the present invention.
[0018] Figure 2 The diagram shows the antibacterial effects of the essential oil of the fragrant round leaf and the fragrant round fruit on different bacterial species; wherein a is Staphylococcus aureus, b is Bacillus subtilis, c is Staphylococcus epidermidis, d is Escherichia coli, e is Pseudomonas aeruginosa, f is Candida albicans, g is Candida tropicalis, h is Malassezia furfur, and i is Trichophyton rubrum. DETAILED DESCRIPTION
[0019] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.
[0020] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.
[0021] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.
[0022] The fragrant roundworm of the present invention is introduced from Jingjiang, Jiangsu Province to Danzhou, Hainan Province for planting in the southern medicinal resource garden of the Institute of Tropical Crops Variety Resources, Chinese Academy of Tropical Agricultural Sciences. The leaves and fruits of the fragrant roundworm are collected from the resource garden. It is identified as the fragrant roundworm Citrus wilsonii Tanaka of the Rutaceae family by Yu Fulai, a researcher at the Institute of Tropical Crops Variety Resources, Chinese Academy of Tropical Agricultural Sciences. The samples are stored in the Southern Medicinal and Health Research Center of the Institute.
[0023] Example 1 - Preparation of Fragrant Bean Essential Oil
[0024] (1) Extraction of essential oil from sweet osmanthus leaves: fresh sweet osmanthus leaves were placed in a ventilated place to dry in the shade for 3 days, crushed into 40 mesh to obtain sweet osmanthus leaf powder, and then the sweet osmanthus leaf powder and deionized water were mixed at a solid-liquid ratio of 1:10 g / ml, and then placed in an essential oil extraction equipment for extraction at a distillation temperature of 220°C for 3 hours to obtain sweet osmanthus leaf essential oil.
[0025] (2) Extraction of essential oil from safflower: fresh safflower slices were placed in a ventilated place to dry for 3 days, and then crushed into 40 mesh to obtain safflower powder. The safflower powder and deionized water were then mixed at a solid-liquid ratio of 1:10 and placed in an essential oil extraction device for extraction at a distillation temperature of 220°C for 3 hours to obtain safflower essential oil.
[0026] Example 2-Bacterial inhibition test of fennel essential oil
[0027] 1.1 Materials, strains, and reagents
[0028] Materials: the essential oil of the fragrant round leaf and the essential oil of the fragrant round fruit prepared in Example 1.
[0029] Bacterial species: Staphylococcus aureus (ATCC6538), Bacillus subtilis (ATCC6633), Staphylococcus epidermidis (ATCC12228), Escherichia coli (ATCC25922), Pseudomonas aeruginosa (ATCC15442), Candida albicans (ATCC10231) were purchased from Guangdong Huankai Biotechnology Co., Ltd.; Candida tropicalis (ATCC750), Malassezia furfur (ATCC44344), Trichophyton rubrum (ATCC MYA-4438) were purchased from Beina Chuanglian Biotechnology Co., Ltd.
[0030] Reagents: MH agar, MH broth, YM agar, YM broth, nutrient medium (NA), nutrient broth (NB), modified Dixon agar (MD), 2693 modified Dixon (mDixon), Tween 40, comprehensive PDA medium, and PDB medium were all purchased from Qingdao Haibo Biotechnology Co., Ltd.; Tween 80 and dimethyl sulfoxide (DMSO) were purchased from Sinopharm Chemical Reagent Co., Ltd.; levofloxacin and fluconazole were purchased from Beijing Solebao Technology Co., Ltd.; ketoconazole was purchased from Shanghai Yuanye Biotechnology Co., Ltd.
[0031] 1.2 Instruments and Equipment
[0032] The ZNHW digital display electric heating mantle and 2L / 29# volatile oil extractor were purchased from Shanghai Menrongxing Biotechnology Co., Ltd.; the Sartarius CPA225D electronic analytical balance was purchased from Sartorius (Beijing) Scientific Instrument Co., Ltd.; the SW-CJ-1F clean bench was purchased from Suzhou Guangyuan Purification Technology Co., Ltd.; the SpectraMax iD3 multifunctional microplate reader was purchased from Meigu Molecular Instrument (Shanghai) Co., Ltd.; the BM2100 optical microscope was purchased from Zhejiang Xinfeng Medical Equipment Co., Ltd.; the DNP-9052BS-Ⅲ electric constant temperature incubator and HZQ-X160 constant temperature shaker were purchased from Shanghai Shenmiao Medical Equipment Manufacturing Co., Ltd.; and the LX-B50L portable pressure steam sterilizer was purchased from Hefei Huatai Medical Equipment Co., Ltd.
[0033] 1.3 Determination of antibacterial activity of fennel essential oil
[0034] 1.3.1 Determination of inhibition zone
[0035] The filter paper method was used to determine the antibacterial effect of the essential oils of the leaves and fruits of the fragrant round fruit by measuring the diameter of the inhibition zone. In the clean bench, 100 μL of bacterial suspension (monoclonal bacteria were cultured to the logarithmic growth phase, and the colony count was diluted to 10) was taken with a pipette. 6 cfu / mL bacterial suspension) was evenly coated on the surface of the culture dish (d=9cm) for standby use; then 20 μL of fragrant round leaf essential oil and fragrant round fruit essential oil were respectively drawn and dropped on a sterile filter paper (d=7mm), and the paper was attached to the surface of the coated culture medium after absorption, and corresponding marks were made, with sterile water as a negative control, levofloxacin as a positive control for bacteria, and fluconazole, ketoconazole, and terbinafine as positive controls for corresponding fungi. The bacterial culture conditions were cultured at 37°C for 24h, and the fungi were cultured at 28°C for 48h. After the culture was completed, the diameter of the inhibition zone was measured using a vernier caliper using the cross method, and the average value was taken as the measurement result, and three parallels were set for each treatment group.
[0036] The results were determined according to the "Pharmacological Experimental Methods" standard: the diameter of the inhibition zone > 20 mm is extremely sensitive, the diameter of the inhibition zone is 15-20 mm is highly sensitive, the diameter of the inhibition zone is 11-15 mm is moderately sensitive, and the diameter of the inhibition zone is ≤ 10 mm is low sensitive. The results are shown in Table 1 and Figure 2 shown.
[0037] Table 1 Diameters of inhibition zones of different bacterial species by essential oils from scutellaria baicalensis leaves and scutellaria baicalensis fruit
[0038]
[0039]
[0040] Note: - means no inhibition zone. The positive control concentration is levofloxacin (20μg / ml); ketoconazole (20mg / ml) is the positive control for Candida albicans and Candida tropicalis, fluconazole (3mg / ml) is the positive control for Malassezia furfur, and terbinafine hydrochloride (8mg / ml) is the positive control for Trichophyton rubrum. Different lowercase letters in the same row indicate differences between different samples of the same species.
[0041] As shown in Table 1 and Figure 2 As shown in the results, the essential oil of the fragrant round leaf has an antibacterial effect on 9 kinds of bacteria and fungi, and its antibacterial activity is much better than that of the fragrant round fruit essential oil. Among them, the fragrant round leaf essential oil is extremely sensitive to 3 Gram-positive bacteria and 4 fungi, with an inhibition zone of more than 30 mm, especially the best antibacterial effect on the fungi Candida albicans and Trichophyton rubrum, with an inhibition zone greater than 90 mm; for Gram-negative bacteria, the fragrant round leaf essential oil is moderately sensitive to Escherichia coli and Pseudomonas aeruginosa. The antibacterial effect of the fragrant round fruit essential oil on fungi is better than that on bacteria, and it is moderately sensitive to the bacteria Escherichia coli, with an inhibition zone of 14.15±0.54 mm, and it is lowly sensitive or resistant to other bacteria; the fragrant round fruit essential oil is extremely sensitive to the fungi Candida tropicalis and Trichophyton rubrum, with inhibition zones of 33.83±3.46 mm and 24.75±0.67 mm, respectively, and it is highly sensitive to other fungi. At the same time, it can be seen from the table that the antibacterial effect of the essential oil of the fragrant round leaf and the fragrant round fruit on Gram-negative bacteria is weak or resistance appears, which may be related to the complex outer membrane structure of Gram-negative bacteria. The outer membrane is composed of lipopolysaccharide, which can effectively limit the penetration of hydrophobic molecules and provide certain protection for cells to avoid being invaded by chemicals and enzymes.
[0042] 13.2 Determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
[0043] The micro-broth dilution method was used for determination. Dimethyl sulfoxide (DMSO) and Tween 80 were used to dissolve the essential oil of the fragrant round leaf into the highest concentration sample solution, and the final concentrations of Tween 80 and DMSO in the sample solution were ensured to be ≤2% and ≤1%, respectively. Under sterile conditions, 100 μL of liquid culture medium was added to each well of the first three rows A / B / C of the 96-well plate, and then 100 μL of the highest concentration sample solution was added to the first well of the first three rows, and then the sample solution and the liquid culture medium were fully mixed by blowing with a pipette. In this way, each row of wells was diluted from left to right until 100 μL was taken from the last well and discarded, and finally 100 μL / well of the diluted bacterial suspension (10 6 cfu / mL), and three parallels were set for each sample; the negative control was Tween 80 and DMSO diluted with water to a final concentration of Tween 80 and DMSO ≤2% and ≤1% respectively, and no sample solution was added; the positive control was only added with culture medium and bacterial suspension. The 96-well plate was then placed in an incubator and cultured at 37°C for 24 hours. After the culture was completed, the absorbance value OD of the 96-well plate was measured using an ELISA reader 600 , calculate the inhibition rate, and the sample concentration with an average inhibition rate of 80% is the MIC value of the tested bacteria. Finally, the agar medium plate method was used, and 100 μL of each well with a concentration greater than MIC was applied to the corresponding solid culture medium, and cultured according to the culture conditions of different strains, and the growth of the bacteria was observed. The lowest sample concentration when the number of colonies on the plate was ≤5 was the MBC value, and three parallels were set for each treatment group. The results are shown in Table 2 below.
[0044] Table 2 MIC and MBC test results of scutellaria baicalensis leaf essential oil and scutellaria baicalensis fruit essential oil against different strains
[0045]
[0046] Note: - means no effect, the bacterial positive drug is levofloxacin (20ug / ml); ketoconazole (20mg / ml) is the positive control for Candida albicans and Candida tropicalis, fluconazole (3mg / ml) is the positive control for Malassezia furfur; terbinafine hydrochloride (8mg / ml) is the positive control for Trichophyton rubrum.
[0047] From the results in Table 2, it can be seen that the MIC and MBC values of the essential oil of the fragrant round leaves against bacteria and fungi are 0.1563-2.5 mg / ml, among which the effect on fungi is the best, with MIC and MBC values of 0.1563-0.625 mg / ml, especially the best antibacterial effect on Trichophyton rubrum, with MIC and MBC values of 0.1563 mg / ml, followed by tropical Candida and Malassezia furfur, and the worst is Pseudomonas aeruginosa among Gram-negative bacteria. The effect of the essential oil of the fragrant round fruit on fungi is also better than that on bacteria, with MIC values of 0.5859-9.375 mg / ml and MBC values of 1.172-50 mg / ml. Among them, the antibacterial effect on Trichophyton rubrum is also the best, with a MIC value of 0.5859 mg / ml and an MBC value of 1.172 mg / ml, followed by Candida albicans and tropical Candida, and no antibacterial effect on Staphylococcus aureus, Bacillus subtilis and Pseudomonas aeruginosa. It can be seen that the antibacterial effect of the fragrant round leaf essential oil is far better than that of the fragrant round fruit essential oil, and the antibacterial and bactericidal effects of the fragrant round leaf essential oil on 9 kinds of bacteria are not very different, showing a broad-spectrum antibacterial and bactericidal effect, while the antibacterial and bactericidal effects of the fragrant round fruit essential oil on 9 kinds of bacteria are quite different, and the antibacterial effect on fungi is better.
[0048] In summary, the essential oils of the leaves of Ampelopsis ternata and Ampelopsis ternata exhibit broad-spectrum and significant antibacterial activity at relatively low concentrations, and can be used in the preparation of antibacterial drugs for skin pathogens.
[0049] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.
Claims
1. Application of safflower essential oil in the preparation of antibacterial drugs for inhibiting skin pathogens.
2. The use according to claim 1, characterized in that: The galangal essential oil is galangal leaf essential oil and / or galangal fruit essential oil.
3. The use according to claim 2, characterized in that: The extraction method of the essential oil of sweet safflower leaf is as follows: fresh sweet safflower leaf is placed in a ventilated place to dry in the shade for 3-4 days, crushed into 40-60 mesh to obtain sweet safflower leaf powder, and then the sweet safflower leaf powder and deionized water are mixed at a solid-liquid ratio of 1:10-12 g / ml, and then placed in an essential oil extraction device for extraction at a distillation temperature of 200-250° C. for 2-4 hours to obtain the sweet safflower leaf essential oil.
4. The use according to claim 2, characterized in that: The extraction method of the fragrant round fruit essential oil is as follows: fresh fragrant round fruit slices are placed in a ventilated place to dry in the shade for 3-4 days, crushed into 40-60 mesh to obtain fragrant round fruit powder, and then the fragrant round fruit powder and deionized water are mixed at a solid-liquid ratio of 1:10-12 g / ml, and then placed in an essential oil extraction device for extraction at a distillation temperature of 200-250° C. for 2-4 hours to obtain the fragrant round fruit essential oil.
5. The use according to claim 1, characterized in that: The skin pathogens include bacteria and / or fungi.
6. The use according to claim 5, characterized in that: The application of the fragrant walnut essential oil in preparing antibacterial drugs for bacterial and / or fungal skin pathogens.
7. The use according to claim 5, characterized in that: The bacteria are one or more of Staphylococcus aureus, Bacillus subtilis, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa.
8. The use according to claim 5, characterized in that: The fungus is one or more of Candida albicans, Candida tropicalis, Malassezia furfur, and Trichophyton rubrum.