Application of traditional Chinese medicine composition volatile oil in preparation of medicine for inhibiting staphylococcus aureus

Abstract

The invention relates to an application of traditional Chinese medicine composition volatile oil in preparation of a medicine for inhibiting staphylococcus aureus. The traditional Chinese medicine composition volatile oil is prepared from the following raw materials: notopterygium root, rheum officinale, radix bupleuri, rhizoma atractylodis, asarum and fructus evodiae. The volatile oil product prepared by the invention has an excellent effect of inhibiting staphylococcus aureus, and the effect can be promoted to be played to the maximum extent by extracting in a specific extraction mode.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and relates to application of volatile oil of a traditional Chinese medicine composition in preparing a medicine for inhibiting Staphylococcus aureus. Background Art

[0002] Staphylococcus aureus is an important pathogen of humans. It is a Gram-positive facultative anaerobic bacterium and is ubiquitous in nature. It can be found in water, air, and dust. Under a microscope, Staphylococcus aureus colonies are arranged in spherical beads. It has no spores or flagella and can produce toxins. It is the most common pathogen of human purulent infections. It can also cause local purulent infections in wounds, pneumonia, pericarditis and other infections, causing great harm to the human body.

[0003] Traditional Chinese medicine has good antibacterial and antifungal effects, and has the advantage of low toxicity and side effects in the treatment of bacterial or fungal infections. Traditional Chinese medicine compositions can enhance the body's immunity while playing an antibacterial role, and have a better bactericidal and anti-infective effect. How to select suitable traditional Chinese medicines, prepare a traditional Chinese medicine composition against Staphylococcus aureus, effectively prevent and treat bacterial infections, and improve people's quality of life has become increasingly important.

[0004] In summary, in order to improve the anti-Staphylococcus aureus effect of a traditional Chinese medicine composition, the present invention provides an anti-Staphylococcus aureus traditional Chinese medicine composition, a preparation method and use thereof. Summary of the invention

[0005] In view of the deficiencies of the prior art, the object of the present invention is to provide an application of volatile oil of a traditional Chinese medicine composition in the preparation of a drug for inhibiting Staphylococcus aureus.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides an application of a volatile oil of a traditional Chinese medicine composition in the preparation of a drug for inhibiting Staphylococcus aureus, wherein the raw materials for preparing the volatile oil of the traditional Chinese medicine composition include Notopterygium wilfordii, Rhubarb, Bupleurum chinense, Atractylodes lancea, Asarum and Evodia rutaecarpa.

[0008] The present invention creatively finds that the mixed volatile oil product prepared with rhubarb, atractylodes, evodia rutaecarpa, asarum, bupleurum and notopterygium wilfordii as raw materials has excellent efficacy in inhibiting Staphylococcus aureus, which is specifically manifested in having a strong antibacterial effect. Compared with Staphylococcus aureus not treated with volatile oil, Staphylococcus aureus treated with volatile oil has differences in multiple metabolic pathways, mainly including secondary biosynthetic metabolism, biofilm-related genes, energy metabolism and other pathways. The results of electron microscopy show that the volatile oil has a destructive effect on both the cell wall and cell membrane of Staphylococcus aureus.

[0009] In a second aspect, the present invention provides an application of a volatile oil of a traditional Chinese medicine composition in the preparation of a Staphylococcus aureus inhibitor, wherein the raw materials for preparing the volatile oil of the traditional Chinese medicine composition include Notopterygium wilfordii, Radix et Rhizoma Rhei, Radix Bupleuri, Rhizoma Atractylodis Macrocephalae, Rhizoma Asarum and Fructus Evodiae.

[0010] According to the research results of the present invention, the volatile oil of the Chinese herbal composition can inhibit the growth and reproduction of Staphylococcus aureus at the in vitro level, that is, the volatile oil of the Chinese herbal composition can be made into a simple experimental preparation for inhibiting the growth of bacteria. The Staphylococcus aureus inhibitor claimed by the present invention is not used to eliminate the cause or lesion, that is, it is an application in the preparation of a product for inhibiting bacterial growth for a non-therapeutic purpose.

[0011] Preferably, the raw materials for preparing the volatile oil of the traditional Chinese medicine composition include, by mass, 1-3 parts of rhubarb, 1-3 parts of atractylodes, 1-3 parts of evodia rutaecarpa, 1-3 parts of asarum, 1-3 parts of bupleurum and 1-3 parts of notopterygium wilfordii.

[0012] The mass proportion of the rhubarb can be selected from 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, etc., the mass proportion of atractylodes can be selected from 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, etc., the mass proportion of evodia rutaecarpa can be selected from 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, etc., the mass proportion of asarum can be selected from 1 The mass portions of Radix Bupleuri can be selected as 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts etc. The mass portions of Radix Notopterygii can be selected as 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described one by one here.

[0013] Preferably, the volatile oil of the traditional Chinese medicine composition is prepared by a method comprising the following steps:

[0014] (1) mixing the prepared raw materials and performing ultrasound-microwave pretreatment;

[0015] (2) The pretreated raw material is extracted with CO2 supercritical fluid to obtain the product.

[0016] Preferably, the power of the ultrasound is 50-150W, for example 50W, 60W, 70W, 80W, 90W, 100W, 110W, 120W, 130W, 140W, 150W, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0017] Preferably, the power of the microwave is 100-200 W, for example 100 W, 110 W, 120 W, 130 W, 140 W, 150 W, 160 W, 170 W, 180 W, 190 W, 200 W, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0018] Preferably, the temperature of the ultrasound-microwave pretreatment is 30-60° C. and the time is 1-3 h.

[0019] The temperature can be selected as 30℃, 32℃, 34℃, 36℃, 38℃, 40℃, 42℃, 44℃, 46℃, 48℃, 50℃, 52℃, 54℃, 56℃, 58℃, 60℃, etc., and the time can be 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, 2.2h, 2.4h, 2.6h, 2.8h, 3h, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0020] Preferably, the extraction temperature is 35-45°C, the pressure is 25-35MPa, and the time is 0.5-2h.

[0021] The temperature can be selected from 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, etc., the pressure can be selected from 25MPa, 26MPa, 27MPa, 28MPa, 29MPa, 30MPa, 31MPa, 32MPa, 33MPa, 34MPa, 35MPa, etc., and the time can be selected from 0.5h, 0.6h, 0.7h, 0.8h, 0.9h, 1h, 1.1h, 1.2h, 1.3h, 1.4h, 1.5h, 1.6h, 1.7h, 1.8h, 1.9h, 2h, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be described one by one here.

[0022] Preferably, the extraction comprises a first separation and a second separation.

[0023] Preferably, the temperature of the first separation is 35-45°C and the pressure is 4-8 MPa.

[0024] The temperature can be selected as 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, etc., and the pressure can be selected as 4MPa, 4.5MPa, 5MPa, 5.5MPa, 6MPa, 6.5MPa, 7MPa, 7.5MPa, 8MPa, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be elaborated here one by one.

[0025] Preferably, the temperature of the second separation is 30-40°C and the pressure is 4-8 MPa.

[0026] The temperature can be selected as 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, etc., and the pressure can be selected as 4MPa, 4.5MPa, 5MPa, 5.5MPa, 6MPa, 6.5MPa, 7MPa, 7.5MPa, 8MPa, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be described here one by one.

[0027] Preferably, the flow rate of the CO2 supercritical fluid is 350-400kg / h, for example 350kg / h, 355kg / h, 360kg / h, 365kg / h, 370kg / h, 375kg / h, 380kg / h, 385kg / h, 390kg / h, 395kg / h, 400kg / h, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0028] Preferably, the dosage form of the drug comprises a spray or an aerosol.

[0029] Preferably, the drug further comprises acceptable pharmaceutical excipients.

[0030] Preferably, the pharmaceutical excipients include any one of a diluent, a filler, a pH adjuster, an antioxidant or a buffer, or a combination of at least two of them.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The present invention creatively finds that the mixed volatile oil product prepared with rhubarb, atractylodes, evodia rutaecarpa, asarum, bupleurum and notopterygium wilfordii as raw materials has excellent efficacy in inhibiting Staphylococcus aureus, which is specifically manifested in having a strong antibacterial effect. Compared with Staphylococcus aureus not treated with volatile oil, Staphylococcus aureus treated with volatile oil has differences in multiple metabolic pathways, mainly including secondary biosynthetic metabolism, biofilm-related genes, energy metabolism and other pathways. The results of electron microscopy show that the volatile oil has a destructive effect on both the cell wall and cell membrane of Staphylococcus aureus. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is the total ion current diagram of essential oil sample.

[0034] Figure 2 It is a stacked bar chart of the percentage of each component.

[0035] Figure 3 This is the result of scanning electron microscopy.

[0036] Figure 4 This is the transmission electron microscopy result.

[0037] Figure 5 This is the volcano chart analysis result.

[0038] Figure 6 This is a graph showing the difference in gene expression.

[0039] Figure 7 This is the COG result diagram. DETAILED DESCRIPTION

[0040] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0041] Example 1

[0042] This embodiment provides a traditional Chinese medicine composition, wherein the raw materials for preparing the traditional Chinese medicine composition include, by mass, 1 part of Notopterygium wilfordii, 1 part of Radix Rhei, 1 part of Radix Bupleuri, 1 part of Atractylodes lancea, 1 part of Asarum and 1 part of Evodia rutaecarpa.

[0043] Its preparation method is:

[0044] (1) The raw materials were crushed and subjected to ultrasound-microwave pretreatment at 40° C. for 2 h. The conditions for ultrasound-microwave pretreatment were: ultrasound power 100 W, microwave power 150 W;

[0045] (2) extracting the pretreated Chinese medicine with a CO2 supercritical fluid extraction device, wherein the flow rate of the CO2 supercritical fluid is 380 kg / h, the extraction temperature is 40° C., the pressure is 30 MPa, and the time is 1 hour; the temperature of separation I (first separation) of the supercritical fluid extraction device is 40° C., the pressure is 6 MPa, and a first separation product is obtained; the temperature of separation II (second separation) of the supercritical fluid extraction device is 40° C., the pressure is 6 MPa, and a second separation product is obtained; the first separation product;

[0046] (3) The first separated product and the second separated product are combined to obtain.

[0047] Example 2

[0048] The present embodiment provides a traditional Chinese medicine composition, wherein the raw materials for preparing the traditional Chinese medicine composition include, by weight, 1 part of Notopterygium wilfordii, 3 parts of Radix Rhei, 3 parts of Radix Bupleuri, 3 parts of Atractylodes lancea, 1 part of Asarum and 1 part of Evodia rutaecarpa.

[0049] Its preparation method is:

[0050] (1) The raw materials were crushed and subjected to ultrasound-microwave pretreatment at 30° C. for 1 h. The conditions for ultrasound-microwave pretreatment were: ultrasound power 150 W, microwave power 200 W;

[0051] (2) extracting the pretreated Chinese medicine using a CO2 supercritical fluid extraction device, wherein the flow rate of the CO2 supercritical fluid is 350 kg / h, the extraction temperature is 35° C., the pressure is 25 MPa, and the time is 2 h; the temperature of separation I (first separation) of the supercritical fluid extraction device is 35° C., the pressure is 4 MPa, and a first separation product is obtained; the temperature of separation II (second separation) of the supercritical fluid extraction device is 30° C., the pressure is 4 MPa, and a second separation product is obtained; the first separation product;

[0052] (3) The first separated product and the second separated product are combined to obtain.

[0053] Example 3

[0054] This embodiment provides a traditional Chinese medicine composition, wherein the raw materials for preparing the traditional Chinese medicine composition include, by mass, 3 parts of Notopterygium wilfordii, 1 part of Rhubarb, 1 part of Bupleurum chinense, 1 part of Atractylodes lancea, 3 parts of Asarum and 3 parts of Evodia rutaecarpa.

[0055] Its preparation method is:

[0056] (1) The raw materials were crushed and subjected to ultrasonic-microwave pretreatment at 60° C. for 3 h. The conditions for the ultrasonic-microwave pretreatment were: ultrasonic power 50 W, microwave power 100 W;

[0057] (2) extracting the pretreated Chinese medicine with a CO2 supercritical fluid extraction device, wherein the flow rate of the CO2 supercritical fluid is 400 kg / h, the extraction temperature is 45° C., the pressure is 35 MPa, and the time is 0.5 h; the temperature of separation I (first separation) of the supercritical fluid extraction device is 45° C. and the pressure is 8 MPa, to obtain a first separation product; the temperature of separation II (second separation) of the supercritical fluid extraction device is 40° C. and the pressure is 8 MPa, to obtain a second separation product; the first separation product;

[0058] (3) The first separated product and the second separated product are combined to obtain.

[0059] Example 4

[0060] This embodiment provides a Chinese medicine composition, which differs from Embodiment 1 only in that step (1) in the preparation method is "crushing the preparation raw materials, and ultrasonically pretreating them at 40° C. for 2 h with an ultrasonic power of 150 W", and the component content of the Chinese medicine composition and other steps of the preparation method remain unchanged.

[0061] Example 5

[0062] This embodiment provides a Chinese medicine composition, which differs from Embodiment 1 only in that step (1) in the preparation method is "crushing the preparation raw materials, pre-treating them with microwaves at 40° C. for 2 h, and the microwave power is 200 W", and the component content of the Chinese medicine composition and the other steps of the preparation method remain unchanged.

[0063] Example 6

[0064] This embodiment provides a Chinese medicine composition, which is different from Embodiment 1 only in that the preparation method does not contain step (1), and the content of the ingredients of the Chinese medicine composition and the other steps of the preparation method remain unchanged.

[0065] Comparative Example 1

[0066] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain Notopterygium wilfordii, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0067] The preparation method is similar to that of Example 1.

[0068] Comparative Example 2

[0069] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain rhubarb, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0070] The preparation method is similar to that of Example 1.

[0071] Comparative Example 3

[0072] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain Bupleurum chinense, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0073] The preparation method is similar to that of Example 1.

[0074] Comparative Example 4

[0075] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain Atractylodes lancea, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0076] The preparation method is similar to that of Example 1.

[0077] Comparative Example 5

[0078] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain Asarum, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0079] The preparation method is similar to that of Example 1.

[0080] Comparative Example 6

[0081] This comparative example provides a Chinese medicine composition, which differs from Example 1 only in that it does not contain Evodia rutaecarpa, and its reduced mass is proportionally distributed to the mass of other preparation raw materials.

[0082] The preparation method is similar to that of Example 1.

[0083] Test Example 1

[0084] GC-MS analysis

[0085] 20 μL of the volatile oil sample prepared in Example 1 was taken and placed in a 10 mL headspace gas phase bottle. The GC-MS analysis method is shown in Table 1, and the results are shown in Table 2.

[0086] Table 1

[0087]

[0088] Table 2

[0089]

[0090]

[0091]

[0092] After GC-MS analysis, the total ion current of the sachet volatile oil sample was obtained (such as Figure 1 A total of 43 components were identified from the volatile oil samples (see Table 2). Among the identified components, they are mainly terpenes and alkanes, in addition to a small number of esters and ketones. In the relative content analysis of each component ( Figure 2 ), it was found that phellandrene, β-ocimene, γ-terpinene, 1-chloro-5-methylhexane, β-pinene, ethyl acetate and other components were high in content, and the relative content accounted for more than 5%. The identified components were attributed and it was found that β-elemene came from Atractylodes and Evodia rutaecarpa, β-ocimene came from Rhubarb and Evodia rutaecarpa, and 3-canene, β-myrcene and β-pinene came from Notopterygium wilfordii.

[0093] Test Example 2

[0094] Yield test

[0095]

[0096] Table 3

[0097] Group Yield (%) Example 1 5.58 Example 2 4.81 Example 3 4.93 Example 4 3.37 Example 5 3.85 Example 6 2.17

[0098] It can be seen from the data in Table 3 that the yield of volatile oil obtained by the extraction method described in the present application is relatively high, and pretreatment can increase the yield of volatile oil.

[0099] Test Example 3

[0100] Antibacterial rate

[0101] Preparation of Staphylococcus aureus bacterial solution: Pick a single colony of Staphylococcus aureus Newman and inoculate it into 5 mL TSB medium. After shaking and culturing at 37°C and 200 rpm until the stable period (24 h), 5 μL of the above bacterial solution was inoculated into 5 mL NB (1:1000 dilution), shaken and cultured for 3 h, and diluted 1:10 to make the working bacterial solution, and the CFU was counted.

[0102] Take 1 mL of the volatile oil of the traditional Chinese medicine composition prepared in the above Examples 1-6 and Comparative Examples 1-6, mix them with 1 mL of Staphylococcus aureus bacterial solution, and add TSB medium to a total volume of 10 mL. At the same time, a blank control group without traditional Chinese medicine volatile oil is set up, which contains 1 mL of the prepared bacterial solution and 9 mL of NB medium.

[0103] Samples were taken from the above groups after 24 hours and placed in plates. The plates were placed in 37°C for anaerobic culture for 3 days, and the number of viable bacteria (CFU / mL) on each plate was recorded.

[0104]

[0105] Table 4

[0106]

[0107]

[0108] Test Example 4

[0109] Electron microscopy observation

[0110] Test method:

[0111] Preparation of volatile oil working solution: 200 μL DMSO and 200 μL volatile oil prepared in Example 1 were pipetted and mixed evenly, 100 μL was added to 8000 μL NB culture medium for dilution, and recorded as 100 MIC.

[0112] Preparation of bacterial solution: Pick a single colony of Staphylococcus aureus Newman strain and inoculate it in 6 mL NB. After shaking and culturing at 37°C and 200 rpm for 24 h, take 60 μL of the above bacterial solution and inoculate it into 6 mL NB (1:100 dilution). Inoculate 10 tubes in total and shake and culture at 37°C and 200 rpm for 3 h.

[0113] Cultivation with drug addition: Take 8 shaking bacterial tubes, add bacterial solution and volatile oil working solution according to the table below, two tubes per group, mark them, and culture at 37℃, 200rpm shaking for 5 hours.

[0114] Table 5

[0115] Group Bacterial liquid volume Volatile oil volume control 5mL 0 1MIC 4.95mL 50μL 2MIC 4.9mL 100μL 5MIC 4.75mL 250μL

[0116] Bacteria collection:

[0117] (1) Transmission electron microscopy samples: Centrifuge at 5000 rpm for 5 min to collect the precipitate, add 400 μL of electron microscopy fixative, and place in a 4°C refrigerator after 2 h at room temperature;

[0118] (2) SEM sample collection: After centrifugation, discard the supernatant, rinse gently twice with PBS, add 400 μL of SEM fixative, incubate at room temperature for 2 h, and then place in a 4°C refrigerator.

[0119] Electron microscopy fixative was purchased from Shanghai Weiao Biotechnology Co., Ltd.

[0120] The fixed samples were observed under transmission electron microscope and scanning electron microscope. Figure 3-4 As shown, the transmission electron microscopy results showed that the bacteria in the culture medium control group had a complete morphology, with clear boundaries between the cell wall and the cell membrane, a darker cell wall color, and evenly distributed cytoplasm; the bacterial cell wall in the 1MIC treatment group was thickened and the color became lighter; the boundary between the cell wall and the cell membrane was blurred, and the cell membrane was concave; there were ruptured bacteria; the bacterial morphology in the 2MIC treatment group was further destroyed, the cell wall and the cell membrane were separated, and vacuoles appeared; there were many transparent bacteria and ruptured bacteria debris; the bacterial morphology in the 5MIC treatment group was further severely destroyed, a large number of bacterial cell walls and cell membranes were separated, and vacuoles appeared; there were a large number of transparent bacteria and ruptured bacteria debris. Conclusion: It is suggested that volatile oil has a destructive effect on both bacterial cell walls and cell membranes.

[0121] Test Example 5

[0122] Sequencing

[0123] Preparation of bacterial solution: Pick a single colony of Staphylococcus aureus Newman strain and inoculate it into 5 mL NB medium. After shaking and culturing at 37°C and 200 rpm for 24 h, 1 mL of the above bacterial solution was inoculated into 100 mL NB medium (1:100 dilution), cultured overnight, and CFU counts were performed. The bacterial solution concentration was 8.3×10 8 CFU / 100μL.

[0124] Preparation of volatile oil working solution: 400 μL DMSO and 400 μL volatile oil obtained in Example 1 were pipetted and mixed evenly, and 10 μL was taken and mixed with 40.95 mL NB culture medium.

[0125] Drug-added culture: Take 3 sterilized 250mL conical flasks, add 43.75mL NB culture medium, 6.25mL volatile oil working solution, and 43μL bacterial solution to each, and mix well; take another 3 sterilized 250mL conical flasks, add 50mL NB culture medium and 43μL bacterial solution to each, mix well, and culture at 200rpm and 37℃ for 5h.

[0126] Bacteria collection: After the cultured bacterial solution was centrifuged at 4000rpm for 6min, the supernatant was removed to collect the bacteria, stored in a -80℃ refrigerator, and then sent to the company for sequencing.

[0127] The results are as follows Figure 5-7 As shown in the figure, after volatile oil treatment, 81 genes of Staphylococcus aureus were down-regulated and 73 genes were up-regulated. KEGG results showed that the experimental groups had differences in multiple metabolic pathways, mainly in secondary biosynthesis metabolism, biofilm-related genes, energy metabolism and other pathways.

[0128] The applicant declares that the present invention uses the above-mentioned embodiments to illustrate the application of a volatile oil of a Chinese medicine composition of the present invention in the preparation of a drug for inhibiting Staphylococcus aureus, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0129] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0130] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. Use of a volatile oil of a traditional Chinese medicine composition in the preparation of a drug for inhibiting Staphylococcus aureus, characterized in that: The raw materials for preparing the volatile oil of the traditional Chinese medicine composition include angelica sinensis, rhubarb, bupleurum, atractylodes, asarum and evodia rutaecarpa.

2. Use of a volatile oil of a traditional Chinese medicine composition in the preparation of a Staphylococcus aureus inhibitor, characterized in that: The raw materials for preparing the volatile oil of the traditional Chinese medicine composition include angelica sinensis, rhubarb, bupleurum, atractylodes, asarum and evodia rutaecarpa.

3. The use according to claim 1 or 2, characterized in that: The raw materials for preparing the volatile oil of the traditional Chinese medicine composition include, by weight, 1-3 parts of Notopterygium wilfordii, 1-3 parts of Radix Rhei, 1-3 parts of Radix Bupleuri, 1-3 parts of Atractylodes macrocephala, 1-3 parts of Asarum and 1-3 parts of Evodia rutaecarpa.

4. The use according to any one of claims 1 to 3, characterized in that: The volatile oil of the Chinese medicine composition is prepared by a method comprising the following steps: (1) mixing the prepared raw materials and performing ultrasound-microwave pretreatment; (2) The pretreated raw material is extracted with CO2 supercritical fluid to obtain the product.

5. The use according to claim 4, characterized in that: The power of the ultrasound is 50-150W; Preferably, the power of the microwave is 100-200W; Preferably, the temperature of the ultrasound-microwave pretreatment is 30-60° C. and the time is 1-3 h.

6. The use according to claim 4 or 5, characterized in that: The extraction temperature is 35-45°C, the pressure is 25-35MPa, and the time is 0.5-2h.

7. The use according to any one of claims 4 to 6, characterized in that: The extraction comprises a first separation and a second separation; Preferably, the temperature of the first separation is 35-45°C and the pressure is 4-8MPa; Preferably, the temperature of the second separation is 30-40°C and the pressure is 4-8 MPa.

8. The use according to any one of claims 4 to 7, characterized in that: The flow rate of the CO2 supercritical fluid is 350-400kg / h.

9. The use according to claim 1, characterized in that: The dosage form of the drug includes spray or aerosol; Preferably, the drug further comprises acceptable pharmaceutical excipients.

10. The use according to claim 9, characterized in that: The pharmaceutical excipients include any one of a diluent, a filler, a pH adjuster, an antioxidant or a buffer, or a combination of at least two of them.

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