A plant essential oil composition having anti-candida activity and a preparation method and application thereof

Abstract

This invention discloses a plant essential oil composition with anti-Candida activity and its preparation method, which consists of a heavy oil component of Sichuan pepper and a light oil component of dried ginger in a volume ratio of 1:8 to 2:1. Research in this invention has found that, after specific volume ratios, particularly when the heavy oil component of Sichuan pepper or the light oil component of dried ginger is in a volume ratio of 1:8, it exhibits a synergistic effect against Candida albicans, Candida tropicalis, and Candida glabrata; when the heavy oil component of Sichuan pepper or the light oil component of dried ginger is in a volume ratio of 2:1, it exhibits a very good synergistic effect against Candida parapsilosis. This invention allows the heavy oil component of Sichuan pepper and the light oil component of dried ginger to be formulated into an anti-Candida drug in a specific volume ratio.

Description

Technical Field

[0001] This invention relates to a plant essential oil, specifically to a plant essential oil composition with anti-Candida activity, its preparation method, and its application. Background Technology

[0002] Plant essential oils mainly refer to the volatile components derived from the flowers, leaves, stems, seeds, fruits, roots, or bark of plants. They possess biological activities such as anti-Candida, antioxidant, and anti-tumor properties, and have promising application prospects in the pharmaceutical and food industries.

[0003] The pericarp of Sichuan pepper contains volatile oil, the main components of which are limonene, cumyl alcohol, and geraniol. It also contains phytosterols and various unsaturated organic acids. Sichuan pepper has a fragrant aroma that can increase appetite. It also has blood pressure-lowering effects, as it dilates blood vessels. Furthermore, Sichuan pepper has anthelmintic properties; drinking Sichuan pepper water can expel parasites.

[0004] Dried ginger is pungent and hot in nature, and enters the spleen, stomach, heart, and lung meridians. It has the effects of warming the middle jiao and dispelling cold, restoring yang and unblocking the meridians, drying dampness and resolving phlegm, and warming the lungs and transforming phlegm. It is mainly used to treat symptoms such as cold pain in the epigastrium and abdomen, vomiting, diarrhea, collapse of yang, cough and wheezing due to cold phlegm, and cold-dampness arthralgia.

[0005] There are currently no reports on the application of a combination of heavy oil from Sichuan peppercorns and light oil from dried ginger in the fight against Candida albicans. Summary of the Invention

[0006] Purpose of the invention: The purpose of this invention is to screen out the optimal volume ratio of heavy oil from Sichuan pepper and light oil from dried ginger through a large number of experiments to achieve a synergistic effect in the anti-Candida application.

[0007] Technical solution: To achieve the above objectives, the technical solution adopted by this invention is as follows:

[0008] A plant essential oil composition with anti-Candida activity, which is composed of Sichuan pepper heavy oil component and dried ginger light oil component.

[0009] As a preferred embodiment, the volume ratio of the heavy oil component of Sichuan pepper and the light oil component of dried ginger in the above-mentioned plant essential oil composition with anti-Candida activity is 1:256 to 64:1.

[0010] As a further preferred embodiment, the volume ratio of the heavy oil component of Sichuan pepper and the light oil component of dried ginger in the above-mentioned plant essential oil composition with anti-Candida activity is 1:32 to 32:1.

[0011] As a more preferred embodiment, the volume ratio of the heavy oil component of Sichuan pepper and the light oil component of dried ginger in the above-mentioned plant essential oil composition with anti-Candida activity is 1:8 to 2:1.

[0012] As a particularly preferred embodiment, the volume ratio of the Sichuan pepper heavy oil component and the dried ginger light oil component in the above-described plant essential oil composition with anti-Candida activity is 1:8 or 2:1.

[0013] The plant essential oil composition with anti-Candida activity described in this invention, wherein...

[0014] (1) The Sichuan pepper heavy oil component is prepared by the following method:

[0015] (1.1) Extraction of Sichuan pepper essential oil: Take Sichuan pepper medicinal material, crush it, add water, soak it, then extract it by steam distillation, centrifuge it, collect the Sichuan pepper essential oil, and store it in the dark and cool.

[0016] (1.2) Extraction of Sichuan pepper heavy oil: Sichuan pepper essential oil is put into a molecular distillation feed bottle, liquid nitrogen is put into the cold trap, the refrigeration device is turned on, distillation is performed, and the Sichuan pepper heavy oil component is collected.

[0017] (2) The light ginger oil component was prepared by the following method:

[0018] (2.1) Extraction of ginger essential oil: Take dried ginger, crush it, add water, extract by steam distillation, centrifuge, collect the ginger essential oil, and store it in the dark and cool.

[0019] (2.2) Refining of ginger essential oil: Take ginger essential oil and put it into a molecular distillation feed bottle. Put liquid nitrogen into the cold trap, turn on the refrigeration device, distill, and collect the light oil component of ginger.

[0020] As a preferred embodiment, the above-described plant essential oil composition with anti-Candida activity, wherein...

[0021] (1) The Sichuan pepper heavy oil component is prepared by the following method:

[0022] (1.1) Extraction of Sichuan pepper essential oil: Take Sichuan pepper medicinal material, crush it, add 12 times the amount of water, soak for 1 hour, then extract by steam distillation for 6 hours, centrifuge, collect Sichuan pepper essential oil, and store at 4℃ away from light;

[0023] (1.2) Extraction of Sichuan pepper heavy oil: Sichuan pepper essential oil was put into a molecular distillation feed bottle, liquid nitrogen was put into the cold trap, the refrigeration device was turned on, the temperature of the reflux water device was set to 5℃, the scraper speed was 240r / min, the material flow rate was 1mL / min, the distillation temperature was 50℃ and the distillation pressure was 80Pa, and the Sichuan pepper heavy oil component was collected.

[0024] (2) The light ginger oil component was prepared by the following method:

[0025] (2.1) Extraction of ginger essential oil: Take dried ginger, crush it, add 20 times the amount of water, steam distill for 4 hours, centrifuge, collect the ginger essential oil, and store it at 4℃ away from light.

[0026] (2.2) Refining of ginger essential oil: Take ginger essential oil and put it into a molecular distillation feed bottle. Fill the cold trap with liquid nitrogen, turn on the refrigeration device, set the temperature of the reflux water device to 5℃, the scraper speed to 240r / min, the material flow rate to 1mL / min, the distillation temperature to 90℃ and the distillation pressure to 100Pa, and collect the light oil component of ginger.

[0027] The present invention relates to the application of the plant essential oil composition in the preparation of a drug with anti-Candida activity. The Candida species include Candida albicans, Candida tropicalis, Candida glabrata, Candida subglabrata, or Candida krusei.

[0028] This invention, through screening experiments using different proportions of Sichuan pepper heavy oil and dried ginger light oil to detect anti-Candida species, found that when combined in specific proportions, it can achieve a very good synergistic effect against Candida albicans, Candida tropicalis, Candida glabrata, and Candida parapsilosis.

[0029] This invention enables the preparation of plant essential oil compositions with pharmaceutically acceptable carriers into suppositories, tablets, granules, capsules, pills, sprays, powders, ointments, mixtures, oral liquids, or powder injections.

[0030] A drug with anti-Candida activity, characterized in that it comprises the plant essential oil composition described in this invention.

[0031] Beneficial effects:

[0032] This invention has found that individual components of Sichuan pepper heavy oil or dried ginger light oil have a certain antibacterial effect against Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, or Candida krusei. When formulated in specific volumes, particularly when the individual components of Sichuan pepper heavy oil or dried ginger light oil are in a volume ratio of 1:8, they exhibit a synergistic effect against Candida albicans, Candida tropicalis, and Candida glabrata. In particular, when the individual components of Sichuan pepper heavy oil or dried ginger light oil are in a volume ratio of 2:1, they show a very good synergistic effect against Candida parapsilosis. Although the MIC of individual Sichuan pepper heavy oil or dried ginger light oil components against Candida krusei is low, the combination of the two does not show a significant synergistic effect against Candida krusei. This invention allows for the formulation of anti-Candida drugs by combining Sichuan pepper heavy oil and dried ginger light oil in a specific volume ratio. Attached Figure Description

[0033] Figure 1 This is a graph showing the MIC determination of the heavy oil component of Sichuan pepper against Candida albicans.

[0034] Figure 2 This is a graph showing the MIC determination of the heavy oil component of Sichuan pepper against Candida tropicalis.

[0035] Figure 3 This is a graph showing the MIC determination of the heavy oil component of Sichuan pepper against Candida glabrata.

[0036] Figure 4 This is a graph showing the MIC determination of the heavy oil component of Sichuan pepper against Candida krusei.

[0037] Figure 5 This is a graph showing the MIC determination of the heavy oil component of Sichuan pepper against Candida glabrata.

[0038] Figure 6 This is a graph showing the MIC determination of the light ginger oil component against Candida albicans.

[0039] Figure 7 This is a graph showing the MIC determination of the light ginger oil component against Candida tropicalis.

[0040] Figure 8 This is a graph showing the MIC determination of the light ginger oil component against Candida glabrata.

[0041] Figure 9 This is a graph showing the MIC determination of the light oil component of dried ginger against Candida krusei.

[0042] Figure 10 This is a graph showing the MIC determination of the light oil component of dried ginger against Candida glabrata.

[0043] Figure 11 The MIC determination diagram for the compound of Sichuan pepper heavy oil and dried ginger light oil against Candida albicans.

[0044] Figure 12 The MIC determination diagram for the compound of Sichuan pepper heavy oil and dried ginger light oil for Candida tropicalis.

[0045] Figure 13 The MIC determination diagram for the combination of Sichuan pepper heavy oil and dried ginger light oil for Candida glabrata.

[0046] Figure 14 The MIC determination diagram for the compound of Sichuan pepper heavy oil and dried ginger light oil against Candida krusei.

[0047] Figure 15 The MIC determination diagram for the compound of Sichuan pepper heavy oil and dried ginger light oil against Candida parapsilosis. Detailed Implementation

[0048] The present invention will be further described below with reference to specific embodiments, but these embodiments should not be construed as limiting the present invention.

[0049] Example 1

[0050] 1. Experimental Methods

[0051] 1.1 Extraction of essential oils from Sichuan pepper and dried ginger

[0052] Extraction of Sichuan pepper essential oil: Take a certain amount of Sichuan pepper powder, add 12 times the amount of water, soak for 1 hour, extract by steam distillation for 6 hours, centrifuge, collect the essential oil, and store at 4℃ away from light.

[0053] Extraction of ginger essential oil: Take a certain amount of dried ginger powder, add 20 times the amount of water, steam distill for 4 hours, centrifuge, collect the essential oil, and store at 4℃ away from light.

[0054] 1.2 Refined from Sichuan peppercorn and dried ginger essential oils

[0055] Sichuan pepper essential oil refining: Sichuan pepper essential oil is placed in a molecular distillation feed bottle, liquid nitrogen is placed in the cold trap, the refrigeration device is turned on, the temperature of the reflux water device is set to 5℃, the scraper speed is 240r / min, the material flow rate is 1mL / min, the distillation temperature is 50℃ and the distillation pressure is 80Pa, and the heavy oil component of Sichuan pepper is collected.

[0056] Refining of ginger essential oil: The ginger essential oil is placed in a molecular distillation feed bottle, liquid nitrogen is placed in the cold trap, the refrigeration device is turned on, the temperature of the reflux water device is set to 5℃, the scraper speed is 240r / min, the material flow rate is 1mL / min, the distillation temperature is 90℃ and the distillation pressure is 100Pa, and the light oil component of ginger is collected.

[0057] 1.3 Determination of the minimum inhibitory concentration (MIC) of Sichuan pepper and dried ginger essential oils

[0058] Strain activation: The strain was inoculated onto a sterilized culture medium using the streak method and incubated at 28°C for 48 hours to obtain the activated test strain for later use.

[0059] Pretreatment of test solutions: The Sichuan pepper heavy oil component and the dried ginger light oil component prepared above were filtered through a 0.22 μM filter membrane for sterilization. The Sichuan pepper heavy oil component was diluted to 128 μL / mL and the dried ginger light oil component was diluted to 256 μL / mL with 1% Tween-80 distilled aqueous solution for later use.

[0060] Preparation of the test bacterial suspension: Take a single colony and dilute it with sterile 0.85% physiological saline to a bacterial suspension of 1.8–2.2 McFarland units; dilute the above bacterial suspension 1000 times with RPMI 1640 medium to obtain the ready-to-use bacterial suspension with a concentration of 1×10⁻⁶. 3 ~5×10 3 cfu / mL.

[0061] MIC determination: The MIC was determined by the microbroth dilution method (96-well plate method). 50 μL of sterilized RPMI 1640 medium was added to each well of the 96-well plate; the first column was the positive control, adding 50 μL each of RPMI 1640 medium and the test bacterial solution; the second column was the negative control, adding 50 μL each of RPMI 1640 medium and Tween 80; 50 μL of the test bacterial solution was added to each well of the 3rd - 11th columns, and 50 μL of the essential oil diluted stepwise was added successively to rows A - F. The concentrations of the heavy oil component of Zanthoxylum bungeanum were 32, 16, 8, 4, 2, 1, 0.5, 0.25 μL / mL in sequence, and the concentrations of the light oil component of dried ginger were 64, 32, 16, 8, 4, 2, 1, 0.5 μL / mL in sequence; after the drugs and bacterial solutions were loaded, the plate lid was covered and placed at 28 °C for 48 h. The lowest concentration at which the medium was clear and no bacteria growth was observed with the naked eye was determined as the minimum inhibitory concentration of the drug.

[0062] 1.4 Evaluation of the combined antibacterial activity of the heavy oil component of Zanthoxylum bungeanum and the heavy oil component of dried ginger

[0063] Based on the MIC values of the two essential oils, a series of concentrations of essential oils were prepared. Using the checkerboard method, namely 2MIC, 1 / MIC, 1 / 2MIC, 1 / 4MIC, 1 / 8MIC, 1 / 16MIC, 1 / 32MIC, 1 / 64MIC, and they were combined pairwise. The fractional inhibitory concentration index FICI was used as the judgment basis.

[0064] FICI = MIC A (combined) / MIC A (used alone)+MIC B (combined) / MIC B (used alone)

[0065] The judgment criteria for FICI are as follows: when FICI ≤ 0.5, there is a synergistic effect between the two drugs; when 0.5 < FICI ≤ 1.0, there is an additive effect between the two drugs; when 1.0 < FICI < 4.0, there is no relevant effect between the two drugs; when FICI ≥ 4.0, the two drugs have an antagonistic effect. Based on the FICI judgment criteria, the optimal compound ratio of the drugs was calculated with the test drug combination with the smallest FICI value.

[0066] 2. Experimental results

[0067] 2.1 The MICs of the heavy oil component of Zanthoxylum bungeanum and the light oil component of dried ginger are as shown in Table 1 and Figures 1-10 shown below.

[0068] Table 1 MICs of the heavy oil component of Zanthoxylum bungeanum and the light oil component of dried ginger (μL / mL)

[0069]

[0070] 2.2 Optimal FIC Index of Sichuan pepper heavy oil component and dried ginger light oil component

[0071] 2.2.1 The optimal FIC index for bacterial inhibition is shown in Table 2 below. Figures 11-15 As shown.

[0072] Table 2. FIC Index of the Optimal Combination of Sichuan Pepper Heavy Oil Component and Dried Ginger Light Oil Component

[0073]

[0074] 2.2.2 Results Summary: The optimal volume of the combination of Sichuan pepper heavy oil component and dried ginger light oil component is shown in Table 3 below.

[0075] Table 3

[0076] strain Sichuan pepper enhancement factor Enhanced efficacy of dried ginger Compound effect Optimal volume ratio Candida albicans 4 2 Add 1∶8 Tropical Candida 2 2 Add 1∶8 Candida glabrata 4 2 Add 1∶8 Candida krusei 64 / irrelevant / Candida glabrata 8 8 Collaboration 2∶1 .

[0077] The above experimental results indicate that this invention has found that individual Sichuan pepper heavy oil components or dried ginger light oil components have a certain antibacterial effect against Candida albicans, Candida tropicalis, Candida glabrata, Candida parapsilosis, or Candida krusei. After specific volume formulations, especially when the individual Sichuan pepper heavy oil components or dried ginger light oil components are formulated at a volume ratio of 1:8, they exhibit a synergistic effect against Candida albicans, Candida tropicalis, and Candida glabrata. When the individual Sichuan pepper heavy oil components or dried ginger light oil components are formulated at a volume ratio of 2:1, they show a very good synergistic effect against Candida parapsilosis.

[0078] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of a 2:1 volume ratio of Sichuan pepper heavy oil component and dried ginger light oil component in the preparation of a drug against Candida parapsilosis; The Sichuan pepper heavy oil component is prepared by the following method: (1.1) Extraction of Sichuan pepper essential oil: Take Sichuan pepper medicinal material, crush it, add 12 times the amount of water, soak for 1 hour, then extract by steam distillation for 6 hours, centrifuge, collect Sichuan pepper essential oil, and store at 4℃ away from light; (1.2) Extraction of Sichuan pepper heavy oil: Sichuan pepper essential oil was put into a molecular distillation feed bottle, liquid nitrogen was put into the cold trap, the refrigeration device was turned on, the temperature of the reflux water device was set to 5℃, the scraper speed was 240r / min, the material flow rate was 1mL / min, the distillation temperature was 50℃ and the distillation pressure was 80Pa, and the Sichuan pepper heavy oil component was collected. The aforementioned ginger light oil component is prepared by the following method: (2.1) Extraction of ginger essential oil: Take dried ginger, crush it, add 20 times the amount of water, steam distill for 4 hours, centrifuge, collect the ginger essential oil, and store it at 4℃ away from light; (2.2) Refining of ginger essential oil: Take ginger essential oil and put it into a molecular distillation feed bottle. Fill the cold trap with liquid nitrogen, turn on the refrigeration device, set the temperature of the reflux water device to 5℃, the scraper speed to 240r / min, the material flow rate to 1mL / min, the distillation temperature to 90℃ and the distillation pressure to 100Pa, and collect the light oil component of ginger.

2. The application according to claim 1, characterized in that, A mixture of Sichuan pepper heavy oil and dried ginger light oil in a volume ratio of 2:1, along with a pharmaceutically acceptable carrier, is prepared into suppositories, tablets, granules, capsules, pills, sprays, powders, ointments, mixtures, oral liquids, or powder injections.

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