Method for achieving antibacterial purpose through cooperation of natural plant essential oil and potassium hydrogen persulfate composite salt

Abstract

The invention discloses a method for achieving the antibacterial purpose through cooperation of natural plant essential oil and potassium hydrogen persulfate composite salt, and relates to the technical field of antibacterial treatment. According to the method for synergistically achieving the antibacterial purpose through the natural plant essential oil and the potassium hydrogen persulfate composite salt, the natural plant essential oil with high antibacterial activity is obtained by testing the antibacterial effect of different natural plant essential oil when the natural plant essential oil is independently used, and the natural plant essential oil and the potassium hydrogen persulfate composite salt are compounded through a chessboard method, so that the antibacterial effect of the natural plant essential oil is achieved. The bacteriostatic effect of the compound solution is quantitatively analyzed through part of bacteriostatic concentration indexes, accurate reference data is provided for judgment of the interaction relation between the natural plant essential oil and the potassium hydrogen persulfate compound salt, support is provided for improvement of bacteriostatic efficiency, reduction of the drug use amount and expansion of a bacteriostatic spectrum, and the method has a wide application prospect. The defects of single natural plant essential oil in use are overcome, the cost and the use dosage are reduced, and the natural plant essential oil has a good application prospect in green and healthy livestock and poultry breeding.

Description

Technical Field

[0001] The invention relates to the technical field of antibacterial treatment, in particular to a method for achieving antibacterial purpose by synergistically combining natural plant essential oil and potassium persulfate composite salt. Background Art

[0002] Plant essential oils have attracted great attention as potential substitutes for traditional antibiotics. Plant essential oils are secondary metabolites extracted from the flowers, leaves, fruits and other tissues of plants. Their main components usually include compounds such as alcohols, acids and phenols. The mechanism of action of plant essential oils is complex, making it difficult for pathogens to develop resistance to them. Therefore, they can be used as a class of natural extract antibacterial agents. Up to now, about 300 kinds of plant essential oils have been developed and applied. At the same time, it also has high safety and low biological toxicity, and is not easy to develop drug resistance. Therefore, it has broad application prospects in industries such as biomedicine. It can be used as a potential substitute for antibiotics in green and healthy livestock and poultry farming. The use of plant essential oils to develop natural food preservatives has more advantages in terms of safety and consumer acceptance.

[0003] However, the antibacterial effect of a single plant essential oil is poor, and it is difficult to achieve a good antibacterial effect. There are also problems such as a large amount of a single essential oil and a single odor stimulation. In particular, the cost of natural plant essential oils is relatively high, which limits their wide application. For this reason, a method for synergizing natural plant essential oils with potassium persulfate complex salts to achieve the antibacterial purpose is proposed. Through the combined antibacterial method, not only can the antibacterial efficacy be significantly improved, but it also helps to reduce the amount of drugs used and further expand the antibacterial spectrum, overcome the shortcomings of the use of a single essential oil, and reduce costs and dosages, thereby showing great application potential and prospects in green and healthy livestock and poultry farming. Summary of the invention

[0004] In view of the deficiencies of the prior art, the present invention provides a method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt, which specifically comprises the following steps:

[0005] Preparation of test liquid: Preparation of bacterial suspension: Place the test bacteria in liquid culture medium under sterile conditions, shake and culture at 160r / min and constant temperature of 37℃ on a shaker until the logarithmic growth phase, obtain the bacterial suspension, and then dilute the bacterial suspension to 1×10 8 CFU / mL, stored in a 4°C refrigerator for future use;

[0006] Method determination; experimental data on the inhibition of bacteria by the combination of natural plant essential oil and potassium persulfate compound salt were obtained through experiments;

[0007] Data statistics: GraphPad Prism 8.0 and Origin 2021 software were used for drawing, and SPSS 19.0 statistical analysis software was used for data analysis. P < 0.05 indicated a significant difference, and P < 0.01 indicated an extremely significant difference.

[0008] Obtain conclusion: By comparing the test data, we can obtain the performance data of various natural plant essential oils.

[0009] Preferably, the test bacteria are Staphylococcus aureus and Escherichia coli.

[0010] Preferably, the natural plant essential oils include tea tree essential oil, eucalyptus essential oil, chamomile essential oil, and lavender essential oil.

[0011] Preferably, the antibacterial analysis method also includes an enzyme-labeled analyzer, an electric constant temperature incubator, a high-pressure steam sterilizer and a dual-function gas bath constant temperature oscillator for determination, and the essential oil categories are determined by the above instruments and relevant data are recorded.

[0012] Preferably, the specific determination method includes but is not limited to broth microdilution method, filter paper determination method, time / inhibition curve determination method and chessboard method combined with drug sensitivity test.

[0013] Preferably, when the duration of the antibacterial test is between 4 and 8 hours, the inhibitory effect on the target bacteria is most obvious.

[0014] The present invention provides a method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt. The method has the following beneficial effects:

[0015] The present invention tests the antibacterial effects of different natural plant essential oils and potassium persulfate composite salts when used alone to obtain natural plant essential oils with top antibacterial properties, and compound the natural plant essential oils with potassium persulfate composite salts through a chessboard method, quantitatively analyzes the antibacterial effects of the compounded solutions through a partial antibacterial concentration index, provides accurate reference data for judging the interaction relationship between the natural plant essential oils and the potassium persulfate composite salts, provides support for improving antibacterial efficacy, reducing drug usage, and further expanding the antibacterial spectrum, overcomes the shortcomings of using a single natural plant essential oil, can also reduce costs and usage dosages, and has good application prospects in green and healthy livestock and poultry breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is an analysis flow chart of the present invention;

[0017] Figure 2 This is a time-inhibition curve chart of domestic chamomile essential oil in Escherichia coli in the embodiment of the present invention;

[0018] Figure 3 This is a time-inhibition curve chart of the domestic lavender essential oil in Escherichia coli in the embodiment of the present invention;

[0019] Figure 4 This is a graph showing the time-inhibition curve of the Australian eucalyptus essential oil in Escherichia coli in an embodiment of the present invention;

[0020] Figure 5 This is a graph showing the time-inhibition curve of the Australian tea tree essential oil in Escherichia coli in an embodiment of the present invention;

[0021] Figure 6 It is a time-inhibition curve chart of domestic chamomile essential oil in Staphylococcus aureus in the embodiment of the present invention;

[0022] Figure 7 It is a time-inhibition curve chart of domestic lavender essential oil in Staphylococcus aureus in the embodiment of the present invention;

[0023] Figure 8 It is a graph showing the time-inhibition curve of the Australian eucalyptus essential oil in Staphylococcus aureus in the embodiment of the present invention;

[0024] Fig. 9 This is a graph showing the time-inhibition curve of Australian tea tree essential oil in Staphylococcus aureus in an embodiment of the present invention. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0026] See also Figure 1-9 The embodiment of the present invention provides the following technical solution: a method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt, specifically comprising the following steps:

[0027] Preparation of test liquid: Preparation of bacterial suspension: Place the test bacteria in liquid culture medium under sterile conditions, shake and culture at 160r / min and constant temperature of 37℃ on a shaker until the logarithmic growth phase, obtain the bacterial suspension, and then dilute the bacterial suspension to 1×10 8 CFU / mL, stored in a 4°C refrigerator for future use;

[0028] Method determination; experimental data on the inhibition of bacteria by the combination of natural plant essential oil and potassium persulfate compound salt were obtained through experiments;

[0029] Data statistics: GraphPad Prism 8.0 and Origin 2021 software were used for drawing, and SPSS 19.0 statistical analysis software was used for data analysis. P < 0.05 indicated a significant difference, and P < 0.01 indicated an extremely significant difference.

[0030] Obtain conclusion: By comparing the test data, we can obtain the performance data of various natural plant essential oils.

[0031] In the present invention, the tested bacteria are Staphylococcus aureus and Escherichia coli.

[0032] In the present invention, the natural plant essential oils include tea tree essential oil, eucalyptus essential oil, chamomile essential oil, and lavender essential oil. The selected essential oils include but are not limited to the above essential oils. The purpose is to take a variety of different essential oils for experimental comparison, so as to use them in combination for antibacterial effect.

[0033] In the present invention, the antibacterial analysis method also includes an enzyme-labeled analyzer, an electric constant temperature incubator, a high-pressure steam sterilizer and a dual-function gas bath constant temperature oscillator for determination, and the essential oil categories are determined by the above instruments and relevant data are recorded.

[0034] In the present invention, specific determination methods include but are not limited to microbroth dilution method, filter paper determination method, time / inhibition curve determination method and chessboard method combined with drug sensitivity test.

[0035] In the present invention, when the duration of the antibacterial test is between 4 and 8 hours, the inhibitory effect on the target bacteria is most obvious.

[0036] The present invention discloses for the first time that lavender essential oil and potassium persulfate composite salt exhibit significant additive antibacterial effect on Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in a specific ratio. This discovery not only broadens the application scope of lavender essential oil, but also improves the inhibitory effect on these two common pathogenic bacteria through the synergistic enhancement between lavender essential oil and potassium persulfate composite salt, thus providing a scientific basis for the development of novel, efficient and safe antibacterial products. At the same time, the invention involves the optimization and improvement of the ratio of lavender essential oil and potassium persulfate composite salt, the preparation process, the stability improvement and other aspects, so as to ensure the efficiency, stability and safety of the product in practical applications.

[0037] Furthermore, this patent also explored the combined effect of chamomile essential oil and potassium persulfate complex salt, and found that there is a stronger synergistic antibacterial effect between them, and the inhibitory effect on S. aureus and E. coli far exceeds the use of any one ingredient alone. This discovery deepens our understanding of the interaction between essential oils and chemical disinfectants, and opens up new avenues for the development of antibacterial solutions with stronger bactericidal power and lower irritation.

[0038] Table 2 Evaluation of the antibacterial effect of four highly antibacterial plant essential oils combined with potassium persulfate complex salt

[0039]

[0040]

[0041] As can be seen from Table 2, when the four highly antibacterial plant essential oils are used in combination with potassium persulfate complex salts, the Australian eucalyptus essential oil and potassium persulfate complex salts have antagonistic effects on Staphylococcus aureus and Escherichia coli when used in combination; the Australian tea tree essential oil and potassium persulfate complex salts have no effect on Staphylococcus aureus and Escherichia coli when used in combination; the domestic lavender essential oil and potassium persulfate complex salts have additive effects on Staphylococcus aureus and Escherichia coli when used in combination; the domestic chamomile essential oil and potassium persulfate complex salts have an additive effect when combined to inhibit Escherichia coli, and a synergistic effect when inhibiting Staphylococcus aureus.

[0042] According to the experimental comparison results shown in the chart, we successfully screened out four highly inhibitory essential oils with good antibacterial effects on Staphylococcus aureus (S.aureus) and Escherichia coli (E.coli), namely Australian eucalyptus essential oil, Australian tea tree essential oil, domestic lavender essential oil and domestic chamomile essential oil. Further research found that when these four highly inhibitory essential oils were used in combination with potassium persulfate complex salts, they showed additive or synergistic antibacterial properties. Specifically, lavender essential oil and potassium persulfate complex salts showed additive antibacterial effects when inhibiting S.aureus and E.coli; while the combination of chamomile essential oil and potassium persulfate complex salts showed synergistic antibacterial effects on S.aureus and E.coli, which provided important experimental basis and reference direction for subsequent applications in related fields. .

[0043] In summary, domestic tea tree essential oil, Australian tea tree essential oil, domestic eucalyptus essential oil, Australian eucalyptus essential oil, domestic lavender essential oil, Australian lavender essential oil, domestic chamomile essential oil and Australian chamomile essential oil all have certain antibacterial activity against both Staphylococcus aureus and Escherichia coli pathogens.

[0044] Compared with single plant essential oils, such as the MIC of domestic chamomile essential oil against Staphylococcus aureus and Escherichia coli is 15.63 mg / mL, the MIC of domestic chamomile essential oil combined with potassium persulfate complex salt is reduced to 3.91 mg / mL, showing a stronger antibacterial effect and can effectively reduce the amount of essential oil required.

[0045] The results of the time-inhibition curve test showed that Australian tea tree essential oil, Australian eucalyptus essential oil, domestic chamomile essential oil and domestic lavender essential oil had low inhibition on the target bacteria after 12 hours of action. Plant essential oil shortened or prevented the growth logarithmic phase of the bacteria, causing the bacteria to enter the stable and decay phases in advance, resulting in the inability of the bacteria to divide and proliferate normally to reach the growth number under natural conditions. The reason may be attributed to the complex chemical composition of plant essential oils. According to the different chemical structures of plant essential oils, they can be divided into three categories: aliphatic, aromatic and esters, such as olefins, alcohols, aldehydes, ketones, phenols, monoterpenes, diterpenes and sesquiterpenes. These compounds have the characteristics of low boiling points and are volatile. Therefore, after 12 hours of action, plant essential oils may evaporate in large quantities and lose their antibacterial effect. At the same time, the experiment found that Australian tea tree essential oil, Australian eucalyptus essential oil, domestic chamomile essential oil and domestic lavender essential oil have the strongest inhibition on target bacteria within 4h to 8h of action. If they are used as antibacterial agents in animal husbandry, they can achieve the best antibacterial effect.

[0046] In summary, the minimum inhibitory concentration of plant essential oils, the time-inhibition curve of different concentrations of highly antibacterial plant essential oils, the in vitro drug sensitivity test, and the evaluation of the combined antibacterial effect of plant essential oils and potassium persulfate complex salts showed that tea tree essential oil, eucalyptus essential oil, lavender essential oil and chamomile essential oil from both domestic and Australian sources all showed certain antibacterial effects on Staphylococcus aureus and Escherichia coli, among which Australian tea tree essential oil, Australian eucalyptus essential oil, domestic chamomile essential oil and domestic lavender essential oil had more prominent antibacterial effects. The combination of domestic chamomile essential oil and potassium persulfate complex salt had the best antibacterial effect and had a synergistic effect. The MIC of Staphylococcus aureus and Escherichia coli decreased from 15.63 mg / mL when domestic chamomile essential oil was used alone to 3.91 mg / mL after combined use. The combined effect of domestic chamomile essential oil and potassium persulfate complex salt can provide an effective reference for the research on plant essential oil compound as a natural and safe disinfectant, and promote the healthy and green development of animal husbandry.

Claims

1. A method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt, characterized in that: The specific steps include: Preparation of bacterial suspension: Place the test bacteria in liquid culture medium under sterile conditions, culture on a shaker at 160 r / min and constant temperature of 37°C until the logarithmic growth phase, obtain the bacterial suspension, and then dilute the bacterial suspension to 1×10 8 CFU / mL, stored in a 4°C refrigerator for future use; Method determination; Obtain test data on the inhibition of bacteria by combining natural plant essential oils with potassium persulfate complex salts; Data statistics: GraphPad Prism 8.0 and Origin 2021 software were used for drawing, and SPSS 19.0 statistical analysis software was used for data analysis. P < 0.05 indicated a significant difference, and P < 0.01 indicated an extremely significant difference. Obtain conclusion: By comparing the test data, we can obtain the performance data of various natural plant essential oils.

2. The method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt according to claim 1, characterized in that: The tested bacteria are Staphylococcus aureus and Escherichia coli.

3. The method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt according to claim 2, characterized in that: The natural plant essential oils include tea tree essential oil, eucalyptus essential oil, chamomile essential oil and lavender essential oil.

4. The method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt according to claim 1, characterized in that: The antibacterial analysis method also includes an enzyme-labeled analyzer, an electric constant temperature incubator, a high-pressure steam sterilizer, and a dual-function gas bath constant temperature oscillator for determination. The essential oil categories are determined by the above instruments and relevant data are recorded.

5. The method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt according to claim 1, characterized in that: The specific determination methods include, but are not limited to, broth microdilution method, filter paper determination method, time / inhibition curve determination method and chessboard method combined with drug sensitivity test.

6. The method for achieving antibacterial effect by synergistically combining natural plant essential oil and potassium persulfate composite salt according to claim 5, characterized in that: When the duration of the antibacterial test is between 4 and 8 hours, the inhibitory effect on the target bacteria is most obvious.

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