WO2022068977A2 - Antimicrobial essential oil composition, product comprising same, and preparation method for product thereof - Google Patents

Abstract

Disclosed is an antimicrobial essential oil composition comprising: carvacrol; thymol; and one or more selected from lemon oil, sweet orange oil, daidai peel oil, sweet lemon oil and shaddock oil; one or more selected from the group consisting of Asian peppermint oil, cornmint oil, peppermint oil, and pennyroyal oil; one or more selected from the group consisting of laureth oil and glycine: one or more selected from the group consisting of patchouli oil, sandalwood oil and rosemary oil; one or more selected from the group consisting of geranium oil, pine needle oil, and cedarwood oil; and optionally one or more selected from the group consisting of perilla oil, chrysanthemum oil, and Chinese chaste tree oil. A product containing the antimicrobial essential oil composition, a product containing the antimicrobial essential oil composition, and a preparation method of the product thereof are also disclosed.

Description

Antimicrobial essential oil compositions, articles comprising the same, and methods of making the articles

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims Chinese Invention Patent Application No. 202110482092.1, filed on April 30, 2021, and entitled "Antimicrobial Essential Oil Composition, Products Containing the Same, and Its Preparation Method" and filed on January 19, 2022, entitled Chinese Invention Patent Application No. 202210059971.8 for "Antimicrobial Essential Oil Compositions, Products Containing the Same, and Methods for their Preparation," which is hereby incorporated by reference in its entirety.

technical field

The present application generally relates to an essential oil composition, and more particularly, to a plant essential oil composition with antimicrobial function. The present application also relates to articles of manufacture comprising antimicrobial essential oil compositions and methods of making articles of manufacture comprising antimicrobial essential oil compositions.

Background technique

Many highly pathogenic microbial pathogens cause rapidly onset and spreading infectious diseases. Such infectious diseases can even spread across several provinces, several countries and even the whole world in a short period of time, that is, rapidly develop into a "pandemic". For example, the outbreak of coronavirus disease 2019 (COVID-19, also known as "new coronavirus pneumonia"), which began in late 2019, has been identified by WHO as meeting the characteristics of a "pandemic".

During a pandemic, disinfection is an important task for epidemic prevention and control. At present, synthetic chemical disinfection products (such as ethanol, peracetic acid, chlorine-containing disinfectants, phenolic disinfectants, etc.), ultraviolet rays, heat, etc. are commonly used worldwide to inactivate pathogenic microorganisms. For example, phenolic disinfectants were once one of the main disinfectants in hospitals and played an important role in preventing and controlling the spread of infectious diseases. At high concentrations, phenols can lyse and penetrate the cell wall, causing bacterial proteins to aggregate and precipitate, quickly killing cells; at low concentrations, the enzyme system of bacteria can be inactivated, resulting in cell death. However, such sterilization measures have the risk of inconvenient application, limited use, or the risk of causing secondary hazard pollution to the human body and the environment.

Natural plant-based essential oil products have traditionally been used for thousands of years to treat infectious diseases and prevent and control outbreaks. Phenolic essential oils, in particular, have excellent antibacterial effects. However, this type of essential oil is also toxic and can irritate the skin and mucous membranes, so it is required to be used with caution in practical applications. The antimicrobial activity of other plant essential oils has also been confirmed by many studies. Such plant essential oil products, while inactivating pathogenic microorganisms, are harmless to the human body, are friendly to the environment, and conform to the trend of green environmental protection and sustainable development.

Therefore, there is a need to provide plant essential oil-based products with antimicrobial activity.

We unexpectedly found that by using a combination of a variety of single essential oils, with specific antimicrobial active ingredients, we can obtain a sterilization product with good antimicrobial properties that is harmless to the human body and environmentally friendly.

SUMMARY OF THE INVENTION

In order to solve the above problems, an antimicrobial essential oil composition is provided in an embodiment of the present application, wherein the components of the essential oil composition are all derived from natural plants, and the combination of these components enables the formed essential oil composition to have Good antimicrobial properties.

At least one embodiment of the present application provides an antimicrobial essential oil composition, based on the total weight of the composition, the microbial essential oil composition comprises: (1) 0.0001-80% of one selected from the group consisting of carvacrol and thymol one or more; (2) 0.0001-99.9% of one or more selected from lemon oil, sweet orange oil, tortoise peel oil, white lemon oil and pomelo peel oil; (3) 0.0001-99.9% of One or more selected from Asian peppermint oil, Asian peppermint oil, peppermint oil and peppermint oil; (4) 0.0001-20% of one or more selected from bay leaf oil and glycerin oregano oil (5) 0.0001-20% of one or more selected from patchouli oil, sandalwood oil and rosemary oil; (6) 0.0001-20% of selected from geranium oil, pine needle oil and cedar oil one or more; and (7) optionally, 0.0001-20% of one or more selected from the group consisting of perilla oil, chrysanthemum oil, and vitex leaf oil.

In one aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 2-80% of one or more selected from carvacrol and thymol; (2) 2-50% % selected from one or more of lemon oil, sweet orange oil, tortoise shell oil, white lemon oil and pomelo peel oil; (3) 5-40% selected from Asian peppermint oil, Asian peppermint oil, peppermint oil one or more of peppermint oil and peppermint oil; (4) 2-20% of one or more selected from bay leaf oil and glycero oregano oil; (5) 0.1-5% of One or more selected from patchouli oil, sandalwood oil, and rosemary oil; (6) 2-20% of one or more selected from geranium oil, pine needle oil, and cedar oil; and (7) any Optionally, 2-15% of one or more selected from the group consisting of perilla oil, chrysanthemum oil and vitex leaf oil.

In another aspect, the antimicrobial essential oil composition further optionally comprises one or more selected from the group consisting of: (8) 0.0001-20% honeysuckle oil, based on the total weight of the composition; (9) 0.0001 - 20% Chuanxiong oil; (10) 0.0001-20% cocklebur oil; and (11) 0.0001-20% forsythia oil.

In another aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 5% carvacrol and 5% thymol; (2) 35% lemon oil; (3) 30 (4) 10% bay leaf oil; (5) 0.5% patchouli oil; (6) 5% geranium oil; and (7) 9.5% perilla oil.

In another aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 0.005% carvacrol and 0.005% thymol; (2) 0.1% lemon oil; (3) 99.58 % Asian Peppermint Oil; (4) 0.005% Bay Leaf Oil; (5) 0.25% Patchouli Oil; (6) 0.005% Geranium Oil; and (7) 0.05% Perilla Oil.

In another aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 0.005% carvacrol and 0.005% thymol; (2) 99.5% lemon oil; (3) 0.25% % Asian Peppermint Oil; (4) 0.005% Bay Leaf Oil; (5) 0.18% Patchouli Oil; (6) 0.005% Geranium Oil; and (7) 0.05% Perilla Oil.

In another aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 51.25% carvacrol; (2) 26.25% thymol; (3) 8.75% lemon oil; ( 4) Asian Menthol Oil 7.5%; (5) Bay Leaf Oil 2.5%; (6) Patchouli Oil 0.125%; (7) Geranium Oil 1.25%; and (8) Perilla Oil 2.375%.

In another aspect, based on the total weight of the composition, the antimicrobial essential oil composition comprises: (1) 25% carvacrol and 25% thymol; (2) 2% lemon oil; (3) 10 % Asian Peppermint Oil; (4) 20% Bay Leaf Oil; (5) 0.5% Patchouli Oil; (6) 2% Geranium Oil; (7) 5% Honeysuckle Oil; (9) 4% Chuanxiong oil; (10) 0.3% cocklebur oil; and (11) 0.2% forsythia oil.

In yet another aspect, the microorganism is selected from bacteria, fungi, viruses, mycoplasmas, and combinations thereof.

In yet another aspect, the microorganism is one or more of new coronavirus, influenza virus, rhinovirus, respiratory syncytial virus, enterovirus, adenovirus, herpes virus, coxsackie virus, poliovirus.

In yet another aspect, the microorganism is a novel coronavirus.

In yet another aspect, the microorganism is one or more of Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, catarrhalis, Escherichia coli, Shigella, Staphylococcus albicans, and Candida albicans.

In yet another aspect, the microorganism is Mycoplasma pneumoniae.

At least one embodiment of the present application also provides an article of manufacture comprising the antimicrobial essential oil composition.

In one aspect, the article includes an article capable of reducing or killing microorganisms on one or more of hard surfaces, textiles, skin, mucous membranes, or hair.

On the other hand, the product is selected from the group consisting of face masks, hygienic wipes, air fresheners, oral sprays, hand soaps, shampoos, conditioners, body washes, cosmetics, skin care products, aerogels, bacteriostatic One or more of sprays, surface cleaners and detergents.

In another aspect, the article is a protective article.

In another aspect, the product is one or more selected from the group consisting of civilian masks, medical masks, protective gloves, and protective clothing.

In another aspect, the article is a bead mask.

In another aspect, the article is a food or nutraceutical.

In another aspect, the article is one or more selected from the group consisting of chewing gum, chewing pills, popping beads, gel candies, hard candies, beverages, and dietary supplements.

At least one embodiment of the present application also provides a method for preparing a product comprising an antimicrobial essential oil composition, comprising: uniformly mixing all components of the antimicrobial essential oil composition to prepare an essential oil masterbatch or an essential oil microcapsule or an essential oil Explosive beads, the essential oil master batch or the essential oil microcapsules or the essential oil exploding beads are combined with other components of the product to obtain the product.

In one aspect, the product is a civilian mask or a medical mask, and the combination of the essential oil master batch or the essential oil microcapsules or the essential oil pop beads with other components of the product comprises combining the essential oil master batch or the essential oil microcapsules Or essential oil burst beads are added to the civilian mask or the medical mask.

Description of drawings

In order to illustrate the technical solutions of the embodiments of the present invention more clearly, the accompanying drawings of the embodiments will be briefly introduced below. Obviously, the drawings in the following description only relate to some embodiments of the present disclosure, rather than limit the present disclosure. .

Figure 1 shows the comparison of the inhibition rates of various essential oils of compound compositions to H1N1, H3N, FluB, hRHV and RSV according to at least one embodiment of the present application;

Figure 2 shows the minimum inhibitory concentrations of various essential oils of the present application against a variety of Gram-positive bacteria;

FIG. 3 shows the minimum inhibitory concentrations of various essential oil blends against various Gram-negative bacteria according to at least one embodiment of the present application.

Detailed ways

For the purposes of the following detailed description, it should be understood that the present application may employ various alternative changes and sequences of steps, unless expressly stated to the contrary. Furthermore, except in any operational example, or where otherwise indicated, all numbers representing quantities of ingredients, eg, as used in the specification and claims, should be understood to be modified in all instances by the term "about." Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties to be obtained herein. At least not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, but each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

Furthermore, it should be understood that any numerical range recited herein is intended to include all subranges subsumed therein. For example, a range of "1 to 10" is intended to include (and include) all subranges between said minimum value of 1 and said maximum value of 10, ie having a minimum value equal to or greater than 1 and a minimum value equal to or less than 10 the maximum value of .

In this application, unless expressly stated otherwise, the use of the singular includes the plural and the plural includes the singular. Furthermore, in this application, the use of "or" means "and/or" unless expressly stated otherwise, even though "and/or" may be expressly used in certain circumstances. Furthermore, in this application, the use of "a (a)" or "an (an)" means "at least one" unless expressly stated otherwise. For example, "a" polymer, "a" composition, etc. refers to any one or more of these articles.

In one embodiment of the present application, an antimicrobial essential oil composition is provided. As used herein, the term "antimicrobial" means that the product used has the activity of reducing/killing microorganisms. In the context of this application, "microorganisms" may include bacteria, fungi, viruses, mycoplasmas, etc., such as, but not limited to, coronaviruses (eg, SARS-CoV (SARS virus), MERS-CoV (Middle East respiratory syndrome virus), SARS-CoV-2 (new coronavirus), etc.), hepatitis viruses (such as hepatitis A, B, C), influenza viruses (such as A, B, and avian influenza viruses, etc.), Staphylococcus aureus , Pneumococcus, Typhoid, Candida, Mold, Mycoplasma pneumoniae, etc. In a preferred aspect, the virus in the context of the present application may be a novel coronavirus.

As used herein, the term "essential oils" refers to a general term for volatile fragrance-containing substances obtained by processing and extraction from natural fragrance plants. Generally, plant essential oils are volatile aromas extracted from the flowers, leaves, roots, seeds, fruits, bark, resin, wood hearts and other parts of plants by steam distillation, cold pressing, liposuction or solvent extraction. substance. Plant essential oils often have specific biological activities. In particular, some plant essential oils have good antimicrobial activities, such as antibacterial, antifungal and/or antiviral activities, and can act as effective environmentally friendly antimicrobial agents. Plant essential oils can be divided into single and compound essential oils. Among them, compared with the single essential oil, the compound essential oil contains a variety of effective essential oil components, and through the synergistic effect of each component, significantly better biological activities, such as antimicrobial activity, can be achieved. Essential oil blends are preferably used in this application.

At least one embodiment of the present application provides an antimicrobial essential oil composition, comprising:

(1) one or more selected from carvacrol and thymol, and its amount can be 0.0001-99.99%, preferably 2-80%, more preferably 2-60%, further preferably 6-16%;

(2) one or more selected from lemon oil, sweet orange oil, tortoise peel oil, white lemon oil and pomelo peel oil, the amount of which can be 0.0001-99.99%, preferably 2-80%, more preferably 2 -50%, more preferably 10-50%, further preferably 32-38%;

(3) be selected from one or more in Asian peppermint oil, Asian peppermint oil, peppermint oil and peppermint oil, and its amount can be 0.0001-99.99%, preferably 2-80%, more preferably 5-40% , further preferably 28-35%;

(4) one or more selected from bay leaf oil and gan oregano oil, the amount of which may be 0.0001-20%, preferably 2-20%, more preferably 2-18%, more preferably 2-15%, Further preferred is 8-12%;

(5) One or more selected from patchouli oil, sandalwood oil and rosemary oil, the amount of which may be 0.0001-20%, preferably 0.1-18%, more preferably 0.1-5%, further preferably 0.3-1% %;

(6) one or more selected from geranium oil, pine needle oil and cedar oil, the amount of which can be 0.0001-20%, preferably 2-20%, more preferably 2-18%, further preferably 3-8%,

The above percentages are based on the total weight of the composition.

In one aspect, the antiviral essential oil composition may also optionally include (7) one or more selected from the group consisting of perilla oil, chrysanthemum oil and vitex leaf oil, and the amount may be 0.0001-20%, preferably 2-18%, more preferably 2-15%, further preferably 8-12%, the above percentages are based on the total weight of the composition.

In another aspect, the antiviral essential oil composition may also optionally comprise selected from (8) 0.0001-20% honeysuckle oil; (9) 0.0001-20% chuanxiong oil; (10) 0.0001-20% cocklebur seed oil; and (11) 0.0001-20% of one or more of forsythia oil, the above percentages being based on the total weight of the composition.

For example, but not limited to, a preferred example of the antimicrobial essential oil composition described in this application may comprise: (1) 5% carvacrol and 5% thymol; (2) 35% lemon oil; (3) (4) 10% bay leaf oil; (5) 0.5% patchouli oil; (6) 5% geranium oil; and (7) 9.5% perilla oil, the above percentages are based on total weight of the composition.

For example, but not limited to, another preferred example of the antimicrobial essential oil composition described in this application may comprise: (1) 0.005% carvacrol and 0.005% thymol; (2) 0.1% lemon oil; (3) ) 99.58% Asian Mentha Oil; (4) 0.005% Bay Leaf Oil; (5) 0.25% Patchouli Oil; (6) 0.005% Geranium Oil; and (7) 0.05% Perilla Oil, Percentage Above Based on the total weight of the composition.

For example, but not limited to, yet another preferred example of the antimicrobial essential oil composition described in this application may comprise: (1) 0.005% carvacrol and 0.005% thymol; (2) 99.5% lemon oil; (3) ) 0.25% Asian Menthol Oil; (4) 0.005% Bay Leaf Oil; (5) 0.18% Patchouli Oil; (6) 0.005% Geranium Oil; and (7) 0.05% Perilla Oil, Percentage Above Based on the total weight of the composition.

For example, but not limited to, yet another preferred example of the antimicrobial essential oil composition described herein may comprise: (1) 51.25% carvacrol; (2) 26.25% thymol; (3) 8.75% lemon Oil; (4) Menthol Oil 7.5%; (5) Bay Oil 2.5%; (6) Patchouli Oil 0.125%; (7) Geranium Oil 1.25%; and (8) Perilla 2.375% Oil, the above percentages are based on the total weight of the composition.

For example, but not limited to, yet another preferred example of the antimicrobial essential oil composition described herein may comprise: (1) 25% carvacrol and 25% thymol; (2) 2% lemon oil; (3) 10% Asian Menthol Oil; (4) 20% Bay Leaf Oil; (5) 0.5% Patchouli Oil; (6) 2% Geranium Oil; (7) 5% Honeysuckle Oil; (9) ) 4% chuanxiong oil; (10) 0.3% cocklebur oil; and (11) 0.2% forsythia oil.

The various components in the antimicrobial essential oil composition described in the present application are combined together to achieve excellent antimicrobial properties through synergistic action. For example, the antimicrobial essential oil compositions described herein can exhibit good killing or inhibiting effects on microorganisms selected from bacteria, fungi, viruses, mycoplasmas, or combinations thereof. In one aspect, examples of bacteria described herein can include one or more of Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Catarella, Escherichia coli, Shigella, Staphylococcus albicans, and Candida albicans . In another aspect, examples of viruses described in this application may include one of novel coronavirus, influenza virus, rhinovirus, respiratory syncytial virus, enterovirus, adenovirus, herpes virus, coxsackie virus, poliovirus One or more, for example, the new coronavirus. In yet another aspect, the Mycoplasma described herein can be Mycoplasma pneumoniae.

For example, when using the essential oil composition of the present application to kill the new coronavirus in vitro, the killing efficiency of 3 minutes and 30 minutes of treatment can be higher than 75%, for example, higher than 80%, 85%, 90%, 95% , 98% or even close to 100%. Without wishing to be bound by any theory, the applicant believes that the mechanism by which the essential oil composition kills SARS-CoV-2 may be based, at least in part, on the fact that SARS-CoV-2 has an envelope structure, is a lipophilic (lipophilic) virus, and is theoretically susceptible to being attacked by natural plants Essential oils are adsorbed and enriched, and then inactivated. The applicant's research has found that after the antiviral plant essential oil of the present application is mixed with the new coronavirus stock solution, the new coronavirus can be "extracted" from the stock solution, and then the new coronavirus can be quickly and efficiently inactivated. Complementary and additive activities exist between the various essential oil components selected in this application, which achieves unexpected synergistic work and achieves significantly better antimicrobial properties compared to single essential oils. In addition, the essential oil composition of the present application can also achieve other beneficial effects, including but not limited to, some components in the composition can block the new coronavirus from binding to the ACE2 receptor of respiratory epithelial cells (that is, to achieve blocking/protective effect). ); some ingredients can improve breathing, improve nasal congestion and discomfort caused by early viral upper respiratory tract infection or cold (ie, soothing effect). Moreover, the essential oil composition of the present application is made based on natural plant materials, has a fragrant smell, has many advantages such as low irritation, low corrosion, non-toxic and harmless to human body, and environmental friendliness.

At least one embodiment of the present application also provides an article of manufacture comprising the antimicrobial essential oil composition. Articles of manufacture may include those capable of microbial disinfection of the environment or the human body. In one aspect, the product described in this application that can kill microorganisms in the environment or the human body refers to non-toxic, non-irritating, non-corrosive, environmentally friendly and capable of reducing or killing microorganisms in the environment or on the human body. Products containing harmful microorganisms. It can include, for example, an article capable of reducing or killing microorganisms on one or more of hard surfaces, textiles, skin, mucous membranes, or hair. In the context of this application, the term "hard surface" refers to surfaces of environmental objects other than textile surfaces and human body surfaces, including but not limited to ceilings, walls, floors/tiles, elevators, stairs, handrails, furniture, tableware, vehicles The term "textile" includes but is not limited to clothing, hats, towels, tablecloths, bedding, etc.; the term "skin" refers to human or animal skin; the term "mucosal" refers to human or Oral, ocular, or rectal mucosa of animals; the term "hair" refers to human hair or body hair, or animal hair or feathers. In some examples, an article capable of reducing or killing microorganisms on one or more of hard surfaces, textiles, skin, mucous membranes, or hair can include a product selected from the group consisting of face mask beads, sanitary wipes, air fresheners such as air gel or air spray, etc.), oral spray, personal toiletries (such as hand soap, shampoo, conditioner, body wash, etc.), cosmetics, skin care products, aerogels, antibacterial sprays, surface cleaners (such as furniture one or more of detergents, floor cleaners, kitchen cleaners, toilet cleaners, etc.) and detergents (eg, laundry detergents, dishwashing detergents, etc.).

Additionally or alternatively, the articles described herein may also include protective equipment. For example, protective equipment can include one or more selected from daily clothing (such as underwear, outerwear or socks, etc.), masks (such as civilian masks, medical masks, etc.), protective gloves, protective clothing, for example, explosive beads masks.

Additionally or alternatively, the articles described in this application may also include food or health products. For example, the food or health product may include one or more of chewing gum, chewing pills, popping beads, gel candies, hard candies, soft candies, beverages, and dietary supplements.

At least one embodiment of the present application also provides a method for preparing the antimicrobial essential oil composition as described above, comprising uniformly mixing all components of the antimicrobial essential oil composition. In one aspect, mixing is performed at ambient temperature and pressure.

At least one embodiment of the present application also provides a method for preparing a product comprising an antimicrobial essential oil composition, comprising uniformly mixing all components of the antimicrobial essential oil composition to prepare an essential oil masterbatch or essential oil microcapsules or essential oil popped beads, and The essential oil masterbatch or essential oil microcapsules are combined with other components of the product to obtain the product. For example, the preparation method can include uniformly mixing all the components of the antimicrobial essential oil composition to make essential oil master batch or essential oil microcapsules or essential oil pop beads; and adding the prepared essential oil master batch or essential oil microcapsules or essential oil pop beads into masks (such as civilian masks or medical masks). During use, the essential oil masterbatch or essential oil microcapsules or essential oil popping beads (after being pressed and broken) in the mask thus made will slowly and continuously release the antiviral essential oil composition, thereby preventing the inhalation or discharge through the mask. The pathogenic microorganisms that may be present in the gas (such as air) have a killing effect.

The present application will be further explained in detail below with reference to the examples. However, those skilled in the art will understand that these examples are provided for illustrative purposes only and are not intended to limit the application.

Example

The embodiments of the present application will be described in detail below with reference to the examples, but those skilled in the art will understand that the following examples are only used to illustrate the present application and should not be regarded as limiting the scope of the present application. If the specific conditions are not indicated in the examples, it is carried out according to the conventional conditions or the conditions suggested by the manufacturer. The reagents or instruments used without the manufacturer's indication are conventional products that can be obtained from the market. All amounts listed are described in parts by weight based on the total weight unless otherwise indicated. This application should not be construed as limited to the specific embodiments described.

Preparation Examples 1-13

According to the formulations in the following table 1-2, the components are mixed at normal temperature and pressure to prepare the essential oil compositions of Examples 1-13, wherein Examples 1-7 are the essential oil compositions of the present application, and Examples 8- 13 is a comparative essential oil composition.

Table 1.

[Note]: 1. The numbers in the above table represent the percentage based on the total weight of the composition;

2. "-" indicates that the corresponding component is not included.

Table 2.

[Note]: 1. The numbers in the above table represent the percentage based on the total weight of the composition;

2. "-" indicates that the corresponding component is not included.

Test Example 1

The essential oil composition of Example 1 (labeled as Food Grade Essential Oil (FEO)) and the essential oil composition of Example 3 (labeled as Cosmetic Grade Essential Oil (CEO)) were prepared, respectively.

The cytotoxicity and efficacy of inactivating the novel coronavirus (SARS-Cov-2) of FEO and CEO were tested according to the test method below.

It can be seen from the above results that the essential oil compositions FEO and CEO of the present application have acceptable cytotoxicity and excellent killing effect of the new coronavirus.

Test case 2

The anti-coronavirus (SARS-CoV-2) effects of the essential oil compositions of Examples and Comparative Examples were evaluated at the cellular level in the following manner.

Material

Cells: VeroE6 cells.

Virus: SARS-CoV-2 (South African variant), the titer was TCID ₅₀ =10 ⁻⁵ /100 μL, and the used virus titer was 100 TCID ₅₀ .

Essential oil: The compound essential oil was diluted to 6 concentrations with cell culture medium by double dilution method.

method

(1) Antiviral experiment of the test drug

(1) Test drugs:

Table 2: Compound essential oil cytotoxic drug name, experimental concentration and grouping

Table 3: Drug name, experimental concentration and grouping of compound essential oil cytotoxicity experiments

(2) Sterile 96-well culture plate, add 100 μL of VeroE6 cells at a concentration of 1×10 ⁵ cells/mL to each well, and culture at 37°C and 5% CO ₂ for 24 hours;

(3) Add 100 TCID ₅₀ virus solution 100 μL/well to the experimental group and virus control group of the culture plate, 37°C

5% _CO2 incubator adsorption 2h;

(4) After 2h, discard the cell culture medium in the 96-well culture plate; the test drugs were diluted to the respective concentrations in Table 1, 3 replicate wells for each concentration, and 100 μL/well was added to the above-mentioned drug solution;

(5) Simultaneously set up cell control, blank control (solvent control), virus control (negative control) and comparative drug control;

(6) Cells were incubated at 37°C, 5% _CO2 incubator for 3 days;

(7) The cytopathic changes (CPE) were observed under a light microscope, and the degree of cytopathic changes was recorded according to the following 6-level standards: "-" no lesions appeared; "±" means less than 10% cytopathic; "+" means about 25% cytopathic %; "++" means that about 50% of the cells have lesions; "+++" means that about 75% of the cells have lesions; "++++" means that more than 75% of the cells have lesions. The half effective concentration (IC ₅₀ ) was calculated using the Reed-Muench method or GraphPad Prism 5.0. Criteria for judging the efficacy of drugs: the concentration of CPE with 50% inhibition of the virus is regarded as the effective concentration.

(8) Experimental conditions: The above experimental operations were all completed in the BSL-3 laboratory.

result

1. MTS detects the cytotoxicity of compound essential oils:

When VeroE6 cells grow best, digest the cells with 0.02% EDTA and 0.5% trypsin, adjust the cells to the desired concentration with growth medium, and add the cell liquid to a 96-well cell culture plate, 100 μL/well, at 37°C , 5% CO2 incubator to a dense cell monolayer. The compound essential oil was diluted with medium to different concentrations, respectively 600 μL/L, 300 μL/L, 150 μL/L, 75 μL/L, 37.5 μL/L, 18.75 μL/L; added to the cell culture plate, 100 μL/well. Three replicate wells were made in parallel for each dilution concentration, and a normal cell control was set at the same time. The cells were cultured in a 37°C 5% CO2 incubator, and the cell viability was detected by the MTS method. The specific results are shown in Table 1. The compound essential oils in each group were not toxic to VeroE6 cells at 300 μL/L, so 300 μL/L was selected as the highest experimental concentration in subsequent experiments.

2. CPE observation: After 72 hours of virus infection, the normal cell control group had no CPE, the virus control group had obvious CPE, and different essential oil concentration groups showed different inhibitory effects on CPE. The Reed-Muench method or GraphPad Prism5.0 was used to calculate the half effective concentration (IC ₅₀ ), and the specific results are shown in Table 2. Each group of compound essential oils has antiviral effect, and compared with the comparative example, the embodiment is significantly better than the comparative example.

Test case 3

The in vitro antiviral effects of the essential oil compositions of Examples and Comparative Examples against influenza virus, rhinovirus and respiratory syncytial virus were evaluated at the cellular level in the following manner.

Experimental Materials:

1, test composition: form according to the formula listed in the following table 4, prepare compound essential oil 1-10, wherein, compound essential oil 8 is the essential oil composition of embodiment 2, and compound essential oil 9 is the essential oil composition of embodiment 1 , compound essential oil 10 is the essential oil composition of embodiment 3.

Table 4.

2. Cells: Vero-E6 cells.

3. Virus:

strain name	source
A/PR/8/34(H1N1)	ATCC
A/Aichi/2/68(H3N2)	ATCC
B/Lee/1940(FluB)	ATCC
Human Rhinovirus (hRHV)	ATCC
Respiratory syncytial virus (RSV)	ATCC

experimental method

1. Cell culture and preparation:

1.1 Vero-E6 cell recovery:

The cryopreserved cells were quickly thawed in a 37°C water bath, centrifuged at 800rpm/min for 5 minutes, the cryopreserved solution was removed, 5mL of fresh medium (DMEM, 10% FBS) was added and transferred to a T25 cell culture flask, and the cells were incubated at 37°C with carbon dioxide. Continue to cultivate in the incubator. After 24 hours, observe the cell growth to confirm whether the normal growth is restored.

1.2 Cell passage:

The surface of Vero-E6 cells was washed twice with 5 mL of PBS, and the cells were digested with 1 mL of 0.25% trypsin-EDTA and placed in an incubator. When the cells were observed to become rounded and fall off, 4 mL of fresh medium was added immediately to terminate the digestion. The digested cells were centrifuged at 800 pm/min for 5 minutes and seeded into new T75 culture flasks at a 1:1 ratio.

1.3 Seeding cells:

Vero-E6 cells in logarithmic growth phase were taken, digested with 3 mL of 0.25% trypsin-EDTA, centrifuged and resuspended, then plated into 96-well plates, and placed in a 37°C carbon dioxide incubator for continuous culture.

2. Vero-E6 cytotoxicity test

2.1 Dilution of essential oils:

Take 60 μL of the tested essential oil and add 60 μL of the maintenance medium, mix well, and take 100 μL of the maintenance medium for 10-fold serial dilution in turn.

2.2 Cytotoxicity assay:

After about 20 hours, the density of Vero-E6 reached 90%, discard the upper medium of Vero-E6 cells in the 96-well plate, wash twice with PBS, take the essential oil dilution and add it to the 96-well plate, and set the cell control group (N, Do not add essential oil, use maintenance medium instead). The cell state was visually observed for 72 hours.

3. Virus processing and cell infection

3.1 Mixed treatment of viruses and essential oils:

Take EP tubes and add 200 μL of virus solution, 2 EP tubes for each virus strain, and add 200 μL of essential oil or DMEM solution to the above 2 EP tubes containing virus solutions, each EP tube has a total of 400 μL of mixed solution. After mixing, they were placed at room temperature for 60 minutes, respectively marked as FEO (essential oil plus virus solution mixed for 60 minutes) and Ctrl (DMEM plus virus solution mixed for 60 minutes).

3.2 Dilution of virus essential oil mixture:

Place at room temperature for 60 minutes, take 100 μL from each tube, dilute the virus essential oil mixture 10-fold with virus diluent in turn, and repeat 3 times for each group.

3.3 Cell infection:

In a 96-well plate that has been covered with Vero-E6 cells, 100 μL of essential oil-virus or DMEM-virus dilutions were added to each well, and 6 replicate wells were set for each dilution sample. Set virus positive control wells and normal cell wells, and put them into a 37°C carbon dioxide incubator for continuous cultivation.

3.4 Observation of cytopathic changes:

After 72 hours, the cytopathic conditions were observed under the microscope, the TCID ₅₀ value of each group was calculated according to the Reed-Muench formula, and the inhibition rate of essential oils on the virus was compared.

Distance ratio = (percentage of lesions higher than 50%-50%)/(percentage of lesions higher than 50%-percentage of lesions lower than 50%);

lgTCID50 = distance ratio × log of dilution factor + log of dilution higher than 50% lesion rate

Inhibition rate=(virus titer in control well-virus titer in experimental well)/virus titer in control well×100%

3.5 Experimental conditions:

The above experimental operations were completed in the BSL-3 laboratory.

Experimental results

^1. Cytotoxicity results of the compound essential oil ^: after 72 hours, the cells in the experimental wells died after the essential oil was ^diluted by ^10-1-10-4 ; normal, no significant difference. Therefore, the maximum non-toxic concentration of essential oils to Vero-E6 cells by the naked eye is 10 ^-5 dilution of the essential oil stock solution.

2. The results of the inhibitory effect of essential oils on the virus: 72 hours after virus infection, the normal cell control group had no CPE, the virus control group had obvious CPE, and different essential oil concentration groups showed different inhibitory effects on CPE, and the essential oils in each group were calculated from the virus titers in each group. The virus inhibition rate is shown in Table 5-9 for the specific results. The summary results of the compound essential oil virus inhibition rate are shown in Table 10 and Figure 1. The results show that each group of compound essential oils has a certain antiviral effect, and the compound essential oil 9, 10, and 1 effect Equivalent, better than 7, 8; better than 3, 6; significantly better than 2, 4, 5.

Table 5. Inhibitory effect of compound essential oil on H1N2

Table 6. Inhibitory effect of compound essential oil on H3N2

Table 7. Inhibitory effect of compound essential oil on FluB

Table 8. Inhibitory effect of compound essential oil on hRHV

Table 9. Inhibitory effect of compound essential oil on RSV

Table 10. Comparison of Inhibitory Rates of Compound Essential Oils on H1N1, H3N, FluB, hRHV, RSV

Essential Oil Recipe	H1N1	H3N2	FluB	hRHV	RSV
1	98.68±12.58	95.68±12.58	99.12±3.80	95.28±3.39	98.07±3.65
2	58.72±7.41	58.72±7.41	59.66±6.64	54.30±5.93	53.15±6.64
3	83.12±5.48	73.12±5.48	88.84±12.19	71.09±10.87	76.11±11.9
4	46.00±6.45	56.00±6.45	61.55±11.48	52.51±5.96	46.50±6.52
5	64.84±6.08	64.84±6.08	63.66±6.68	55.99±10.24	50.31±11.2
6	84.68±5.43	84.68±5.43	84.80±7.21	76.72±6.43	78.24±9.97
7	85.65±10.42	85.65±10.42	86.05±3.90	74.27±3.48	83.58±9.61
8	79.12±3.32	79.12±3.32	92.72±5.62	78.25±5.01	83.88±8.19
9	99.83±1.35	98.83±1.35	98.67±4.16	98.49±4.28	97.98±6.67
10	98.72±5.79	96.72±5.79	97.97±2.11	96.13±2.04	97.45±5.49

Test Example 4

The in vitro bacteriostatic effects of the essential oil compositions of Examples and Comparative Examples against various Gram-positive and Gram-negative bacteria were evaluated at the cellular level in the following manner.

Experimental Materials:

1, tested composition: the tested composition used in this test example is identical with the compound essential oil 1-10 used in test example 3, wherein, compound essential oil 8 is the essential oil composition of embodiment 2, and compound essential oil 9 is implemented The essential oil composition of Example 1, compound essential oil 10 is the essential oil composition of Example 3.

2. Standard strains:

Standard strain	source
Streptococcus pneumoniae (No. 31001)	China National Institute for the Control of Pharmaceutical and Biological Products
Staphylococcus aureus (No. 26112)	China National Institute for the Control of Pharmaceutical and Biological Products
Staphylococcus epidermidis (No. 26487)	China National Institute for the Control of Pharmaceutical and Biological Products
Catarella catarrhalis (No. 29103)	China National Institute for the Control of Pharmaceutical and Biological Products
Escherichia coli (Bacteria No. 44155)	China National Institute for the Control of Pharmaceutical and Biological Products
Shigella (No. 51592)	China National Institute for the Control of Pharmaceutical and Biological Products

3. Culture medium: common broth medium, common agar medium and serum agar medium, all prepared according to conventional methods.

experimental method

Three standard gram-positive bacteria (pneumococcus, staphylococcus aureus, and staphylococcus epidermidis) and three gram-negative bacteria (catarrhal, Escherichia coli, dysentery bacillus) were selected, and the compound essential oil was observed in vitro by test tube method. Inhibitory effect on standard strains.

(1) Dilution of essential oil: Dilute 1 ml of essential oil stock solution by 5 times, that is, the concentration of essential oil is 200 μL/ml, and then dilute by doubling dilution method, so that the final concentrations of essential oil liquid are 200, 100, 50, 25, 12.5, 6.25, 3.13 respectively. , 1.56, 0.78 μL/ml in 9 concentrations.

(2) Preparation of bacterial liquid: take the cultures of Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Catarella catarrhalis, Escherichia coli and Shigella dysenteriae cultured for 18h, and dilute them with pH7.0 sodium chloride-peptone buffer solution respectively The number of colonies added to 10 ^-7 -10 ^-5 points.

(3) Test method: The above-mentioned drug sensitive paper sheets containing 1000 CFU of bacterial liquid that were dried in the air were soaked in 8 kinds of compound essential oils of different concentrations for 1 hour, respectively placed in 10 ml of TSB and incubated at 37°C for 48 hours to observe whether the TSB was turbid and inoculated at the same time. Separate the agar plates from each strain and incubate at 37°C for 24h to observe whether there is colony growth. At the same time, a positive control without essential oil liquid and a negative control with essential oil without strain were set, and each was repeated 3 times. The minimum inhibitory concentration (MIC) of the strain was taken as the minimum essential oil concentration for aseptic growth.

Experimental results

Under the conditions of this test, the compound essential oil has different effects on common gram-positive bacteria (pneumococcus, staphylococcus aureus, staphylococcus epidermidis) and gram-negative bacteria (catarrhalis, Escherichia coli, dysentery bacillus). degree of inhibition. It can be seen from the minimum inhibitory concentration results in Table 11, Figure 2, and Figure 3 that Examples 9, 10, and 1 have equivalent effects, better than 3, 6, 7, and 8, and significantly better than 2, 4, and 5.

Table 11. Minimum inhibitory concentration (MIC, μL/ml) of compound essential oil against standard strains in vitro

Test Example 5

According to the method of "Disinfection Technical Specifications" 2002 Edition-2.1.3.4 Air Disinfection Effect Simulation Field Test, the air disinfection effect of the essential oil composition of Example 1 (marked as food grade essential oil (FEO)) against Staphylococcus albus was tested.

Measure 200mL of FEO, add it to the nebulizer, turn on the maximum gear for nebulization for 1 hour, and use the BY-300 air microbiological sampler to sample at an air volume of 28.3 liters/min. The sampling time of the treatment control group was 30 s, the sampling time of the treatment experimental group was 5 min, and the size of the experimental cabin was 20 m ³ . The results of the kill rate experiment were processed to eliminate the influence of the natural extinction factor of microorganisms in the air. The results are shown in Table 12 below.

Table 12.

It can be seen from the above results that the essential oil composition of the present application has a good disinfection effect against Staphylococcus albus in the air.

Test Example 6

With reference to the virus inactivation test method of "Disinfection Technical Specifications" 2002 Edition-2.1.1.10.7, the essential oil composition of Example 1 (marked as food grade essential oil (FEO)) was tested against enterovirus 71, adenovirus, herpes Virus inactivation of viruses, coxsackie virus, influenza A virus H1N1, poliovirus. The results show:

- After treating enterovirus type 71 (ATCC VR-1432, host name: Vero cells) with FEO stock solution for 30 minutes, the average log inactivation value (KL) was 1.36, and the virus inactivation rate was 95.69%;

-After treating the human herpesvirus A5C strain (ATCC VR-2019, host name: Vero cells) with FEO stock solution for 30min, the average logarithm of inactivation (KL)>4.37, and the virus inactivation rate>99.99%;

- After treating adenovirus type 3 (ATCC VR-3, host name: A454 cells) with FEO stock solution for 30 minutes, the average log inactivation value (KL) was 1.61, and the virus inactivation rate was 97.56%;

- After treating Coxsackie virus A21 (ATCC VR-850, host name: Vero cells) with FEO stock solution for 30 minutes, the average log inactivation value (KL) was 1.41, and the virus inactivation rate was 95.97%;

- After treating influenza A virus H1N1: A/PR/8/34 (ATCC VR-1469, host name: MDCK cells) with FEO stock solution for 30min, the average log inactivation value (KL)>4.37, the virus inactivation rate>99.99 %;

- After treating the poliovirus-I vaccine strain (host name: Vero cells) with FEO stock solution for 30 minutes, the average log inactivation value (KL) was 1.24, and the virus inactivation rate was 94.23%;

- The cells of the negative control group grew well, and the experimental results met all the conditions specified in the evaluation.

It can be seen from the above results that the essential oil composition of the present application has good virus inactivation effect against enterovirus 71, adenovirus, herpes virus, coxsackie virus, influenza A virus H1N1 and polio virus.

Test Example 7

With reference to the antibacterial rate detection method of the 2002 edition of "Disinfection Technical Specifications"-2.1.11.3, the essential oil composition (marked as food grade essential oil (FEO)) of Example 1 was tested against Escherichia coli, Staphylococcus aureus, Candida albicans antibacterial rate. The results showed that after treating Escherichia coli, Staphylococcus aureus, and Candida albicans with FEO stock solution for 2 minutes, the bacteriostatic rates were all >99.9%, which met the requirements of "Disinfection Technical Specifications" 2002 Edition - 2.1.11.3.

It can be seen from the above results that the essential oil composition of the present application has a good bacteriostatic effect against Escherichia coli, Staphylococcus aureus and Candida albicans.

Test Example 8

According to the mycoplasma culture method and the detection methods of 2.1.1.7 and 2.1.1.9 of the 2002 edition of "Disinfection Technical Specifications", the essential oil composition of Example 1 (marked as food grade essential oil (FEO)) was tested to inhibit the growth of mycoplasma against Mycoplasma pneumoniae. experiment. The results showed that under the specified experimental conditions, after treating Mycoplasma pneumoniae (ATCC 15531) with FEO stock solution at 25°C for 30 minutes, the logarithmic value of the viable bacteria concentration decreased by >3, and the killing rate was >99%, which had a killing effect on Mycoplasma pneumoniae.

It can be seen from the above results that the essential oil composition of the present application has a good killing effect against Mycoplasma pneumoniae.

Claims (23)

1. An antimicrobial essential oil composition comprising, based on the total weight of the composition:

   (1) 0.0001-80% of one or more selected from carvacrol and thymol;

   (2) 0.0001-99.9% of one or more selected from lemon oil, sweet orange oil, tortoise shell oil, white lemon oil and pomelo peel oil;

   (3) 0.0001-99.9% of one or more selected from Asiamentum mint oil, Asiamentum peppermint oil, peppermint oil and peppermint oil;

   (4) 0.0001-20% of one or more selected from bay leaf oil and glycerin oregano oil;

   (5) 0.0001-20% of one or more selected from patchouli oil, sandalwood oil and rosemary oil;

   (6) 0.0001-20% of one or more selected from the group consisting of geranium oil, pine needle oil and cedar oil; and

   (7) Optionally, 0.0001-20% of one or more selected from the group consisting of perilla oil, chrysanthemum oil and vitex leaf oil.
2. The antimicrobial essential oil composition of claim 1, comprising, based on the total weight of the composition:

   (1) 2-80% of one or more selected from carvacrol and thymol;

   (2) 2-50% of one or more selected from lemon oil, sweet orange oil, tortoise shell oil, white lemon oil and pomelo peel oil;

   (3) 5-40% is selected from one or more in Asian peppermint oil, Asian peppermint oil, peppermint oil and peppermint oil;

   (4) 2-20% of one or more selected from bay leaf oil and glycerin oregano oil;

   (5) 0.1-5% of one or more selected from patchouli oil, sandalwood oil and rosemary oil;

   (6) 2-20% of one or more selected from the group consisting of geranium oil, pine needle oil and cedar oil; and

   (7) Optionally, 2-15% of one or more selected from perilla oil, chrysanthemum oil and vitex leaf oil.
3. The antimicrobial essential oil composition of claim 1 or 2, further optionally comprising one or more selected from the group consisting of:

   (8) 0.0001-20% honeysuckle oil;

   (9) 0.0001-20% Chuanxiong oil;

   (10) 0.0001-20% cocklebur oil; and

   (11) 0.0001-20% forsythia oil.
4. The antimicrobial essential oil composition of any one of claims 1-3, comprising, based on the total weight of the composition:

   (1) 5% carvacrol and 5% thymol;

   (2) 35% lemon oil;

   (3) 30% Asian peppermint oil;

   (4) 10% bay leaf oil;

   (5) 0.5% patchouli oil;

   (6) 5% geranium oil; and

   (7) 9.5% perilla oil.
5. The antimicrobial essential oil composition of any one of claims 1-3, comprising, based on the total weight of the composition:

   (1) 0.005% carvacrol and 0.005% thymol;

   (2) 0.1% lemon oil;

   (3) 99.58% Asian peppermint oil;

   (4) 0.005% bay leaf oil;

   (5) 0.25% patchouli oil;

   (6) 0.005% geranium oil; and

   (7) 0.05% perilla oil.
6. The antimicrobial essential oil composition of any one of claims 1-3, comprising, based on the total weight of the composition:

   (1) 0.005% carvacrol and 0.005% thymol;

   (2) 99.5% lemon oil;

   (3) 0.25% Asian peppermint oil;

   (4) 0.005% bay leaf oil;

   (5) 0.18% patchouli oil;

   (6) 0.005% geranium oil; and

   (7) 0.05% perilla oil.
7. The antimicrobial essential oil composition of claim 1, comprising, based on the total weight of the composition:

   (1) 51.25% carvacrol;

   (2) 26.25% thymol;

   (3) 8.75% lemon oil;

   (4) 7.5% Asian peppermint oil;

   (5) 2.5% bay leaf oil;

   (6) 0.125% patchouli oil;

   (7) 1.25% geranium oil; and

   (8) 2.375% perilla oil.
8. The antimicrobial essential oil composition of any one of claims 1-3, comprising, based on the total weight of the composition:

   (1) 25% carvacrol and 25% thymol;

   (2) 2% lemon oil;

   (3) 10% Asian peppermint oil;

   (4) 20% bay leaf oil;

   (5) 0.5% patchouli oil;

   (6) 2% geranium oil;

   (7) 5% honeysuckle oil;

   (9) 4% Chuanxiong oil;

   (10) 0.3% cocklebur oil; and

   (11) 0.2% forsythia oil.
9. 8. The antimicrobial essential oil composition of any one of claims 1-8, wherein the microorganism is selected from the group consisting of bacteria, fungi, viruses, mycoplasmas, and combinations thereof.
10. The antimicrobial essential oil composition of any one of claims 1-9, wherein the microorganism is a new coronavirus, influenza virus, rhinovirus, respiratory syncytial virus, enterovirus, adenovirus, herpes virus, coxsackie virus , one or more of the polioviruses.
11. The antimicrobial essential oil composition of claim 10, wherein the microorganism is a novel coronavirus.
12. The antimicrobial essential oil composition of any one of claims 1-9, wherein the microorganisms are pneumococcus, staphylococcus aureus, staphylococcus epidermidis, catarrhalis, Escherichia coli, Shigella, Staphylococcus albicans and white One or more of Candida.
13. 9. The antimicrobial essential oil composition of any one of claims 1-9, wherein the microorganism is Mycoplasma pneumoniae.
14. An article of manufacture comprising the antimicrobial essential oil composition of any of claims 1-13.
15. 15. The article of claim 14, wherein the article comprises an article capable of reducing or killing microorganisms on one or more of hard surfaces, textiles, skin, mucous membranes, or hair.
16. The product of claim 14 or 15, wherein the product is selected from the group consisting of face mask popping beads, sanitary wipes, air fresheners, oral spray, hand soap, shampoo, conditioner, body wash, cosmetics, skin care products, air One or more of gels, skin antibacterial sprays, surface cleansers and detergents.
17. 15. The article of claim 14, wherein the article is a protective article.
18. The product of claim 14 or 17, wherein the product is one or more selected from the group consisting of civilian masks, medical masks, protective gloves, and protective clothing.
19. The article of any one of claims 14, 17, and 18, wherein the article is a bead mask.
20. The article of claim 14, wherein the article is a food or a health product.
21. The product of claim 14 or 20, wherein the product is one or more selected from the group consisting of chewing gum, chewing pills, popping beads, gel candies, hard candies, beverages, and dietary supplements.
22. A method of making the article of any of claims 14-21, comprising:

    All components of described antimicrobial essential oil composition are evenly mixed, make essential oil master batch or essential oil microcapsule or essential oil popping beads,

    The product is obtained by combining the essential oil master batch or the essential oil microcapsules or the essential oil popping beads with other components of the product.
23. The method of claim 22, wherein the product is a civilian mask or a medical mask,

    Described combining described essential oil master batch or essential oil microcapsules or essential oil explosive beads with other components of described product comprises adding described essential oil master batch or essential oil microcapsules or essential oil explosive beads to described civilian mask or described medical treatment. in the mask.

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