Plant essential oil amino acid composition and method for producing same

Abstract

The present invention relates to a plant essential oil amino acid composition and a method for producing the same. The composition of the present invention is a composition comprising a plant essential oil and an amino acid, the composition being obtained by mixing the plant essential oil with the amino acid, and the plant essential oil is one or two selected from the group consisting of thymol and carvacrol. The composition of the present invention has a significantly higher melting point and reduced volatility, and therefore has significantly improved stability. The composition of the present invention has improved palatability and reduced die-off rate when used as a growth promoting feed additive for animals.

Description

[Technical field]

[0001] The present invention relates to the technical field of plant essential oil antibiotic substitute products, specifically to a plant essential oil amino acid composition and its preparation method, which can improve plant essential oil stability and reduce volatilization, and the composition has better thermal stability. [Background technology]

[0002] Since 2006, the European Union (EU) has completely banned the use of growth-promoting antibiotic feed additives in livestock and poultry feed, and the current animal health, growth and reproductive performance are all very good. At present, the use of antibiotics in the breeding process in the EU has been significantly reduced, down to 70% of the 2006 level. That is, the use of antibiotics used in treatment has been reduced by 30%, but the health of animals has been greatly improved, which is a great achievement. Since January 1, 2017, the United States has strictly restricted the use of "antibiotics of medical importance to humans" in livestock farming. From 2015 to the present, China has introduced many "antibiotic ban" regulations and policies, sequentially banning the use of a total of eight antibiotics, including lomefloxacin, colistin sulfate and olaquindox, in animal feed and animal husbandry, and in 2020, it has completely banned feed additives using growth-promoting antibiotics in feed. If the use of antibiotics in feed is banned, the antibacterial ingredients of herbs such as carvacrol and thymol components in oregano oil, koji oil, and marjoram oil, which have been used for millennia, will undoubtedly become the preferred antibacterial agents. Carvacrol and thymol have strong bactericidal activity, with a minimum inhibitory concentration (MIC) of only 0.12 to 50 ppm, and can improve the immune capabilities of animals. In addition, due to their unique bactericidal mechanism, bacteria do not develop resistance, and they can be artificially synthesized in large quantities.

[0003] Thymol and carvacrol have antibacterial, antiseptic and antioxidant properties and are widely used in oral hygiene and food additives. The chemical structure of thymol is shown in Formula I, and the chemical structure of carvacrol is shown in Formula II:

[0004] [ka]

[0005] Both are plant essential oils and are the main components (60-80%) of oregano oil extract. Because they have synergistic antibacterial and bactericidal activity, they are rarely used alone and are the main active ingredients in commercially available essential oil antibiotic replacement products. However, thymol and carvacrol have low melting points (50°C for thymol and 2°C for carvacrol), making them thermally unstable and prone to volatilization during production, storage, transportation, and granulation as feed additives, resulting in large losses of active ingredients. Various methods have been developed to improve the stability of thymol and carvacrol during long-term storage and high-temperature granulation. For example, CN101331915A reports the direct mixing of thymol and eugenol with benzoic acid. CN103478232A reports the inclusion mixture of carvacrol and β-cyclodextrin to achieve a sustained release effect. CN107753462A reports the use of a polymer material matrix to protect thymol and carvacrol. However, the above-mentioned existing techniques cannot significantly improve the stability of thymol and carvacrol during long-term storage or high-temperature granulation process.

[0006] Therefore, there is a need to develop new technologies that can address the chemical stability of plant essential oils. Summary of the Invention

[0007] Therefore, the present invention deals with the above-mentioned shortcomings of essential oil antibiotic substitute products, and has studied the development of a product manufactured with high stability and low volatility. It has been found by chance that the composition is formed by using the interaction of sarcosine or proline with thymol or carvacrol, and the melting point and stability of the composition are greatly improved. The present invention has obtained the composition by adding amino acids as specific auxiliary compounds that interact with thymol or carvacrol at the molecular level in the presence of non-covalent bonds. At the same time, the combination of thymol and carvacrol has a remarkable synergistic antibacterial, bactericidal and antioxidant effect. Compared with carvacrol in a liquid state at room temperature, the preparation of a stable solid powder of the composition is more convenient for application in the fields of food additives, pharmaceutical additives and feed additives. In the present invention, it has been found that the melting points of thymol and carvacrol are significantly improved after the formation of the composition (e.g., melting point of thymol-proline 1:1 (molar ratio) composition: 123°C; melting point of carvacrol-proline 1:1 (molar ratio) composition: 133°C), and thus its thermal stability is improved. In addition, the method for producing the composition is simple and reproducible, which is advantageous for processing and improving the physical and chemical properties.

[0008] Therefore, the present invention provides a composition of plant essential oil and amino acid, which is a composition that improves the stability and reduces the volatility of plant essential oil, said plant essential oil can be thymol and / or carvacrol, and said amino acid can be sarcosine and / or proline. The composition of the present invention can significantly improve the melting point and reduce the volatility of plant essential oil, thereby significantly improving the stability. The composition of the present invention has better palatability and lower die-off rate when used as a growth promoting feed additive for animals.

[0009] One object of the present invention is to provide a composition comprising a plant essential oil and an amino acid.

[0010] A second object of the present invention is to provide a method for producing a composition containing a plant essential oil and an amino acid.

[0011] A third object of the present invention is to provide a product containing the above-mentioned composition of plant essential oils and amino acids.

[0012] A fourth object of the present invention is to provide a use of a composition containing the above-mentioned plant essential oil and amino acid in the manufacture of a supplement, food, cosmetic, pharmaceutical, pharmaceutical auxiliary, or feed.

[0013] According to a first aspect of the present invention, there is provided a composition comprising a plant essential oil and an amino acid, said composition being obtainable by mixing a plant essential oil with an amino acid, said plant essential oil being one or two selected from the group consisting of thymol and carvacrol, and said amino acid being one or two selected from the group consisting of L-proline and sarcosine.

[0014] Preferably, the plant essential oil and amino acid are mixed in a molar ratio of 9-1:1-9, preferably 2-1:1-2, for example, the molar ratio may be 1:9, 1:2, 1:1, 2:1, 9:1, etc., but is not limited thereto.

[0015] Preferably, the plant essential oil is a mixture of thymol and carvacrol, in which the molar ratio of thymol to carvacrol may be 9-1:1-9, preferably 2-1:1-2.

[0016] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 1:2.

[0017] In one embodiment, the composition of the invention is obtained by mixing thymol and L-proline in a 1:1 molar ratio.

[0018] In one embodiment, the composition of the present invention is obtained by mixing carvacrol and L-proline in a 1:1 molar ratio.

[0019] In one embodiment, the composition of the present invention is obtained by mixing carvacrol and L-proline in a molar ratio of 2:1.

[0020] In one embodiment, the composition of the present invention is obtained by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:2.

[0021] In one embodiment, the composition of the present invention is obtained by mixing thymol, carvacrol and sarcosine in a molar ratio of 1:1:2.

[0022] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 2:1.

[0023] In one embodiment, the composition of the invention is obtained by mixing thymol and L-proline in a molar ratio of 1:2.

[0024] In one embodiment, the composition of the present invention is obtained by mixing carvacrol and L-proline in a molar ratio of 1:2.

[0025] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 2:1.

[0026] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 1:3.

[0027] In one embodiment, the composition of the present invention is obtained by mixing carvacrol and sarcosine in a molar ratio of 1:2.

[0028] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 1:9.

[0029] In one embodiment, the composition of the present invention is obtained by mixing thymol and sarcosine in a molar ratio of 9:1.

[0030] In one embodiment, the composition of the invention is obtained by mixing thymol and L-proline in a molar ratio of 1:9.

[0031] In one embodiment, the composition of the invention is obtained by mixing thymol and L-proline in a molar ratio of 9:1.

[0032] In one embodiment, the composition of the present invention is obtained by mixing carvacrol and L-proline in a molar ratio of 1:9.

[0033] In one embodiment, the composition of the invention is obtained by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:18.

[0034] In the composition of the present invention, the mixing is preferably achieved by any one of the following methods (1) and (2):

[0035] (1) Recrystallizing the plant essential oil and amino acid in a solvent to obtain a white precipitate, and then drying the precipitate to obtain the composition of the present invention;

[0036] (2) Mixing the plant essential oil and the amino acid, and feeding them into a mixing device to thoroughly contact them by mechanical force, or spray-drying the mixture solution, or processing it through an extruder to obtain a composition containing the plant essential oil and the amino acid after partial or complete reaction.

[0037] In the composition of the present invention, in the above method (1), the solvent includes all solvents that have a certain solubility for the raw materials and do not alter the raw materials, and is preferably one or more selected from the group consisting of water, alcohols, ketones, esters, alkanes, aromatic hydrocarbons, haloalkanes, and the like; more preferably, the solvent is one or more selected from the group consisting of methanol, ethanol, ethyl acetate, acetone, and dichloromethane.

[0038] In the composition of the present invention, in the above method (ii), the grinding device is preferably selected from the group consisting of a ball mill, a grinder, a blender and a stirrer.

[0039] The above-mentioned method (1) and method (2) are simple in operation, easy to control the crystallization process, have a high degree of crystallization, and are reproducible, and can stably obtain a composition containing thymol and / or carvacrol and an amino acid.

[0040] In one preferred embodiment of the present invention, the composition comprising plant essential oils and amino acids comprises thymol and Sarcosine in a molar ratio of 1:2. Preferably, the composition has an X-ray powder diffraction pattern with characteristic peaks at 2θ angles of about 6.4°±0.2°, 12.1°±0.2°, 12.4°±0.2°, 17.7°±0.2°, 20.2°±0.2°, 22.1°±0.2°, 23.0°±0.2°, 23.7°±0.2°, and 25.1°±0.2°. More preferably, the composition has an X-ray powder diffraction pattern essentially as shown in FIG. 1. Preferably, the composition has a differential scanning calorimetry diagram with a characteristic endothermic peak at about 129±2° C.; more preferably, the composition has a differential scanning calorimetry diagram essentially as shown in FIG. 2.

[0041] In one preferred embodiment of the present invention, the composition comprising a plant essential oil and an amino acid is obtained by mixing thymol and L-proline in a molar ratio of 1:1. Preferably, the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of about 6.2°±0.2°, 6.9°±0.2°, 12.5°±0.2°, 13.8°±0.2°, 18.0°±0.2°, 18.4°±0.2°, 18.7°±0.2°, 19.1°±0.2°, 19.3°±0.2°, 21.1°±0.2°, and 24.8°±0.2°. More preferably, the composition has an X-ray powder diffraction pattern essentially as shown in FIG. 3. Preferably, the differential scanning calorimetry diagram of the composition has a characteristic endothermic peak at about 123±2° C.; more preferably, the differential scanning calorimetry diagram has a characteristic endothermic peak at about 123±2° C.; more preferably, the composition has a differential scanning calorimetry diagram essentially as shown in FIG. 4.

[0042] In one preferred embodiment of the present invention, the composition comprising plant essential oil and amino acid is obtained by mixing carvacrol and L-proline in a molar ratio of 1:1. Preferably, the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of about 6.9°±0.2°, 11.5°±0.2°, 13.6°±0.2°, 13.9°±0.2°, 19.0°±0.2°, 19.2°±0.2°, 23.1°±0.2°, and 24.9°±0.2°. More preferably, the composition has an X-ray powder diffraction pattern essentially as shown in FIG. 5. Preferably, the differential scanning calorimetry diagram of the composition has a characteristic endothermic peak at about 133±2° C.; more preferably, the differential scanning calorimetry diagram essentially as shown in FIG. 6.

[0043] In one preferred embodiment of the present invention, the composition comprising plant essential oils and amino acids is obtained by mixing carvacrol and L-proline in a molar ratio of 2:1. Preferably, the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of about 6.5°±0.2°, 11.0°±0.2°, 13.1°±0.2°, 13.2°±0.2°, 15.0°±0.2°, 15.5°±0.2°, 15.8°±0.2°, 17.5°±0.2°, 19.7°±0.2°, 20.5°±0.2°, 21.6°±0.2°, 21.9°±0.2°, 23.6°±0.2°, 25.1°±0.2°, and 26.7°±0.2°. More preferably, the composition has an X-ray powder diffraction pattern essentially as shown in FIG. 7. Preferably, the composition has a differential scanning calorimetry pattern with a characteristic endothermic peak at about 80±2° C.; more preferably, the composition has a differential scanning calorimetry pattern essentially as shown in FIG.

[0044] In one preferred embodiment of the present invention, the composition comprising plant essential oil and amino acid is obtained by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:2. Preferably, the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of about 6.2°±0.2°, 6.8°±0.2°, 12.5°±0.2°, 13.8°±0.2°, 18.4°±0.2°, 18.8°±0.2°, 19.4°±0.2°, 21.3°±0.2°, 24.8°±0.2°, 25.0°±0.2°. More preferably, the composition has an X-ray powder diffraction pattern essentially as shown in FIG. 9. Preferably, the composition has a differential scanning calorimetry diagram with a characteristic endothermic peak at about 125±2° C.; more preferably, the composition has a differential scanning calorimetry diagram essentially as shown in FIG. 10.

[0045] As can be seen by comparison, the X-ray powder diffraction pattern of the composition obtained by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:2 is essentially a superposition of the X-ray powder diffraction patterns of the composition obtained by mixing thymol and L-proline in a molar ratio of 1:1 and the composition obtained by mixing carvacrol and L-proline in a molar ratio of 1:1, and the differential scanning calorimetry diagram was affected by components other than the composition, and its characteristic endothermic peak was different from the characteristic endothermic peak of the corresponding composition.

[0046] According to a second aspect of the present invention, there is provided a method for producing a composition comprising a plant essential oil and an amino acid, the method comprising a step of mixing the plant essential oil and the amino acid to obtain a composition comprising the plant essential oil and the amino acid.

[0047] In the production method of the present invention, the amino acids are preferably one or two selected from the group consisting of L-proline and sarcosine.

[0048] In the production method of the present invention, the plant essential oil and the amino acid are preferably mixed in a molar ratio of 9-1:1-9, preferably 2-1:1-2, for example, the molar ratio may be 1:9, 1:2, 1:1, 2:1, 9:1, etc., but is not limited thereto.

[0049] In the production method of the present invention, preferably, the molar ratio of thymol to carvacrol in the plant essential oil is 9-1:1-9, and preferably 2-1:1-2.

[0050] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 1:2.

[0051] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and L-proline in a 1:1 molar ratio.

[0052] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing carvacrol and L-proline in a 1:1 molar ratio.

[0053] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing carvacrol and L-proline in a molar ratio of 2:1.

[0054] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:2.

[0055] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing thymol, carvacrol and sarcosine in a molar ratio of 1:1:2.

[0056] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 2:1.

[0057] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 1:3.

[0058] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and L-proline in a molar ratio of 1:2.

[0059] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing carvacrol and L-proline in a molar ratio of 1:2.

[0060] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 2:1.

[0061] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing carvacrol and sarcosine in a molar ratio of 1:2.

[0062] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 1:9.

[0063] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and sarcosine in a molar ratio of 9:1.

[0064] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and L-proline in a molar ratio of 1:9.

[0065] In one embodiment, the process of the invention comprises the step of obtaining said composition by mixing thymol and L-proline in a molar ratio of 9:1.

[0066] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing carvacrol and L-proline in a molar ratio of 1:9.

[0067] In one embodiment, the process of the present invention comprises the step of obtaining said composition by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:18.

[0068] In the production method of the present invention, the mixing is preferably achieved by either of the following methods (1) and (2): (1) recrystallizing the plant essential oil and the amino acid in a solvent to obtain a precipitate, and then drying the precipitate to obtain a composition containing the plant essential oil and the amino acid; (2) mixing the plant essential oil and the amino acid, feeding them into a grinding device to thoroughly contact them by mechanical force, or spray-drying the mixture solution, or processing it through an extruder to form a composition containing the plant essential oil and the amino acid after partial or complete reaction.

[0069] In the production method of the present invention, in the above method (1), the solvent includes all solvents that have a certain solubility for the raw materials and do not alter the raw materials, and is preferably one or more selected from the group consisting of water, alcohols, ketones, esters, alkanes, aromatic hydrocarbons, haloalkanes, and the like; more preferably, the solvent is one or more selected from the group consisting of methanol, ethanol, ethyl acetate, acetone, and dichloromethane.

[0070] In the production method of the present invention, in the above-mentioned method (ii), the grinding device is preferably selected from the group consisting of a ball mill, a grinder, a blender, and a stirrer.

[0071] In one embodiment, the plant essential oil and amino acid are added to a ball mill at a shaking frequency of 5-60 Hz, preferably 30 Hz, for a shaking time of 5-60 minutes, preferably 20 minutes, to obtain the composition of the present invention.

[0072] In one embodiment, the plant essential oil and amino acid are added to a grinder, and the grinding time is 1 to 60 minutes, preferably 2 minutes, to obtain the composition of the present invention.

[0073] In the production method of the present invention, the above-mentioned method (1) and method (2) are simple in operation, the crystallization process is easy to control, the degree of crystallization is high, and reproducibility is good, so that a composition containing a plant essential oil and an amino acid can be stably obtained.

[0074] According to a third aspect of the present invention, there is provided a product, such as a supplement, food, cosmetic, pharmaceutical, pharmaceutical aid or feed, comprising the above-mentioned plant essential oil and amino acid composition.

[0075] The product may further include other suitable ingredients required for the product. For example, the food may include a food base and edible food additives that are acceptable for food, such as sweeteners, flavorings, preservatives, flavorings, colorings, etc. The cosmetics may include cosmetically acceptable cosmetic bases and additives, such as solvents, flavorings, preservatives, essences, colorings, etc. The pharmaceuticals may include medicamentous active ingredients and pharmacologically acceptable auxiliaries, such as carriers, diluents, adjuvants, colorings, etc. The feed may include feed bases, such as soybean meal, hay, etc., and feed-acceptable feed auxiliaries, such as sweeteners, flavorings, preservatives, flavorings, colorings, etc. However, the present invention is not limited thereto.

[0076] The above-mentioned product is produced by adding the composition containing plant essential oil and amino acid according to the present invention. Except for adding the composition containing plant essential oil and amino acid according to the present invention, the production method of the above-mentioned product can be the same as that of the conventional method.

[0077] The different ratios described herein are all molar ratios; the term feed employed means any compound, preparation, mixture or composition suitable for or intended to be ingested by an animal. The auxiliary compounds described herein have molecular level interactions with part or all of the main molecule. The thymol and carvacrol described herein apply to natural and synthetic sources.

[0078] Although the present invention has been described in detail above, the above embodiments are merely exemplary in nature and are not intended to limit the present invention. Furthermore, the present specification is not limited by any of the theories described in the prior art or summary of the invention or the following examples.

[0079] Unless otherwise stated, numerical ranges throughout this application include any subranges therein and any numerical value incremented by the smallest subunit of the given value therein. Unless otherwise stated, numerical values ​​throughout this application include small deviations from the given value and have values ​​approximately recited, and represent approximate measures or limits of the range of embodiments having the exact value recited. Except for the working examples provided at the end of the detailed description, the values ​​of parameters (e.g., amounts or conditions) in this application, including the appended claims, are understood to be modified in all cases by the term "about", regardless of whether "about" actually appears before the value. "About" means that the stated value is allowed to be slightly inaccurate (the precision of the value is somewhat close; roughly, or reasonably close to the value; approximate value). If the inaccuracy provided by "about" is not understood in this ordinary sense in the art, "about" as used herein at least indicates the variation that may occur with the ordinary methods of measuring and using these parameters. For example, "about" can include variations of up to 10%, up to 5%, up to 4%, up to 3%, up to 2%, up to 1%, or up to 0.5%, and in some embodiments up to 0.1%.

[0080] Unless otherwise indicated, the terms "comprising," "including," "having," "containing," or other similar terms used throughout this application are open ended terms indicating that a composition or product may contain, in addition to the elements recited herein, other elements not expressly recited but normally inherent in a composition or product. Also, in this specification, the terms "comprising," "including," "having," and "containing" are considered to be specifically disclosed and to include both the closed or semi-closed conjunctions "consisting of" and "consisting essentially of." "Consisting essentially of" means that the elements described herein constitute 95% or more, 97% or more, and in some embodiments 99% or more of the composition or product.

[0081] Beneficial effects The present invention provides a composition that combines thymol and / or carvacrol, the main active ingredient of phenols, with amino acids to realize molecular level interactions, in response to the problem of poor stability and easy volatilization and loss during use of essential oil products. The melting points of compositions 1 to 4 of the present invention were increased by 79, 73, 131 and 78°C, respectively, compared to the spices themselves. The mechanism of the composition of the present invention is to increase the melting points of thymol and carvacrol, which results in a significant reduction in their vapor pressure, thereby suppressing volatilization and improving the stability of spices during storage and processing. For example, composition 5 showed better stability than commercial products both during storage and granulation processing (Test Examples 4 and 5). At the same time, the advantages of the composition provided by the present invention in the use of animal feed additives are that it is difficult to volatilize, has good palatability, improves stability, and is manifested in the quality and quantity of the condition inside the animal's body, fully exerts antibacterial properties, and reduces the impact of die-off rate. The test results of meat ducks showed that it has the effect of significantly reducing the die-off rate. [Brief description of the drawings]

[0082] [Figure 1] 1 is an X-ray powder diffraction (XRPD) pattern of composition 1. [Diagram 2] FIG. 1 is a differential scanning calorimetry (DSC) diagram of composition 1. [Diagram 3] 1 is an X-ray powder diffraction (XRPD) pattern of composition 2. [Figure 4] FIG. 2 is a differential scanning calorimetry (DSC) diagram of composition 2. [Diagram 5] 1 is an X-ray powder diffraction (XRPD) pattern of composition 3. [Figure 6] FIG. 2 is a differential scanning calorimetry (DSC) diagram of composition 3. [Figure 7] 1 is an X-ray powder diffraction (XRPD) pattern of composition 4. [Figure 8] FIG. 1 is a differential scanning calorimetry (DSC) diagram of composition 4. [Figure 9] 1 is an X-ray powder diffraction (XRPD) pattern of composition 5. [Figure 10] FIG. 1 is a differential scanning calorimetry (DSC) diagram of composition 5. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0083] The present invention will be further described below with reference to specific examples, but the present invention is not limited to these.

[0084] All experimental methods used in the following examples are conventional methods unless otherwise specified.

[0085] All materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

[0086] Thymol and carvacrol were purchased from Nanjing Jiulonghui Spices Co., Ltd; sarcosine and L-proline were purchased from Sinopharm Chemical Reagent Co., Ltd.

[0087] The ball mill was purchased from Shanghai Jingxin Industrial Development Co., Ltd.

[0088] The crusher was purchased from Xinlongwei Machinery Parts Co., Ltd.

[0089] Inspection equipment and analysis methods:

[0090] X-ray powder diffraction (XRPD) method, instrument model: Bruker D8 advance, target: Cu Kα (40 kV, 40 mA), peak position was calibrated with a standard sample attached to the instrument before use. Sampling software was Diffrac Plus XRD Commander, and analysis software was MDI Jade 6.0. Samples were tested at room temperature, and samples requiring measurement were placed on an organic glass plate. Distance from sample to detector: 30 cm, scan range: 3-40° (2θ value), scan step: 0.02°, step width: 0.1 sec / step.

[0091] Differential scanning calorimetry (DSC) method, instrument model: TA DSC Q2000, temperature range: 50-200°C, scan rate: 10°C / min, nitrogen flow rate: 50mL / min.

[0092] Thermogravimetric analysis (TGA) method: Netzsch TG 209F3 thermogravimetric analyzer, temperature: 80°C, maintained for 100 minutes, purge gas: 25mL / min.

[0093] The analysis method for thymol and carvacrol content was based on the China Ministry of Agriculture industry standard NY / T 3137-2017.

[0094] Manufacturing Example

[0095] Preparation Example 1: Preparation of Composition 1

[0096] The process for preparing Composition 1 was as follows.

[0097] 5 mmol of thymol and 10 mmol of sarcosine were added to 20 mL of methanol in a molar ratio of 1:2, and stirred at 60°C for 1 h. The mixture was then recrystallized to obtain a white precipitate. The precipitate was filtered and placed in a vacuum desiccator to dry at room temperature to obtain composition 1 formed from thymol and sarcosine, with a yield of 86%.

[0098] Alternatively, 5 mmol of thymol and 10 mmol of sarcosine were added to a ball mill in a molar ratio of 1:2, and the shaking frequency was 30 Hz and the shaking time was 20 minutes to obtain composition 1 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 98%.

[0099] Alternatively, 1 mol thymol and 2 mol sarcosine were added to a grinder in a molar ratio of 1:2, and the grinding time was 2 minutes to obtain composition 1 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 95%.

[0100] The sample prepared in Preparation Example 1 was identified by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the measurement results of the samples obtained in the above three preparation processes were consistent, and the results are shown in Figures 1 and 2. Figure 1 is the X-ray powder diffraction (XRPD) pattern of composition 1; Figure 2 is the differential scanning calorimetry (DSC) pattern of composition 1.

[0101] Preparation Example 2: Preparation of Composition 2

[0102] The preparation of Composition 2 was as follows:

[0103] 10 mmol thymol and 10 mmol L-proline were added to 10 mL ethanol in a molar ratio of 1:1, and stirred at 40°C for 1 h. The mixture was then recrystallized to obtain a white precipitate. The precipitate was filtered and placed in a vacuum desiccator to dry at room temperature to obtain composition 2 formed from thymol and L-proline, with a yield of 92%.

[0104] Alternatively, 10 mmol of thymol and 10 mmol of L-proline were added to a ball mill in a molar ratio of 1:1, and the shaking frequency was 30 Hz and the shaking time was 20 minutes to obtain composition 2 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 98%.

[0105] Alternatively, 1 mmol thymol and 1 mmol L-proline were added to a grinder in a molar ratio of 1:1, and the shaking time was 2 minutes to obtain composition 2 formed from thymol and sarcosine, which was a crystalline powder with good flowability, with a yield of 94%.

[0106] The sample prepared in Preparation Example 2 was identified by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC). The measurement results of the samples obtained in the above three preparation processes were consistent, and the results are shown in Figures 3 and 4. Figure 3 is the X-ray powder diffraction (XRPD) pattern of Composition 2; Figure 4 is the differential scanning calorimetry (DSC) pattern of Composition 2.

[0107] Preparation Example 3: Preparation of Composition 3

[0108] The preparation of Composition 3 was as follows:

[0109] 10 mmol of carvacrol and 10 mmol of L-proline were added to 10 mL of ethanol in a molar ratio of 1:1, and stirred at 40°C for 1 h. The mixture was then recrystallized to obtain a white precipitate. The precipitate was filtered and placed in a vacuum desiccator to dry at room temperature to obtain composition 3 formed from carvacrol and L-proline, with a yield of 94%.

[0110] Alternatively, 10 mmol of carvacrol and 10 mmol of L-proline were added to a ball mill in a molar ratio of 1:1, and the shaking frequency was 30 Hz and the shaking time was 20 minutes to obtain composition 3 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 98%.

[0111] Alternatively, 10 mmol of carvacrol and 10 mmol of L-proline were added to a grinder in a molar ratio of 1:1, and the shaking time was 2 minutes to obtain composition 3 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 95%.

[0112] The sample prepared in Preparation Example 3 was identified by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the measurement results of the samples obtained in the above three preparation processes were consistent, and the results are shown in Figures 5 to 6. Figure 5 is the X-ray powder diffraction (XRPD) pattern of composition 3; Figure 6 is the differential scanning calorimetry (DSC) pattern of composition 3.

[0113] Preparation Example 4: Preparation of Composition 4

[0114] The preparation of Composition 4 was as follows:

[0115] 10 mmol of carvacrol and 5 mmol of L-proline were added to 10 mL of ethanol in a molar ratio of 2:1, and the mixture was stirred at 40°C for 1 h. The mixture was then recrystallized to obtain a white precipitate. The precipitate was filtered and placed in a vacuum desiccator to dry at room temperature to obtain composition 4 formed from carvacrol and L-proline, with a yield of 82%.

[0116] Alternatively, 10 mmol of carvacrol and 5 mmol of L-proline were added to a ball mill in a molar ratio of 2:1, and the shaking frequency was 30 Hz and the shaking time was 20 minutes to obtain composition 4 formed from thymol and sarcosine, which was a crystalline powder with good flowability and a yield of 98%.

[0117] Alternatively, 10 mmol of carvacrol and 5 mmol of L-proline were added to a grinder in a molar ratio of 2:1, and the shaking time was 2 minutes to obtain composition 4 formed from thymol and sarcosine, which was a crystalline powder with good flowability, with a yield of 96%.

[0118] The sample prepared in Preparation Example 4 was identified by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the measurement results of the samples obtained in the above three preparation processes were consistent, and the results are shown in Figures 7 and 8. Figure 7 is the X-ray powder diffraction (XRPD) pattern of composition 4; Figure 8 is the differential scanning calorimetry (DSC) pattern of composition 4.

[0119] Preparation Example 5: Preparation of Composition 5

[0120] The preparation of Composition 5 was as follows:

[0121] 5 mmol thymol, 5 mmol carvacrol and 10 mmol L-proline were added to a ball mill in a molar ratio of 1:1:2, and the shaking frequency was 30 Hz and the shaking time was 20 minutes to obtain composition 5, which was a crystalline powder with good flowability and a yield of 98%.

[0122] Alternatively, 5 mmol of carvacrol, 5 mmol of carvacrol and 10 mmol of L-proline were added to a grinder in a molar ratio of 1:1:2, and the shaking time was 2 minutes to obtain composition 5, which was a crystalline powder with good flowability, with a yield of 95%.

[0123] The sample prepared in Preparation Example 5 was identified by X-ray powder diffraction (XRPD) and differential scanning calorimetry (DSC), and the measurement results of the samples obtained in the above three preparation processes were consistent, and the results are shown in Figures 9 and 10. Figure 9 is the X-ray powder diffraction (XRPD) pattern of Composition 5; Figure 10 is the differential scanning calorimetry (DSC) pattern of Composition 5.

[0124] Test Example

[0125] Test Example 1

[0126] From the DSC diagrams, the melting points of Composition 1, Composition 2, Composition 3 and Composition 4 were obtained and the results are shown in the table below.

[0127] [Table 1]

[0128] Test Example 2

[0129] Thymol and / or carvacrol and L-proline and / or sarcosine were added to a ball mill in the composition shown in Table 2, and the mixture was shaken at a shaking frequency of 30 Hz for a shaking time of 20 minutes to obtain a composition. The composition was maintained at 80°C for 100 minutes and subjected to thermal weight loss analysis (TGA), and the vapor pressure of thymol and / or carvacrol and the composition was calculated according to the following formula:

[0130]

number

[0131] where m is the mass of the sample in time t, -dm / dt is the rate of weight loss per unit area, p is the saturation vapor pressure, α is the vaporization constant, T is the absolute temperature, R is the gas constant, and M is the molecular weight of the evaporated substance. Under vacuum conditions, α=1. In the presence of a protective gas, α can be considered as a constant.

[0132] The higher the vapor pressure, the higher the volatility. From Table 2, it can be seen that the vapor pressure can be significantly reduced by the formation of the composition, thus effectively reducing the volatility, thereby significantly improving their stability during processing and storage. The composition of the present invention significantly reduced the vapor pressure of both thymol and carvacrol, thereby reducing their volatility and improving their stability. Thus, the composition of thymol and / or carvacrol with sarcosine and / or proline can both significantly reduce the vapor pressure of thymol and / or carvacrol, reduce their volatility, thereby improving their stability.

[0133] [Table 2]

[0134] Test Example 3

[0135] This study adopted a single factor design, with a total of four treatments: Treatment 1: no plant essential oil was added to the basal diet; Treatment 2: 30 mg / kg of composition 5 was added to the basal diet; Treatment 3: 60 mg / kg of composition 5 was added to the basal diet; Treatment 4: 120 mg / kg of composition 5 was added to the basal diet.

[0136] 384 one-day-old meat ducks were divided into four treatment groups, with six replicates per treatment group, and 16 meat ducks per replicate. The basic diet was corn-soybean meal type, and the experimental period was 35 days. The meat ducks were reared in two stages, 1-14 days and 15-35 days.

[0137] As shown in Table 3, compared with treatment 1 without the addition of the plant essential oil composition, treatments 2 and 3 showed a significant reduction in die-off rate, and when the amount added reached 120 mg / kg, the die-off rate was 0. Thus, the advantages of using the composition provided by the present invention as an animal feed additive are that it is difficult to volatilize, has good palatability, improves stability, and is reflected in the quality and quantity of the animal's internal conditions, fully exerts antibacterial properties, and reduces the impact of die-off rate. The test results on meat ducks showed the effect of a significant reduction in die-off rate.

[0138] [Table 3]

[0139] Test Example 4

[0140] The accelerated stability advantage of Composition 5 was investigated.

[0141] From the test samples: Composition 5 prepared in Preparation Example 5 above and a commercially available mixed essential oil additive coating product A (containing thymol 25%; carvacrol 25%).

[0142] Under conditions of 40°C / 75% RH, on day 0 the contents of thymol and carvacrol in Composition 5 were 25.7% / 25.5%, and in Product A additive were 26.1% / 26.2%, respectively. After 2 months the contents of thymol and carvacrol in Composition 5 were 25.6% / 25.2%, and in Product A additive were 20.6% / 20.7%, respectively.

[0143] In a 2-month accelerated experiment, the retention rate of composition 5 was 99.2%, whereas thymol and carvacrol were both unstable in product A, with a retention rate of 79.0%, indicating that the composition of the present invention has better storage stability.

[0144] [Table 4]

[0145] Test Example 5

[0146] The stability advantage of Composition 5 during the granulation process was investigated.

[0147] From test samples: Composition 5 prepared in Preparation Example 5 above and a commercially available mixed essential oil additive product B (containing 8.5% thymol and 8.5% carvacrol).

[0148] Referring to the granulation processing of meat duck feed according to the Chinese duck feed standard, the essential oil retention rate of product B was 53%, while that of composition 5 was 72.3%, which showed that the essential oil of the composition of the present invention has better processing stability.

[0149] [Table 5]

[0150] Test Example 6

[0151] This study adopted a single factor design, with a total of four treatments: Treatment 1: no plant essential oil was added to the basal diet; Treatment 2: 30 mg / kg of composition 5 was added to the basal diet; Treatment 3: 60 mg / kg of composition 5 was added to the basal diet; Treatment 4: 120 mg / kg of composition 5 was added to the basal diet.

[0152] 640 1-day-old chickens were divided into 4 treatment groups, with 8 replicates per treatment group, and 20 chickens per replicate. The basic diet was corn and soybean meal type, and the experimental period was 42 days. The chickens were raised in two stages, 1-21 days and 22-42 days.

[0153] As shown in Table 6, treatments 2 to 4 significantly increased body weight compared to treatment 1 without the addition of the plant essential oil composition, with a 6% increase in body weight at an addition level of 30 mg / kg. Thus, the advantages of using the composition provided by the present invention as an animal feed additive are that it is less volatile, has good palatability, and improves stability, and is manifested in the quality and quantity of the animal's internal conditions, fully exerting the antibacterial and weight gain effects. The test results for meat chickens showed a significant increase in body weight.

[0154] [Table 6]

[0155] In the data in the same row, lowercase letters on the shoulders indicate significant differences (p<0.05), while the same or no letters indicate no significant differences (p>0.05).

[0156] Test Example 7

[0157] This study adopted a single factor design, with a total of four treatments: Treatment 1: no plant essential oil was added to the basal diet; Treatment 2: 30 mg / kg of composition 5 was added to the basal diet; Treatment 3: 60 mg / kg of composition 5 was added to the basal diet; Treatment 4: 120 mg / kg of composition 5 was added to the basal diet.

[0158] A 28-day feeding study was conducted on 96 weaned piglets, divided into four treatment groups with four replicates per treatment group and six pigs per replicate. Body weight, feed intake, and daily diarrhea were recorded on days 1 and 28, respectively, and feed-to-body-weight ratios and diarrhea rates were calculated.

[0159] As shown in Table 7, at the end of the 28-day trial, treatment group 3, which was treated with 60 mg / kg, showed a significant increase in body weight and a significant decrease in feed-to-body weight ratio compared to treatment 1, which did not receive the plant essential oil composition, with a 15% increase in body weight and a 22% decrease in feed-to-body weight ratio.

[0160] [Table 7]

[0161] In the data in the same row, lowercase letters on the shoulders indicate significant differences (p<0.05), while the same or no letters indicate no significant differences (p>0.05).

[0162] The above are merely specific embodiments of the present invention, and the scope of protection of the present invention is not limited thereto. Any modifications or replacements that a person skilled in the art can think of without any creative effort within the technical scope of the present invention as disclosed in this specification should be considered to be included in the scope of protection of the present invention.

Claims

1. A composition comprising an antibacterial agent and an amino acid, The composition comprises a co-crystal or co-crystal powder of the antibacterial agent and the amino acid obtained by mixing the antibacterial agent and the amino acid, wherein the antibacterial agent is thymol, carvacrol, or a mixture thereof, and the amino acid is L-proline, sarcosine, or a mixture thereof.

2. The composition of claim 1, wherein the antibacterial agent and the amino acid are mixed in a molar ratio of 9-1:1-9.

3. The composition described in claim 1, wherein the antibacterial agent and amino acid are mixed in a molar ratio of 2-1:1-2.

4. 2. The composition of claim 1, wherein the antimicrobial agent is a mixture of thymol and carvacrol, the molar ratio of thymol to carvacrol being 9-1:1-9.

5. The composition described in claim 1, wherein the antibacterial agent is a mixture of thymol and carvacrol, and the molar ratio of thymol to carvacrol is 2-1:1-2.

6. 2. The composition of claim 1, wherein the composition is obtained by mixing thymol and sarcosine in a molar ratio of 1:2, and the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of 6.4°±0.2°, 12.1°±0.2°, 12.4°±0.2°, 17.7°±0.2°, 20.2°±0.2°, 22.1°±0.2°, 23.0°±0.2°, 23.7°±0.2°, and 25.1°±0.2°.

7. 2. The composition of claim 1, wherein the composition is obtained by mixing thymol and L-proline in a 1:1 molar ratio, and the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of 6.2°±0.2°, 6.9°±0.2°, 12.5°±0.2°, 13.8°±0.2°, 18.0°±0.2°, 18.4°±0.2°, 18.7°±0.2°, 19.1°±0.2°, 19.3°±0.2°, 21.1°±0.2°, and 24.8°±0.2°.

8. 2. The composition of claim 1, wherein the composition is obtained by mixing carvacrol and L-proline in a 1:1 molar ratio, and the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of 6.9°±0.2°, 11.5°±0.2°, 13.6°±0.2°, 13.9°±0.2°, 19.0°±0.2°, 19.2°±0.2°, 23.1°±0.2°, and 24.9°±0.2°.

9. 2. The composition of claim 1, wherein the composition is obtained by mixing carvacrol and L-proline in a molar ratio of 2:1, and the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of 6.5°±0.2°, 11.0°±0.2°, 13.1°±0.2°, 13.2°±0.2°, 15.0°±0.2°, 15.5°±0.2°, 15.8°±0.2°, 17.5°±0.2°, 19.7°±0.2°, 20.5°±0.2°, 21.6°±0.2°, 21.9°±0.2°, 23.6°±0.2°, 25.1°±0.2°, and 26.7°±0.2°.

10. 2. The composition of claim 1, wherein the composition is obtained by mixing thymol, carvacrol and L-proline in a molar ratio of 1:1:2, and the X-ray powder diffraction pattern of the composition has characteristic peaks at 2θ angles of 6.2°±0.2°, 6.8°±0.2°, 12.5°±0.2°, 13.8°±0.2°, 18.4°±0.2°, 18.8°±0.2°, 19.4°±0.2°, 21.3°±0.2°, 24.8°±0.2°, and 25.0°±0.2°.

11. A method for producing the composition according to any one of claims 1 to 10, comprising the step of mixing the antibacterial agent and the amino acid to obtain a composition comprising the antibacterial agent and the amino acid, The mixing is carried out by the following method (1) and method (2): Method (1): recrystallizing the antibacterial agent and the amino acid in a solvent to obtain a precipitate, and drying the precipitate to obtain a composition containing the antibacterial agent and the amino acid; Method (2): Mixing the antibacterial agent and the amino acid, adding them to a grinding device to thoroughly contact them by mechanical force, or spray-drying the mixture solution, or processing it through an extruder to obtain a composition containing the antibacterial agent and the amino acid; The manufacturing method according to any one of the above items.

12. The method of claim 11, wherein in the method (ii), the antibacterial agent and the amino acid are added to a ball mill and vibrated at a vibration frequency of 5 to 60 Hz for a vibration time of 5 to 60 minutes to obtain a composition containing the antibacterial agent and the amino acid, or the antibacterial agent and the amino acid are added to a grinder and ground for a grinding time of 1 to 60 minutes to obtain a composition containing the antibacterial agent and the amino acid.

13. An article of manufacture comprising a composition according to any one of claims 1 to 10.

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