CN112868657A - Application of ethyl maltol as fungal inhibitor - Google Patents
Application of ethyl maltol as fungal inhibitor Download PDFInfo
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- CN112868657A CN112868657A CN202110072116.6A CN202110072116A CN112868657A CN 112868657 A CN112868657 A CN 112868657A CN 202110072116 A CN202110072116 A CN 202110072116A CN 112868657 A CN112868657 A CN 112868657A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
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Abstract
The ethyl maltol serving as a fungus inhibitor can be used as a fungus bacteriostatic agent to be applied to preparation of plant pesticides and plant seed coating agents, and can also be used as a fungus bacteriostatic agent to be added into a traditional Chinese medicine liquid preparation or a semi-fluid preparation, so that the growth and reproduction of fungi including moulds can be effectively inhibited, and plant fungus diseases can be reduced as safe plant pesticides; the mildew and deterioration of seeds are inhibited, the mildew and deterioration probability of the seeds is reduced or the mildew and deterioration of the seeds are avoided, and the germination rate of the seeds can be improved; the natural aroma-improving function is exerted, meanwhile, aroma-improving, mildew-proof and corrosion prevention are integrated, the stability of the traditional Chinese medicine liquid preparation and the semi-fluid preparation is enhanced, the use of other toxic preservatives can be replaced or reduced, the use variety of the additives is reduced, and the potential safety risk of the additives, particularly the preservatives, is reduced.
Description
Technical Field
The invention relates to an application of ethyl maltol, in particular to an application of ethyl maltol as a fungus bacteriostatic agent, and especially an application of ethyl maltol as a fungus bacteriostatic agent in plant pesticides, seed coatings and traditional Chinese medicine liquid preparations.
Background
Ethyl maltol (ethyl maltol), chemical name 3-hydroxy-2-ethyl-4-pyridone, relative molecular mass 140.11, white crystalline powder, melting point 89-93 ℃, readily soluble in hot water. Heretofore, it has been one of the usual safe food flavoring additives that can be used for flavoring foods. National standard for food safety of GB 2760-2014-food additive use standard appendix b.2.3 lists ethyl maltol as a permitted use synthetic flavor for food (FEMA number: 3487). The US food Standard specifies a food flavoring limit for ethyl maltol of 100mg/L (BATF-27CFR § 240.1051).
At present, most of domestic plant pesticides are phenols, aldehydes or antibiotics, so that toxicity and irritation are inevitably brought while growth and propagation of microorganisms are effectively inhibited, and certain influence is brought to the environment.
The seed coating technology has good effect on preventing and treating crop diseases and insect pests, has obvious effects of protecting and promoting seedlings, increasing yield, enhancing stress resistance and the like, is favorable for protecting the environment, and can improve economic and ecological benefits. The seed coating technology has wide application prospect in the aspect of traditional Chinese medicine cultivation and breeding, and is particularly suitable for some seeds with late ripening stage or needing fresh preservation, such as golden cypress, immature bitter orange and the like. At present, seed coatings mainly comprise bactericides and insecticides, such as quintozene, captan, thiabendazole, diniconazole and the like, have certain disease prevention and treatment effects on underground seeds, and have certain adverse effects on seed germination and safety.
The bacteriostatic agent (preservative) is used as one of important auxiliary material additives of a traditional Chinese medicine liquid preparation or a semi-fluid preparation (paste preparation), and plays an important role in preventing food or medicine from being polluted by microorganisms and ensuring the quality of the product. Generally, most of the traditional Chinese medicine preparations, particularly animal and plant traditional Chinese medicine preparations, contain substances which are easily contaminated by microorganisms and go moldy, such as saccharides, proteins, mucilaginous substances and the like, particularly traditional Chinese medicine preparations taking water as a solvent, including various traditional Chinese medicine liquid preparations and semi-fluid preparations, preservatives (such as sodium benzoate and the like) are often added, and the pharmacopoeia of the people's republic of China (2015 edition), the U.S. pharmacopoeia (USP41), the European pharmacopoeia (EP9) and the like make specific regulations on the use of preservatives in medicines. Because all preservatives have certain toxicity, the addition amount is limited and specified in various food standards and Chinese pharmacopoeia (2015 edition), for example, the amount of sodium benzoate in 0181 mixture specified in Chinese pharmacopoeia appendix is not more than 0.3%.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects in the prior art and provide a safe fungus bacteriostatic agent which can inhibit the growth and reproduction of conventional mildew microorganisms, reduce plant fungal diseases, inhibit the mildew and deterioration of seeds or medicaments, reduce the mildew and deterioration of the seeds, improve the germination rate of the seeds, enhance the stability of traditional Chinese medicine liquid preparations and semi-fluid preparations, reduce or avoid potential hazards, and is prepared from ethyl maltol.
One of the technical schemes adopted by the invention for solving the technical problems is that the application of ethyl maltol as a fungus bacteriostatic agent in the preparation of plant pesticides specifically comprises the following steps:
(1) weighing ethyl maltol, dissolving the ethyl maltol in double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(2) and (3) sterilizing the ethyl maltol solution under high pressure, and filling the ethyl maltol solution into a plastic bottle to obtain the ethyl maltol plant pesticide.
Further, in the step (1), the amount of the double distilled water used is 400 to 600mL/g ethyl maltol, preferably 500mL/g ethyl maltol.
Further, in the step (2), the temperature of the high-pressure sterilization is 100-140 ℃, and preferably 120-125 ℃; the pressure of the high-pressure sterilization is 0.1-0.5 PP, preferably 0.3-0.4 PP; the autoclaving time is 20-60 min, preferably 30-40 min.
A method for applying ethyl maltol plant pesticide to plants comprises spraying the ethyl maltol plant pesticide onto the plants.
Further, the amount of the ethyl maltol plant pesticide spray is 50-75L/mu of plant; the time interval of spraying the ethyl maltol plant pesticide is 7-10 days per time.
The second technical scheme adopted for solving the technical problems is that the application of ethyl maltol as a fungal bacteriostatic agent in preparing a seed coating medicament specifically comprises the following steps:
(3) weighing ethyl maltol, dissolving the ethyl maltol in double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(4) and (3) sterilizing the ethyl maltol solution under high pressure, and filling the ethyl maltol solution into a spray bottle to obtain the ethyl maltol seed coating agent.
Further, in the step (1), the usage amount of the double distilled water is 400-600 mL/g ethyl maltol; preferably 500mL/g ethyl maltol. The concentration of the ethyl maltol is too low to achieve the bacteriostatic effect, and the concentration of the ethyl maltol is too high to cause insufficient dissolution of the ethyl maltol, thereby causing waste.
Further, in the step (2), the temperature of the high-pressure sterilization is 100-140 ℃, and preferably 120-125 ℃; the pressure of the high-pressure sterilization is 0.1-0.5 PP, preferably 0.3-0.4 PP; the autoclaving time is 20-60 min, preferably 30-40 min.
The method for coating the seeds by applying the ethyl maltol seed coating agent comprises the following steps: and spraying the ethyl maltol seed coating agent on seeds, uniformly mixing, and drying to obtain the finished product.
Further, the spraying amount of the ethyl maltol seed coating agent is 200-600 mL/kg of seeds, preferably 400mL/kg of seeds; the drying temperature is 10-35 ℃, and preferably 30 ℃.
The third technical scheme adopted for solving the technical problems is that the ethyl maltol is used as a fungus bacteriostatic agent in the preparation of a traditional Chinese medicine liquid preparation or a semi-fluid liquid medicine, the ethyl maltol is subjected to ultraviolet sterilization treatment, and then the ethyl maltol subjected to sterilization treatment is added into the traditional Chinese medicine liquid preparation or the semi-fluid liquid medicine which is subjected to sterilization treatment and is uniformly mixed.
Further, the time of ultraviolet sterilization treatment is 20-40min, preferably 30 min.
Furthermore, the addition amount of the ethyl maltol is 1-3 g/L of the traditional Chinese medicine liquid preparation or the semi-fluid liquid medicine, and preferably 1.5-2 g/L of the traditional Chinese medicine liquid preparation or the semi-fluid liquid medicine. The addition amount of the ethyl maltol is too much and does not meet the addition standard of the bacteriostatic agent in the traditional Chinese medicine liquid medicament, for example, the addition amount of the common sodium benzoate is usually 3g/L, and the bacteriostatic effect is not obvious when the addition amount of the ethyl maltol is too little. The difference from the amount added as flavoring additive is that the standard of addition of ethyl maltol as flavoring agent is 1 g/L.
The working principle of the ethyl maltol as the fungus inhibitor is that the ethyl maltol can better play the inhibiting role when directly contacting with the fungus, and the action mechanism of the ethyl maltol is that the ethyl maltol can effectively inhibit the hypha and the spore germination of the fungus. Under the culture conditions of the same concentration of ethyl maltol, the same inoculation amount and the same temperature, the ethyl maltol has an inhibition effect on aspergillus niger hyphae inoculated to the conical flask, the inhibition effect is close to sodium benzoate, but the inhibition effect on aspergillus niger fungus blocks inoculated to the flat dish is not obvious, and the aspergillus niger hyphae also comprise 6 common fungi such as penicillium citrinum and the like.
Compared with the prior art, the invention has the beneficial effects that: the ethyl maltol is used as a fungus bacteriostatic agent to prepare plant pesticide and seed coating agent or used as a traditional Chinese medicine liquid preparation and a traditional Chinese medicine semi-fluid preparation, can effectively inhibit the growth and reproduction of conventional mildew microorganisms, reduce plant fungus diseases, reduce the mildew and deterioration probability of seeds or avoid the mildew and deterioration of the seeds, and can play the inherent functions of flavoring in the liquid preparation, combine flavoring and corrosion prevention into a whole, improve the germination rate of the seeds, enhance the stability of the traditional Chinese medicine liquid preparation, replace or reduce the use of other toxic preservatives, reduce the use variety of additives and reduce the potential safety risk of the additives, particularly the preservatives.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1
The embodiment is an application of ethyl maltol as a fungal bacteriostatic agent in preparing ethyl maltol plant pesticide, and the method comprises the following steps:
(1) weighing 2g of ethyl maltol, dissolving the ethyl maltol in 1000mL of double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(2) and (3) autoclaving the ethyl maltol solution for 30min at the temperature of 121 ℃ and the pressure of 0.3PP, and filling the ethyl maltol solution into a plastic bottle to obtain the ethyl maltol plant pesticide.
Use of: the amount of pesticide spraying of the ethyl maltol plants is 60L/mu of plants; the time interval of spraying the ethyl maltol plant pesticide is 7 days/time.
Example 2
The embodiment is an application of ethyl maltol as a fungus bacteriostatic agent in preparing a bacteriostatic agent, namely an ethyl maltol seed coating medicament, and the method comprises the following steps:
(3) weighing 0.2g of ethyl maltol, dissolving with 100mL of double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(4) and (3) autoclaving the ethyl maltol solution at the temperature of 121 ℃ and the pressure of 0.3PP for 30min, and filling the solution into a spray bottle to obtain the bacteriostatic agent ethyl maltol seed coating agent.
Use of: and spraying the obtained bacteriostatic agent ethyl maltol seed coating agent on seeds according to the using amount of 400mL/kg of seeds, uniformly mixing, spreading, and drying at 30 ℃ to obtain the seeds coated with the bacteriostatic agent ethyl maltol seed coating.
Example 3
This example is the application of ethyl maltol as a fungal bacteriostatic in a liquid preparation of traditional Chinese medicine: weighing 2g of ethyl maltol, and performing ultraviolet sterilization for 30min to obtain sterile ethyl maltol; then mixing the sterile ethyl maltol with 1000mL of traditional Chinese medicine liquid or semifluid liquid medicine uniformly, sterilizing at 121 ℃ for 30min, and encapsulating to obtain the traditional Chinese medicine liquid preparation of ethyl maltol with the bacteriostatic agent.
The following are the experimental results of the ethyl maltol plant pesticide test of example 1, the ethyl maltol seed coating agent test of example 2, and the application effect test of the liquid preparation of ethyl maltol with the bacteriostatic agent added in example 3.
A. Ethyl maltol as a plant pesticide
The test method comprises the following steps: selecting leaves in the same growth period, and applying pesticides to the leaves respectively by using carbendazim solution with the same concentration and ethyl maltol as a fungal bacteriostat according to the method of the example 1 to respectively obtain the treated leaves. The leaves obtained by respectively spraying pesticides on the carbendazim solution and the ethyl maltol with the same concentration are sequentially used as a control group and an ethyl maltol group, and the leaves without any pesticide are used as a blank group. A number of disposable sealed bags containing 20 ml of sterile water were prepared in advance in the laboratory. 30 plants are respectively sampled in the same treatment and different plants, and are placed in the same sterilized sealing bag to be brought back.
And (3) testing the viable count: fully contacting the leaves with sterile water, uniformly shaking the liquids of the control group, the blank group and the ethyl maltol group respectively, transferring 1mL of the liquids respectively into a test tube filled with 9mL of sterile water, and shaking in a vortex mode to uniformly disperse the medicine sample. Then transferring 1mL of the dispersed medicine sample into a test tube filled with 9mL of sterile water, blowing, mixing, and making into 10-1、10-2、10-33 concentrations of bacterial suspension. Respectively take the above 10-1、10-2、10-3Concentration tubes, 50. mu.L of each inoculum were plated, plates were prepared, 3 replicates per concentration, inverted at 25 ℃ for 48h, and colony counts were determined separately, and the results are shown in Table 1.
TABLE 1 colony count test results of control group, blank group and ethyl maltol group (n ═ 3)
Therefore, the ethyl maltol is used as an effective and only active component to prepare the plant pesticide, so that the growth and the propagation of conventional mildew microorganisms can be effectively inhibited, and the plant fungal diseases are reduced.
B. Ethyl maltol as coating of traditional Chinese medicine immature bitter orange seeds
The test method comprises the following steps: healthy, full and uniform immature bitter orange seeds harvested in the current year are selected, and coating treatment is respectively carried out on the immature bitter orange seeds with carbendazim solution with the same concentration, water and ethyl maltol serving as a bacteriostatic agent according to the method in the embodiment 2, so that the treated seeds are respectively obtained. And (3) respectively coating the carbendazim solution, water and ethyl maltol with the same concentration to obtain seeds which are sequentially used as a control group, a blank group and an ethyl maltol group.
1. Testing of seed germination rate: respectively sowing a control group, a blank group and an ethyl maltol group in a plastic germination box, selecting a mixed matrix of vermiculite and turf (1: 1) as a matrix, placing the matrix in an illumination incubator for culture, wherein the illumination intensity is 12000lx, the illumination period is 12h/12h (day/night), the relative humidity is 70-75%, the temperature is 26-28 ℃, and the seed germination rate is tested, and the test results are shown in table 2.
Table 2 test results of seed germination rates of control group, blank group and ethyl maltol group (n ═ 3)
2. Testing of germination spore inhibition rate: the control group, the blank group and the ethyl maltol group were placed in the center of a 90mm petri dish, and 2% water agar was placed in the petri dish, and were incubated at 27 ℃ for 48 hours in the dark. Respectively collecting with scotch tapeSporeThe tape was placed on a glass slide, 0.1mL of lactic acid and glycerol (lactic acid: glycerol: water: 1:2:1, V: V) were added, and all spores and germinating conidia of the control group, blank group and ethyl maltol group were calculated under 100-fold microscope to perform a test of the germinating spore inhibition ratio, and the test results are shown in table 3. The germination spore inhibition rate calculation formula is as follows: IR% (conidium germination rate of drug-untreated group-conidium germination rate of drug-treated group)/conidium germination rate of drug-untreated group.
Table 3 test results of germination spore inhibition rates of control group, blank group and ethyl maltol group (n ═ 3)
Therefore, the ethyl maltol is used as an effective and only active component to prepare the seed coating, so that the germination rate of the seeds can be improved, the germination of germination spores in the seeds can be effectively inhibited, and the disease inhibiting effect can be more effectively achieved. C. Ethyl maltol as bacteriostatic preservative of traditional Chinese medicine liquid preparation
The test method comprises the following steps: selecting the traditional Chinese medicine liquid preparations with the same concentration and the same amount, and respectively processing the traditional Chinese medicine liquid preparations with the same amount of sodium benzoate and ethyl maltol according to the method of the example 3 to respectively obtain the traditional Chinese medicine liquid preparations. Standing for 14 days, and respectively treating sodium benzoate with the same mass, without any substance, and with ethyl maltol to obtain traditional Chinese medicine liquid preparations which are sequentially used as a control group, a blank group and an ethyl maltol group.
And (3) testing the viable count: detection in control, blank and ethyl maltol groups by plate countingNumber of viable bacteria. After shaking up the drug samples of the control group, the blank group and the ethyl maltol group, 1mL of the drug samples were transferred to a test tube containing 9mL of sterile water, and the drug samples were uniformly dispersed by vortex shaking. Then transferring 1mL of the dispersed medicine sample into a test tube filled with 9mL of sterile water, blowing, mixing, and making into 10-1、10-2、10-3、10-4、10-55 test tubes in total. Respectively take the above 10-1、10-3、10-5Each concentration tube was coated with 50. mu.L of each inoculum, and plates were prepared, each concentration was repeated 3 times, and the plate was cultured in an inverted state at 25 ℃ for 48 hours, and the number of colonies was measured, respectively, and the results are shown in Table 4.
TABLE 4 colony count test results of control group, blank group and ethyl maltol group (n ═ 3)
Therefore, the ethyl maltol serving as the antibacterial preservative is applied to the traditional Chinese medicine liquid preparation, the inherent functions of aroma enhancement are exerted, the growth and the propagation of the conventional mildew microorganisms can be effectively inhibited, and the storage life of the traditional Chinese medicine liquid preparation is prolonged.
Claims (14)
1. Use of ethyl maltol as a fungal inhibitor.
2. Application of ethyl maltol as a fungus inhibitor in preparation of plant pesticides.
3. Use of ethyl maltol as a fungal inhibitor in the preparation of a plant pesticide according to claim 2, comprising the steps of:
(1) weighing ethyl maltol, dissolving the ethyl maltol in double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(2) and (3) sterilizing the ethyl maltol solution under high pressure, and filling the ethyl maltol solution into a plastic bottle to obtain the ethyl maltol plant pesticide.
4. The use of ethyl maltol as a fungal bacteriostatic agent in the preparation of plant pesticides according to claim 3, wherein in the step (1), the amount of the double distilled water is 400-600 mL/g ethyl maltol, preferably 500mL/g ethyl maltol; in the step (2), the temperature of the high-pressure sterilization is 100-140 ℃, and preferably 120-125 ℃; the pressure of the high-pressure sterilization is 0.1-0.5 PP, preferably 0.3-0.4 PP; the autoclaving time is 20-60 min, preferably 30-40 min.
5. A method of applying ethyl maltol plant pesticide to plants as claimed in claim 3 or 4, wherein said ethyl maltol plant pesticide is sprayed onto the plants.
6. The method of applying ethyl maltol plant pesticide to plants as claimed in claim 5, wherein the ethyl maltol plant pesticide is sprayed at an amount of 50-75L per acre of plant; the time interval of spraying the ethyl maltol plant pesticide is 7-10 days per time.
7. Use of ethyl maltol as a fungal inhibitor in the preparation of a seed coating medicament.
8. Use of ethyl maltol as a fungal inhibitor in the preparation of a seed coating medicament according to claim 7, comprising the steps of:
(1) weighing ethyl maltol, dissolving the ethyl maltol in double distilled water, and uniformly stirring to obtain an ethyl maltol solution;
(2) and (3) sterilizing the ethyl maltol solution under high pressure, and filling the ethyl maltol solution into a spray bottle in a filling mode to obtain the ethyl maltol seed coating agent.
9. The use of ethyl maltol as a fungal bacteriostatic agent in preparing a seed coating medicament according to claim 8, wherein in the step (1), the amount of the double distilled water is 400-600 mL/g ethyl maltol; preferably 500mL/g ethyl maltol; in the step (2), the temperature of the high-pressure sterilization is 100-140 ℃, and preferably 120-125 ℃; the pressure of the high-pressure sterilization is 0.1-0.5 PP, preferably 0.3-0.4 PP; the autoclaving time is 20-60 min, preferably 30-40 min.
10. A method of coating seeds with the ethyl maltol seed coating composition of claim 8 or 9, wherein the ethyl maltol seed coating composition is sprayed onto the seeds, mixed until uniform, and dried.
11. The method for coating seeds with the ethyl maltol seed coating agent according to claim 10, wherein the spraying amount of the ethyl maltol seed coating agent is 200-600 mL/kg of seeds, preferably 400mL/kg of seeds of traditional Chinese medicine; the drying temperature is 10-35 ℃, and preferably 30 ℃.
12. Use of ethyl maltol as a fungal inhibitor in a liquid or semi-liquid formulation of a traditional Chinese medicine.
13. The method of claim 12, wherein the ethyl maltol is first treated by UV sterilization, and then the ethyl maltol is added to the sterilized liquid preparation or semi-liquid solution, and then mixed uniformly.
14. Method for the use of ethyl maltol as a fungal bacteriostatic agent in liquid and semi-liquid formulations of traditional Chinese medicines according to claim 13, wherein the time of the uv-sterilization treatment is 20-40min, preferably 30 min; the addition amount of the ethyl maltol is 1-3 g/L of the traditional Chinese medicine liquid preparation or the semi-fluid liquid medicine, and preferably 1.5-2 g/L of the traditional Chinese medicine liquid preparation or the semi-fluid liquid medicine.
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Application publication date: 20210601 |