CN113925061A - Preparation method and application of magnetic nano ferrite material - Google Patents
Preparation method and application of magnetic nano ferrite material Download PDFInfo
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- CN113925061A CN113925061A CN202111372091.8A CN202111372091A CN113925061A CN 113925061 A CN113925061 A CN 113925061A CN 202111372091 A CN202111372091 A CN 202111372091A CN 113925061 A CN113925061 A CN 113925061A
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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
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/16—Heavy metals; Compounds thereof
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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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
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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
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
- A01N59/16—Heavy metals; Compounds thereof
- A01N59/20—Copper
Abstract
The invention discloses a preparation method and application of a magnetic nano ferrite material, which comprises the following steps: s1: 2.4mM ferric chloride and 1.2mM cupric chloride are added into ethylene glycol to form a transparent solution; s2: then adding NaAc and sodium citrate into the transparent solution, and uniformly dispersing to form a mixed solution; s3: and transferring the mixed solution into a 25mL stainless steel autoclave, cooling at room temperature after the reaction is finished, washing the black solid product with ultrapure water and ethanol, and drying in a vacuum drying oven to finish the preparation. The magnetic nano ferrite material prepared by the invention is based on magnetic ferroferric oxide, copper oxide and elementary copper are compounded, the generation of ROS is improved by utilizing the electron migration between compounds, so that the bactericidal activity is improved, the bacteriostatic activity is 10 times of that of Bordeaux mixture, the seed germination rate of plants, the chlorophyll content and the root activity are obviously improved by the released iron ions and copper ions, and the potential application value of the magnetic nano ferrite material can replace Bordeaux mixture to be used for agricultural disease treatment.
Description
Technical Field
The invention belongs to the technical field of inorganic nano antibacterial materials, and particularly relates to a preparation method and application of a magnetic nano ferrite material.
Background
The main ingredient of the commercially available bacteriostatic agent such as Bordeaux mixture, Lvdebao, Shuangxiaoling and other common medicaments is Cu2+，Cu2+The bactericidal composition is taken as a disposable medicine for daily life, has the advantages of high bactericidal efficiency, small side effect on human bodies, safety, reliability and the like, and is widely applied to the fields of bacteria prevention and control in agriculture, forestry and animal husbandry, medicine for daily life and the like.
Since Cu2+As trace elements of plants, the trace elements are important elements forming roots, stems, leaves and fruits of plants, and trace Cu2+Promoting plant development, Cu2+The bacteriostatic mechanism of (A) is as follows: plants require higher amounts of Cu than microorganisms, but when Cu is present in the environment2 +Cu at a concentration above the minimum inhibitory limit of the bacteria and below the phytotoxic concentration2+After entering the inside of the microorganism through the copper ion channel, the copper ion channel is dissociated in cytoplasm to change the osmotic pressure balance inside and outside the bacteria and destroy the organelles of the bacteria, so that the copper ion channel can not only sterilize but also promote the growth of plants in the concentration range.
The Reactive Oxygen Species (ROS) as an important bactericide can cause irreversible damage to bacterial organelles, liquid environment required by metabolism, protein molecules and the like and does not generate drug resistance.
Currently, only the commercially available copper ion or iron ion related compounds are available as materials for inhibiting bacteria and promoting plant growth. The conventional bacteriostatic agent sold in the market is low in bacteriostatic activity, difficult to recover, low in promotion influence on plant seed germination, root activity and chlorophyll, and more in composite components. Therefore, we propose a preparation method and application of magnetic nano ferrite material to solve the problems mentioned in the above background art.
Disclosure of Invention
The present invention aims at providing a preparation method of magnetic nano ferrite material and its application, so as to solve the problems proposed in the background art.
In order to achieve the purpose, the invention provides the following technical scheme: a preparation method of a magnetic nano ferrite material comprises the following steps:
s1: 2.4mM ferric chloride and 1.2mM cupric chloride are added into ethylene glycol to form a transparent solution;
s2: then adding NaAc and sodium citrate into the transparent solution, and performing ultrasonic dispersion uniformly to form a mixed solution;
s3: and transferring the mixed solution into a polytetrafluoroethylene inner container of a 25mL stainless steel autoclave, cooling the reaction kettle at room temperature after the reaction is finished, washing a black solid product with ultrapure water and ethanol, and drying in a vacuum drying oven to finish the preparation.
In the step S3, the reaction temperature is 200 ℃, and the reaction time is 10 h.
The number of washing times of the black solid washed with ultrapure water and ethanol in the step S3 is 3 to 4.
In the step S3, the drying temperature of the vacuum drying oven is 60 ℃, and the drying time is 12 h.
The invention also provides an application of the magnetic nano ferrite material, which comprises the application of the magnetic nano ferrite material prepared by the preparation method in bacteriostasis and plant growth promotion.
Compared with the prior art, the invention has the beneficial effects that: the invention provides a preparation method and application of a magnetic nano ferrite material, namely magnetic nano Fe3O4The synthesis method of the/CuO/Cu composite material is simpler, and the external magnetism is addedCan be recycled under the action of field because of the Fe2+，Fe3+，Cu2+Cu has different oxidation-reduction potentials to enable electrons to have a certain migration rate, the electrons are combined with a medium solution to generate ROS, the ROS has bactericidal performance and seed germination promoting performance, the antibacterial activity of the ROS is 10 times of that of Bordeaux mixture, and trace Fe released by a magnetic nano ferrite material (compared with the Bordeaux mixture alone) is released2+And Cu2+Can promote root growth and increase chlorophyll content.
The magnetic nano ferrite material prepared by the invention is based on magnetic ferroferric oxide, copper oxide and elementary copper are compounded, the generation of ROS is improved by utilizing the electron migration between compounds, the bactericidal activity is further improved, and the released iron ions and copper ions can promote the growth of plants.
Fe3O4The CuO/Cu has strong bacteriostasis and promotes plant growth when the concentration is 200 mu g/mL, and the potential application value can replace Bordeaux mixture to be used for agricultural disease treatment. Novel spherical nano Fe3O4The application of the/CuO/Cu material is 10 times that of the commercial Bordeaux solution.
Drawings
FIG. 1 is a schematic diagram of a transmission electron microscope of the magnetic nano ferrite material of the present invention;
FIG. 2 is a schematic diagram of the X-ray photoelectron spectroscopy of the magnetic nano ferrite material of the present invention;
FIG. 3 is a schematic diagram of X-ray diffraction spectrum analysis of the magnetic nano ferrite material of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a preparation method of a magnetic nano ferrite material as shown in figures 1-3, which comprises the following steps:
s1: 2.4mM ferric chloride and 1.2mM cupric chloride are added into ethylene glycol to form a transparent solution;
s2: then adding NaAc and sodium citrate into the transparent solution, and performing ultrasonic dispersion uniformly to form a mixed solution;
s3: and transferring the mixed solution into a polytetrafluoroethylene inner container of a 25mL stainless steel autoclave, cooling the reaction kettle at room temperature after the reaction is finished, washing a black solid product with ultrapure water and ethanol, and drying in a vacuum drying oven to finish the preparation.
In the step S3, the reaction temperature is 200 ℃, and the reaction time is 10 h.
The number of washing times of the black solid washed with ultrapure water and ethanol in the step S3 is 3 to 4.
In the step S3, the drying temperature of the vacuum drying oven is 60 ℃, and the drying time is 12 h.
The invention also provides an application of the magnetic nano ferrite material, which comprises the application of the magnetic nano ferrite material prepared by the preparation method in bacteriostasis and plant growth promotion.
Firstly, bacteriostatic experiment:
the bacteriostatic activity of the magnetic nano ferrite material is monitored by salmonella, ralstonia solanacearum, potato dry rot pathogen and the like, the salmonella and the ralstonia solanacearum are exponential phase bacteria activated at 37 ℃ (the fungi are at 28 ℃), and biological materials such as an LB culture medium (a potato culture medium for the fungi), ultrapure water, normal saline, a phosphate buffer solution and the like are sterilized in an autoclave (121 ℃,20min) and are cultured in a biochemical incubator.
Secondly, filter paper diffusion experiment:
the solution is prepared by dispersing the sterile magnetic nano ferrite material in sterilized ultrapure water, and the concentration of the solution is 100 mu g/mL, 200 mu g/mL, 300 mu g/mL and 400 mu g/mL in a gradient manner. Diluting overnight activated bacteria with sterile physiological saline to 5 × 107CFU (colony-forming)/mL, uniformly spreading 100 μ L on sterilized solid LB culture medium, placing 8 μ L of antibacterial solution with different materials on the sterilized LB culture medium, culturing for 12h, and making 6 groups of observations in parallel. After the optimal bacteriostatic concentration is obtained, the bacteriostatic activity of the commercial Bordeaux mixture is used as a reference to compare the bacteriostatic activity of the composite material.
Thirdly, colony counting experiment:
adding magnetic nano ferrite material to 5 x 105And (3) in the CFU/mL bacterial suspension, the final mass concentration is 300 mug/mL, the mixture is respectively mixed for 5min, 10min, 20min and 40min, 10 muL of supernatant obtained by magnetic separation is uniformly coated on a sterilized solid LB culture medium for culture for 12h, 6 groups of observation results are parallelly made, and the bacteriostasis efficiency (n) is as follows:
in the formula: n represents the bacteriostasis efficiency, B0 is the number of colonies in the reference, and B is the number of colonies containing different materials as bacteriostasis results.
Fourthly, bacterial growth curve monitoring method experiment:
the specific influence of the magnetic nano ferrite material on the growth stage of the bacteria is not detected, and the bacteria are detected in the adaptation stage and the logarithmic phase by using micro calorimetric analysis. The bacteriostatic activity of the material is analyzed by the change of the released heat intensity in the stationary phase and the decay phase, and the higher the released heat of the bacteria is, the stronger the growth activity of the bacteria is. For this purpose, the material and the inoculated liquid LB medium were mixed to make a 5mL solution with a final bacterial concentration of 5X 107CFU, material concentration 300 μ g/mL, growth and heat release intensity of bacteria at 37 deg.C (fungi at 28 deg.C).
Fifth, bacterial PI staining method experiment:
to test the integrity of the cell membrane disruption of bacterial cells, Propidium Iodide (PI) was used as the DNA stain. Normal bacteria, which do not allow PI to permeate without color due to the intact cell membrane, also, if red color appears, it represents that the bacteria are damaged or dead, the cell membrane is destroyed, and PI stain enters the inside of the bacteria and DNA acts to show red fluorescence. For this purpose, the mixture was mixed in 50. mu.L of Propidium Iodide (PI) at 50. mu.g/mL in the dark for 15min, washed 3 times with phosphate buffer in a centrifuge at 13000r/min, and the bacterial damage was observed under a fluorescent inverted microscope.
Sixthly, toxicity experiment of the mammalian cells:
HDMEM of 10% standard fetal bovine serum, and the medium was changed every day, the cells were cultured in an environment of 37 ℃ with a carbon dioxide concentration of 5% and a humidity of 95%. Specifically, human mammary epithelial cells at a normal concentration of 2000 cells/well were added to a 96-well plate and subjected to adhesion culture overnight, the medium was removed and 100. mu.L of medium containing materials at different concentrations was added, after 3 days of culture, 25. mu.L of MTT solution (5mg/mLinPBS) was added and cultured for 2 hours, the supernatant was removed and 100. mu.L of DMSO was added to dissolve the Formazan crystals, the medium was sealed and cultured overnight, and the toxicity w of the materials to the cells was evaluated by monitoring the absorption peak with a spectrophotometer. The calculation formula is as follows:
w-OD (experimental)/OD (control) 100%.
Seventhly, plant toxicology experiment:
1. seed germination experiment:
taking full mung bean seeds, and using 10% (v/v) NaClO4Soaking and sterilizing for 10min, washing with distilled water for several times, soaking with distilled water at 4 deg.C overnight, uniformly spreading the soaked seeds in culture dishes (20 strains per culture dish), repeating for three times (initial semen Phaseoli Radiati seeds), adding ultrapure water and mixed solution of Bordeaux mixture with certain concentration and target material into the culture dishes, germinating in a greenhouse at 25 deg.C, and observing germination result after 36 hr.
2. Root activity and chlorophyll content test experiments:
adding nutrient solution into the initial mung bean seeds, culturing at 25 ℃ until the mung bean seeds germinate, transferring the germinated seedlings to the nutrient solution and mixing with a material with a certain concentration gradient, placing the mixture in a climatic incubator at 25 ℃, wherein the humidity is 60-70%, the illumination time is 16h/d, and the illumination intensity is 150 mu mol/sm2Root growth activity and chlorophyll content were tested 14 days after treatment.
Root activity test: monitoring the root growth activity of the mung bean sprouts by using a triphenyltetrazolium chloride (TTC) method, taking 0.5g of mung bean sprout roots, sequentially adding 5mL of LTTC solution (4g/L) and 5mL of phosphate buffer solution (0.07mol/L), uniformly mixing, and culturing at 37 ℃ in a dark environmentCulturing for 2h, adding 2mLH2SO4(1M) stopping the reaction, taking out the root system, wiping the root system clean, putting the root system into a graduated test tube, adding 10mL of methanol, taking supernatant fluid at 25 ℃ until the root tip is completely whitened, and monitoring the reduction amount of tetrazole by a spectrophotometry.
3. Testing of chlorophyll content:
grinding 0.15g of the above leaves with liquid nitrogen for 20min to obtain powder, soaking in 20mL of a mixture of acetone and ethanol (v/v is 1: 1) for 24h, collecting supernatant, and measuring the content of chlorophyll with a spectrophotometer.
In summary, compared with the prior art, fig. 1 is a schematic diagram of a transmission electron microscope of a magnetic nano ferrite material, and it is proved that Fe3O4the/CuO/Cu is a nano spherical material, and the material contains Fe proved by X-ray photoelectron spectroscopy3O4CuO and Cu, the material is proved to be spinel type Fe by X-ray diffraction3O4The bacteriostatic activity of the material at 100 mu g/mL is 10 times of that of Bordeaux mixture with the same concentration, and the bacteriostatic time result proves that the bacteriostatic rate of the material within 30min is 99.9 percent, so that the material can kill bacteria and damage the cell walls of the bacteria as shown by calorimetric analysis and PI dyeing results. Toxicity studies on papillary cells showed that IC50298 μ g/mL, the material showed that it increased the seed germination rate, promoted root growth and increased chlorophyll content in the seed germination, root activity, and chlorophyll content tests.
The magnetic nano ferrite material prepared by the invention is magnetic nano Fe3O4The composite material of/CuO/Cu has simple synthesis method and can be recycled under the action of an external magnetic field because of the Fe2+，Fe3+，Cu2+The electron has certain migration rate due to different oxidation-reduction potentials with Cu, the electron is combined with a medium solution to generate ROS, the ROS has bactericidal property and seed germination promoting performance, the antibacterial activity of the ROS is 10 times of that of Bordeaux mixture, and trace Fe released by a magnetic nano ferrite material (compared with the Bordeaux mixture alone) is released2+And Cu2+Can promote root growth and chlorophyll contentThe amount is increased.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.
Claims (5)
1. A preparation method of a magnetic nano ferrite material is characterized by comprising the following steps: the method comprises the following steps:
s1: 2.4mM ferric chloride and 1.2mM cupric chloride are added into ethylene glycol to form a transparent solution;
s2: then adding NaAc and sodium citrate into the transparent solution, and performing ultrasonic dispersion uniformly to form a mixed solution;
s3: and transferring the mixed solution into a polytetrafluoroethylene inner container of a 25mL stainless steel autoclave, cooling the reaction kettle at room temperature after the reaction is finished, washing a black solid product with ultrapure water and ethanol, and drying in a vacuum drying oven to finish the preparation.
2. The preparation method and the application of the magnetic nano ferrite material according to claim 1 are characterized in that: in the step S3, the reaction temperature is 200 ℃, and the reaction time is 10 h.
3. The preparation method and the application of the magnetic nano ferrite material according to claim 1 are characterized in that: the number of washing times of the black solid washed with ultrapure water and ethanol in the step S3 is 3 to 4.
4. The preparation method and the application of the magnetic nano ferrite material according to claim 1 are characterized in that: in the step S3, the drying temperature of the vacuum drying oven is 60 ℃, and the drying time is 12 h.
5. The application of the magnetic nano ferrite material is characterized by comprising the application of the magnetic nano ferrite material prepared by the preparation method of any one of claims 1 to 4 in bacteriostasis and plant growth promotion.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116267989A (en) * | 2022-09-09 | 2023-06-23 | 兰州大学 | Nanometer iron antibacterial agent for fusarium trilineum |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116267989A (en) * | 2022-09-09 | 2023-06-23 | 兰州大学 | Nanometer iron antibacterial agent for fusarium trilineum |
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Application publication date: 20220114 |