CN115918652A - Preparation method and application of halloysite nanotube coated drug-loaded naphthylacetic acid and pyraclostrobin nanoparticles - Google Patents
Preparation method and application of halloysite nanotube coated drug-loaded naphthylacetic acid and pyraclostrobin nanoparticles Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention discloses a preparation method of a nano-drug of Halloysite Nanotubes (HNTs) simultaneously coated with Naphthalalene Acetic Acid (NAA) and pyraclostrobilus fat (pyraclosabine), which comprises the following steps: (1) preparation of HNTs solution: preparing a solution from halloysite nanotubes and deionized water in proportion, and performing ultrasonic treatment in an ice-water bath; (2) preparing naphthylacetic acid and pyraclostrobin solution: respectively dissolving naphthylacetic acid and pyraclostrobin by using a method similar to that in the step (1), and performing ultrasonic treatment in an ice-water bath; (3) preparation of NAA-pyraclotrobine @ HNTs: mixing the naphthylacetic acid solution and the pyraclostrobin solution subjected to ultrasonic treatment with the HNTs solution respectively in proportion, and preparing the NAA-pyraclostatin @ HNTs nano-drug by a one-pot method by utilizing the difference of the osmotic potentials of small molecules. The nano-medicament prepared by the invention has the advantages of no toxicity, slow release, more prominent effect and the like.
Description
Technical Field
The invention belongs to the technical field of halloysite nanotube drug sustained release materials, and particularly relates to a halloysite nanotube-coated drug-loaded nano-drug of naphthylacetic acid and pyraclostrobin, and a preparation method and application thereof.
Background
Halloysite Nanotubes (HNTs) are nano-carriers with tubular structures formed by silicate minerals under the long-term weathering action, the length of the tubular structures is generally 500-1000nm, the tubes are hollow structures, and the inner diameter of the tubular structures is generally 15-100nm. Small molecular substances can be brought into the nanotube through osmosis, and long retention effect can be achieved due to the long length of the small molecular substances, so that the small molecular substances are effectively accumulated in the nanotube; secondly, a large number of hydroxyl groups and silicon-oxygen groups exist on the surface of the material, so that the material can effectively adsorb small molecular substances, and is an ideal natural adsorbing material.
In recent years, nano-carriers gradually become an important means for drug use, and the drug loaded on HNTs has a longer hollow structure and a large surface area, so that the release speed of the drug can be effectively controlled, the drug slow-release effect is achieved, the irritation of the drug is reduced, the adaptability of animals and plants to the drug is improved, the drug application frequency and the drug application amount can be reduced, and the drug is safer and more effective. At present, nano-carriers are applied to many fields, wherein HNTs play an important role in nano-drug research due to the characteristics of nature, safety, hollowness, good biocompatibility, low cost and the like. However, in the application process, the problems of low loading efficiency, poor slow release effect and the like still exist. Therefore, the key point of loading the drug by the nano-carrier is to improve the drug loading efficiency and increase the sustained release effect.
alpha-Naphthalene acetic acid (alpha-NAA) as a broad-spectrum plant growth regulator with the chemical formula of C 12 H 10 O 2 The molecular weight is 224.3, and the naphthylacetic acid has an alpha type and a beta type, wherein the alpha type is more active than the beta type. It can promote plant cell proliferation and division, induce the formation of adventitious root and raise the activity of root system, so as to help plant resist drought, salt and alkali. The application range is wide, the effect is good, the pesticide is widely used in the market, the best application mode is foliage spraying, but the growth and development of plants can be inhibited when the concentration is higher.
Pyrazolyl ether bacterioester (pyraclosterine) is a strobilurin fungicide with chemical formula C 19 H 18 CIN 3 O 4 And a molecular weight of 387.817. Has good effect on treating crop diseases caused by fungi, and in addition, the pyraclostrobin also has certain effect on plant stress resistance.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a preparation method for obtaining NAA @ HNTs and pyraclosabine @ HNTs nano-drugs by using nano-carriers HNTs to carry NAA or pyraclosabine, the nano-drugs prepared by the method are coated with HNTs, and the nano-drugs have the advantages of lasting curative effect, reduction of toxic and side effects, slow release of the drugs under the synergistic action of the carriers and the like.
In order to achieve the purpose, the invention adopts the technical scheme that: a preparation method for obtaining NAA @ HNTs and pyraclostatic @ HNTs nano-drugs by using nano-carrier HNTs to carry NAA or pyraclostatic mycolipid is characterized by comprising the following steps: (1) preparation of HNTs solution: dissolving HNTs in deionized water, and then putting the solution in an ice-water bath for ultrasonic treatment; (2) preparing NAA and pyraclostrobin solution: dissolving NAA completely, and carrying out ultrasonic treatment on the NAA solution in a beaker in an ice water bath; completely dissolving pyraclostrobin, and carrying out ultrasonic treatment on the pyraclostrobin solution in a beaker in an ice water bath; (3) preparation of NAA @ HNTs, pyraclostrobine @ HNTs: preparing the HNTs solution prepared in the step (1) with the NAA and pyraclostrobin solution prepared in the step (2) respectively according to a proportion, and preparing the NAA @ HNTs and pyraclostrobine @ HNTs by a one-pot method.
Further, in the step (1), HNTs are dissolved in deionized water, and then the HNTs solution in the beaker is subjected to ultrasonic treatment for 1-2 hours in a water bath at the ultrasonic power of 100-200W. So as to obtain the HNTs which are fully dissolved and uniformly dispersed.
Further, in the step (2), firstly, NAA and pyraclostrobin solution are respectively prepared, wherein after NAA is completely dissolved by acetone, deionized water is added into a beaker, and the NAA solution in the beaker is subjected to ultrasonic treatment for 1-2 hours in a water bath at the ultrasonic power of 100-200W. (ii) a And (3) utilizing acetonitrile to completely dissolve the pyraclostrobin, then adding deionized water into a beaker, and carrying out ultrasonic treatment on the mixture for 1-2h in a water bath at the ultrasonic power of 100-200W. .
Further, in the step (3), HNTs and NAA are proportionally prepared, react for 48 hours at the room temperature at the rotating speed of 100-300 r/min, then the solution is collected, centrifuged at 6000-8000 r/min for 5-10min, the supernatant is discarded, the precipitate is washed by absolute ethyl alcohol, frozen in a refrigerator at-80 ℃, and then a freeze dryer is used for vacuumizing and drying to obtain a sample; preparing HNTs and pyraclostrobin according to a certain proportion, reacting for 24-72h at a rotation speed of 100-300 r/min at room temperature, collecting the solution, centrifuging for 5-10min at 6000-8000 r/min, discarding the supernatant, washing the precipitate with absolute ethyl alcohol, freezing in a refrigerator at-80 ℃, and performing vacuum drying by a freeze dryer to obtain a sample.
Further, in the step (2), the purity of NAA is 99%; the purity of pyraclostrobin is 98%.
Further, the ratio of HNTs to NAA is 1:0.5-1.5; the ratio of HNTs to pyraclostrobin is 1:0.3-1.
Another object of the invention is: the invention also provides a nano-drug obtained by loading NAA or pyraclostrobin on nano-carrier HNTs to obtain NAA @ HNTs and pyraclostrobine @ HNTs, which is characterized in that: the preparation method is adopted to prepare the compound.
Another object of the invention is: the invention also provides a nano-drug obtained by loading NAA or pyraclostrobin on nano-carrier HNTs to obtain NAA @ HNTs and pyraclostrobine @ HNTs, which is characterized in that: application in preparing nanometer medicine of animal and plant.
Another object of the invention is: the invention also provides a drug sustained release method, which is characterized by comprising the step of obtaining the nano-drugs of NAA @ HNTs and pyraclocombining @ HNTs by utilizing nano-carriers HNTs to carry NAA or pyraclostrobin as claimed in claim 7.
Another object of the invention is: the invention also provides a fungal therapeutic agent, which is characterized by comprising the nano-medicament obtained by using the nano-carrier HNTs to carry pyraclostrophene resin to obtain pyraclosterone @ HNTs.
By adopting the scheme, the nano-carrier HNTs is used for carrying NAA or pyraclostrobin to obtain the nano-drugs of NAA @ HNTs and pyraclostatic @ HNTs, namely the HNTs has unique position in drug loading due to the advantages of unique hollow structure, large specific surface area, large amount of hydroxyl groups on the surface and the like. The NAA has obvious effect in plant stress resistance and small molecular weight, so that the NAA can be loaded by a nano carrier; in addition, pyraclostrobin has broad-spectrum bactericidal effect and can be adsorbed on HNTs by utilizing the characteristic of small molecular weight. In a word, NAA and pyraclostrobin can be loaded on HNTs to regulate and control the drug release speed, so that a safer and more effective drug delivery mode is provided, and a new idea is provided for complex and various drug therapies.
The invention is further described below with reference to the accompanying drawings.
Drawings
FIG. 1 is a Scanning Electron Microscope (SEM) image of the nano-drug NAA @ HNTs in example 1;
FIG. 2 is a standard curve diagram of NAA in the nano-drug NAA @ HNTs in example 1;
FIG. 3 is the slow release profile of the nano-drug NAA @ HNTs in example 1;
FIG. 4 shows the effect analysis of NAA @ HNTs as a nano-drug by using the growth phenotype difference of different processed national wheat 22 in example 1;
FIG. 5 shows the effect analysis of the NAA @ HNTs as a nano-drug by using the growth trend change of the national approved commercial 22 in example 1;
FIG. 6 is a Scanning Electron Microscope (SEM) image of the nanopharmaceutical pyraclotrobine @ HNTs in example 2;
FIG. 7 is a slow release profile of the NanoTadicine pyraclostrine @ HNTs in example 2;
FIG. 8 shows the effect analysis of the NanoTadicine pyraclostrine @ HNTs in example 2 using the growth phenotype change of Country wheat 22;
FIG. 9 shows the effect analysis of the NAA @ HNTs as a nano-drug in example 1 of example 2, using the growth trend change of national approved wheat 22;
FIG. 10 is a Scanning Electron Microscope (SEM) image of the nano-drug NAA-pyraclostrobine @ HNTs in example 3;
FIG. 11 is a graph showing the effect of NAA-pyraclostrine @ HNTs as a nano-drug in example 3 using the growth phenotype difference of different treated domestic wheat 22.
Detailed Description
The reagents and instruments used in the specific embodiment of the invention are as follows:
halloysite Nanotubes (HNTs) were purchased from qian xiaofeng nanomaterial science and technology ltd, jiang su;
alpha-Naphthylacetic acid (alpha-NAA) was purchased from Dai Biochemical Co., ltd, bao chicken;
pyraclostrobin (pyraclosterone) was purchased from biotech ltd, austria;
shanju Ji 22 is purchased from Shandong research agriculture fine breed Co., ltd;
acetonitrile was purchased from Tianjin Bohai chemical reagent GmbH;
acetone was purchased from Jin Weier chemical ltd, tianjin;
the magnetic stirrer adopts RCT digital of the moxa card, germany;
the centrifuge adopts TGL-16M of Hunan instrument centrifuge instruments ltd;
the vacuum freeze drier adopts LGJ-10 of Beijing pine source Huaxing science and technology development Limited company;
the ultra-low temperature refrigerator adopts THERMO Scientific form 900 series, USA;
the ultraviolet visible spectrophotometer adopts an Alpha-1 series of Shanghai Pu Yuan instruments ltd;
scanning Electron Microscopy (SEM) used was a Gemini SEM 300 from Zeiss (Zeiss) in germany;
example 1
1. The preparation method utilizes nano-carrier HNTs to carry NAA to obtain NAA @ HNTs, namely the preparation of the NAA @ HNTs, and comprises the following specific steps:
step one, preparing halloysite nanotubes into an opalescent solution, namely preparing HNTs solution
(1) The halloysite nanotube is obtained from Jiangsu pioneer nanometer materials science and technology limited, has purity of more than 95%, diameter of 50-300nm, length of 1-10um, and molecular formula of Al 2 Si 2 O 5 (OH) 4 ·2H 2 O。
(2) Dissolving 10mgHNTs in 10mL of aqueous solution, stirring for preliminary dissolution to obtain a milky solution, and performing ultrasonic treatment on the milky solution at the ultrasonic power of 100W in an ice-water bath at room temperature for 1h to obtain the HNTs solution with better solubility and dispersibility.
Step two, preparation of colorless solution of alpha-naphthylacetic acid, namely preparation of alpha-NAA solution
(1) alpha-Naphthylacetic acid was obtained from Senri Biochemical Co., ltd, baoji, having a purity of 99%, and having the chemical formula C 12 H 10 O 2 The molecular weight is 224.3.
(2) Adding 0.5mL of acetone into 10mg of alpha-NAA to obtain colorless transparent liquid, adding 10mL of aqueous solution, and performing ultrasonic treatment on the solution for 2 hours at room temperature in an ice-water bath with ultrasonic power of 100W to obtain the alpha-NAA solution with better solubility and dispersibility.
Step three, loading alpha-naphthylacetic acid into halloysite nanotubes, namely preparation of NAA @ HNTs
The preparation of the NAA @ HNTs is completed by one-pot method of HNTs solution and alpha-NAA solution.
And (3) mixing the solutions prepared in the first step and the second step according to the mass ratio of HNTs to alpha-NAA of 1:1, putting 20ml of mixed solution into a beaker, reacting for 48 hours at the room temperature at the rotation speed of 200 revolutions per minute, collecting the solution, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, washing the precipitate with absolute ethyl alcohol, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, collecting the precipitate, freezing in a refrigerator at-80 ℃, and vacuumizing and drying by a freeze dryer to obtain the sample.
2. Test characterization experiment:
1. the prepared NAA @ HNTs are subjected to scanning electron microscope test and characterization experiments as follows:
respectively dissolving 1mgNAA @ HNTs in 5ml of deionized water, and after the solution is fully dissolved, taking a proper amount of liquid to be conventionally dripped on a silicon wafer for natural airing or drying test or directly conventionally adhering powder on a conductive adhesive for testing. As shown in FIG. 1, it was observed that the dispersibility of NAA @ HNTs was good and the hollow structure of HNTs was clearly observed.
2. The test characterization experiments performed on the NAA @ HNTs prepared in example 1 were as follows:
preparing 10mg/L, 5mg/L, 2.5mg/L, 1.25mg/L, 0.625mg/L and 0.3125mg/L of standard solution by NAA, respectively, performing ultrasonic treatment for 1-2h under the ultrasonic power of ice-water bath of 100W, measuring the absorbance at 281nm by using an ultraviolet spectrophotometer, and drawing a standard curve, as shown in FIG. 2.
3. The test and characterization experiment of the slow release effect of the prepared NAA @ HNTs is as follows:
5mg of NAA @ HNTs prepared in example 1 was added with 10ml of deionized water to prepare a solution, which was put into a dialysis bag with cut-off molecular weight of 3500 and put into a centrifuge tube with 50ml of deionized water. Under the condition of 25 ℃, the ultraviolet absorption at 281nm is measured at 0h, 2h, 4h, 6h, 8h, 10h, 12h, 24h, 48h and 72h respectively, the content of NAA in NAA @ HNTs of a sample is calculated by utilizing a standard curve of NAA, and a slow release curve graph is drawn, as shown in figure 3, the slow release effect is demonstrated.
4. The prepared NAA @ HNTs helps the stress resistance of plants to detect:
preparing a NaCl solution with the concentration of 200mM/L and simulating salt stress.
II, taking the NAA @ HNTs prepared in the example 1, weighing 1mg of NAA @ HNTs, adding 1L of deionized water, preparing a 1mg/L NAA @ HNTs solution,
III, adding the prepared solution in the II into wheat (national examined wheat 22) stressed by 200 mM/LNacl. Another group of clear water contrast is arranged; a set of 200mM/LNacl controls. By the sixth day of culture, the growth condition of wheat stressed by 200mM/L NaCl after adding NAA @ HNTs can be obviously observed to be better, as shown in figure 4, a) the 200mM/L NaCl treatment group; b) 200mM/L NaCl +1mg/L NAA @ HNTs treated group. The growth changes of the 3 groups of wheat at 0, 4, 8, 12 and 14 days were recorded, respectively, as shown in fig. 5, demonstrating that naa @ hnts can help wheat to resist salt stress.
Example 2
1. The preparation method comprises the following specific steps of preparing pyraclosterone @ HNTs by using a nanocarrier HNTs to carry pyraclosterone mycolipid, namely the preparation method of pyraclosterone @ HNTs comprises the following steps:
step one, preparing halloysite nanotubes into opalescent solution, namely preparation of HNTs solution
(1) The halloysite nanotube is obtained from Jiangsu pioneer nanometer materials science and technology limited, has purity of more than 95%, diameter of 50-300nm, length of 1-10um, and molecular formula of Al 2 Si 2 O 5 (OH) 4 ·2H 2 O。
(2) Dissolving 10mgHNTs in 10mL of aqueous solution, stirring for preliminary dissolution to obtain a milky solution, and performing ultrasonic treatment on the milky solution at the ultrasonic power of 100W in an ice-water bath at room temperature for 1h to obtain the HNTs solution with better solubility and dispersibility.
Step two, preparing the pyraclostrobin into solution by using acetonitrile, namely preparing the pyraclostrobin solution
(1) Pyraclostrobin is obtained from Ecklonia cava Biotech limited, with purity of more than 98%, and has chemical formula C 19 H 18 CIN 3 O 4 And a molecular weight of 387.817.
(2) Adding 1mL of acetonitrile into 8mg of pyraclostrobin, adding 8mL of water solution, and performing ultrasonic treatment for 2 hours at room temperature in an ice-water bath with ultrasonic power of 100W to obtain the pyraclostrobin solution with better solubility and dispersibility.
Step three, loading pyraclostrobin into halloysite nanotubes, namely preparation of pyraclosterine @ HNTs
Preparation of pyraclostrobine @ HNTs was accomplished by mixing a solution of HNTs with a solution of pyraclostrobin.
And (3) mixing the solutions prepared in the first step and the second step according to the mass ratio of HNTs to pyraclostrobin of 1: preparing according to the proportion of 0.8, putting 20ml of prepared mixed solution into a beaker, reacting for 48 hours at the room temperature at the rotating speed of 200 revolutions per minute, collecting the solution, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, washing the precipitate with absolute ethyl alcohol once, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, collecting the precipitate, freezing in a refrigerator at the temperature of minus 80 ℃, and vacuumizing and drying by a freeze dryer to obtain a sample.
2. Test characterization experiment:
1. scanning electron microscopy tests and characterization experiments were performed on pyraclostrobine @ hnts prepared in example 2 as follows:
dissolving 1mg of Pyraclostrobine @ HNTs in 5ml of deionized water, and after the materials are fully dissolved, taking a proper amount of liquid and conventionally dripping the liquid on a silicon wafer for natural airing or drying test or directly and conventionally adhering powder on a conductive adhesive for testing. As shown in FIG. 6, the dispersibility of Pyraclostrobine @ HNTs was found to be good, and the hollow structure of HNTs was clearly observed.
2. The slow release effect of the prepared pyraclosterobine @ HNTs is tested and characterized as follows:
5mg of pyraclosterone @ HNTs prepared in example 2 was added with 10ml of deionized water to prepare a solution, the solution was put into a dialysis bag with a cut-off molecular weight of 3500, and the dialysis bag was centrifuged with 50ml of deionized water, and the absorbance at 272nm was measured at 25 ℃ for 2h, 4h, 6h, 8h, 10h, 12h, 24h, 48h and 72h, respectively, and the content of pyraclosterone bacterioester in the pyraclosterone @ HNTs was calculated using a standard curve of pyraclosterone bacterioester, and a sustained release curve was plotted, as shown in FIG. 7, indicating that it had a sustained release effect.
4. The prepared pyraclostrobine @ HNTs helps the stress resistance of plants to be detected:
preparing a NaCl solution with the concentration of 200mM/L and simulating salt stress.
II, taking the pyraclosterone @ HNTs prepared in the example 1, weighing 3mg of pyraclosterone @ HNTs, adding 1L of deionized water, and preparing a pyraclosterone @ HNTs solution with the concentration of 3 mg/L.
III, adding the solution prepared in the step II into wheat (national audition wheat 22) stressed by 200mM/LNacl, setting clear water and 200mM/LNacl control, culturing for six days, and observing the phenotype, wherein a) the 200mM/L NaCl treatment group is shown in figure 8; b) 200mM/L Nacl +3mg/L pyraclostrobine @ HNTs treatment group. The growth changes of the 3 groups of wheat were recorded at 0, 4, 8, 12, 14 days, respectively, as shown in fig. 9, demonstrating that pyraclostrobine @ hnts can help wheat to resist salt stress.
Example 3
1. The preparation method comprises the following steps of simultaneously loading pyraclostrobin and naphthylacetic acid by using nano-carrier HNTs to obtain NAA-pyraclosterone @ HNTs, namely the preparation of the NAA-pyraclosterone @ HNTs, and comprises the following specific steps:
step one, preparing halloysite nanotubes into an opalescent solution, namely preparing HNTs solution
(1) The halloysite nanotube is obtained from Jiangsu pioneer nanometer materials science and technology limited, has purity of more than 95%, diameter of 50-300nm, length of 1-10um, and molecular formula of Al 2 Si 2 O 5 (OH) 4 ·2H 2 O。
(2) Dissolving 10mgHNTs in 10mL of aqueous solution, stirring for preliminary dissolution to obtain a milky solution, and performing ultrasonic treatment on the milky solution at the ultrasonic power of 100W in an ice-water bath at room temperature for 1h to obtain the HNTs solution with better solubility and dispersibility.
Step two, preparing naphthylacetic acid and pyraclostrobin solution
(1) Adding 1mL of acetonitrile into 8mg of pyraclostrobin, adding 8mL of water solution, and performing ultrasonic treatment for 2 hours at room temperature in an ice-water bath with ultrasonic power of 100W to obtain the pyraclostrobin solution with better solubility and dispersibility.
(2) Adding 0.5mL of acetone into 10mg of alpha-NAA to obtain colorless transparent liquid, adding 10mL of aqueous solution, and performing ultrasonic treatment on the solution for 2 hours at room temperature in an ice-water bath with ultrasonic power of 100W to obtain the alpha-NAA solution with better solubility and dispersibility.
Step three, loading pyraclostrobin and naphthylacetic acid into halloysite nanotubes, namely preparation of NAA-pyraclosterine @ HNTs
Namely, the preparation of NAA-pyraclotrobine @ HNTs was carried out by mixing a solution of HNTs with a solution of pyraclostrobin and then carrying out the one-pot method.
And (3) mixing the solutions prepared in the first step and the second step according to the mass ratio of HNTs to pyraclostrobin to naphthylacetic acid of 1:0.8: preparing at the ratio of 0.2, putting 20ml of prepared mixed solution into a beaker, reacting for 48 hours at the room temperature at the rotation speed of 200 revolutions per minute, collecting the solution, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, washing the precipitate with absolute ethyl alcohol, centrifuging for 10 minutes at 8000 revolutions per minute, discarding the supernatant, collecting the precipitate, freezing in a refrigerator at-80 ℃, and vacuumizing and drying by a freeze dryer to obtain the sample.
2. Test characterization experiment:
1. scanning electron microscopy characterization experiments were performed on NAA-pyraclotrobine @ HNTs prepared in example 2 as follows:
dissolving 1mg NAA-pyraclotrobine @ HNTs in 5ml deionized water, and after fully dissolving, taking a proper amount of liquid to drop on a silicon wafer conventionally for natural airing or drying test or directly sticking powder on a conductive adhesive conventionally for test. As shown in FIG. 10, it was observed that the dispersion of the NAA-pyraclotrobine @ HNTs was good, and the hollow structure of HNTs was clearly observed.
2. The prepared pyraclostrobine @ HNTs helps the stress resistance of plants to be detected:
preparing a NaCl solution with the concentration of 200mM/L and simulating salt stress.
II, taking the NAA-pyraclosterobine @ HNTs prepared in the example 1, weighing 1.5mg, adding 1L of deionized water, and preparing a NAA-pyraclosterobine @ HNTs solution with the concentration of 1.5 mg/L.
III, adding the prepared solution in the II into wheat (national examined wheat 22) stressed by 200mM/LNacl, setting clear water and 200mM/LNacl control, and obviously differentiating phenotypes when culturing for the fifth day, as shown in figure 11, a) 200mM/L NaCl treatment groups; b) 200mM/L NaCl +1.5mg/L NAA-Pyraclostrobine @ HNTs treated group. The NAA-pyraclosterobine @ HNTs can help wheat to resist salt stress and have obvious effect.
The present invention is not limited to the above embodiments, and those skilled in the art can implement the present invention in other various embodiments according to the disclosure of the present invention, or can make simple changes or modifications according to the design structure and idea of the present invention, and fall into the protection scope of the present invention.
Claims (10)
1. A preparation method of a halloysite nanotube coated drug-loaded naphthylacetic acid and pyraclostrobin nano-drug is characterized by comprising the following steps: (1) preparation of HNTs solution: preparing a solution of halloysite nanotubes with deionized water, and carrying out ultrasonic treatment on the HNTs solution prepared in a beaker in an ice-water bath; (2) preparing naphthylacetic acid and pyraclostrobin solution: grinding naphthylacetic acid into finer powder by using a mortar, preparing into a solution, and putting into an ice water bath for ultrasonic treatment; grinding pyraclostrobin into finer powder in a mortar, preparing into solution, and placing into ice water bath for ultrasonic treatment; (3) preparation of NAA @ HNTs, pyraclostrobine @ HNTs, NAA-pyraclosterobine @ HNTs: mixing the HNTs solution after ultrasonic treatment with naphthylacetic acid and pyraclostrobin solution in proportion, and preparing NAA @ HNTs, pyraclosterone @ HNTs and NAA-pyraclosterone @ HNTs by a one-pot method.
2. The method for preparing the nano-drug by using the halloysite nanotube coated drug-loaded naphthylacetic acid and pyraclostrobin as claimed in claim 1, wherein in the step (1), the HNTs are dissolved by deionized water and then subjected to ultrasound for 1-2 hours in a water bath under the ultrasound power of 100-200W.
3. The method for preparing the nano-drug by using the halloysite nanotube coated drug-loaded naphthylacetic acid and pyraclostrobin nano-drug as the claimed in claim 1, wherein in the step (2), a naphthylacetic acid solution is prepared, the naphthylacetic acid is ground to be finer and then placed into a beaker, acetone or an ethanol solution (the solubility of the naphthylacetic acid in the water solution is low) is used for dissolving the naphthylacetic acid, then deionized water is added, the prepared naphthylacetic acid solution is subjected to ultrasonic treatment for 1-2 hours at an ice-water bath ultrasonic power of 100-200W, and the temperature is controlled to be 20-40 ℃; preparing a pyraclostrobin solution, putting the ground pyraclostrobin in a beaker, adding an acetonitrile solution to dissolve the pyraclostrobin solution, adding deionized water into the pyraclostrobin turbid solution, and carrying out ultrasonic treatment for 1-2h at an ice-water bath ultrasonic power of 100-200W.
4. The method for preparing nano-drugs by coating drug-loaded naphthylacetic acid and pyraclostrobin with halloysite nanotubes according to claim 1, wherein in the step (3), NAA @ HNTs, pyraclostrobine @ HNTs, NAA and Pyraclostrobine in NAA-Pyraclostrobine @ HNTs are prepared by ultrasonic treatment in advance in the steps (1) and (2). Mixing HNTs and NAA in proportion, reacting for 24-72h at the rotation speed of 100-300 r/min and at room temperature, collecting solution, centrifuging at 6000-8000 r/min for 5-10min, discarding supernatant to obtain purer NAA @ HNTs nano-drug, washing precipitate with absolute ethyl alcohol, washing off unloaded NAA, centrifuging at 8000 r/min for 5-10min, discarding supernatant, collecting precipitate, freezing in a-80 ℃ refrigerator, sealing with a sealing film, perforating at the sealing film, and freeze-drying and storing by a freeze dryer; mixing HNTs and pyraclostrobin in proportion, reacting for 48h at the room temperature at the rotation speed of 100 r/min, collecting the solution, centrifuging at 6000-8000 r/min for 5-10min, discarding the supernatant, washing the precipitate with absolute ethanol, washing off the unloaded pyraclostrobin, centrifuging at 6000-8000 r/min for 5-10min, discarding the supernatant, collecting the precipitate, freezing in a refrigerator at-80 ℃, sealing with a sealing membrane, pricking a hole at the sealing membrane, and freeze-drying and storing by using a freeze dryer; mixing HNTs with pyraclostrobin and naphthylacetic acid in proportion, reacting at 100 r/min at room temperature for 48h, collecting solution, centrifuging at 6000-8000 r/min for 5-10min, discarding supernatant, washing precipitate with anhydrous ethanol, washing off unloaded pyraclostrobin, centrifuging at 6000-8000 r/min for 5-10min, discarding supernatant, collecting precipitate, freezing in-80 deg.C refrigerator, sealing with sealing membrane, perforating at the sealing membrane, and freeze-drying with freeze dryer.
5. The method for preparing nano-drugs by using halloysite nanotubes to coat drug-loaded naphthylacetic acid and pyraclostrobin according to any one of claims 1 to 4, wherein in the step (3), the purity of NAA is 99% and the purity of pyraclostrobin is 98%.
6. The method for preparing nano-drugs by coating drug-loaded naphthylacetic acid and pyraclostrobin with halloysite nanotubes according to any one of claims 1 to 4, wherein the mass ratio of HNTs to naphthylacetic acid is 1:0.5-1.5 (mg/mg); the mass ratio of HNTs to pyraclostrobin is 1:0.3-1 (mg/mg); the mass ratio of HNTs to pyraclostrobin to naphthylacetic acid is 1:0.3-1:0.1-0.5.
7. A nano-drug containing HNTs coated with NAA or pyraclostrobin is characterized in that: prepared by the preparation method of any one of claims 1 to 5.
8. A nano-drug containing HNTs coated with NAA or pyraclostrobin is characterized in that: the application in stress resistance and sterilization of plants.
9. A nano-drug comprising the nano-drug carrying NAA coated with HNTs nano-carrier of claim 7.
10. A nano-drug comprising the nano-drug of claim 7 coated with drug-loaded pyraclostrobin with HNTs.
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| CN106212450A (en) * | 2016-07-21 | 2016-12-14 | 河南师范大学 | A kind of modified halloysite nanotubes composite sustained-release pesticides and preparation method thereof |
| CN108849952A (en) * | 2017-05-10 | 2018-11-23 | 江苏龙灯化学有限公司 | A kind of pyraclostrobin water-dispersible solid composition and preparation method thereof |
| CN109704868A (en) * | 2019-02-28 | 2019-05-03 | 南开大学 | A kind of novel membrane-biological membrane inhibits the sustainable release method of molecule |
| CN112006019A (en) * | 2020-09-11 | 2020-12-01 | 浙江奚态生物科技有限公司 | Preparation method of halloysite-based nano drug-loaded material |
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| CN106212450A (en) * | 2016-07-21 | 2016-12-14 | 河南师范大学 | A kind of modified halloysite nanotubes composite sustained-release pesticides and preparation method thereof |
| CN108849952A (en) * | 2017-05-10 | 2018-11-23 | 江苏龙灯化学有限公司 | A kind of pyraclostrobin water-dispersible solid composition and preparation method thereof |
| CN109704868A (en) * | 2019-02-28 | 2019-05-03 | 南开大学 | A kind of novel membrane-biological membrane inhibits the sustainable release method of molecule |
| CN112006019A (en) * | 2020-09-11 | 2020-12-01 | 浙江奚态生物科技有限公司 | Preparation method of halloysite-based nano drug-loaded material |
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