CN106942256A - A kind of new preparation method for containing thiacloprid and the protein nano ball of pheromones - Google Patents
A kind of new preparation method for containing thiacloprid and the protein nano ball of pheromones Download PDFInfo
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- 239000003016 pheromone Substances 0.000 title claims abstract description 31
- HOKKPVIRMVDYPB-UVTDQMKNSA-N (Z)-thiacloprid Chemical compound C1=NC(Cl)=CC=C1CN1C(=N/C#N)/SCC1 HOKKPVIRMVDYPB-UVTDQMKNSA-N 0.000 title claims abstract description 22
- 239000005940 Thiacloprid Substances 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 11
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 11
- 239000011807 nanoball Substances 0.000 title claims 6
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 35
- 108010088751 Albumins Proteins 0.000 claims abstract description 20
- 102000009027 Albumins Human genes 0.000 claims abstract description 20
- 239000007864 aqueous solution Substances 0.000 claims abstract description 13
- 239000002077 nanosphere Substances 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- FGPPDYNPZTUNIU-UHFFFAOYSA-N pentyl pentanoate Chemical compound CCCCCOC(=O)CCCC FGPPDYNPZTUNIU-UHFFFAOYSA-N 0.000 claims description 6
- 238000004945 emulsification Methods 0.000 claims description 5
- 239000002105 nanoparticle Substances 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims 2
- 238000002156 mixing Methods 0.000 claims 2
- 239000000203 mixture Substances 0.000 claims 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 235000007164 Oryza sativa Nutrition 0.000 claims 1
- 239000011805 ball Substances 0.000 claims 1
- 238000001816 cooling Methods 0.000 claims 1
- 239000006185 dispersion Substances 0.000 claims 1
- 239000012153 distilled water Substances 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 235000009566 rice Nutrition 0.000 claims 1
- 238000007789 sealing Methods 0.000 claims 1
- 238000002604 ultrasonography Methods 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 9
- 229940079593 drug Drugs 0.000 abstract description 9
- 239000000575 pesticide Substances 0.000 abstract description 8
- 241000607479 Yersinia pestis Species 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 2
- 239000002699 waste material Substances 0.000 abstract 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 8
- 229940098773 bovine serum albumin Drugs 0.000 description 8
- 241000238631 Hexapoda Species 0.000 description 6
- 239000012154 double-distilled water Substances 0.000 description 4
- 238000009775 high-speed stirring Methods 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 3
- 231100000053 low toxicity Toxicity 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- 239000012888 bovine serum Substances 0.000 description 2
- 230000005847 immunogenicity Effects 0.000 description 2
- 239000002917 insecticide Substances 0.000 description 2
- 239000003094 microcapsule Substances 0.000 description 2
- 231100000956 nontoxicity Toxicity 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 239000005906 Imidacloprid Substances 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001055 chewing effect Effects 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- YWTYJOPNNQFBPC-UHFFFAOYSA-N imidacloprid Chemical compound [O-][N+](=O)\N=C1/NCCN1CC1=CC=C(Cl)N=C1 YWTYJOPNNQFBPC-UHFFFAOYSA-N 0.000 description 1
- 229940056881 imidacloprid Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
Classifications
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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
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/40—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides
-
- 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
- A01N25/10—Macromolecular compounds
-
- 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
- A01N25/12—Powders or granules
-
- 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
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Agronomy & Crop Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Toxicology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明公开了一种新型包载噻虫啉和信息素的蛋白纳米球的制备方法。所述噻虫啉和信息素溶解于乙腈做油相，将油相逐滴加入白蛋白水溶液中，乳化一段时间，冷干得到白蛋白‑噻虫啉‑信息素纳米球的粉末制剂。该方法将药物和信息素同时载入白蛋白中，在增加载药量的同时，通过信息素的引诱，更好的实现纳米药物对害虫的触杀作用。纳米科技的利用可以使农药产品微型化，独特的缓释功能减少药物资源的浪费，降低成本，显著地提高药效，粉末制剂便于运输，方便配置。The invention discloses a preparation method of a novel protein nanosphere loaded with thiacloprid and pheromone. The thiacloprid and pheromone are dissolved in acetonitrile to make an oil phase, and the oil phase is added dropwise into an albumin aqueous solution, emulsified for a period of time, and lyophilized to obtain a powder preparation of albumin-thiacloprid-pheromone nanospheres. In this method, drugs and pheromones are loaded into albumin at the same time. While increasing the amount of drugs loaded, the contact effect of nano-medicines on pests can be better realized through the lure of pheromones. The use of nanotechnology can miniaturize pesticide products. The unique slow-release function reduces the waste of drug resources, reduces costs, and significantly improves drug efficacy. Powder preparations are easy to transport and configure.
Description
技术领域technical field
本发明公开了一种新型包载噻虫啉和信息素的蛋白纳米球的制备方法，特别是牛血清白蛋白同时装载农药噻虫啉和昆虫信息素戊酸戊酯的过程。属于纳米技术和新型农药领域。The invention discloses a preparation method of a novel protein nanosphere loaded with thiacloprid and a pheromone, in particular a process in which bovine serum albumin is simultaneously loaded with the pesticide thiacloprid and the insect pheromone amyl valerate. It belongs to the fields of nanotechnology and new pesticides.
背景技术Background technique
噻虫啉是继吡虫啉之后由拜耳公司开发的另一个新型氯代烟碱类杀虫剂，具有高效、低毒、广谱作用，同时兼具胃毒、触杀及内吸活性，是防治刺吸式和咀嚼式口器害虫的高效药剂。目前较为新型的农药制剂有微胶囊粉剂和微胶囊水悬浮剂等，粒径较大，水溶性差，制备复杂，不便于农业害虫的防治，且运输困难、不易长期存放等诸多问题。Thiacloprid is another new type of chloronicotinoid insecticide developed by Bayer after imidacloprid. It has high efficiency, low toxicity and broad-spectrum effects. It also has stomach toxicity, contact and systemic activities. High-efficiency insecticide for insect and chewing mouthparts pests. At present, relatively new pesticide preparations include microcapsule powder and microcapsule water suspension concentrate, etc., which have large particle size, poor water solubility, complicated preparation, inconvenient control of agricultural pests, difficult transportation, and difficult long-term storage.
白蛋白具有安全无毒、无免疫原性、可生物降解及生物相容性好等优点，是一种理想的给药系统载体材料。具有的可生物降解、无毒、无抗原性和生物利用度高等特点。单纯的白蛋白载药不能引诱害虫并大范围消灭虫害，信息素可以刺激虫子信息接收神经元的敏感性，从而增强对虫子的引诱力。一种新型牛血清白蛋白-噻虫啉-信息素纳米农药可以大范围的引诱并杀灭虫子，有效地消灭虫害。且易被吞食和吸收，更好地发挥触杀和胃毒作用，实现纳米生物农药的高效、低毒、安全。Albumin has the advantages of safety, non-toxicity, non-immunogenicity, biodegradability and good biocompatibility, and is an ideal carrier material for drug delivery systems. It has the characteristics of biodegradable, non-toxic, non-antigenic and high bioavailability. Simple albumin-loaded drugs cannot attract pests and eliminate pests on a large scale. Pheromone can stimulate the sensitivity of insect information receiving neurons, thereby enhancing the attractiveness to insects. A novel bovine serum albumin-thiacloprid-pheromone nano-pesticide can lure and kill insects in a wide range and effectively eliminate insect pests. And it is easy to be swallowed and absorbed, and it can better play the role of contact killing and stomach poisoning, and realize the high efficiency, low toxicity and safety of nano-biological pesticides.
发明内容Contents of the invention
本发明的目的是提供一种新型包载噻虫啉和信息素的蛋白纳米球的制备方法，该方法采用的新型白蛋白纳米制备技术是在不改变白蛋白结构的前提下，将药物和信息素同时载入其中。The purpose of the present invention is to provide a new method for preparing protein nanospheres loaded with thiacloprid and pheromones. The new albumin nano-preparation technology used in the method is to combine drugs and information without changing the albumin structure. elements are loaded into it at the same time.
本发明的上述目的通过以下技术方案实现：牛血清白蛋白包裹噻虫啉的过程采用了乳化固化法，白蛋白纳米粒作为一种药物载体，无免疫原性、生物相容性好。The above object of the present invention is achieved through the following technical scheme: the process of encapsulating thiacloprid with bovine serum albumin adopts an emulsification and solidification method, and albumin nanoparticles are used as a drug carrier, which has no immunogenicity and good biocompatibility.
所述的新型包载噻虫啉和信息素的蛋白纳米球的过程，噻虫啉和信息素溶解于乙腈做油相，将油相逐滴加入白蛋白水溶液中，乳化一段时间，冷干得到白蛋白-噻虫啉-信息素纳米球的粉末制剂，具体步骤如下：In the process of the novel protein nanospheres loaded with thiacloprid and pheromones, thiacloprid and pheromones are dissolved in acetonitrile to make the oil phase, and the oil phase is added dropwise into the albumin aqueous solution, emulsified for a period of time, and lyophilized to obtain The powder preparation of albumin-thiacloprid-pheromone nanospheres, the specific steps are as follows:
（1）取一定量的白蛋白溶于双蒸水，50oC水浴30min，过滤，得到白蛋白水溶液，封口待用；(1) Dissolve a certain amount of albumin in double-distilled water, bathe in 50 o C water for 30 minutes, filter to obtain albumin aqueous solution, and seal it for later use;
（2）另取噻虫啉溶于乙腈，超声5-20min，配制成2-15mg/mL的噻虫啉-乙腈有机溶液，封口待用；(2) Another thiacloprid was dissolved in acetonitrile, ultrasonicated for 5-20 minutes, and prepared into a 2-15 mg/mL thiacloprid-acetonitrile organic solution, sealed for use;
（3）有机溶剂与信息素戊酸戊酯混合，得到混合油相，封口待用；(3) The organic solvent is mixed with the pheromone pentyl valerate to obtain a mixed oil phase, which is sealed for use;
（4）将白蛋白水溶液调节转速1000 rpm，高速搅拌下逐滴加入混合油相，乳化40min，低温冷却5-10min，然后超声5min，得到白蛋白-噻虫啉-信息素纳米球的粉末制剂。(4) Adjust the rotation speed of the albumin aqueous solution to 1000 rpm, add dropwise to the mixed oil phase under high-speed stirring, emulsify for 40 minutes, cool at low temperature for 5-10 minutes, and then sonicate for 5 minutes to obtain the powder preparation of albumin-thiacloprid-pheromone nanospheres .
所述的新型包载噻虫啉和信息素的蛋白纳米球的形貌见图1。The morphology of the novel protein nanospheres loaded with thiacloprid and pheromone is shown in FIG. 1 .
步骤（2）中的乙腈也可用氯仿、乙醇、甲醇等多种有机溶剂替代。The acetonitrile in step (2) can also be replaced by various organic solvents such as chloroform, ethanol, and methanol.
步骤（2）中的超声时间5-20min，所得到的纳米粒粒径均匀，分散性好。The ultrasonic time in step (2) is 5-20 minutes, and the obtained nanoparticles have uniform particle size and good dispersibility.
步骤（3）中的乳化过程时间30-60min，时间太短乳化不充分，太长会导致更多的聚集。The emulsification process time in step (3) is 30-60min. If the time is too short, the emulsification will not be sufficient. If the time is too long, more aggregation will result.
本发明具有以下优点：The present invention has the following advantages:
（1）白蛋白本身具有可生物降解、无毒、无抗原性和生物利用度高等特点。(1) Albumin itself has the characteristics of biodegradability, non-toxicity, non-antigen and high bioavailability.
（2）包裹信息素的药物可以引诱害虫，更好地发挥触杀和胃毒作用。(2) Drugs wrapped with pheromones can lure pests and better exert contact and stomach poisoning effects.
（3）白蛋白纳米粒的载药量大，具有控释能力，实现纳米生物农药的高效、低毒、安全。(3) Albumin nanoparticles have a large drug loading capacity and have controlled release capabilities, realizing the high efficiency, low toxicity and safety of nano-biological pesticides.
（4）本发明不需要特殊的化学试剂，工艺简单，设备要求低，绿色环保，降低制造成本。(4) The present invention does not require special chemical reagents, has a simple process, requires low equipment requirements, is environmentally friendly, and reduces manufacturing costs.
附图说明Description of drawings
图1为本发明制备的新型农药牛血清白蛋白-噻虫啉-信息素纳米粒子的场发射电镜形貌。Fig. 1 is the field emission electron microscope morphology of the novel pesticide bovine serum albumin-thiacloprid-pheromone nanoparticles prepared by the present invention.
具体实施方式detailed description
实施例一Embodiment one
（1）取0.05g牛血清白蛋白溶于4mL双蒸水中，配制成水相，50oC水浴30min，得到澄清透明牛血清白蛋白水溶液；(1) Dissolve 0.05g of bovine serum albumin in 4mL of double-distilled water to prepare the water phase, and bathe in 50 o C water for 30 minutes to obtain a clear and transparent aqueous solution of bovine serum albumin;
（2）另取0.05mg噻虫啉，加入0.5mL乙腈超声溶解5min，配制成0.1mg/mL的噻虫啉-乙腈油相；(2) Take another 0.05 mg of thiacloprid, add 0.5 mL of acetonitrile for ultrasonic dissolution for 5 minutes, and prepare a 0.1 mg/mL thiacloprid-acetonitrile oil phase;
（3）有机溶剂与0.5mL戊酸戊酯混合，得到混合油相；(3) The organic solvent is mixed with 0.5mL amyl valerate to obtain a mixed oil phase;
（4）将白蛋白水溶液调节转速1000 rpm，高速搅拌下逐滴加入混合油相，乳化40min，低温冷却5-10min，然后超声5min，得到白蛋白-噻虫啉-信息素纳米球的粉末制剂。(4) Adjust the rotation speed of the albumin aqueous solution to 1000 rpm, add dropwise to the mixed oil phase under high-speed stirring, emulsify for 40 minutes, cool at low temperature for 5-10 minutes, and then sonicate for 5 minutes to obtain the powder preparation of albumin-thiacloprid-pheromone nanospheres .
实施例二Embodiment two
（1）取0.1g牛血清白蛋白溶于10mL双蒸水中，配制成水相，50oC水浴30min，得到澄清透明牛血清白蛋白水溶液；(1) Dissolve 0.1g of bovine serum albumin in 10mL of double-distilled water to prepare the water phase, and bathe in 50 o C water for 30 minutes to obtain a clear and transparent bovine serum albumin aqueous solution;
（2）另取0.05mg噻虫啉，加入0.5mL乙腈超声溶解5min，配制成0.1mg/mL的噻虫啉-乙腈油相；(2) Take another 0.05 mg of thiacloprid, add 0.5 mL of acetonitrile for ultrasonic dissolution for 5 minutes, and prepare a 0.1 mg/mL thiacloprid-acetonitrile oil phase;
（3）有机溶剂与0.5mL戊酸戊酯混合，得到混合油相；(3) The organic solvent is mixed with 0.5mL amyl valerate to obtain a mixed oil phase;
（4）将白蛋白水溶液调节转速1000 rpm，高速搅拌下逐滴加入混合油相，乳化30min，低温冷却5-10min，然后超声5min，得到白蛋白-噻虫啉-信息素纳米球的粉末制剂。(4) Adjust the rotation speed of the albumin aqueous solution to 1000 rpm, add dropwise to the mixed oil phase under high-speed stirring, emulsify for 30 minutes, cool at low temperature for 5-10 minutes, and then sonicate for 5 minutes to obtain the powder preparation of albumin-thiacloprid-pheromone nanospheres .
实施例三Embodiment three
（1）取0.05g牛血清白蛋白溶于4mL双蒸水中，配制成水相，50oC水浴30min，得到澄清透明牛血清白蛋白水溶液；(1) Dissolve 0.05g of bovine serum albumin in 4mL of double-distilled water to prepare the water phase, and bathe in 50 o C water for 30 minutes to obtain a clear and transparent aqueous solution of bovine serum albumin;
（2）另取0.05mg噻虫啉，加入0.1mL乙腈超声溶解5min，配制成0.05mg/mL的噻虫啉-乙腈油相；(2) Take another 0.05 mg of thiacloprid, add 0.1 mL of acetonitrile for ultrasonic dissolution for 5 minutes, and prepare a 0.05 mg/mL thiacloprid-acetonitrile oil phase;
（3）有机溶剂与1mL戊酸戊酯混合，得到混合油相；(3) The organic solvent is mixed with 1 mL amyl valerate to obtain a mixed oil phase;
（4）将白蛋白水溶液调节转速1000 rpm，高速搅拌下逐滴加入混合油相，乳化40min，低温冷却5-10min，然后超声5min，得到白蛋白-噻虫啉-信息素纳米球的粉末制剂。(4) Adjust the rotation speed of the albumin aqueous solution to 1000 rpm, add dropwise to the mixed oil phase under high-speed stirring, emulsify for 40 minutes, cool at low temperature for 5-10 minutes, and then sonicate for 5 minutes to obtain the powder preparation of albumin-thiacloprid-pheromone nanospheres .
Claims (4)
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