Device and method for preparing water-in-oil nano-emulsion

Abstract

The invention relates to the technical field of nanometer, in particular to a preparation device and method.The preparation device of the water-in-oil nanometer emulsion comprises a stirring cylinder, a base, blades, a sleeve, an adjusting assembly, a scraping plate, a filter screen and a transmission mechanism, a mixing cylinder is arranged on one side of the stirring cylinder, and a rotating rod is installed in the stirring cylinder; the preparation device comprises a rotating rod, a stirring cylinder is installed on the rotating rod, a base is installed at the bottom of the stirring cylinder, blades are installed on the outer wall of the rotating rod at equal intervals, through holes are formed in the multiple blades at equal intervals, and the outer wall of the rotating rod is sleeved with a sleeve. Through holes are formed inside, liquid can conveniently pass through, resistance is reduced, meanwhile, the angles of the blades are adjustable, and the angles of the blades can be changed according to mixing requirements in the stirring process through meshing of the racks and the gears, so that the stirring effect is optimized, and more efficient mixing is achieved.

Description

Technical Field

[0001] The invention relates to the field of nanotechnology, and in particular to a device and method for preparing water-in-oil nanoemulsion. Background Art

[0002] With the long-term use of chemical pesticides in the control of hawthorn spider mites in apple orchards, the problem of plant disease and pest resistance has become increasingly serious, and it also poses a major threat to human health and environmental safety. To meet this challenge, it is particularly important to develop new, efficient and environmentally friendly pest control technologies. In recent years, RNA interference (RNAi) technology has shown great potential in the field of pest control due to its high specificity and harmless and environmentally friendly characteristics to humans and animals. In particular, double-stranded RNA (dsRNA), as the core of RNAi technology, achieves effective inhibition or killing of pests by specifically degrading the homologous mRNA of target pests. However, the problem of dsRNA being easily degraded in the natural environment and difficult to deliver effectively has severely limited its widespread application in agriculture. To solve these problems, researchers have explored a variety of nanocarrier technologies. Although these technologies have improved the stability and delivery efficiency of dsRNA to a certain extent, the complex preparation process, high cost, uncertainty in long-term stability and biocompatibility, and bottlenecks in large-scale production have hindered their practical application in large-scale agricultural production. Liquid white wax has unique adhesion, deposition and penetration capabilities, which can significantly accelerate the speed at which the active ingredients enter the target body and prolong their retention time on the target. At the same time, as a non-polar oil phase, liquid white wax has excellent chemical stability and is not easily oxidized or degraded, and can provide a reliable physical barrier for dsRNA. This barrier can effectively protect dsRNA from environmental factors such as light, humidity and temperature fluctuations, thereby significantly extending its shelf life. In addition, the non-polar properties of liquid wax can inhibit the activity of nucleases in the aqueous phase, thereby further preventing dsRNA degradation; compared with other oil phases, the non-polar environment and good chemical stability of liquid wax play an important role in improving the stability of dsRNA; Liquid paraffin, with its chemical inertness, non-polarity and excellent shielding ability, can effectively isolate the influence of external factors (such as hydrolysis, nuclease, light and oxidative stress), further improving the stability of dsRNA; therefore, in the fields of agriculture and biotechnology, liquid white wax and liquid paraffin have become ideal oil phase choices for protecting dsRNA due to their low cost, easy access and safety; At present, although some devices and methods for preparing water-in-oil nanoemulsions have been proposed, these devices often have problems such as complex structure, inconvenient operation, high maintenance cost, and low preparation efficiency. For example, some devices use a single stirring method, making it difficult to achieve sufficient mixing of the oil phase, water phase, and surfactant. At the same time, during the production of water-in-oil nanoemulsions, bubbles are generated, and these devices also have deficiencies in removing bubbles, resulting in a large number of bubbles in the prepared emulsion, which affects the quality and stability of the emulsion.

[0003] Therefore, it is necessary to provide a new device and method for preparing water-in-oil nanoemulsions to solve the above technical problems. Summary of the Invention

[0004] To solve the above technical problems, the present invention provides a device and method for preparing water-in-oil nanoemulsions.

[0005] The device and method for preparing water-in-oil nanoemulsions provided by the present invention include: a stirring tank, a base, blades, a sleeve, an adjusting assembly, a scraper, a filter screen, and a transmission mechanism. A mixing tank is provided on one side of the stirring tank. A rotating rod is installed inside the stirring tank. A base is installed at the bottom of the stirring tank. Blades are equidistantly installed on the outer wall of the rotating rod. Through holes are equidistantly opened inside several of the blades. A sleeve is sleeved on the outer wall of the rotating rod. An adjusting assembly is installed between the sleeve and the blades. The adjusting assembly is used to adjust the angle of the blades. Scrapers are symmetrically provided on the inner wall of the stirring tank. A filter screen is provided at the top of the sleeve. Filter holes are equidistantly opened inside the filter screen. A transmission mechanism is installed between the filter screen and the rotating rod. The transmission mechanism drives the filter screen to move up and down to remove the bubbles generated by stirring inside the stirring tank.

[0006] Preferably, the adjusting assembly includes: racks, gears, and elliptical grooves. Several of the blades are rotatably connected to the rotating rod. Gears are fixedly connected to one end of several of the blades close to the rotating rod. Elliptical grooves are equidistantly opened on the outer wall of the sleeve. Racks are fixedly connected to the inner walls of several of the elliptical grooves. Several of the racks are meshed with the corresponding gears.

[0007] Preferably, an electric push rod is fixedly connected inside the base. The output end of the electric push rod is fixedly connected to a moving frame. The bottom of the moving frame is slidably connected to the inner wall of the base. The bottom end of the sleeve is rotatably connected to a connecting ring. The inner wall of the connecting ring is rotatably connected to the outer wall of the rotating rod. Cross bars are fixedly connected to both sides of the connecting ring. Vertical bars are fixedly connected to one end of the two cross bars away from the connecting ring. Cylinders are symmetrically fixedly connected inside the base. Springs are fixedly connected to the inner walls of the two cylinders. One end of the two springs is fixedly connected to the bottom end of the corresponding vertical bar. The two vertical bars are slidably connected to the inner walls of the corresponding cylinders.

[0008] Preferably, drums are rotatably connected to the middle parts of the two cross bars. The two ends of the moving frame are designed as trapezoidal bosses, and the outer walls of the two drums are in contact with the outer walls of the corresponding trapezoidal bosses of the moving frame.

[0009] Preferably, the transmission mechanism includes: a reciprocating lead screw, a slider, a limiting rod and a limiting hole. A motor is fixedly connected to the inside of the base, the output end of the motor is fixedly connected to the bottom end of the rotating rod, the top end of the rotating rod is fixedly connected to the reciprocating lead screw, the top of the reciprocating lead screw is fitted with a slider, the outer wall of the slider is fixedly connected to the filter screen, limiting holes are symmetrically formed inside the slider, limiting rods are symmetrically fixedly connected to the inner wall of the stirring cylinder, and the two limiting rods are slidably connected to the inner walls of the corresponding limiting holes.

[0010] Preferably, sliding avoidance grooves are symmetrically formed in the middle of the sleeve, a scraping plate is fixedly connected to the position of the rotating rod close to the sliding groove, an exhaust valve is installed at the top of the stirring cylinder, a feeding pipe is installed at the bottom of the stirring cylinder, and a feeding hopper is communicated with the top of the mixing cylinder.

[0011] Preferably, a hose I is communicated with the bottom of the mixing cylinder, one end of the hose I is communicated with the top of the stirring cylinder, and a peristaltic pump I is installed in the middle of the hose I. The peristaltic pump I is fixedly connected to the base.

[0012] Preferably, a liquid storage cylinder is arranged on the other side of the stirring cylinder. A hose II is communicated with the top of the liquid storage cylinder, one end of the hose II is communicated with the top of the stirring cylinder, and a peristaltic pump II is installed in the middle of the hose II. The peristaltic pump II is fixedly connected to the top of the liquid storage cylinder.

[0013] Preferably, the preparation method of the water-in-oil nanoemulsion comprises the following steps: A water-in-oil nanoemulsion using liquid paraffin to encapsulate V-ATPase A dsRNA, the raw materials are composed of 40% surfactant by mass percentage (including 52.2% Tween 80, 46.8% Span 80 and 1% isooctyl salicylate), 20% dsRNA solution with a concentration of 1000 ng / μL - 10000 ng / μL, and 40% liquid paraffin; S1: Mix Tween 80, Span 80 and isooctyl salicylate and stir until it becomes clear, as the surfactant; S2: Mix the liquid paraffin with the surfactant. During the mixing process, the surfactant is added drop by drop to the liquid paraffin and stirred while adding until it becomes uniform. S3: Then, quantitatively add the aqueous dsRNA solution drop by drop. After vibrating on a vortex mixer for 5 minutes, place it in an ultrasonic crusher and ultrasonicate for more than 10 minutes. At this time, a water-in-oil nanoemulsion of liquid paraffin encapsulating V-ATPase A dsRNA is prepared. The obtained preparation is nanoscale, with a particle size of about 150 nanometers, good stability, significantly improving the control effect on the target spider mite Tetranychus viennensis while enhancing the stability of dsRNA and extending the degradation time of dsRNA in nature.

[0014] Compared with the related technologies, the device and method for preparing the water-in-oil nanoemulsion provided by the present invention have the following beneficial effects: Efficient mixing mechanism: Multi-angle blade design: The blades are not only equidistantly installed on the rotating rod, but also have through holes inside to facilitate the passage of liquid and reduce resistance. At the same time, the blade angle is adjustable. Through the meshing of the rack and the gear, the blades can change the angle according to the mixing requirements during the stirring process, thereby optimizing the stirring effect and achieving more efficient mixing. In the horizontal state, the blades are more likely to form a horizontal stirring force, which helps in horizontal mixing. The inclined blades can stir more effectively in the vertical direction, helping to break the layering phenomenon and improve the mixing efficiency. Strong shear force and turbulence: The motor drives the rotating rod and the blades to rotate, generating strong shear force and turbulence, effectively breaking the interfacial tension between the materials and promoting the rapid mixing of the oil phase, water phase, and surfactant. Precisely controlled bubble removal: The filter screen moves up and down: The transmission mechanism drives the filter screen to move up and down in the stirring cylinder through a reciprocating lead screw, effectively breaking the bubbles formed during the stirring process and discharging the gas in the broken bubbles in time through the exhaust valve installed at the top of the stirring cylinder, avoiding the accumulation of bubbles in the emulsion and affecting the stability and quality of the emulsion. The limiting rod and the limiting hole ensure the smooth movement of the slider and prevent shaking. The scraper closely adheres to the inner wall of the stirring cylinder to ensure that after the stirring and mixing process is completed, the mixed liquid can be easily discharged from the stirring cylinder, avoiding residue and waste. Description of the drawings

[0015] Figure 1 It is a schematic structural diagram of the device and method for preparing the water-in-oil nanoemulsion provided by the present invention; Figure 2 For Figure 1 The schematic structural diagram of the cross-section of the stirring cylinder shown; Figure 3 For Figure 2 The schematic structural diagram of the blade shown; Figure 4 For Figure 3 The schematic structural diagram of the moving frame shown; Figure 5 is Figure 3 a schematic structural diagram of the adjusting assembly shown; Figure 6 is Figure 3 a schematic structural diagram of the transmission mechanism shown.

[0016] Reference numerals in the figure: 1, stirring cylinder; 2, mixing cylinder; 3, rotating rod; 4, base; 5, blade; 6, sleeve; 7, scraper; 8, filter screen; 9, rack; 10, gear; 11, elliptical groove; 12, electric push rod; 13, moving frame; 14, connecting ring; 15, cross bar; 16, vertical bar; 17, cylinder; 18, tension spring; 19, roller; 20, reciprocating lead screw; 21, slider; 22, limiting rod; 23, limiting hole; 24, motor; 25, feed hopper; 26, hose 1; 27, peristaltic pump 1; 28, exhaust valve; 29, liquid storage cylinder; 30, hose 2; 31, peristaltic pump 2; 32, blanking pipe. Specific embodiments

[0017] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0018] The specific implementation of the present invention will be described in detail below with reference to specific embodiments.

[0019] Please refer to Figures 1 to 6, A device and method for preparing a water-in-oil nanoemulsion. The device and method for preparing the water-in-oil nanoemulsion include: a stirring cylinder 1, a base 4, blades 5, a sleeve 6, an adjusting assembly, a scraper 7, a filter screen 8, and a transmission mechanism. A mixing cylinder 2 is provided on one side of the stirring cylinder 1. A rotating rod 3 is installed inside the stirring cylinder 1. A base 4 is installed at the bottom of the stirring cylinder 1. Blades 5 are equidistantly installed on the outer wall of the rotating rod 3. Through holes are equidistantly opened inside several of the blades 5. A sleeve 6 is sleeved on the outer wall of the rotating rod 3. An adjusting assembly is installed between the sleeve 6 and the blades 5 for adjusting the angles of the blades 5. Scrapers 7 are symmetrically provided on the inner wall of the stirring cylinder 1. A filter screen 8 is provided at the top of the sleeve 6. Filter holes are equidistantly opened inside the filter screen 8. A transmission mechanism is installed between the filter screen 8 and the rotating rod 3 to drive the filter screen 8 to move up and down for removing bubbles generated inside the stirring cylinder 1 due to stirring. Slide avoidance grooves are symmetrically opened in the middle of the sleeve 6. The position of the rotating rod 3 close to the chute is fixedly connected to the scraper 7. An exhaust valve 28 is installed at the top of the stirring cylinder 1. A discharge pipe 32 is installed at the bottom of the stirring cylinder 1. A feed hopper 25 is communicated with the top of the mixing cylinder 2. A hose 26 is communicated with the bottom of the mixing cylinder 2. One end of the hose 26 is communicated with the top of the stirring cylinder 1. A peristaltic pump 27 is installed in the middle of the hose 26. The peristaltic pump 27 is fixedly connected to the base 4. A liquid storage cylinder 29 is provided on the other side of the stirring cylinder 1. A hose 30 is communicated with the top of the liquid storage cylinder 29. One end of the hose 30 is communicated with the top of the stirring cylinder 1. A peristaltic pump 31 is installed in the middle of the hose 30. The peristaltic pump 31 is fixedly connected to the top of the liquid storage cylinder 29.

[0020] It should be noted that: The design of the scraper 7 assists the blades 5 in mixing the liquid inside the stirring cylinder 1 when the motor 24 rotates forward, and assists the discharge pipe 32 in discharging the liquid inside the stirring cylinder 1 when the motor 24 rotates in reverse. The scraper 7 is in contact with the inner wall of the stirring cylinder 1. When the liquid is fully mixed and discharged, the liquid mixture adhering to the inner wall of the stirring cylinder 1 can be scraped off.

[0021] Please refer to Figure 3 and Figure 5, the adjusting assembly includes: a rack 9, a gear 10, and an elliptical groove 11. A plurality of the vanes 5 are all rotatably connected to the rotating rod 3. One end of each of the plurality of vanes 5 close to the rotating rod 3 is fixedly connected to a gear 10. The outer wall of the sleeve 6 is equidistantly provided with elliptical grooves 11. The inner walls of the plurality of elliptical grooves 11 are all fixedly connected to racks 9. The plurality of racks 9 are meshed with the corresponding gears 10. An electric push rod 12 is fixedly connected to the inside of the base 4. The output end of the electric push rod 12 is fixedly connected to a moving frame 13. The bottom of the moving frame 13 is slidably connected to the inner wall of the base 4. The bottom end of the sleeve 6 is rotatably connected to a connecting ring 14. The inner wall of the connecting ring 14 is rotatably connected to the outer wall of the rotating rod 3. Both sides of the connecting ring 14 are fixedly connected to cross bars 15. One end of each of the two cross bars 15 away from the connecting ring 14 is fixedly connected to a vertical rod 16. Cylinders 17 are symmetrically fixedly connected to the inside of the base 4. The inner walls of the two cylinders 17 are all fixedly connected to tension springs 18. One end of each of the two tension springs 18 is fixedly connected to the bottom end of the corresponding vertical rod 16. The two vertical rods 16 are slidably connected to the inner walls of the corresponding cylinders 17. The middle parts of the two cross bars 15 are all rotatably connected to rollers 19. The two ends of the moving frame 13 are designed as trapezoidal bosses. The outer walls of the two rollers 19 are in contact with the outer walls of the corresponding trapezoidal bosses of the moving frame 13; It should be noted that: the top end of the connecting ring 14 is rotatably connected to the sleeve 6, the inner wall of the connecting ring 14 is rotatably connected to the outer wall of the rotating rod 3, and both sides of the connecting ring 14 are fixedly connected to the cross bars 15, so that the connecting ring 14 is not affected by the motion states of other parts and is only allowed to move up and down along the outer wall of the rotating rod 3; Please refer to Figure 3 and Figure 6 , the transmission mechanism includes: a reciprocating lead screw 20, a slider 21, a limiting rod 22, and a limiting hole 23. A motor 24 is fixedly connected to the inside of the base 4. The output end of the motor 24 is fixedly connected to the bottom end of the rotating rod 3. A reciprocating lead screw 20 is fixedly connected to the top end of the rotating rod 3. A slider 21 is fitted and installed at the top of the reciprocating lead screw 20. The outer wall of the slider 21 is fixedly connected to the filter screen 8. Limiting holes 23 are symmetrically opened inside the slider 21. Limiting rods 22 are symmetrically fixedly connected to the inner wall of the stirring cylinder 1. The two limiting rods 22 are slidably connected to the inner walls of the corresponding limiting holes 23; It should be noted that: while the rotating rod 3 is rotating, the reciprocating lead screw 20 at its top end is also rotating. The reciprocating lead screw 20 is in threaded cooperation with the slider 21 to drive the slider 21 to reciprocate up and down inside the stirring cylinder 1. The limiting rods 22 symmetrically fixedly connected to the inside of the stirring cylinder 1 are slidably connected to the inner walls of the limiting holes 23 inside the slider 21, so that the slider 21 can move more smoothly when moving up and down. The outer wall of the slider 21 is fixedly connected to the filter screen 8. The up and down movement of the filter screen 8 can break the bubbles generated during the stirring process and further discharge the gas through the exhaust valve 28 at the top of the stirring cylinder 1; Please refer toFigures 1 to 6 , the preparation method of the water-in-oil nanoemulsion comprises the following steps: A water-in-oil nanoemulsion using liquid paraffin to encapsulate V-ATPase A dsRNA, the raw materials include 40% surfactant by mass percentage (including 52.2% Tween 80, 46.8% Span 80 and 1% isooctyl salicylate), 20% dsRNA solution with a concentration of 1000 ng / μL - 10000 ng / μL, and 40% liquid paraffin; S1: Mix Tween 80, Span 80, and isooctyl salicylate and stir until it becomes clear, serving as the surfactant; S2: Mix liquid paraffin with the surfactant. During the mixing process, the surfactant is added drop by drop to the liquid paraffin while stirring until it becomes uniform; S3: Then add a quantitative dsRNA aqueous solution drop by drop, vibrate on a vortex mixer for 5 minutes, and then put it into an ultrasonic crusher and ultrasonicate for more than 10 minutes. At this time, a water-in-oil nanoemulsion with liquid paraffin encapsulating V-ATPase A dsRNA is prepared; It should be noted that: The preparation of the 1% water-in-oil nanoemulsion A of V-ATPase is specifically the following steps: 1. At room temperature, mix tween-80, span-80, and isooctyl salicylate according to the mass ratio of 52.2%, 46.8%, and 1% and stir for 10 minutes. When the color becomes clear, it serves as the surfactant; 2. Mix liquid paraffin with the surfactant according to a mass ratio of 1:1. During the mixing process, the surfactant is added drop by drop to the liquid paraffin while stirring until it becomes uniform; 3. Then add 20% of the dsRNA aqueous solution with a concentration of 1000 ng / μL drop by drop and vibrate on a vortex mixer until it becomes uniform; 4. Crush the prepared water-in-oil primary emulsion at an ultrasonic power of 195 W for 10 minutes; at this time, the emulsion is in a white semi-transparent state; The preparation of the 0.5% water-in-oil nanoemulsion B of V-ATPase is specifically the following steps: 1. At room temperature, mix tween-80, span-80, and isooctyl salicylate according to the mass ratio of 52.2%, 46.8%, and 1% and stir for 10 minutes. When the color becomes clear, it serves as the surfactant; 2. Mix liquid paraffin with the surfactant according to a mass ratio of 1:1. During the mixing process, the surfactant is added drop by drop to the liquid paraffin while stirring until it becomes uniform; 3. Then add 20% of the dsRNA aqueous solution with a concentration of 1000 ng / μL drop by drop and vibrate on a vortex mixer until it becomes uniform; 4. Break the prepared water-in-oil primary emulsion at an ultrasonic power of 195 W for 10 minutes; at this time, the emulsion is in a white semi-transparent state; Preparation of water-in-oil nanoemulsion C of 0.1% V-ATPase, specifically the following steps: 1. At room temperature, mix tween-80, span-80, and isooctyl salicylate according to a mass ratio of 52.2%, 46.8%, and 1% and stir for 10 minutes. When the color is clear, it is used as a surfactant; 2. Mix liquid paraffin wax and the surfactant according to a mass ratio of 1:1. During the mixing process, the surfactant is added drop by drop to the liquid paraffin wax, and it is stirred until it is in a uniform state while adding; 3. Then add 20% of the dsRNA aqueous solution with a concentration of 1000 ng / μL drop by drop and vibrate it on a vortex mixer until it is uniform; 4. Break the prepared water-in-oil primary emulsion at an ultrasonic power of 195 W for 10 minutes; at this time, the emulsion is in a white semi-transparent state; After diluting nanoemulsion A by 1000 times, the particle size is 140 nm; after diluting nanoemulsion B by 1000 times, the particle size is 150 nm; after diluting nanoemulsion C by 1000 times, the particle size is 175 nm; Through the ingenious water-in-oil nanoemulsion preparation technology, using liquid paraffin oil as the oil phase, the dsRNA targeting the V-ATPase A subunit of Tetranychus viennensis is successfully encapsulated. This encapsulation method not only significantly increases the mortality rate of target pests and advances the death time, but also effectively protects dsRNA from enzymatic hydrolysis and photodegradation, thus significantly extending its effective period in nature; Compared with traditional dsRNA nano-carriers, the preparation proposed in this patent is simpler in the processing process and lower in cost. Liquid paraffin oil provides a reliable physical barrier for dsRNA with its unique adhesion, deposition, and penetration capabilities, as well as excellent chemical stability, effectively isolating the interference of external factors such as enzymes and ultraviolet rays. This not only improves the stability and bioavailability of dsRNA, but also opens up a new path for the wide application of RNAi technology in the field of agricultural pest control; In summary, through the innovative water-in-oil nanoemulsion preparation technology, this patent successfully realizes the synergistic protection effect of liquid paraffin oil on dsRNA pesticides, provides an effective solution to the stability and delivery problems faced by dsRNA in practical applications, and has important theoretical significance and practical value.

[0022] The working principle of the water-in-oil nanoemulsion preparation device and method provided by the present invention is as follows: Surfactant preparation: Tween 80, Span 80 and isooctyl salicylate are conveyed into the mixing cylinder 2 through the feed hopper 25. The stirring device in the mixing cylinder 2 is started, and these three components are mixed and stirred until they become clear, and are reserved as surfactants; Mixing of the surfactant and liquid paraffin wax: Peristaltic pump 1 27 starts to work, and the surfactant is added drop by drop into the stirring cylinder 1 filled with liquid paraffin wax through hose 1 26; at the same time, motor 24 is started, and the forward rotation of motor 24 drives the rotating rod 3 to rotate clockwise, and the blades 5 rotatably connected thereto rotate in the stirring cylinder 1, generating shear force and turbulence, and promoting the mixing of liquid paraffin wax and the surfactant solution; Electric push rod 12 is started, and the moving frame 13 (designed with trapezoidal bosses at both ends) fixedly connected to its output end pushes the cross bar 15 upward. The trapezoidal bosses at both ends of the moving frame 13 contact the outer wall of the roller 19 in the middle of the corresponding cross bar 15, further reducing the friction during pushing. The movement of the cross bar 15 drives the vertical rod 16 to slide upward along the inner wall of the cylinder 17 and stretch the tension spring 18. At the same time, the connecting ring 14 fixedly connected to the cross bar 15 also moves upward and pushes the sleeve 6 to slide upward on the rotating rod 3; The upward movement of the sleeve 6 causes the rack 9 on the inner wall of the elliptical groove 11 equidistantly opened on its outer wall to move upward. The gear 10 at one end of the blade 5 close to the rotating rod 3 meshes with the rack 9 on the inner wall of the elliptical groove 11 on the outer wall of the sleeve 6. Therefore, the movement of the rack 9 drives the gear 10 to rotate, thereby changing the angle of the blade 5. The change in the angle of the blade 5 can adjust the intensity and direction of stirring, so that the liquid in the stirring cylinder 1 can be more fully mixed. The through holes equidistantly opened inside the blade 5 help the liquid to pass through, further promoting mixing, and at the same time reducing the resistance of the blade 5 in the liquid, making the stirring process smoother and reducing energy consumption; When the electric push rod 12 contracts, due to the pulling force of the tension spring 18, the vertical rod 16 moves downward, thereby driving the cross bar 15 and the connecting ring 14 to move downward, and then the sleeve 6 moves downward. Through the meshing connection of the rack 9 and the gear 10, the angle of the blade 5 is restored to the initial state; Bubble removal: While the rotating rod 3 is rotating, the reciprocating lead screw 20 at its top is also rotating. The reciprocating lead screw 20 cooperates with the slider 21 through the thread, driving the slider 21 to move up and down reciprocally in the stirring cylinder 1. The limiting rods 22 symmetrically and fixedly connected inside the stirring cylinder 1 slide on the inner wall of the limiting holes 23 inside the slider 21, making the slider 21 move more smoothly when moving up and down. The outer wall of the slider 21 is fixedly connected with a filter screen 8. The up and down movement of the filter screen 8 can break the bubbles generated during the stirring process and further discharge the gas through the exhaust valve 28 at the top of the stirring cylinder 1; Addition of the dsRNA aqueous solution: After the surfactant is completely added to the liquid white wax in a predetermined ratio of 1:1 and stirred evenly, the peristaltic pump II 31 is started; The peristaltic pump II 31 adds the dsRNA aqueous solution inside the liquid storage cylinder 29 drop by drop into the stirring cylinder 1 through the hose II 30, and continues to stir to ensure that the dsRNA aqueous solution is fully mixed with the oil phase and the surfactant; Final mixing and emulsification: After the stirring is completed, the mixed liquid in the stirring cylinder 1 is discharged through the discharge pipe 32. At this time, the motor 24 is started to rotate in the reverse direction and the speed is reduced. The blade 5 is in an inclined state under the action of the adjustment assembly, which helps the discharged mixed liquid. The scraper 7 fixedly connected to the middle of the rotating rod 3 fits with the inner wall of the stirring cylinder 1, scrapes off the mixture adhering to the inner wall of the stirring cylinder 1 and discharges it through the discharge pipe 32 for the next treatment; The discharged mixture is placed on a vortex instrument and vibrated for 5 minutes to further promote the uniformity and stability of the mixture; Then the mixture is placed in an ultrasonic crusher and ultrasonically treated for more than 10 minutes. The ultrasonic treatment can break larger oil droplets and water droplets to form smaller and more uniform water-in-oil nanoemulsions.

[0023] The above are only the embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. Any equivalent structure or equivalent process transformation made by using the specification and drawings of the present invention, or directly or indirectly applied in other related technical fields, shall be equally included in the patent protection scope of the present invention.

Claims

1. A device and method for preparing a water-in-oil nanoemulsion, characterized in that: include: A stirring cylinder (1), wherein a mixing cylinder (2) is provided on one side of the stirring cylinder (1), and a rotating rod (3) is installed inside the stirring cylinder (1); A base (4), wherein the bottom of the mixing tank (1) is provided with a base (4); Blades (5), the outer wall of the rotating rod (3) is provided with blades (5) at equal intervals, and through holes are provided in the interior of a plurality of the blades (5) at equal intervals; A sleeve (6), wherein the outer wall of the rotating rod (3) is sleeved with the sleeve (6); An adjustment component is installed between the sleeve (6) and the blade (5), and the adjustment component is used to adjust the angle of the blade (5); A scraper (7), wherein the inner wall of the mixing tank (1) is symmetrically provided with a scraper (7); A filter screen (8), wherein the top of the sleeve (6) is provided with a filter screen (8), and filter holes are provided at equal intervals inside the filter screen (8); A transmission mechanism is installed between the filter screen (8) and the rotating rod (3), and the transmission mechanism drives the filter screen (8) to move up and down to remove bubbles generated by stirring inside the stirring tank (1).

2. The preparation device and method of the water-in-oil nanoemulsion according to claim 1, characterized in that: The adjustment component comprises: a rack (9), a gear (10) and an elliptical groove (11); a plurality of blades (5) are rotatably connected to the rotating rod (3); one end of a plurality of blades (5) close to the rotating rod (3) is fixedly connected to the gear (9); an outer wall of the sleeve (6) is provided with elliptical grooves (11) at equal intervals; inner walls of a plurality of the elliptical grooves (11) are fixedly connected to the racks (10); and a plurality of the racks (10) are meshingly connected to corresponding gears (9).

3. The preparation device and method of the water-in-oil nanoemulsion according to claim 1, characterized in that: The base (4) is fixedly connected with an electric push rod (12) inside, the output end of the electric push rod (12) is fixedly connected with a moving frame (13), the bottom of the moving frame (13) is slidably connected to the inner wall of the base (4), the bottom end of the sleeve (6) is rotatably connected with a connecting ring (14), the inner wall of the connecting ring (14) is rotatably connected to the outer wall of the rotating rod (3), both sides of the connecting ring (14) are fixedly connected with cross bars (15), one end of the two cross bars (15) away from the connecting ring (14) is fixedly connected with a vertical bar (16), the base (4) is symmetrically fixedly connected with a cylinder (17), the inner walls of the two cylinders (17) are fixedly connected with tension springs (18), one end of the two tension springs (18) is fixedly connected to the bottom end of the corresponding vertical bar (16), and the two vertical bars (16) are slidably connected to the inner wall of the corresponding cylinder (17).

4. The preparation device and method of the water-in-oil nanoemulsion according to claim 3, characterized in that: The middle parts of the two cross bars (15) are rotatably connected to rollers (19), and the two ends of the movable frame (13) are designed as trapezoidal bosses, and the outer walls of the two rollers (19) are in contact with the outer walls of the corresponding trapezoidal bosses of the movable frame (13).

5. The preparation device and method of the water-in-oil nanoemulsion according to claim 1, characterized in that: The transmission mechanism comprises: a reciprocating screw (20), a slider (21), a limiting rod (22) and a limiting hole (23); a motor (24) is fixedly connected inside the base (4); an output end of the motor (24) is fixedly connected to the bottom end of the rotating rod (3); the top end of the rotating rod (3) is fixedly connected to the reciprocating screw (20); a slider (21) is mounted on the top of the reciprocating screw (20); an outer wall of the slider (21) is fixedly connected to the filter screen (8); the limiting holes (23) are symmetrically arranged inside the slider (21); the limiting rods (22) are symmetrically fixedly connected to the inner wall of the mixing cylinder (1); and two limiting rods (22) are slidably connected to the inner walls of the corresponding limiting holes (23).

6. The preparation device and method of the water-in-oil nanoemulsion according to claim 1, characterized in that: The sleeve (6) is symmetrically provided with a slide avoidance groove in the middle, the rotating rod (3) is fixedly connected to the scraper (7) near the slide groove, the top of the mixing cylinder (1) is provided with an exhaust valve (28), the bottom of the mixing cylinder (1) is provided with a feed pipe (32), and the top of the mixing cylinder (2) is connected with a feed hopper (25).

7. The preparation device and method of water-in-oil nanoemulsion according to claim 1, characterized in that: The bottom of the mixing tank (2) is connected to a hose 1 (26), one end of which is connected to the top of the mixing tank (1), and a peristaltic pump 1 (27) is installed in the middle of the hose 1 (26), which is fixedly connected to the base (4).

8. The device and method for preparing water-in-oil nanoemulsion according to claim 1, characterized in that: A liquid storage cylinder (29) is provided on the other side of the mixing cylinder (1). The top of the liquid storage cylinder (29) is connected to a second hose (30). One end of the second hose (30) is connected to the top of the mixing cylinder (1). A second peristaltic pump (31) is installed in the middle of the second hose (30). The second peristaltic pump (31) is fixedly connected to the top of the liquid storage cylinder (29).

9. The method for preparing the water-in-oil nanoemulsion according to any one of claims 1 to 8, characterized in that: The following steps are involved: The invention discloses an oil-in-water nanoemulsion for encapsulating V-ATPase A dsRNA by using liquid white wax, wherein the raw materials are 40% by weight of surfactants including (52.2% of Tween 80, 46.8% of Span 80 and 1% of isooctyl salicylate), 20% of dsRNA solution with a concentration of 1000ng / μL-10000ng / μL, and 40% of liquid white wax; S1: Tween 80, Span 80 and 2-ethylhexyl salicylate are mixed and stirred until a clear state, which serve as surfactants; S2: mixing the liquid wax with the surfactant, adding the surfactant dropwise into the liquid wax during the mixing process while stirring until the mixture is uniform; S3: A quantitative dsRNA aqueous solution is then added dropwise, and after vibrating on a vortex for 5 minutes, the solution is placed in an ultrasonic disruptor and ultrasonicated for more than 10 minutes. At this time, a water-in-oil nanoemulsion of V-ATPase A dsRNA encapsulated by liquid white wax is prepared.

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