Preparation device and preparation method for cattle superovulation hormone

Abstract

The invention provides a preparation device and a preparation method for cattle superovulation hormone, and relates to the technical field of gonadotropin preparation, the preparation device comprises a mounting rack, two rotating racks are fixed on the vertical surface of the mounting rack, a centrifugal assembly is arranged between the two rotating racks, the centrifugal assembly comprises a central disc, and the central disc is fixed on the mounting rack. The centrifugal assembly is composed of two centrifugal cylinders which do not interfere with each other, the centrifugal cylinder at the upper end is used for centrifugal rotation, the centrifugal cylinder at the lower end is used for centrifugal rotation, the centrifugal cylinder at the upper end is used for centrifugal rotation, and the centrifugal cylinder at the lower end is used for centrifugal rotation. The centrifugal cylinder at the lower end is used for standing and layering, supernatant is conveniently taken out, meanwhile, the cylindrical protective film arranged in the centrifugal cylinder can prevent residual liquid from being left in the centrifugal cylinder, sediment is conveniently discharged, and the centrifugal cylinder can be continuously used by replacing the cylindrical protective film.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of gonadotropin preparation, in particular to a preparation device and method for bovine superovulation hormone. BACKGROUND

[0002] Gonadotropin in biopharmaceutical manufacturing is a genetically engineered drug and vaccine manufacturing product. Human chorionic gonadotropin is a glycoprotein hormone secreted by the syncytiotrophoblast cells of the placenta during pregnancy, which is composed of two different subunits, alpha and beta, connected by a non-covalent bond. Therefore, HCG is generally extracted from pregnant women's urine. During the production, secretion and metabolism of hormones, HCG molecules undergo various changes such as fragmentation and dissociation, thus existing in various molecular forms in blood and urine. The pharmacological action of HCG is mainly similar to that of luteinizing hormone, and has little effect on follicle maturation. It can promote follicle maturation and ovulation, and make the ruptured follicle transform into a corpus luteum. It can promote the secretion of progesterone. For males, it has the effect of interstitial cell hormone, especially the activity of testicular interstitial cells. It can produce androgen, promote the development and maturation of sexual organs and secondary sexual characteristics, and promote the production of sperm. Currently, the main process steps for extracting HCG from pregnant women's urine are as follows: after collecting the pregnant women's urine, sodium benzoate or kaolin is used for adsorption to prepare HCG urine concentrate. Then, organic solvent is used for extraction to obtain HCG crude product, which is further purified to obtain HCG fine product.

[0003] In the existing technology, when purifying the HCG crude product, multiple steps of dissolution, mixing, centrifugation and filtration are required. In the centrifugation step, the conventional centrifugal equipment needs to wait for the mixed liquid to separate after use, and then the supernatant is taken out. The waiting time in this process causes the centrifugal equipment to be unable to continue to be used, and the subsequent cleaning of the centrifugal equipment also takes time, which is low in efficiency. SUMMARY

[0004] In view of the deficiencies of the prior art, the present application aims to provide a preparation device and method for bovine superovulation hormone to solve the problems raised in the background art. The present application has a novel structure, and the centrifugal assembly is composed of two centrifugal cylinders that do not interfere with each other. The upper centrifugal cylinder is used for centrifugal rotation, and the lower centrifugal cylinder is used for static layering, which facilitates the taking out of the supernatant. At the same time, the cylindrical protective film built-in the centrifugal cylinder can avoid residual liquid left in the centrifugal cylinder, facilitate the discharge of the precipitate, and the centrifugal cylinder can be continuously used by replacing the cylindrical protective film.

[0005] In order to achieve the above object, the present application is realized by the following technical scheme: A preparation device for bovine superovulation hormone, comprising a mounting frame, two rotating frames are fixed on the vertical surface of the mounting frame, a centrifugal assembly is arranged between the two rotating frames, the centrifugal assembly comprises a center disc, centrifugal cylinders are rotatably installed on the top and bottom of the center disc, a moving ring is slidably sleeved on the outer wall of the centrifugal cylinder, a cylindrical protective film is sleeved on the inner wall of the centrifugal cylinder, the outer end of the cylindrical protective film is clamped inside the moving ring, a top cover covers one end of the centrifugal cylinder away from the center disc, a pull ring is fixed on the outer surface of the top cover, a drainage port is fixed on one side of the top cover, a rubber plug is sealingly inserted into the drainage port, a collection disc is placed at the position corresponding to the centrifugal cylinder at the bottom of the mounting frame, a collection barrel is placed on the other side of the collection disc, a suction pump is installed on the top of the collection barrel, the suction pump is fixedly installed on the side of the rotating frame, a rotating disc is arranged on the top of the centrifugal cylinder, vertical plates are fixed on both sides of the rotating disc, and the vertical plates are movably connected with the outer wall of the centrifugal cylinder.

[0006] Further, the centrifugal assembly further comprises a rotating ring, rotating rings are rotatably installed on the top and bottom of the center disc, the rotating rings are fixedly connected with the two groups of centrifugal cylinders, bottom plates are arranged on the upper end and lower end of the center disc, and the bottom plates slide along the inside of the corresponding centrifugal cylinders.

[0007] Further, a slide rail strip is fixed on the inner wall of the centrifugal cylinder, the bottom plate slides along the slide rail strip, a plurality of return springs are fixed between the bottom plate and the center disc in a equidistant annular manner.

[0008] Further, an L-shaped pipe is fixed at the center of the center disc, one end of the L-shaped pipe penetrates out from the side of the center disc, the other end of the L-shaped pipe penetrates through the inside of the bottom plate and the cylindrical protective film, the suction pump is provided with a connecting port at the position corresponding to the L-shaped pipe penetrating out from the lower end centrifugal cylinder, and a one-way valve is arranged on the end of the L-shaped pipe penetrating through the cylindrical protective film.

[0009] Further, four vertical rods are slidably installed on the outer side of the centrifugal cylinder in an equidistant manner, a slide plate is arranged on the outer side of the moving ring at the position corresponding to the vertical rod, the slide plate is slidably sleeved on the vertical rod, a ring groove is formed in the outer side of the moving ring, and the slide plate slides along the inside of the ring groove.

[0010] Further, a plurality of clamping plates are rotatably installed on the top of the moving ring through a rotating shaft and a torsion spring, and the outer end of the cylindrical protective film is clamped between the clamping plates and the moving ring.

[0011] Further, a buckle plate is rotatably installed on the top of the vertical rod through a rotating shaft and a torsion spring, an arc-shaped groove is formed in the top cover at the position corresponding to the buckle plate, a bottom protrusion of the buckle plate slides along the arc-shaped groove, a filter plate is fixed on the inner wall of the top cover, and the filter plate is sealingly inserted into the inside of the centrifugal cylinder.

[0012] Further, the top of the vertical rod is fixed with a convex plate, the vertical plate is provided with a clamping groove corresponding to the position of the convex plate, and the convex plate is slidingly clamped in the clamping groove, the top of the mounting frame is fixed with a top frame, and the rotating disc is rotatably installed on the top frame, the top frame is fixed with a first motor at the position corresponding to the center of the rotating disc, and the output end of the first motor is fixedly connected with the rotating disc.

[0013] Further, the front end of the rotating frame is slidingly clamped with a circular ring, the side surface of the circular ring is fixedly connected with the central disc, and the outlet end of the L-shaped pipe penetrates through the center of the circular ring.

[0014] A preparation method of a bovine superovulation hormone, the preparation method comprising the following steps:

[0015] (1) adding a buffer solution into the low-quality HCG and stirring to mix the solution, then adding the mixed solution into an upper centrifugal cylinder of a centrifugal assembly for centrifugal treatment, after centrifugation of the solution for 15-20 min, rotating the upper centrifugal cylinder to the lower layer to precipitate the solution, and collecting the supernatant after the solution is stratified;

[0016] (2) taking out the remaining solution in the lower centrifugal cylinder, adding a buffer solution into the remaining solution and stirring to mix, then adding the mixed solution into the upper centrifugal cylinder for secondary centrifugal treatment, and collecting the supernatant after centrifugation in the same way;

[0017] (3) adding the collected supernatants twice into a collection barrel for combination, then adding artificial zeolite and stirring, and after mixing, using a Buchner funnel to filter the supernatant, and repeatedly washing with distilled water to obtain a semi-finished product solution;

[0018] (4) loading the semi-finished product solution into a chromatography column, adding an ethanol solution for elution, collecting a protein elution peak part, and adding an ethanol solution, after standing for 6 h, centrifuging, discarding the supernatant and collecting the precipitate;

[0019] (5) dissolving the collected precipitate with a buffer solution, continuously adding the dissolved precipitate solution into the chromatography column, starting collection when the ultraviolet analyzer at the column mouth shows that protein solution flows out, after the solution is completely added into the column, continuously adding a phosphate buffer solution until the ultraviolet analyzer shows that no protein flows out, and then stopping collection, adding ethanol, standing for 6 h, removing the supernatant, collecting the precipitate, dehydrating the precipitate with anhydrous ethanol, and vacuum drying to obtain high-quality HCG.

[0020] The beneficial effects of the present application are as follows:

[0021] 1. The present application is characterized in that the upper and lower groups of centrifugal cylinders are arranged without interference, the upper centrifugal cylinder can be rotated with the rotating disc, and the lower centrifugal cylinder can be treated by static layering, the bottom of the centrifugal cylinder is supported by the bottom plate to keep it on a plane.

[0022] 2. The L-shaped pipe passes through the circular ring, so it does not interfere with the rotation of the centrifugal assembly, the upper end of the L-shaped pipe is sealed by the suction pump, and the lower end of the L-shaped pipe is connected to the connecting port of the suction pump, so that the supernatant in the lower centrifugal cylinder is pumped out and collected in the collection barrel for subsequent mixing.

[0023] 3. After the centrifugal cylinder rotates to the lower end, the supernatant after static treatment is pumped out through the L-shaped pipe, the sealing plug of the drain hole is opened, the remaining solution is collected in the collection disc, and secondary centrifugation is performed, then the sliding plate is moved along the ring groove, the vertical rod and the buckle plate are driven to slide along the arc-shaped groove, the bottom of the vertical rod is actually connected to the outer wall of the centrifugal cylinder in a conventional convex strip sliding connection mode, and springs are arranged between adjacent vertical rods to keep the four vertical rods equidistantly distributed, the two vertical rods are brought together to seal the open end of the cylindrical protective film, and the opening of the top cover is facilitated to take out the cylindrical protective film for separate collection and treatment of the internal precipitate.

[0024] 4. After the mixed liquid is added to the cylindrical protective film in the centrifugal cylinder, the cylindrical protective film gradually descends with the accumulation of weight, and the bottom plate slides downward along the sliding rail to compress the return spring, and the moving ring slides upward along the vertical rod, the top cover is wrapped around the open end of the centrifugal cylinder and the cylindrical protective film, the buckle plate is buckled in the arc-shaped groove to vertically limit the top cover, and the top cover is prevented from opening during centrifugal rotation.

[0025] 5. In the present application, one end of the cylindrical protective film is inserted into the centrifugal cylinder and adheres to the inner wall of the centrifugal cylinder, the bottom is in contact with the bottom plate, and the position corresponding to the L-shaped pipe is drilled for the L-shaped pipe to pass out, the outer end of the cylindrical protective film is sleeved on the periphery of the centrifugal cylinder and clamped by the moving ring and the clamping plate, and the depth of the cylindrical protective film can be adjusted by sliding in the vertical direction of the vertical rod to adjust the content of different mixed liquids.

[0026] 6. The present application replaces the upper and lower two sets of centrifuge cylinders by artificial overturning, and the side and bottom of the mounting frame give the centrifuge cylinder enough rotating space and do not interfere with the rotation, in the process of centrifuge cylinder rotation, the convex plate is inserted into the vertical plate clamping groove in the vertical direction, the convex plate and the clamping groove are arc-shaped, and the arc is the same as the rotation track of the circular ring, after the convex plate is clamped with the vertical plate, the upper centrifuge cylinder is driven to rotate by the first motor, and the mixed liquid in the centrifuge cylinder is centrifuged, because the front and rear ends of the clamping groove are open, the rotation of the centrifuge cylinder is not interfered, and the lower centrifuge cylinder can also be connected with the vertical plate after rotating to the upper layer, and then the centrifuge cylinder is driven by the rotating disc.

[0027] 7. Compared with the prior art, the centrifuge assembly is composed of two non-interfering centrifuge cylinders, the upper centrifuge cylinder is used for centrifugal rotation, and the lower centrifuge cylinder is used for static layering, so that the supernatant can be easily taken out, and the cylindrical protective film built in the centrifuge cylinder can avoid residual liquid in the centrifuge cylinder, facilitate the discharge of the precipitate, and the centrifuge cylinder can be continuously used by replacing the cylindrical protective film. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 It is a flowchart of a preparation method of a bovine superovulation hormone according to the present application;

[0029] Figure 2 It is a schematic diagram of the overall structure of a preparation device for a bovine superovulation hormone according to the present application;

[0030] Figure 3 It is a schematic diagram of the front structure of a preparation device for a bovine superovulation hormone according to the present application;

[0031] Figure 4 It is a schematic diagram of the side structure of a central disc of a preparation device for a bovine superovulation hormone according to the present application;

[0032] Figure 5 It is a schematic diagram of the upper end structure of a top cover of a preparation device for a bovine superovulation hormone according to the present application;

[0033] Figure 6 It is a schematic diagram of the bottom structure of a top cover of a preparation device for a bovine superovulation hormone according to the present application;

[0034] Figure 7 It is a schematic diagram of the separation of a cylindrical protective film and a centrifuge cylinder of a preparation device for a bovine superovulation hormone according to the present application;

[0035] Figure 8 It is a schematic diagram of the connection of a moving ring and a vertical rod of a preparation device for a bovine superovulation hormone according to the present application;

[0036] Figure 9The application is a centrifugal cylinder and center disc connection diagram for preparing a device of a superovulation hormone for cattle.

[0037] In the figure: 1, mounting frame; 11, rotating frame; 12, collection disc; 13, collection barrel; 14, suction pump; 15, ring; 2, top frame; 21, first motor; 22, rotating disc; 23, vertical plate; 3, centrifugal assembly; 31, center disc; 32, centrifugal cylinder; 33, top cover; 34, cylindrical protective film; 35, vertical rod; 36, rotating ring; 37, L-shaped tube; 38, moving ring; 39, pull ring; 310, drain port; 311, arc-shaped groove; 312, convex plate; 313, buckle plate; 314, filter plate; 315, clamping plate; 316, ring groove; 317, sliding plate; 318, bottom plate; 319, return spring; 320, sliding rail strip. DETAILED DESCRIPTION

[0038] In order to make the technical means, creative features, purposes and effects of the application easy to understand, the application is further described below in combination with specific embodiments.

[0039] Please refer to Figures 1 to 9 The application provides a technical solution:

[0040] A preparation method of a superovulation hormone for cattle, the preparation method comprising the following steps:

[0041] (1) adding a buffer solution into low-quality HCG and stirring to mix the solution, then adding the mixed solution into an upper centrifugal cylinder of a centrifugal assembly for centrifugal treatment, after centrifuging the solution for 15-20 min, rotating the upper centrifugal cylinder to the lower layer, precipitating the solution, and collecting the supernatant after the solution is layered;

[0042] (2) taking out the remaining solution of the lower centrifugal cylinder, adding a buffer solution into the remaining solution and stirring to mix, then adding the mixed solution into the upper centrifugal cylinder for secondary centrifugal treatment, after centrifuging, rotating the centrifugal cylinder to the lower end to stand, and collecting the supernatant in the same way;

[0043] (3) adding the collected supernatants twice into a collection barrel to combine, then adding artificial zeolite and stirring, after mixing, using a Buchner funnel to filter the supernatant, and repeatedly washing with distilled water to obtain a semi-finished product solution;

[0044] (4) loading the semi-finished product solution into a chromatography column, adding an ethanol solution for elution, collecting a protein elution peak part, adding an ethanol solution, standing for 6 h, centrifuging, discarding the supernatant, and collecting the precipitate;

[0045] (5) Dissolve the collected precipitate with buffer solution and continuously add the dissolved precipitate solution to the chromatography column. When the UV analyzer at the column inlet shows that protein solution is flowing out, start collecting. After all the solution is added to the column, when the liquid level is equal to the column bed level, continuously add phosphate buffer solution until the UV analyzer shows no protein flowing out, then stop collecting and add ethanol. Let stand for 6 hours, then remove the supernatant, collect the precipitate, dehydrate the precipitate with anhydrous ethanol, and vacuum dry to obtain high-quality HCG.

[0046] An apparatus for preparing bovine superovulation hormone includes a mounting frame 1. Two rotating frames 11 are fixed on the vertical surface of the mounting frame 1. A centrifuge assembly 3 is provided between the two rotating frames 11. The centrifuge assembly 3 includes a central disk 31. Centrifuge cylinders 32 are rotatably mounted on the top and bottom of the central disk 31. A movable ring 38 is slidably sleeved on the outer wall of the centrifuge cylinder 32. A cylindrical protective film 34 is sleeved on the inner wall of the centrifuge cylinder 32. The outer end of the cylindrical protective film 34 is clamped inside the movable ring 38. A top cover 33 covers the end of the centrifuge cylinder 32 away from the central disk 31. A pull ring 39 is fixed on the outer surface of the top cover 33, and a drain outlet 310 is fixed on one side of the top cover 33 located on the pull ring 39. A rubber plug is sealed inside the drain outlet 310. A collection tray 12 is placed at the bottom of the mounting frame 1, directly opposite the centrifuge cylinder 32. A collection bucket 13 is placed on the other side of the collection tray 12, and a suction pump 14 is installed on the top of the collection bucket 13. The suction pump 14 is fixedly installed on the side of the rotating frame 11. A turntable 22 is provided on the top of the centrifuge cylinder 32. Vertical plates 23 are fixed on both sides of the turntable 22, and the vertical plates 23 are movably engaged with the outer wall of the centrifuge cylinder 32. When using the device, the solution mixed once is added into the upper centrifuge cylinder 32. The turntable 22 and the centrifuge cylinder 32 work together to drive the mixture to rotate. Then the centrifuge cylinder 32 is rotated to the lower layer. After a period of time, the supernatant is taken out. The remaining solution is collected and added to the buffer solution for a second centrifugation. Finally, the supernatants taken out twice are mixed together for subsequent preparation processes.

[0047] In the scheme, the centrifugal assembly 3 further comprises a rotating ring 36, the top and bottom of the center disc 31 are rotatably installed with the rotating ring 36, the rotating ring 36 is fixedly connected with the two groups of centrifugal cylinders 32, the upper end and the lower end of the center disc 31 are provided with a bottom plate 318, and the bottom plate 318 slides along the inside of the corresponding centrifugal cylinder 32, the inner wall of the centrifugal cylinder 32 is fixedly provided with a sliding rail strip 320, and the bottom plate 318 slides along the sliding rail strip 320, a plurality of return springs 319 are fixedly arranged between the bottom plate 318 and the center disc 31 at equal intervals in a ring shape, through the arrangement of the center disc 31 and the rotating ring 36, the upper and lower two groups of centrifugal cylinders 32 do not interfere with each other, the upper centrifugal cylinder 32 can be cooperated with the rotating disc 22 to perform centrifugal rotation, and the lower centrifugal cylinder 32 can be placed and layered, the bottom plate 318 supports the bottom of the centrifugal cylinder 32, and keeps the bottom of the centrifugal cylinder 32 on a plane.

[0048] In the embodiment, the center of the center disc 31 is fixedly provided with an L-shaped pipe 37, one end of the L-shaped pipe 37 penetrates from the side of the center disc 31, the other end of the L-shaped pipe 37 rotatably penetrates the inside of the bottom plate 318 and the cylindrical protective film 34, the suction pump 14 is provided with a connecting port corresponding to the position of the L-shaped pipe 37 penetrating out of the lower centrifugal cylinder 32, one end of the L-shaped pipe 37 penetrating the cylindrical protective film 34 is provided with a one-way valve, the L-shaped pipe 37 penetrates the circular ring 15, so that the rotation interference of the centrifugal assembly 3 is avoided, meanwhile, the upper end of the L-shaped pipe 37 is sealed by the suction pump 14, the lower end of the L-shaped pipe 37 is connected with the connecting port of the suction pump 14, the supernatant in the lower centrifugal cylinder 32 is sucked out by the suction pump 14 and discharged into the collecting barrel 13 for collection, so as to facilitate subsequent mixing.

[0049] The outer side of the centrifugal cylinder 32 is equidistantly slidably connected with four vertical rods 35, the outer side of the moving ring 38 is provided with sliding plates 317 corresponding to the positions of the vertical rods 35, the sliding plates 317 are slidably connected with the vertical rods 35, the outer side of the moving ring 38 is provided with an annular groove 316, the sliding plates 317 slide along the inner side of the annular groove 316, the top of the moving ring 38 is rotatably connected with a plurality of clamping plates 315 through a rotating shaft and a torsion spring, the outer end of the cylindrical protective film 34 is clamped between the clamping plates 315 and the moving ring 38, the top of the vertical rod 35 is rotatably connected with a buckle plate 313 through a rotating shaft and a torsion spring, the top cover 33 is provided with an arc-shaped groove 311 corresponding to the position of the buckle plate 313, the bottom of the buckle plate 313 is slidably connected with the arc-shaped groove 311, the inner wall of the top cover 33 is fixedly connected with a filter plate 314, the filter plate 314 is sealingly inserted into the inner side of the centrifugal cylinder 32, one end of the cylindrical protective film 34 is inserted into the inner side of the centrifugal cylinder 32 and is in close contact with the inner wall of the centrifugal cylinder 32, the bottom is in contact with the bottom plate 318, and the position corresponding to the L-shaped pipe 37 is provided with a hole for the L-shaped pipe 37 to pass through, the outer end of the cylindrical protective film 34 is sleeved on the outer periphery of the centrifugal cylinder 32 and is clamped by the moving ring 38 and the clamping plate 315, and the depth of the cylindrical protective film 34 can be adjusted by sliding along the vertical direction of the vertical rod 35, so as to adjust the content of different mixed liquids, after the mixed liquid is added into the cylindrical protective film 34 in the centrifugal cylinder 32, the cylindrical protective film 34 gradually descends along with the accumulation of weight, the bottom plate 318 is slid downward along the sliding rail 320 to compress the reset spring 319, the moving ring 38 is slid upward along the vertical rod 35, the top cover 33 is wrapped around the opening end of the centrifugal cylinder 32 and the cylindrical protective film 34, the buckle plate 313 is buckled in the arc-shaped groove 311 to limit the vertical position of the top cover 33, so as to prevent the top cover 33 from being opened during the centrifugal rotation, after the centrifugal cylinder 32 rotates to the lower end, the supernatant after standing is pumped out through the L-shaped pipe 37, the sealing plug of the drain hole is opened, the remaining solution is collected in the collection disc 12, and secondary centrifugation is performed, then the sliding plate 317 is moved along the annular groove 316 to slide the vertical rod 35 and the buckle plate 313 along the arc-shaped groove 311, the bottom of the vertical rod 35 is actually connected with the outer wall of the centrifugal cylinder 32 in a conventional convex strip sliding connection mode, springs are arranged between adjacent vertical rods 35 to keep the four vertical rods 35 equidistantly distributed, the two vertical rods 35 are brought together, so as to seal the opening end of the cylindrical protective film 34, the top cover 33 is opened, the cylindrical protective film 34 is taken out, the inside of the cylindrical protective film 34 is collected and treated separately.

[0050] The top of the vertical rod 35 is fixed with a convex plate 312, the vertical plate 23 is provided with a clamping groove corresponding to the position of the convex plate 312, and the convex plate 312 is slidingly clamped in the clamping groove. The top of the mounting frame 1 is fixed with a top frame 2, and the rotating disc 22 is rotatably installed on the top frame 2. The first motor 21 is fixed on the upper end of the top frame 2 corresponding to the position of the center of the rotating disc 22, and the output end of the first motor 21 is fixedly connected with the rotating disc 22. The front end of the rotating frame 11 is slidingly clamped with a circular ring 15, and the side surface of the circular ring 15 is fixedly connected with the center disc 31. The outlet end of the L-shaped pipe 37 passes through the center of the circular ring 15, and the center disc 31 is rotatably installed between the rotating frames 11 through the circular ring 15. The upper and lower two groups of centrifugal cylinders 32 can be replaced and used by artificial overturning. At the same time, the side surface and the bottom of the mounting frame 1 give the centrifugal cylinder 32 enough rotating space and do not interfere with the rotation. In the process of rotating the centrifugal cylinder 32, the convex plate 312 is inserted into the vertical direction clamping groove of the vertical plate 23. The convex plate 312 and the clamping groove are arc-shaped and are inserted into each other. The arc is the same as the rotating track of the circular ring 15. After the convex plate 312 is clamped with the vertical plate 23, the rotating disc 22 is driven by the first motor 21 to rotate the centrifugal cylinder 32 at the upper end, so as to centrifuge the mixed liquid in the inside. Because the front and rear ends of the clamping groove are open, the rotation of the centrifugal cylinder 32 is not interfered. At the same time, the lower centrifugal cylinder 32 can also be connected with the vertical plate 23 after rotating to the upper layer, so as to drive the centrifugal cylinder 32 alone through the rotating disc 22.

[0051] When using the device, the once-mixed solution is added to the inside of the upper centrifuge cylinder 32, and the mixed solution is driven to rotate by the cooperation of the rotating disc 22 and the centrifuge cylinder 32, and then the centrifuge cylinder 32 is rotated to the lower layer, and after a period of time, the supernatant is taken out, the remaining solution is collected and added to the buffer for secondary centrifugation, and finally the two times of supernatant taken out are mixed together for subsequent preparation process. The central disc 31 is installed between the rotating frames 11 by rotating the ring 15, and the upper and lower two sets of centrifuge cylinders 32 can be replaced and used by manually turning over. The side and bottom of the mounting frame 1 give enough rotating space to the centrifuge cylinder 32, and there is no rotating interference. In the process of rotating the centrifuge cylinder 32, the convex plate 312 is inserted into the vertical plate 23 clamping groove in the vertical direction, and the convex plate 312 and the clamping groove are arc-shaped, and the arc is the same as the rotating track of the ring 15. After the convex plate 312 is clamped with the vertical plate 23, it is driven by the first motor 21, and the rotating disc 22 drives the centrifuge cylinder 32 at the upper end to rotate, and the mixed solution inside is centrifuged. Because the front and rear ends of the clamping groove are open, the rotation of the centrifuge cylinder 32 is not interfered. At the same time, the centrifuge cylinder 32 at the lower end can also be connected with the vertical plate 23 after rotating to the upper end, and then the centrifuge cylinder 32 is driven by the rotating disc 22. One end of the cylindrical protective film 34 is inserted into the inside of the centrifuge cylinder 32 and is attached to the inner wall of the centrifuge cylinder 32. The bottom is in contact with the bottom plate 318. The position corresponding to the L-shaped pipe 37 is drilled to allow the L-shaped pipe 37 to pass out. The outer end of the cylindrical protective film 34 is sleeved outside the centrifuge cylinder 32 and is clamped by the moving ring 38 and the clamping plate 315. It can be adjusted by sliding in the vertical direction of the vertical rod 35 to adjust the depth of the cylindrical protective film 34. It is adjusted according to the content of the mixed solution. After the mixed solution is added to the cylindrical protective film 34 in the centrifuge cylinder 32, the cylindrical protective film 34 gradually descends with the accumulation of weight, and the bottom plate 318 slides downward along the sliding rail 320 to compress the return spring 319. At the same time, the moving ring 38 slides upward along the vertical rod 35. The top cover 33 is wrapped around the opening end of the centrifuge cylinder 32 and the cylindrical protective film 34. The buckle plate 313 is buckled in the arc-shaped groove 311 to realize the vertical limiting of the top cover 33 to prevent the top cover 33 from opening during centrifugal rotation. After the centrifuge cylinder 32 rotates to the lower end, the supernatant after standing is drawn out through the L-shaped pipe 37, the sealing plug of the drain hole is opened, and the remaining solution is collected in the collection disc 12 for secondary centrifugation. Then the sliding plate 317 is moved along the ring groove 316, driving the vertical rod 35 and the buckle plate 313 to slide along the arc-shaped groove 311. The bottom of the vertical rod 35 is actually connected to the outer wall of the centrifuge cylinder 32 in a conventional convex strip sliding connection manner, and springs are provided between adjacent vertical rods 35 to keep the four vertical rods 35 equally distributed. The two vertical rods 35 are brought together to seal the opening end of the cylindrical protective film 34, and the top cover 33 is opened to facilitate the removal of the cylindrical protective film 34 for separate collection and treatment of the internal precipitate. The L-shaped pipe 37 passes through the ring 15, so it will not cause rotating interference of the centrifuge assembly 3, and the upper end of the L-shaped pipe 37 is sealed by the suction pump 14.The L-shaped pipe 37 at the lower end is connected with the connecting port of the suction pump 14, and the supernatant in the centrifugal cylinder 32 at the lower end is extracted through the suction pump 14 and discharged into the collecting barrel 13 for collection, facilitating subsequent mixing.

[0052] The above shows and describes the basic principles and main features of the present application and the advantages of the present application. It is obvious for those skilled in the art that the present application is not limited to the details of the above exemplary embodiments, and the present application can be implemented in other specific forms without departing from the spirit or essential characteristics of the present application.

[0053] In addition, it should be understood that although the present specification is described in terms of embodiments, not every embodiment contains only one independent technical solution, and the description of the specification is only for the sake of clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that those skilled in the art can understand.

Claims

1. An apparatus for preparing bovine superovulation hormone, comprising a mounting frame (1), characterized in that: Two rotating frames (11) are fixed on the vertical surface of the mounting frame (1). A centrifugal assembly (3) is provided between the two rotating frames (11). The centrifugal assembly (3) includes a central disk (31). Centrifugal cylinders (32) are rotatably mounted on the top and bottom of the central disk (31). A movable ring (38) is slidably sleeved on the outer wall of the centrifugal cylinder (32). A cylindrical protective film (34) is sleeved on the inner wall of the centrifugal cylinder (32). The outer end of the cylindrical protective film (34) is clamped inside the movable ring (38). A top cover (33) covers the end of the centrifugal cylinder (32) away from the central disk (31). A pull rod is fixed on the outer surface of the top cover (33). A ring (39) and a top cover (33) are fixed to one side of the ring (39) with a drain outlet (310). A rubber plug is inserted into the drain outlet (310) for sealing. A collection tray (12) is placed at the bottom of the mounting frame (1) directly opposite the centrifuge cylinder (32). A collection bucket (13) is placed on the other side of the collection tray (12). A suction pump (14) is installed on the top of the collection bucket (13). The suction pump (14) is fixedly installed on the side of the rotating frame (11). A turntable (22) is provided on the top of the centrifuge cylinder (32). Vertical plates (23) are fixed on both sides of the turntable (22). The vertical plates (23) are movably engaged with the outer wall of the centrifuge cylinder (32).

2. The apparatus for preparing bovine superovulation hormone according to claim 1, characterized in that: The centrifugal assembly (3) also includes a rotating ring (36). The rotating ring (36) is rotatably installed on the top and bottom of the central disk (31). The rotating ring (36) is fixedly connected to two sets of centrifugal cylinders (32). The upper and lower ends of the central disk (31) are provided with a base plate (318), and the base plate (318) slides along the inside of the corresponding centrifugal cylinder (32).

3. The apparatus for preparing bovine superovulation hormone according to claim 2, characterized in that: The centrifuge tube (32) is fixed with a slide rail (320) on its inner wall, and the bottom plate (318) slides along the slide rail (320). Multiple return springs (319) are fixed in a ring at equal intervals between the bottom plate (318) and the central plate (31).

4. The apparatus for preparing bovine superovulation hormone according to claim 3, characterized in that: An L-shaped tube (37) is fixed at the center of the central disk (31). One end of the L-shaped tube (37) passes through the side of the central disk (31), and the other end of the L-shaped tube (37) rotates through the bottom plate (318) and the inside of the cylindrical protective film (34). The suction pump (14) is provided with a connection port at the position of the L-shaped tube (37) that passes through the lower centrifuge cylinder (32). A one-way valve is provided at the end of the L-shaped tube (37) that passes through the cylindrical protective film (34).

5. The apparatus for preparing bovine superovulation hormone according to claim 4, characterized in that: Four vertical rods (35) are equidistantly slidably installed on the outer side of the centrifuge tube (32). A sliding plate (317) is provided on the outer side of the moving ring (38) corresponding to the position of the vertical rods (35), and the sliding plate (317) is slidably sleeved on the vertical rods (35). An annular groove (316) is opened on the outer side of the moving ring (38), and the sliding plate (317) slides along the inside of the annular groove (316).

6. The apparatus for preparing bovine superovulation hormone according to claim 5, characterized in that: The top of the movable ring (38) is rotatably mounted with multiple clamping plates (315) via a rotating shaft and a torsion spring, and the outer end of the cylindrical protective film (34) is clamped between the clamping plates (315) and the movable ring (38).

7. The apparatus for preparing bovine superovulation hormone according to claim 6, characterized in that: The top of the vertical rod (35) is rotatably mounted with a buckle plate (313) via a pivot and a torsion spring. The top cover (33) has an arc-shaped groove (311) corresponding to the buckle plate (313), and the bottom protrusion of the buckle plate (313) slides along the arc-shaped groove (311). A filter plate (314) is fixed to the inner wall of the top cover (33), and the filter plate (314) is sealed and inserted into the centrifuge cylinder (32).

8. The apparatus for preparing bovine superovulation hormone according to claim 7, characterized in that: The top of the vertical rod (35) is fixed with a protruding plate (312). The vertical plate (23) has a slot corresponding to the position of the protruding plate (312), and the protruding plate (312) is slidably engaged in the slot. The top of the mounting frame (1) is fixed with a top frame (2), and the turntable (22) is rotatably mounted on the top frame (2). The upper end of the top frame (2) is fixed with a first motor (21) corresponding to the center of the turntable (22), and the output end of the first motor (21) is fixedly connected to the turntable (22).

9. The apparatus for preparing bovine superovulation hormone according to claim 8, characterized in that: The front end of the rotating frame (11) is slidably connected to a ring (15), the side of the ring (15) is fixedly connected to the central disk (31), and the outlet end of the L-shaped tube (37) passes through the center of the ring (15).

10. A method for preparing bovine superovulation hormone, comprising using an apparatus for preparing bovine superovulation hormone as described in any one of claims 1 to 9, characterized in that: The preparation method includes the following steps: (1) Add buffer solution to low-quality HCG and stir to mix the solution. Then add the mixed solution to the upper centrifuge tube of the centrifuge assembly for centrifugation. After centrifuging the solution for 15-20 minutes, rotate the upper centrifuge tube to the lower layer to precipitate the solution until the solution separates into layers and collect the supernatant. (2) Remove the remaining solution from the lower centrifuge tube, add buffer solution to the remaining solution and stir to mix. Then add the mixture to the upper centrifuge tube for a second centrifugation. After centrifugation, rotate the centrifuge tube to the lower end and let it stand in the same way to collect the supernatant. (3) Combine the supernatant collected twice in the collection bucket, then add artificial zeolite and stir. After mixing, filter the supernatant with a Buchner funnel and wash repeatedly with distilled water to obtain a semi-finished solution. (4) Load the semi-finished product solution into the chromatography column, add ethanol solution for elution, collect the protein elution peak, add ethanol solution, let stand for 6 hours, centrifuge, discard the supernatant and collect the precipitate. (5) Dissolve the collected precipitate with buffer solution and continuously add the dissolved precipitate solution to the chromatography column. When the UV analyzer at the column inlet shows that protein solution is flowing out, start collecting. After all the solution is added to the column, when the liquid level is equal to the column bed level, continuously add phosphate buffer solution until the UV analyzer shows no protein flowing out, then stop collecting and add ethanol. Let stand for 6 hours, then remove the supernatant, collect the precipitate, dehydrate the precipitate with anhydrous ethanol, and vacuum dry to obtain high-quality HCG.

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