Synthesis system and synthesis process of monosialotetrahexosyl ganglioside GM1

Abstract

The present invention relates to a monosialotetrahexosyl ganglioside GM1 synthesis system and a synthesis process thereof, and belongs to the technical field of monosialotetrahexosyl ganglioside GM1 synthesis, the synthesis system comprises: a cylinder movably arranged and provided with a liquid inlet; the cover body is used for opening and closing the liquid inlet; the filtering assembly is used for filtering the mixed liquid; the liquid separation assembly is rotationally mounted in the cylinder body; pipettes are uniformly mounted on the pipette tray, and the liquid separation assembly is used for sub-packaging the filtered mixed liquid into the pipettes; the liquid outlet and exhaust assembly is used for opening and closing the pipette and exhausting gas in the pipette; and the controller is in communication connection with the liquid separation assembly and the liquid outlet and exhaust assembly and is used for controlling the rotation of the liquid separation assembly, controlling the liquid outlet and exhaust assembly to open or close the pipette and exhausting gas in the pipette. The method has the beneficial effects that the contact time between the sample and external air and the like is reduced as much as possible in the transfer process, and some components sensitive to air are prevented from changing.

Description

Technical Field

[0001] The present application relates to the technical field of the synthesis of monosialotetrahexosylganglioside GM1, and in particular to a synthesis system and a synthesis process of monosialotetrahexosylganglioside GM1. Background Art

[0002] The main synthesis processes of monosialotetrahexosylganglioside GM1 are as follows: 1. Extraction and purification from animal brain tissue Acetone treatment: Freeze the animal brain tissue (such as pig brain) and remove the meninges and blood vessels. Add acetone and crush and stir, then filter and dry to obtain acetone-insoluble dry powder.

[0003] Total lipid extraction: Use a mixed solution of chloroform-methanol-water (volume ratio of 5:5:1) as the extracting solution, add 1 kg of acetone insoluble dry powder to 10 L of extracting solution, add the acetone insoluble dry powder to the extracting solution and stir evenly. After extraction, centrifuge to remove the insoluble matter and collect the supernatant.

[0004] Folch distribution: Add distilled water to the supernatant and mix thoroughly. For the first time, add distilled water in an amount of 15% of the volume of the supernatant, then let it stand to separate into layers and take the upper layer. Then add distilled water in an amount of 10% of the volume of the lower layer solution to the remaining lower layer solution, let it stand to separate into layers and take the upper layer. Repeat 2-4 times, combine the upper layer solutions, and remove the methanol by rotary evaporation under reduced pressure from the combined upper layer solutions until the volume of the solution no longer decreases to obtain a GM1 aqueous solution, which is then desalted and concentrated to obtain a crude GM1 extract.

[0005] Column chromatography purification: The crude GM1 extract is first subjected to weak anion exchange chromatography, then to gel exclusion chromatography, and finally to lyophilization to obtain purified GM1.

[0006] 2. Enzyme-catalyzed hydrolysis and synthesis Extraction and purification of GM1: the same as the steps for extraction and purification from animal brain tissue.

[0007] Hydrolysis of purified GM1: The purified GM1 is subjected to a hydrolysis reaction catalyzed by SCDase, the pH value of the reaction system is controlled at 5.8, surfactant TDC, metal ion Ca²⁺ and SCDase are added to the system, the mass volume concentration of TDC is controlled at 0.07-0.7%, the concentration of Ca²⁺ is controlled at 2.5-100mM, and the concentration of SCDase is controlled at more than 4000U per mol of substrate, so that the fatty acid module of the polyhydroxy fatty amine in the sphingosine part of the purified GM1 is hydrolyzed and removed, and then centrifuged to remove the precipitate, and the supernatant is purified with a Sep-PakC18 solid phase extraction column, first eluting TDC with 65% methanol, and finally eluting Lyso-GM1 with 85% methanol, and the eluted Lyso-GM1 is evaporated to remove methanol and then freeze-dried to finally obtain purified Lyso-GM1.

[0008] Synthesis of GM1 with specific fatty acid module: The purified Lyso-GM1 and carbon 18 or carbon 20 fatty acids are reacted under SCDase catalysis conditions, the pH value of the reaction system is controlled at 7.5, DME and SCDase are added to the system, the mass percentage concentration of DME is controlled at 10%, the concentration of SCDase is controlled at 150-200U, the concentration of Lyso-GM1 is controlled at 0.75-10mM, and the mass percentage of fatty acid is 1-10mg / ml. After synthesis, the nanofiltration method is first used for desalination, and then it is purified with a Sep-PakC18 solid phase extraction column. The desalted reaction mixture is passed through a 500mg Sep-PakC18 solid phase extraction column, and then the flow-through liquid is passed through a 5g Sep-PakC18 solid phase extraction column, washed with 50ml of water, and then washed with 50% methanol. Then, the GM1 with a specific fatty acid module is washed with 100% methanol, and finally evaporated, concentrated and freeze-dried to obtain purified GM1 with a specific fatty acid module.

[0009] 3. Detergent extraction method Extraction: Weigh the animal brain tissue, homogenize it, use a detergent solution with a weight percentage of 1%-10% as the extraction solvent, the volume ratio of the homogenized animal brain tissue to the extraction solvent is 1:0.5-5, stir and extract at 4°C-60°C, remove the precipitate by centrifugation or filtration, the precipitate can be extracted multiple times in the same way until the ganglioside is completely extracted, and combine the multiple extracts.

[0010] Concentration: The extract is concentrated to obtain a crude monosialotetrahexosylganglioside extract, which can be further made into a dry powder, i.e. a crude ganglioside extract.

[0011] Purification: The crude extract or crude extract is further purified by DEAE-Sephadex A-25 and porous silica gel IATR to obtain monosialotetrahexosylganglioside with a purity greater than 95%, preferably 98%.

[0012] 4. Column chromatography purification Mobile phase preparation: Mix the organic solvent and the buffer solution in a volume ratio of 0.5-8:1, and adjust the pH value to 2.5-5.5 using an aqueous NaOH solution or an aqueous phosphoric acid solution to obtain the mobile phase.

[0013] Sample dissolution: Use an appropriate amount of mobile phase to fully dissolve the crude GM1 and then filter to obtain a crude solution.

[0014] Elution on the column: Use an injection pump to pump the crude product solution into the dynamic axial compression preparation column equipped with hydrophobic filler, and then elute with the mobile phase. Collect components GM1A and GM1B at the corresponding elution time and stage, and evaporate the solvent separately to obtain their corresponding single-component samples, or mix the collected eluents and evaporate them to obtain high-purity GM1.

[0015] Regardless of which of the above processes is used to synthesize GM1, centrifugation of the mixed solution is involved. Before centrifugation, the mixed solution needs to be transferred from the pre-treatment to the centrifugal equipment. During the transfer of the mixed solution, the mixed solution is easily exposed to the outside air, which causes changes in the air-sensitive components in the mixed solution, thereby affecting the subsequent preparation of GMI. Summary of the invention

[0016] In order to minimize the contact time of the sample with the outside air and the like during the transfer process and prevent some air-sensitive components from changing, the present application provides a synthesis system and a synthesis process of monosialotetrahexosylganglioside GM1.

[0017] In the first aspect, the present application provides a synthesis system of monosialotetrahexosylganglioside GM1, which adopts the following technical scheme: A synthesis system of monosialotetrahexosylganglioside GM1, comprising: The cylinder is movably arranged and has a liquid inlet; A cover body, rotatably connected to the cylinder body, and used for opening and closing the liquid inlet; A filter assembly is installed in the cylinder and is used to filter the mixed liquid; A liquid dispensing assembly, rotatably mounted in the cylinder; A pipette tray, with a plurality of pipettes evenly mounted thereon, wherein the liquid dispensing assembly is used to dispense the filtered mixed liquid into the pipettes; A liquid outlet and exhaust assembly, mounted on the cylinder, for opening and closing the pipette and exhausting the gas in the pipette; The controller is in communication connection with the liquid separation component and the liquid outlet and exhaust component, and is used to control the rotation of the liquid separation component, control the liquid outlet and exhaust component to open or close the pipette and exhaust the gas in the pipette.

[0018] By adopting the above technical solution, after the mixed liquid is discharged into the cylinder, the cylinder is closed to make the cylinder sealed, filtered by the filter component, exhausted by the liquid outlet exhaust component, and separated by the liquid separation component, so that each pipette is filled with the mixed liquid, and then the cylinder is driven to move to the separation device. After moving into place, the mixed liquid in each pipette is discharged into the separation device; because the cylinder is always in a closed state during the movement of the cylinder, the contact time between the sample and the outside air is reduced, and changes in some air-sensitive components are avoided.

[0019] Optionally, the liquid outlet and exhaust assembly includes: A liquid discharge pipe, consisting of a curved section and a vertical section, wherein the vertical section is coaxial with the pipette, one end of the curved section is connected to the vertical section, and the other end is connected to the pipette near the bottom; A liquid outlet one-way valve connected to the bottom of the vertical section; An exhaust check valve is coaxially rotatably connected to the pipette, and an air outlet is provided near the top of the pipette, and the exhaust check valve can be connected to the air outlet; A liquid-blocking and air-exhausting block is slidably connected in the pipette and is used to open or isolate the connection between the curved section and the pipette; a surface of the liquid-blocking and air-exhausting block close to the pipette is an inclined surface and is inclined toward the curved section; A driving component is installed on the cylinder and is used to drive the sliding of the liquid blocking and exhausting block. The driving component is communicatively connected with the controller.

[0020] By adopting the above technical scheme, when the liquid-stopping and exhaust block is located at the connection point between the curved section and the pipette, the mixed liquid is stored in the pipette, and when the liquid-stopping and exhaust block moves to the bottom of the pipette, the curved section is connected with the pipette, the mixed liquid enters the curved section, and enters the vertical section from the curved section to be discharged; the inclined surface of the liquid-stopping and exhaust block is conducive to the mixed liquid entering the curved section; in addition, when it is necessary to exhaust the inside of the pipette, the exhaust check valve can be rotated, and when the exhaust check valve is connected with the air outlet, the gas is discharged by the exhaust check valve, and the external gas will not enter the inside of the pipette through the exhaust check valve; when the mixed liquid is dispensed into the pipette, the exhaust check valve can be rotated so that the exhaust check valve is no longer connected with the air outlet, thereby preventing the mixed liquid from being discharged from the exhaust check valve to the outside.

[0021] Optionally, the driving component includes: A driving gear ring, rotatably connected to the pipetting plate; A driving rod, one end of which is threadedly connected to the liquid blocking and exhausting block; A driving slave gear is mounted on the other end of the driving rod through a spline and is capable of meshing with the driving ring gear; A driving cylinder, a cylinder body of which is mounted on the transfer plate, and a piston rod is slidingly connected to the end surface of the driving slave gear, and the driving cylinder is communicatively connected to the controller.

[0022] By adopting the above technical solution, when it is necessary to discharge the mixed liquid in a pipette, the driving cylinder corresponding to the pipette can be controlled to contract, so that the driving slave gear engages with the driving ring gear, so that when the driving ring gear rotates, it only drives the rotation of the driving slave gear, thereby realizing the movement of the liquid blocking and exhausting block.

[0023] Optionally, the filtering component includes: A conical filter screen is installed in the cylinder; The conical collecting hopper is fixedly connected to the conical filter screen and is used to collect the filtered mixed liquid. The bottom end of the conical collecting hopper is connected to the liquid separation component.

[0024] By adopting the above technical solution, after the mixed liquid enters the cylinder, it is filtered by the conical filter screen, gathered in the conical collecting bucket, and then enters the liquid separation component.

[0025] Optionally, the filtering component further includes: A cleaning brush is fixedly connected to the inner wall of the cylinder and abuts against the conical filter; the conical filter is rotatably connected to the cylinder; A liquid baffle plate, a slag discharge hole is opened on the side wall of the cylinder, and the liquid baffle plate is slidably connected to the cylinder and is used to open and close the slag discharge hole.

[0026] By adopting the above technical solution, when the conical filter screen rotates, the cleaning brush assists in cleaning the conical filter screen to reduce the degree of blockage of the conical filter screen.

[0027] Optionally, the liquid separation component includes: A liquid separation main pipe is connected to the bottom of the conical collecting bucket and is connected to a liquid separation valve, and the liquid separation valve is in communication with the controller; The liquid dispensing branch pipe is communicated with the liquid dispensing main pipe and abuts against the top of the pipette.

[0028] By adopting the above technical solution, when the conical collecting bucket rotates, the liquid separation main pipe drives the liquid separation branch pipe to rotate, so that the filtered mixed liquid enters each pipette through which the liquid separation branch pipe passes.

[0029] Optionally, the synthesis system further comprises: The liquid collection and recovery component is installed on the transfer plate, the transfer plate is conical, and the liquid collection and recovery component is used to recover the excess mixed liquid in the transfer plate into each of the pipettes.

[0030] By adopting the above technical solution, since the liquid dispensing branch rotates when dispensing liquid to the pipette, part of the mixed liquid will fall onto the pipette plate, or the mixed liquid in the pipette will overflow. Therefore, the pipette plate is conically set to facilitate the collection of excess mixed liquid; then, when needed, it is recovered into each pipette through the liquid collection recovery component to avoid waste.

[0031] Optionally, the liquid collection recovery component includes: A liquid pump, the inlet of which is connected to the lowest point of the pipetting plate, and the outlet of which is connected to a liquid discharge pipe; the liquid pump is in communication with the controller; The drainage branch pipe has one end connected to each of the pipettes near the top, and the other end connected to the drainage main pipe.

[0032] By adopting the above technical solution, when recovery is required, the liquid pump is controlled to start, and the liquid pump pumps the mixed liquid in the pipetting tray into each pipette.

[0033] Optionally, the synthesis system further comprises: An auxiliary gear ring, mounted on the cover body; An auxiliary motor is mounted on the cylinder, and the output shaft is mounted with a main gear via a spline; the auxiliary motor is in communication connection with the controller; An opening and closing gear is rotatably connected to the cylinder, and the main gear can mesh with the opening and closing gear; the auxiliary gear ring meshes with the opening and closing gear; A rotating rod, fixedly connected to the conical filter screen and rotatably connected to the cylinder; A rotating gear is coaxially fixedly connected to the rotating rod, and the main gear can mesh with the rotating gear; An electromagnet, coaxially fixedly connected to the output shaft of the auxiliary motor; The iron sheet is coaxially fixedly connected to the main gear; when the electromagnet is not energized, the main gear is meshed with the opening and closing gear; after the electromagnet is energized, the iron sheet is attracted, and the main gear is meshed with the rotating gear.

[0034] By adopting the above technical solution, when liquid is introduced, the main gear is meshed with the rotating gear, so that the auxiliary motor drives the rotation of the conical filter; after the liquid is introduced, the electromagnet is powered off, so that the main gear is meshed with the opening and closing gear, so that the auxiliary motor drives the rotation of the auxiliary gear ring to drive the rotation of the cover body, so that the cover body closes the liquid inlet.

[0035] In the second aspect, the present application provides a synthesis process of monosialotetrahexosylganglioside GM1, which adopts the following technical scheme: A synthesis process of monosialotetrahexosylganglioside GM1, comprising: Filter and collect the mixed solution after pretreatment; Expel air from each pipette; Dispense the mixed solution into each of the pipettes; Closing the liquid inlet of the cylinder and driving the cylinder to move; After the cylinder moves to the separation device, controlling the pipette to discharge the mixed liquid to the separation device; The mixed liquid collected in the transfer tray is recovered into the transfer pipette to continue to discharge the mixed liquid into the separation device.

[0036] By adopting the above technical solution, after the mixed liquid is discharged into the cylinder, the cylinder is closed to make it sealed, and after filtering, exhausting and liquid separation, each pipette is filled with the mixed liquid, and then the cylinder is driven to move to the separation device. After moving into place, the mixed liquid in each pipette is discharged into the separation device; since the cylinder is always in a closed state during the movement of the cylinder, the contact time between the sample and the outside air is reduced, thereby avoiding changes in some air-sensitive components.

[0037] In summary, the present application has at least the following beneficial effects: 1. The purpose of setting the cylinder, cover, filter assembly, liquid separation assembly and liquid outlet exhaust assembly is to close the cylinder after the mixed liquid is discharged into the cylinder, so that the cylinder is sealed, and the mixed liquid is filtered by the filter assembly, exhausted by the liquid outlet exhaust assembly, and separated by the liquid separation assembly, so that each pipette is filled with the mixed liquid, and then the cylinder is driven to move to the separation device. After moving into place, the mixed liquid in each pipette is discharged into the separation device; because the cylinder is always in a closed state during the movement of the cylinder, the contact time between the sample and the outside air is reduced, and changes in some air-sensitive components are avoided.

[0038] 2. The purpose of setting the pipette plate to be conical and the liquid collection and recovery component is that since the liquid dispensing branch rotates when dispensing liquid to the pipette, part of the mixed liquid will fall onto the pipette plate, or the mixed liquid in the pipette will overflow. Therefore, the pipette plate is set in a conical shape to facilitate the collection of excess mixed liquid; then, when needed, it can be recovered into each pipette through the liquid collection and recovery component to avoid waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 It is a structural schematic diagram of the pipetting-related structure of the present application; Figure 2 It is a schematic diagram of the overall structure behind the hidden cylinder; Figure 3 It is a structural schematic diagram of the relevant structure on the pipetting plate; Figure 4 is a schematic diagram of the cross-sectional structure of a pipette; Figure 5 It is a control structure block diagram of the synthesis system of the present application; Figure 6 It is a flowchart of the synthesis process of this application.

[0040] Explanation of the reference numerals: 100, cylinder; 101, cover; 102, auxiliary motor; 103, main gear; 104, auxiliary gear ring; 105, opening and closing gear; 200, filter assembly; 210, conical filter screen; 211, rotating rod; 212, rotating gear; 220, conical collecting bucket; 230, cleaning brush; 240, liquid baffle plate; 250, electric push rod; 300, liquid separation assembly; 310, liquid separation main pipe; 311, liquid separation valve; 320, liquid separation branch pipe; 400, transfer plate; 410, transfer pipe; 500, liquid outlet exhaust assembly; 510, discharge pipe; 520, liquid outlet one-way valve; 530, exhaust one-way valve; 531, transfer rod The motor; 532, exhaust main gear; 533, exhaust slave gear; 540, liquid blocking exhaust block; 550, driving component; 551, driving gear ring; 552, driving rod; 553, driving slave gear; 554, driving cylinder; 555, driving motor; 556, driving main gear; 581, liquid flow main gear; 582, liquid flow slave gear; 583, liquid flow block; 584, liquid flow hole; 585, liquid closing arc gear ring; 600, liquid collection and recovery component; 610, liquid suction pump; 620, liquid discharge main pipe; 630, liquid discharge branch pipe; 701, electromagnet; 702, iron sheet; 703, controller; 704, liquid level sensor; 705, prompter. DETAILED DESCRIPTION

[0041] To make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the following will be combined with the appended drawings of the embodiments of the present invention. Figure 1 -Attached Figure 6 , the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0042] The first embodiment of the present application discloses a synthesis system of monosialotetrahexosylganglioside GM1. Figure 1 and Figure 2 As one embodiment of the synthesis system, the synthesis system may include a barrel 100, a filtering component 200, a liquid separation component 300, a pipetting plate 400, a liquid outlet and exhaust component 500 and a liquid collection and recovery component 600.

[0043] Among them, the top of the cylinder 100 is provided with a liquid inlet, and a cover 101 is rotatably connected to the cylinder 100, and the cover 101 is used to open or seal the liquid inlet. The filter assembly 200 is installed in the cylinder 100, and is used to filter the mixed liquid entering the cylinder 100, and the liquid separation assembly 300 is rotatably installed in the cylinder 100. The pipette plate 400 is installed in the cylinder 100, and the pipette 410 is evenly installed on the circumference of the pipette plate 400, and the liquid separation assembly 300 is used to dispense the filtered mixed liquid into the pipette 410. The liquid outlet exhaust assembly 500 is installed on the cylinder 100, and is used to open and close the pipette 410 and exhaust the gas in the pipette 410. In addition, the pipette plate 400 is conically arranged to collect excess mixed liquid; the liquid collection recovery assembly 600 is installed on the pipette plate 400, and is used to recover the excess mixed liquid into the pipette 410.

[0044] It should be noted that the cylinder 100 can be driven to move by an external driving mechanism (such as a walking mechanism, etc.) The external driving mechanism is not described in detail in this application and is a conventional structure.

[0045] Reference Figure 1 In order to realize the rotation of the cover body 101, an auxiliary motor 102 is installed on the cylinder body 100, and the output shaft of the auxiliary motor 102 is installed with a main gear 103 through a spline. An auxiliary gear ring 104 is fixedly connected to the cover body 101, and an opening and closing gear 105 is rotatably connected to the cylinder body 100, and the opening and closing gear 105 can mesh with the main gear 103 and mesh with the auxiliary gear ring 104.

[0046] Reference Figure 1 and Figure 2 , the filter assembly 200 may include a conical filter screen 210, a conical collection bucket 220, a cleaning brush 230 and a liquid baffle plate 240. The conical filter screen 210 is rotatably connected to the cylinder 100, and the conical collection bucket 220 is inverted on the conical filter screen 210 and fixedly connected to the conical filter screen 210. The cleaning brush 230 is obliquely installed on the inner wall of the cylinder 100 and abuts against the surface of the conical filter screen 210. A slag discharge hole is provided at the cylinder 100 near the abutment between the conical filter screen 210 and the inner wall of the cylinder 100, and the liquid baffle plate 240 is slidably connected to the cylinder 100 to open and close the slag discharge hole. An electric push rod 250 may be installed on the outer wall of the cylinder 100, and the telescopic end of the electric push rod 250 is fixedly connected to the liquid baffle plate 240 to drive the sliding of the liquid baffle plate 240.

[0047] After the mixed liquid enters the cylinder 100, the conical filter 210 filters the mixed liquid, and the conical collection bucket 220 collects the filtered mixed liquid. When removing slag, the liquid baffle plate 240 is driven to slide, and the liquid baffle plate 240 opens the slag discharge hole. When the conical filter 210 is driven to rotate, the cleaning brush 230 cleans the conical filter 210.

[0048] In addition, a rotating rod 211 is fixedly connected to the end of the conical filter screen 210 , and a rotating gear 212 is coaxially fixedly connected to the rotating rod 211 . The rotating gear 212 can mesh with the main gear 103 .

[0049] In order to realize the movement of the main gear 103 on the output shaft of the auxiliary motor 102, an electromagnet 701 is coaxially fixedly connected to the output shaft of the auxiliary motor 102, and an iron sheet 702 is coaxially fixedly connected to the end face of the main gear 103. When the electromagnet 701 is powered on, the iron sheet 702 is attracted to make the main gear 103 mesh with the rotating gear 212. When the electromagnet 701 is powered off, the main gear 103 meshes with the opening and closing gear 105. In addition, in other embodiments, in order to facilitate the resetting of the main gear 103, a reset spring can be installed on the output shaft to assist the resetting of the main gear 103.

[0050] The liquid separation assembly 300 may include a liquid separation main pipe 310 and a liquid separation branch pipe 320. The liquid separation main pipe 310 is connected to the bottom of the conical collecting bucket 220, and a liquid separation valve 311 is connected to the liquid separation main pipe 310. The liquid separation branch pipe 320 is connected to the liquid separation main pipe 310 and abuts against the top of the pipette 410. When the conical collecting bucket 220 rotates, the liquid separation main pipe 310 is driven to rotate, so that the mixed liquid passes through the liquid separation branch pipe 320 and enters each pipette 410.

[0051] Reference Figure 3 and Figure 4 The liquid outlet and exhaust assembly 500 may include a liquid discharge pipe 510 , a liquid outlet check valve 520 , an exhaust check valve 530 , a liquid blocking and exhaust block 540 , and a driving component 550 .

[0052] Among them, the liquid discharge pipe 510 is composed of a curved section and a vertical section. The vertical section is coaxially arranged with the pipette 410. One end of the curved section is connected to the vertical section, and the other end is connected to the pipette 410 near the bottom. The liquid outlet one-way valve 520 is connected to the bottom of the vertical section. An air outlet hole is provided near the top of the pipette 410, and the exhaust one-way valve 530 is coaxially rotatably connected to the pipette 410 and can be connected to the air outlet hole. The liquid-blocking exhaust block 540 is slidably connected to the pipette 410, and is used to open and close or isolate the connection between the curved section and the pipette 410; one side of the liquid-blocking exhaust block 540 close to the pipette 410 is set as an inclined surface, and is inclined toward the curved section to assist the mixed liquid to flow to the curved section. The driving component 550 is installed on the cylinder 100, and is used to drive the sliding of the liquid-blocking exhaust block 540.

[0053] In order to realize the rotation of the exhaust one-way valve 530, a rotating motor 531 is installed on the pipette plate 400, and the output shaft of the rotating motor 531 is coaxially fixedly connected with the exhaust main gear 532, and the exhaust slave gear 533 is coaxially fixedly connected to the pipette 410, and the exhaust one-way valve 530 is installed on the exhaust slave gear 533, and the exhaust slave gear 533 is meshed with the exhaust main gear 532.

[0054] The driving component 550 may include a driving ring gear 551, a driving rod 552, a driving slave gear 553 and a driving cylinder 554. The driving ring gear 551 is rotatably connected to the pipetting plate 400, and each pipette 410 corresponds to a driving rod 552, a driving slave gear 553 and a driving cylinder 554. One end of the driving rod 552 is threadedly connected to the corresponding liquid blocking and exhausting block 540, and the driving slave gear 553 is installed on the other end of the corresponding driving rod 552 through a spline and can be meshed with the driving ring gear 551. The cylinder body of the driving cylinder 554 is installed on the pipetting plate 400, and the piston rod is slidably connected to the end surface of the corresponding driving slave gear 553.

[0055] In order to realize the rotation of the driving gear ring 551 , a driving motor 555 is installed on the cylinder 100 . The output shaft of the driving motor 555 is coaxially fixedly connected with a driving main gear 556 , and the driving main gear 556 is meshed with the driving gear ring 551 .

[0056] The liquid collection recovery assembly 600 may include a liquid suction pump 610, a liquid discharge main pipe 620 and a liquid discharge branch pipe 630. The inlet of the liquid suction pump 610 is connected to the lowest point of the liquid transfer plate 400, and the outlet is connected to the liquid discharge main pipe 620. One end of the liquid discharge branch pipe 630 is respectively connected to each liquid transfer tube 410 near the top and is located above the exhaust check valve 530, and the other end is connected to the liquid discharge main pipe 620.

[0057] In addition, a main liquid-flowing gear 581 is coaxially fixedly connected to the output shaft of the rotating motor 531, and a liquid-flowing slave gear 582 is coaxially fixedly connected to one end of the driving rod 552 that passes through the pipette 410, and the liquid-flowing slave gear 582 is meshed with the main liquid-flowing gear 581. A liquid-flowing block 583 is coaxially fixedly connected to the inlet of the pipette 410, and the liquid-flowing block 583 is provided with a liquid-flowing hole 584, and the liquid-distributing branch pipe 320 enters the pipette 410 through the liquid-flowing hole 584. A liquid-closing arc gear ring 585 is rotatably connected to the liquid-flowing block 583, and the liquid-closing arc gear ring 585 is meshed with the liquid-flowing slave gear 582, and is used to open and close the liquid-flowing hole 584. When the rotating motor 531 drives the exhaust check valve 530 to communicate with the air outlet, the liquid-closing arc gear ring 585 closes the liquid-flowing hole 584.

[0058] It should also be noted that, referring to Figure 5The synthesis system further includes a controller 703, a liquid level sensor 704 and a prompter 705; wherein the liquid level sensor 704 is installed in the transfer plate 400, and is used to detect the liquid level of the mixed liquid collected in the transfer plate 400. When the liquid level reaches the liquid level threshold, the controller 703 sends a prompt signal to the prompter 705, and the prompter 705 responds to the prompt information to prompt to stop the liquid inlet. In addition, the controller 703 is also connected to the auxiliary motor 102, the electric push rod 250, the electromagnet 701, the liquid separation valve 311, the rotating motor 531, the driving cylinder 554, the driving motor 555 and the liquid pump 610; to control the corresponding device actions according to the set program.

[0059] It should be noted that the gear rings in the present application have hidden teeth and hidden splines.

[0060] The implementation principle of this embodiment is: The pre-treatment is connected to the liquid inlet of the cylinder 100, and the mixed liquid enters the cylinder 100, is filtered through the conical filter screen 210, and enters the conical collecting bucket 220; the exhaust check valve 530 is rotated to connect the exhaust check valve 530 with the air outlet, and then the liquid blocking exhaust block 540 is driven to move toward the top of the pipette 410, so that the internal gas of the pipette 410 is discharged through the exhaust check valve 530, and then the exhaust check valve 530 is rotated to close the air outlet, and the liquid blocking exhaust block 540 is driven to move, so that the discharge pipe 510 and the pipette 410 are connected. 0 partition; open the liquid separation valve 311; then drive the rotating rod 211 to rotate, so that the filtered mixed liquid enters the pipette 410, and the excess mixed liquid enters the pipette plate 400 for collection; after the liquid level reaches the liquid level threshold, stop the liquid intake; then move the cylinder 100 to the separation device, and then drive the corresponding liquid blocking and exhausting block 540 to move, so that the discharge pipe 510 is connected with the pipette 410, so that the mixed liquid is discharged into the separation device; in addition, the liquid pump 610 can be controlled to operate to draw the mixed liquid into the pipette 410.

[0061] Based on the above embodiments, the second embodiment of the present application discloses a synthesis process of monosialotetrahexosylganglioside GM1. Figure 6 , the synthesis process includes S1-S6: S1, filtering and collecting the mixed solution after pretreatment; S2, exhausting the gas in each pipette 410; S3, dispensing the mixed solution into each pipette 410; S4, closing the liquid inlet of the cylinder 100 and driving the cylinder 100 to move; S5, after the cylinder 100 moves to the separation device, the pipette 410 is controlled to discharge the mixed liquid to the separation device; S6, recovering the mixed liquid collected in the transfer tray 400 into the transfer tube 410, so as to continue to discharge the mixed liquid into the separation device.

[0062] The above are all preferred embodiments of the present application, and are not intended to limit the protection scope of the present application in sequence. Any feature disclosed in this specification (including the abstract and drawings), unless otherwise stated, can be replaced by other equivalent or alternative features with similar purposes. That is, unless otherwise stated, each feature is only an example of a series of equivalent or similar features.

Claims

1. A synthesis system of monosialotetrahexosylganglioside GM1, characterized in that: include: The cylinder (100) is movably arranged and is provided with a liquid inlet; A cover body (101) is rotatably connected to the cylinder body (100) and is used to open and close the liquid inlet; A filter assembly (200) is installed in the cylinder (100) and is used to filter the mixed liquid; A liquid separation component (300) is rotatably mounted in the cylinder (100); A pipetting plate (400) is evenly mounted with a plurality of pipettes (410), wherein the liquid dispensing assembly (300) is used to dispense the filtered mixed liquid into the pipettes (410); A liquid outlet and air exhaust component (500), mounted on the barrel (100), and used for opening and closing the pipette (410) and exhausting the gas in the pipette (410); The controller (703) is connected to the liquid separation component (300) and the liquid outlet and exhaust component (500) for controlling the rotation of the liquid separation component (300), controlling the liquid outlet and exhaust component (500) to open or close the pipette (410) and exhaust the gas in the pipette (410).

2. A synthesis system of monosialotetrahexosylganglioside GM1 according to claim 1, characterized in that: The liquid outlet and exhaust assembly (500) comprises: A liquid discharge tube (510) is composed of a curved section and a vertical section, wherein the vertical section is coaxial with the pipette (410), one end of the curved section is connected to the vertical section, and the other end is connected to a portion of the pipette (410) near the bottom; A liquid outlet one-way valve (520) connected to the bottom of the vertical section; An exhaust check valve (530) is coaxially rotatably connected to the pipette (410); an exhaust hole is provided near the top of the pipette (410); and the exhaust check valve (530) can be in communication with the exhaust hole; a liquid-blocking and air-exhausting block (540) slidably connected inside the pipette (410) and used to open or isolate the connection between the curved section and the pipette (410); a surface of the liquid-blocking and air-exhausting block (540) close to the pipette (410) is an inclined surface and is inclined toward the curved section; A driving component (550) is mounted on the cylinder (100) and is used to drive the sliding movement of the liquid blocking and exhausting block (540). The driving component (550) is communicatively connected with the controller (703).

3. A synthesis system of monosialotetrahexosylganglioside GM1 according to claim 2, characterized in that: The driving component (550) comprises: A driving gear ring (551) rotatably connected to the transfer plate (400); A driving rod (552), one end of which is threadedly connected to the liquid blocking and exhausting block (540); A driving slave gear (553) is mounted on the other end of the driving rod (552) via a spline and is capable of meshing with the driving ring gear (551); A driving cylinder (554) has a cylinder body mounted on the transfer plate (400), and a piston rod is slidably connected to the end surface of the driving slave gear (553). The driving cylinder (554) is communicatively connected to the controller (703).

4. The synthesis system of monosialotetrahexosylganglioside GM1 according to claim 1, characterized in that: The filtering assembly (200) comprises: A conical filter screen (210) installed in the cylinder (100); The conical collecting hopper (220) is fixedly connected to the conical filter screen (210) and is used to collect the filtered mixed liquid. The bottom end of the conical collecting hopper (220) is in communication with the liquid separation assembly (300).

5. A synthesis system of monosialotetrahexosylganglioside GM1 according to claim 4, characterized in that: The filtering assembly (200) further comprises: A cleaning brush (230) is fixedly connected to the inner wall of the barrel (100) and abuts against the conical filter (210); the conical filter (210) is rotatably connected to the barrel (100); A liquid baffle plate (240), a slag discharge hole is provided on the side wall of the cylinder (100), and the liquid baffle plate (240) is slidably connected to the cylinder (100) and is used to open and close the slag discharge hole.

6. A synthesis system of monosialotetrahexosylganglioside GM1 according to claim 4, characterized in that: The liquid separation component (300) comprises: A liquid separation main pipe (310) is connected to the bottom of the conical collecting bucket (220) and is connected to a liquid separation valve (311), and the liquid separation valve (311) is in communication with the controller (703); The liquid dispensing branch pipe (320) is in communication with the liquid dispensing main pipe (310) and abuts against the top of the pipette (410).

7. The synthesis system of monosialotetrahexosylganglioside GM1 according to claim 1, characterized in that: The synthesis system also includes: The liquid collection recovery component (600) is installed on the transfer plate (400), the transfer plate (400) is arranged in a conical shape, and the liquid collection recovery component (600) is used to recover the excess mixed liquid in the transfer plate (400) into each of the pipettes (410).

8. The synthesis system of monosialotetrahexosylganglioside GM1 according to claim 7, characterized in that: The liquid collection recovery component (600) comprises: A liquid extraction pump (610), the inlet of which is connected to the lowest point of the liquid transfer plate (400), and the outlet of which is connected to a liquid discharge main pipe (620); the liquid extraction pump (610) is in communication connection with the controller (703); The drainage branch pipe (630) has one end connected to each of the pipettes (410) near the top, and the other end connected to the drainage main pipe (620).

9. The synthesis system of monosialotetrahexosylganglioside GM1 according to claim 4, characterized in that: The synthesis system also includes: An auxiliary gear ring (104) mounted on the cover body (101); An auxiliary motor (102) is mounted on the cylinder (100), and the output shaft is mounted with a main gear (103) via a spline; the auxiliary motor (102) is communicatively connected with the controller (703); An opening and closing gear (105) is rotatably connected to the cylinder (100); the main gear (103) can mesh with the opening and closing gear (105); and the auxiliary gear ring (104) meshes with the opening and closing gear (105); A rotating rod (211) fixedly connected to the conical filter screen (210) and rotatably connected to the cylinder (100); A rotating gear (212) is coaxially fixedly connected to the rotating rod (211), and the main gear (103) is capable of meshing with the rotating gear (212); An electromagnet (701) is coaxially fixedly connected to the output shaft of the auxiliary motor (102); The iron sheet (702) is coaxially fixedly connected to the main gear (103); when the electromagnet (701) is not energized, the main gear (103) meshes with the opening and closing gear (105); after the electromagnet (701) is energized, the iron sheet (702) is attracted, and the main gear (103) meshes with the rotating gear (212).

10. A process for synthesizing monosialotetrahexosylganglioside GM1, characterized in that: Based on the synthesis system of monosialotetrahexosylganglioside GM1 as described in any one of claims 1 to 9, the synthesis process comprises: Filter and collect the mixed solution after pretreatment; Exhausting the gas in each pipette (410); Dispensing the mixed solution into each of the pipettes (410); Closing the liquid inlet of the cylinder (100) and driving the cylinder (100) to move; After the cylinder (100) moves to the separation device, controlling the pipette (410) to discharge the mixed liquid to the separation device; The mixed liquid collected in the transfer tray (400) is recovered into the transfer pipette (410) to continue discharging the mixed liquid into the separation device.

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