Efficient extraction and separation integrated system and method for bioactive substances

Abstract

The efficient extraction and separation integrated system comprises a main shell, a liquid supply chamber and a plurality of partition plates, a cracking chamber is formed between every two adjacent partition plates, a cracking tank and a combination tank are installed in each cracking chamber from top to bottom, the cracking tanks are communicated with the combination tanks, and the combination tanks are communicated with the main shell. A conical discharging bin of an integrated structure is arranged at the bottom of the combining pipe, a sunken part which is sunken upwards is arranged at the bottom of the conical discharging bin, and the magnetic device is arranged outside the sunken part. Cross contamination between samples and environmental pollution can be avoided, efficient capture, thorough washing and sufficient elution of magnetic beads are ensured by combining the unique conical cavity design and the movable magnetic device, so that target products with higher purity and more stable yield are obtained, operation is easy and convenient, reliability is high, a user only needs to add the samples and consumables, and the cost is low. The technical requirements for operators are reduced, and the system structure is simpler and higher in reliability compared with a large robot workstation.

Description

Technical Field

[0001] This invention relates to the field of bio-extraction technology, specifically to an integrated system and method for the efficient extraction and separation of bioactive substances. Background Technology

[0002] In fields such as disease prevention and control, food safety testing, forensic evidence identification, genomics, and drug development, the efficient and high-purity extraction of bioactive substances such as DNA, RNA, and proteins from complex biological samples (such as blood, tissues, microorganisms, and food matrices) is a crucial first step in subsequent PCR, sequencing, and immunoassay analysis.

[0003] Currently used extraction methods include, but are not limited to: boiling, centrifugation column extraction, solution extraction (such as the phenol-chloroform method), and magnetic bead extraction. Among these, the magnetic bead method is becoming increasingly popular due to its ease of automation and the elimination of the need for high-speed centrifugation. However, existing magnetic bead extraction technologies and equipment still have several shortcomings: Discretization of processes: Most devices or reagent kits still require the transfer of liquids between different tubes, which is cumbersome and can easily lead to cross-contamination between samples and aerosol contamination.

[0004] Efficiency bottleneck: Despite the availability of automated instruments, their throughput is often limited by the speed of the robotic arm and the format of the well plate, resulting in long processing times when handling large batches of samples.

[0005] Purity and yield issues: Traditional magnetic bead capture methods (such as bottom adsorption in 96-well plates) may have problems such as uneven magnetic bead adsorption, incomplete washing, and liquid residue, which affect the purity and yield of the final product.

[0006] System complexity: Fully automated extractors are typically complex in structure, large in size, and expensive.

[0007] Therefore, there is an urgent need in this field for an integrated system and method that is highly integrated, has a high throughput, can minimize the risk of human intervention and cross-contamination, and can ensure high extraction purity and yield. Summary of the Invention

[0008] (a) Technical problems to be solved This invention provides an integrated system and method for the efficient extraction and separation of bioactive substances that is compact, easy to operate, highly efficient, and of good purity, and is particularly suitable for large-scale sample processing, aiming to solve the problems mentioned in the background art.

[0009] (II) Technical Solution To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides an integrated system for the efficient extraction and separation of bioactive substances, comprising: The main shell, from top to bottom, comprises: The liquid supply chamber is used to receive samples and add lysis buffer, washing buffer, and elution buffer; Multiple partitions are provided, and a pyrolysis chamber is formed between adjacent partitions. A pyrolysis tank and a bonding tank are installed from top to bottom in the pyrolysis chamber. The pyrolysis tank and the bonding tank are connected. A conical discharge bin with an integral structure is provided at the bottom of the bonding tube. The bottom of the conical discharge bin has an upwardly recessed part. Elution chamber, used to collect target substances; A magnetic device is disposed outside the recess. The device includes an openable permanent magnet that can be controlled to move closer to or away from the outer wall of the recess to capture and release magnetic beads to form a target substance. A fluid drive and control module, which is connected to each chamber of the main housing via pipelines, is used to add pyrolysis fluid, washing fluid, eluent, and to transfer mixed liquids between chambers; The heating module and the oscillation module are integrated on the pyrolysis vessel to provide the thermal energy and mechanical oscillation required for pyrolysis.

[0010] As a preferred technical solution of this application, an annular capture groove is formed between the recessed portion and the inner wall of the conical feeding bin, and the inner walls of the combined tank, the conical feeding bin and the capture groove are treated with a superhydrophobic coating.

[0011] As a preferred technical solution of this application, the fluid drive and control module includes a microfluidic pump, a central controller, an inlet pipe, an outlet pipe, and multiple two-way solenoid valves. The microfluidic pump is connected to the outlet of the outlet pipe. The inlet pipe delivers washing liquid and elution liquid to the corresponding bonding tanks through multiple filling pipes. An outlet is provided at the bottom of the recess, and a straight pipe is inserted into the outlet. The lower end of the straight pipe is connected to the outlet pipe. The pyrolysis tank and the bonding tank are connected through two-way solenoid valves. The filling pipes are connected to the two-way solenoid valves. The flow path switching of the two-way solenoid valves and the movement of the magnetic device are controlled collaboratively by the central controller.

[0012] As a preferred technical solution of this application, the magnetic device includes two arc-shaped magnetic frames, which are permanent magnets. The two arc-shaped magnetic frames are respectively arranged on both sides of the outer side of the recess. A pull rod is fixedly installed on the outer side of the arc-shaped magnetic frames. The partition plate has a groove at the position of the magnetic device. Two sliders are slidably installed in the groove. The surface of the slider has threaded holes that pass through both sides. A threaded rod is rotatably installed in the groove. The threaded rod has threaded surfaces with opposite directions at the middle and both ends. The two threaded surfaces of the threaded rod are threadedly connected to the two sliders respectively. One end of the pull rod is fixedly connected to the corresponding slider.

[0013] As a preferred technical solution of this application, a sampling tube is connected to the bottom of the capture tank, the lower end of the sampling tube is inserted into the elution chamber, and a centrifuge tube is threadedly installed at the lower end of the sampling tube.

[0014] As a preferred technical solution of this application, a notch is provided on one side of the partition, and a sprocket fixedly connected to the corresponding threaded rod is provided in the notch. Multiple sprockets are connected by chain drive, one of which is fixedly connected to the output shaft of the drive motor. Pressure rollers are rolled above and below the remaining sprockets, and the pressure rollers are rotatably installed in the notch.

[0015] As a preferred technical solution of this application, multiple sample injection chambers are fixedly installed in the liquid supply chamber, and the sample injection chambers are connected to the corresponding pyrolysis tanks.

[0016] Secondly, the present invention also provides a method for integrating the extraction and separation of bioactive substances using any of the systems described above, comprising the following steps: (a) The sample to be processed and magnetic beads coated with specific affinity ligands are added into the lysis vessel through the sample injection chamber. The heating module and oscillation module are started to carry out full lysis, so that the target substance is released and binds to the magnetic beads. (b) The mixture in the pyrolysis tank is transferred to the bonding tank via the fluid drive and control module; (c) Control the magnetic device to move to the working position and capture the magnetic beads with the target material attached to them into the capture tank; (d) The microfluidic pump draws the waste liquid through a straight pipe into the drain pipe and discharges the waste liquid. (e) Control the magnetic device to keep it in working condition, pump multiple washing liquids sequentially through the liquid inlet pipe 7 and discard them through the straight pipe to complete the washing process; (f) Control the magnetic device to move away from the working position, add eluent to the bonding tank to resuspend the magnetic beads to dissociate the target substance; (g) Control the magnetic device to move to the working position again, capture the magnetic beads, and transfer the eluent containing the purified target substance to the capture tank. Then, discharge it through the sampling tube to the centrifuge tube for collection to complete the extraction.

[0017] As a preferred technical solution of this application, in step (f), the eluent is a buffer solution with low ionic strength or a specific pH value, and the elution temperature is controlled by the heating module and the oscillation module.

[0018] (III) Beneficial Effects Extremely high integration and anti-contamination capability: All critical steps are completed within a single housing, avoiding cross-contamination between samples and environmental pollution, making it particularly suitable for high-sensitivity applications such as PCR.

[0019] Exceptional purity and yield: The unique conical chamber design combined with the movable magnetic device ensures efficient magnetic bead capture, thorough washing, and sufficient elution, resulting in a target product with higher purity and more stable yield.

[0020] High throughput and high efficiency: Parallel processing design and highly automated processes greatly increase the amount of samples processed per unit time, solving the bottleneck problem of low efficiency in traditional methods.

[0021] Easy to operate and highly reliable: Users only need to add samples and consumables and start with one click, which reduces the technical requirements for operators and reduces human error. The system structure is simpler and more reliable than large robot workstations. Attached Figure Description

[0022] Figure 1 This is a schematic diagram of a highly efficient integrated system for the extraction and separation of bioactive substances. Figure 2 This is a structural diagram of the pyrolysis chamber of the present invention; Figure 3 This is a structural diagram of the magnetic device of the present invention; Figure 4 This is a cross-sectional view of the can combined with the present invention; Figure 5 This is a front view of the main housing of the present invention; Figure 6 This is a flowchart of a method for the efficient extraction and separation of bioactive substances.

[0023] In the picture: 1. Main shell; 11. Liquid supply chamber; 12. Pyrolysis chamber; 13. Elution chamber; 14. Partition; 141. Notch; 2. Pyrolysis vessel; 21. Oscillation module; 22. Heating module; 23. Two-way solenoid valve; 24. Filling tube; 25. Sampling tube; 3. Binding vessel; 31. Conical feed hopper; 32. Recess; 33. Capture tank; 34. Liquid outlet; 35. Straight tube; 4. Centrifuge tube; 5. Magnetic device; 51. Arc-shaped magnetic frame; 52. Pull rod; 53. Slider; 54. Threaded rod; 55. Sprocket; 56. Drive motor; 57. Chain; 58. Pressure roller; 6. Sample inlet chamber; 7. Liquid inlet pipe; 8. Liquid outlet pipe. Detailed Implementation

[0024] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] See Figure 1 and Figure 5 As shown, the present invention provides an integrated system for the efficient extraction and separation of bioactive substances, comprising: The main housing 1 is injection molded from medical-grade polymer material (such as polypropylene). It has a working window on its front side, which can be closed with a cover (the working window and cover are not shown in the figure for easy observation of the internal structure). Its interior, from top to bottom, includes: The liquid supply chamber 11 is used to receive samples and add lysis buffer, washing buffer and elution buffer; Multiple partitions 14 are used, with lysis chambers 12 formed between adjacent partitions 14. A lysis vessel 2 and a binding vessel 3 are installed from top to bottom within each lysis chamber 12, and the lysis vessel 2 and binding vessel 3 are connected. The lysis vessel 2 is used to add lysis buffer (typically containing guanidine hydrochloride, ionizing salt, etc.) to disrupt the cell / nuclear membrane, releasing the target substance (such as DNA) into the solution. Simultaneously, surface-modified magnetic beads are added. Under high temperature and high salt (ionizing salt) conditions, these ionizing salt ions disrupt the water molecule structure, causing hydrophilic biomolecules such as DNA / RNA to become hydrophobic, thereby binding to the hydrophobic surface or specific functional groups (such as silanol groups) of the magnetic beads and adsorbing onto the surface of the magnetic beads. Impurities such as proteins and cell debris remain in the solution. The bottom of the connecting tube 3 is provided with an integral conical feeding bin 31. The bottom of the conical feeding bin 31 is provided with an upwardly recessed part 32. This structure is conducive to the magnetic beads being gathered to the point of minimum volume under the action of the magnetic field, so as to minimize the loss of magnetic beads when discarding waste liquid and facilitate more thorough removal of waste liquid. Elution chamber 13 is used to collect the target substance; A magnetic device 5, disposed outside the recess 32, includes an openable permanent magnet that can be controlled to approach or move away from the outer wall of the recess 32 to capture and release magnetic beads to form the target substance. This device is not a fixed magnetic rod or sleeve, but a movable mechanical structure with a built-in permanent magnet. It can be precisely "approached" or "moved away" from the outer wall of the recess 32 through program control. When magnetic beads need to be captured, the magnet approaches, and the strong magnetic field tightly attracts the beads to the bottom of the recess 32. When the magnetic beads need to be resuspended for washing or elution, the magnet quickly moves away, the magnetic field weakens or disappears, and the magnetic beads can easily disperse in the liquid, ensuring the sufficiency and uniformity of the reaction. The fluid drive and control module is connected to each chamber of the main housing 1 through pipelines. It is used to add pyrolysis fluid, washing fluid, eluent, and transfer mixed liquid between chambers. Its operation and the magnetic device 5 are coordinated and time-controlled by a central controller (such as a PLC or microcontroller). For example: liquid addition → magnet moving away → oscillation and mixing → magnet moving closer → static adsorption → waste liquid extraction. The whole process is fully automatic and requires no manual intervention. The heating module 22 and the oscillation module 21 are integrated on the pyrolysis tank 2 to provide the thermal energy and mechanical oscillation required for pyrolysis. The heating module 22 can be an electric heating tube or a heating plate in the prior art, and the oscillation module 21 can be an ultrasonic oscillation device or a motor oscillation device in the prior art.

[0026] See Figure 4 As shown, as a preferred technical solution of this application, an annular capture groove 33 is formed between the recessed portion 32 and the inner wall of the conical feeding bin 31. The inner walls of the combined tank 3, the conical feeding bin 31 and the capture groove 33 are treated with a superhydrophobic coating to reduce liquid residue.

[0027] Optionally, the fluid drive and control module includes a microfluidic pump, a central controller, an inlet pipe 7, an outlet pipe 8, and multiple two-way solenoid valves 23. The microfluidic pump is connected to the outlet of the outlet pipe 8. The inlet pipe 7 delivers washing liquid and elution liquid to the corresponding bonding tank 3 through multiple filling pipes 24. An outlet 34 is provided at the bottom of the recess 32, and a straight pipe 35 is inserted into the outlet 34. The lower end of the straight pipe 35 is connected to the outlet pipe 8. The pyrolysis tank 2 and the bonding tank 3 are connected through the two-way solenoid valves 23. The filling pipes 24 are connected to the two-way solenoid valves 23. The flow path switching of the two-way solenoid valves 23 and the movement of the magnetic device 5 are controlled by the central controller.

[0028] See Figure 3 As shown, in a preferred embodiment of this application, the magnetic device 5 includes two arc-shaped magnetic frames 51, each being a permanent magnet. The two arc-shaped magnetic frames 51 are respectively positioned on both sides of the outer surface of the recess 32. A pull rod 52 is fixedly installed on the outer side of each arc-shaped magnetic frame 51. The partition 14 has a groove at the location of the magnetic device 5, and two sliders 53 are slidably installed within the groove. The surface of each slider 53 has threaded holes penetrating both sides. A threaded rod 54 is rotatably installed within the groove. The threaded rod 54 has threaded surfaces in opposite directions at its center and both ends. The two threaded surfaces of the threaded rod 54 are threadedly connected to the two sliders 53. One end of the pull rod 52 is fixedly connected to the corresponding slider 53. By rotating the threaded rod 54, the two sliders 53 can move closer or further away, thereby causing the two arc-shaped magnetic frames 51 to move closer or further away from the recess 32.

[0029] Optionally, a sampling tube 25 is connected to the bottom of the capture tank 33. The lower end of the sampling tube 25 is inserted into the elution chamber 13. A centrifuge tube 4 is threadedly installed at the lower end of the sampling tube 25. When the target substance enters the centrifuge tube 4, the centrifuge tube 4 can be removed to facilitate subsequent extraction work.

[0030] See Figure 2 As shown, in a preferred embodiment of this application, a notch 141 is provided on one side of the partition 14. A sprocket 55 is fixedly connected to the corresponding threaded rod 54 in the notch 141. Multiple sprockets 55 are connected by a chain 57. One of the sprockets 55 is fixedly connected to the output shaft of the drive motor 56. Pressure rollers 58 are rolled above and below the remaining sprockets 55. The pressure rollers 58 are rotatably installed in the notch 141. The pressure rollers 58 can effectively prevent the chain 57 from slipping off the sprockets 55.

[0031] Optionally, a plurality of sample inlet chambers 6 are fixedly installed in the liquid supply chamber 11, and the sample inlet chambers 6 are connected to the corresponding pyrolysis vessel 2.

[0032] Example 1: Extraction of genomic DNA from whole blood samples Preparation: Connect the lysis buffer, washing buffer I, II and elution buffer to the corresponding tubing beforehand.

[0033] Sample addition and lysis: Add 200 μL of whole blood sample and 20 μL of protein kinase K, along with 500 μL of lysis buffer and 50 μL of magnetic bead suspension to the lysis vessel. Start the heating / shaking module and incubate at 56°C with shaking for 10 minutes to fully lyse the cells, release DNA, and bind to the magnetic beads.

[0034] Transfer and capture: The program controls the pump valve to draw all the pyrolysis mixture into the binding tank. Then, the magnetic device 5 is brought close to the working position so that the arc-shaped magnetic frame 51 is tightly attached to the recess 32. After standing for 1 minute, the magnetic beads are firmly attracted.

[0035] Discarding waste liquid and washing: Aspirate and discard the lysis waste liquid, keep the magnet in the adsorption state, and pump in 700 μL of washing buffer I and 700 μL of washing buffer II (usually a salt solution containing ethanol) in sequence. The purpose of this buffer is to maintain the salt concentration conditions for DNA to bind to the magnetic beads, and at the same time dissolve and remove residual proteins, salt ions, enzyme inhibitors and other impurities. After each addition, move the magnet away briefly and gently shake to mix for 5 seconds, then re-adsorb and drain the waste liquid to ensure thorough cleaning of impurities.

[0036] Elution and Collection: After discarding the final wash buffer, remove the magnet and add 100 μL of preheated 65°C elution buffer (TE buffer) to binding vessel 3. Vortex for 3 minutes to allow the DNA to fully dissociate from the magnetic beads. Then, bring the magnet close to the magnetic beads again, and finally transfer all the elution buffer containing the purified DNA to a clean centrifuge tube 4.

[0037] Completed: Remove centrifuge tube 4 and use the DNA for downstream experiments. The purity of the obtained DNA (A260 / A280) was between 1.8 and 2.0 as detected by Nanodrop and Qubit, indicating a high yield. Furthermore, agarose gel electrophoresis showed no RNA or protein contamination.

[0038] Example 2: Extraction of high-quality genomic DNA from plant leaves Objective: Plant tissues typically contain a large amount of secondary metabolites such as polysaccharides, polyphenols, and pigments. These substances are highly susceptible to co-precipitation with DNA during extraction, forming a viscous and insoluble mixture that severely inhibits downstream enzymatic reactions (such as PCR and restriction endonuclease digestion).

[0039] Sample: 20 mg of fresh Arabidopsis thaliana leaves.

[0040] The reagents are as follows: PL lysate: A plant-specific lysate containing high concentrations of CTAB (hexadecyltrimethylammonium bromide) and β-mercaptoethanol; Washing buffers I and II: Similar to Example 1, but the pH and salt concentration can be adjusted appropriately to optimize the removal of plant impurities; Elution buffer: TE buffer; Magnetic beads: silanol magnetic beads.

[0041] Experimental group (using the system and method of this invention): Preparation and sample loading: Place the plant leaves into lysis vessel 2, add 400 μL of lysis buffer PL and 10 μL of β-mercaptoethanol (additional), and add 50 μL of magnetic bead suspension.

[0042] Lysis and binding: Activate the heating / oscillation module and incubate vigorously at 65°C for 15 minutes to completely lyse the plant cell wall and inhibit polyphenol oxidation.

[0043] Transfer and capture: The mixture is transferred to the binding vessel 3 and the magnetic device 5 is controlled to capture the magnetic bead-DNA complex.

[0044] Washing: Discard the dark brown lysis waste liquid (containing a large amount of polyphenols and polysaccharides). Under the action of a magnetic field, wash twice with 700 μL washing buffer I and II in sequence. The dried waste liquid is colorless and transparent, indicating that the impurities have been effectively removed.

[0045] Elution and collection: Add 50 μL of elution buffer preheated to 65°C, mix thoroughly, and elute to obtain a clear and transparent DNA solution.

[0046] Comparative examples (using a traditional centrifuge column method kit): Following the instructions of a commercial plant DNA extraction kit: grind the leaves, lyse in a 65°C water bath, centrifuge, transfer the supernatant to a centrifuge column, wash by multiple centrifugations, and finally elute by centrifugation. The results are compared in the table below: Table 1 Comparison of test results

[0047] The integrated system and efficient washing process of this invention can effectively overcome the challenge of complex inhibitors in samples, and the obtained DNA is significantly better than that of traditional methods in terms of purity and yield, while completely avoiding tedious manual operations.

[0048] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. An integrated system for the efficient extraction and separation of bioactive substances, characterized in that, include: The main shell, from top to bottom, comprises: The liquid supply chamber is used to receive samples and add lysis buffer, washing buffer, and elution buffer; Multiple partitions are provided, and a pyrolysis chamber is formed between adjacent partitions. A pyrolysis tank and a bonding tank are installed from top to bottom in the pyrolysis chamber. The pyrolysis tank and the bonding tank are connected. The bottom of the bonding tank is provided with an integral conical feeding bin. The bottom of the conical feeding bin is provided with an upwardly recessed part. Elution chamber, used to collect target substances; A magnetic device is disposed outside the recess. The device includes an openable permanent magnet that can be controlled to move closer to or away from the outer wall of the recess to capture and release magnetic beads to form a target substance. A fluid drive and control module, which is connected to each chamber of the main housing via pipelines, is used to add pyrolysis fluid, washing fluid, eluent, and to transfer mixed liquids between chambers; The heating module and the oscillation module are integrated on the pyrolysis vessel to provide the thermal energy and mechanical oscillation required for pyrolysis.

2. The integrated system for efficient extraction and separation of bioactive substances according to claim 1, characterized in that, An annular trapping groove is formed between the recessed portion and the inner wall of the conical feeding hopper. The inner walls of the combined tank, the conical feeding hopper, and the trapping groove are treated with a superhydrophobic coating.

3. The integrated system for efficient extraction and separation of bioactive substances according to claim 1, characterized in that, The fluid drive and control module includes a microfluidic pump, a central controller, an inlet pipe, an outlet pipe, and multiple two-way solenoid valves. The microfluidic pump is connected to the outlet of the outlet pipe. The inlet pipe delivers washing liquid and elution liquid to the corresponding bonding tanks through multiple filling pipes. An outlet is provided at the bottom of the recess, and a straight pipe is inserted into the outlet. The lower end of the straight pipe is connected to the outlet pipe. The pyrolysis tank and the bonding tank are connected through two-way solenoid valves. The filling pipes are connected to the two-way solenoid valves. The flow path switching of the two-way solenoid valves and the movement of the magnetic device are controlled collaboratively by the central controller.

4. The integrated system for efficient extraction and separation of bioactive substances according to claim 1, characterized in that, The magnetic device includes two arc-shaped magnetic frames, which are permanent magnets. The two arc-shaped magnetic frames are respectively located on both sides of the outside of the recess. A pull rod is fixedly installed on the outside of the arc-shaped magnetic frames. The partition plate has a groove at the location of the magnetic device. Two sliders are slidably installed in the groove. The surface of the slider has threaded holes that pass through both sides. A threaded rod is rotatably installed in the groove. The threaded rod has threaded surfaces in opposite directions at the middle and two ends. The two threaded surfaces of the threaded rod are threadedly connected to the two sliders respectively. One end of the pull rod is fixedly connected to the corresponding slider.

5. The integrated system for efficient extraction and separation of bioactive substances according to claim 2, characterized in that, The bottom of the capture tank is connected to a sampling tube, the lower end of which is inserted into the elution chamber, and a centrifuge tube is threadedly installed at the lower end of the sampling tube.

6. The integrated system for efficient extraction and separation of bioactive substances according to claim 4, characterized in that, The partition has a notch on one side, and a sprocket fixedly connected to the corresponding threaded rod is provided in the notch. Multiple sprockets are connected by chain drive. One of the sprockets is fixedly connected to the output shaft of the drive motor. Pressure rollers are rolled above and below the remaining sprockets and are installed in the notch.

7. The integrated system for efficient extraction and separation of bioactive substances according to claim 1, characterized in that, Multiple sample inlet chambers are fixedly installed in the liquid supply chamber, and the sample inlet chambers are connected to the corresponding pyrolysis vessels.

8. An integrated method for the efficient extraction and separation of bioactive substances, characterized in that, The integrated system for efficient extraction and separation of bioactive substances as described in any one of claims 1-7 is characterized by comprising the following steps: (a) The sample to be processed and magnetic beads coated with specific affinity ligands are added into the lysis vessel through the sample injection chamber. The heating module and oscillation module are started to carry out full lysis, so that the target substance is released and binds to the magnetic beads. (b) The mixture in the pyrolysis tank is transferred to the bonding tank via the fluid drive and control module; (c) Control the magnetic device to move to the working position and capture the magnetic beads with the target material into the capture tank; (d) The microfluidic pump draws the waste liquid through a straight pipe into the drain pipe and discharges the waste liquid. (e) Control the magnetic device to keep it in working condition, pump multiple washing liquids in sequence through the liquid inlet pipe and discard them through the straight pipe to complete the washing process; (f) Control the magnetic device to move away from the working position, add eluent to the bonding tank to resuspend the magnetic beads to dissociate the target substance; (g) Control the magnetic device to move to the working position again, capture the magnetic beads, and transfer the eluent containing the purified target substance to the capture tank. Then, discharge it through the sampling tube to the centrifuge tube for collection to complete the extraction.

9. The integrated method for efficient extraction and separation of bioactive substances according to claim 8, characterized in that, In step (f), the eluent is a buffer solution with low ionic strength or a specific pH value, and the elution temperature is controlled by the heating module and the oscillation module.

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