A serum lipidomics pretreatment device and method

Abstract

The application discloses a serum lipidomics pretreatment device and method, relates to the technical field of serum lipidomics pretreatment, and comprises a rack and a pipetting module, a switch cover module, a gun head box, sample cryopreservation tubes, a first heating and refrigeration oscillation module, a first reaction tube, a reagent storage module, a second heating and refrigeration oscillation module, a second reaction tube, a nitrogen blowing module and a software control module arranged on the rack; the pipetting module is used for sucking and transferring liquid; the switch cover module is used for opening and closing the tube cover of the sample tube and / or the reaction tube; the first reaction tube is arranged on the first heating and refrigeration oscillation module, and the second reaction tube is arranged on the second heating and refrigeration oscillation module; the nitrogen blowing module is used for nitrogen blowing and drying and concentrating the sample; the test error caused by human operation is reduced, the cumbersome manual operation is reduced, the harm of organic solvents to operators is reduced, the serum lipidomics sample pretreatment efficiency is improved, and the detection data quality is improved.

Description

Technical Field

[0001] This invention relates to the field of serum lipidomics preprocessing technology, specifically to a serum lipidomics preprocessing device and method. Background Technology

[0002] Lipids are a large class of compounds in organisms that are poorly soluble in water but readily soluble in organic solvents, characterized by their structural diversity and wide variety. As a form of energy storage, lipids participate in the regulation of various life activities, such as cell growth, development, substance transport, and signal transduction. Increasing research indicates that defects in lipid synthesis and metabolism are closely related to the development of many chronic diseases (diabetes, hypertension), tumors, and other diseases. Because of the extremely important functions of lipids, following genomics and proteomics, lipid research was proposed as an independent discipline—lipomics—in 2003. Lipidomics aims to study lipids in organisms using various methods, determine their basic structures, study their functions, and ultimately reveal the relationship between lipid synthesis and metabolism and physiological processes in organisms.

[0003] The analytical workflow for lipidomics in biological samples includes sample collection, sample pretreatment, lipid molecule isolation and identification, data processing, and result analysis. Sample pretreatment is the foundation and crucial step in lipid analysis, directly affecting the stability and accuracy of the final experimental results, and is also one of the most time-consuming steps in the entire process. Researchers need to develop sample pretreatment methods based on information such as the concentration range, type, and structure of lipid molecules. Blood is one of the most commonly used samples for lipid analysis, but because it contains proteins, organic matter, and salts, which can interfere with lipid isolation and identification, it places higher demands on sample pretreatment. The pretreatment process includes multiple steps such as pipetting, mixing, protein precipitation, liquid-liquid extraction, nitrogen blowing concentration, and reconstitution, which can effectively remove interference from proteins and other impurities, improving the accuracy and recovery rate of the experiment. Factors such as the precision of liquid transfer, the uniformity of sample mixing, and cross-contamination between samples can affect the stability and accuracy of experimental results. Currently, the pretreatment equipment for serum lipidomics analysis is not highly automated, and some operations still require manual operation.

[0004] Therefore, it is necessary to develop and design a serum lipidomics pretreatment device and method to address the above-mentioned shortcomings, which is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] To address the aforementioned issues, this invention provides a serum lipidomics pretreatment device and method. This fully automated pretreatment device integrates liquid transfer, sample and reaction tube capping, oscillation mixing, liquid-liquid extraction, and nitrogen blowing concentration. It reduces testing errors introduced by human operation, minimizes tedious manual operations, and mitigates the harm of organic solvents to operators, thereby improving the efficiency of serum lipidomics sample pretreatment and the quality of detection data.

[0006] To achieve the above objectives, the present invention provides the following solution: A serum lipidomics pretreatment device includes a frame and a pipetting module, a cap-opening module, a pipette tip box, a sample cryopreservation tube, a first heating-cooling-oscillation module, a first reaction tube, a reagent storage module, a second heating-cooling-oscillation module, a second reaction tube, a nitrogen blowing module, and a software control module disposed on the frame. The pipetting module, the cap switching module, and the nitrogen blowing module can all move along the transverse, longitudinal, and height directions of the frame. The pipetting module is used to aspirate and transfer liquids, and the cap switching module is used to open and close the caps of sample tubes and / or reaction tubes. The first reaction tube is disposed on the first heating and cooling oscillation module, and the second reaction tube is disposed on the second heating and cooling oscillation module; The nitrogen blowing module is used to dry and concentrate the sample with nitrogen gas. The software control module is communicatively connected to the pipetting module, the cap switching module, the first heating-cooling-oscillation module, the second heating-cooling-oscillation module, and the nitrogen blowing module, and is used to edit and control each module to perform cap switching, pipetting, oscillation mixing, liquid-liquid extraction, and nitrogen blowing concentration operations in the serum lipidomics pretreatment process.

[0007] Preferably, the frame is provided with transverse moving guide rails corresponding to the pipetting module, the cap switching module and the nitrogen blowing module, respectively. A longitudinal moving guide rail is slidably arranged on the transverse moving guide rail, and a height moving guide rail is slidably arranged on the longitudinal moving guide rail. The pipetting module, the cap switching module and the nitrogen blowing module are respectively arranged on the corresponding height moving guide rail.

[0008] Preferably, the pipetting module includes a pipetting support frame, at least one pipetting channel disposed on the pipetting support frame, a piston disposed inside the pipetting channel, and a drive device for driving the piston to move inside the pipetting channel.

[0009] Preferably, the pipetting channel is slidably connected to the pipetting support frame, and a spacing adjustment device is provided between adjacent pipetting channels to adjust the spacing between adjacent pipetting channels.

[0010] Preferably, the spacing adjustment device is a guide groove spacing mechanism, which includes a spacing plate disposed on the pipetting support frame, a fan-shaped groove on the spacing plate corresponding to the pipetting channel, and a sliding block disposed in the groove. The pipetting channel is fixedly connected to the corresponding sliding block, and the sliding block is provided with a driving mechanism for driving the sliding block to move along the groove.

[0011] Preferably, the switch cover module includes a rotary motor, a drive shaft, and a chuck. The output shaft of the rotary motor is connected to the chuck through the drive shaft. The rotary motor is used to drive the chuck to rotate around the axis of the drive shaft. The chuck is provided with grippers.

[0012] Preferably, the nitrogen blowing module includes a nitrogen blowing needle, a proportional valve connected to the nitrogen blowing needle via a pipeline, and a nitrogen storage device connected to the proportional valve.

[0013] Preferably, both the first heating and cooling oscillation module and the second heating and cooling oscillation module include an oscillation module and a heating and cooling module disposed on the oscillation module. The oscillation module includes a rotor motor, an eccentric mechanism, an oscillation plate, a linear guide rail, and a guide rail base. The output shaft of the rotor motor is connected to the eccentric mechanism, and the eccentric mechanism is connected to the oscillation plate to convert the rotational motion of the rotor motor into the reciprocating motion of the oscillation plate in the horizontal direction. The oscillation plate is slidably mounted on the guide rail base via the linear guide rail, and the linear guide rail is used to constrain the oscillation plate to slide reciprocally in the horizontal direction. The heating and cooling module includes a semiconductor refrigeration chip, a temperature sensor disposed on the semiconductor refrigeration chip, and an outer frame for fixing the semiconductor refrigeration chip. The first reaction tube and the second reaction tube are respectively placed on the corresponding semiconductor refrigeration chips.

[0014] Preferably, the frame is also equipped with a waste collection device for collecting discarded gun heads.

[0015] This invention also discloses a method for preprocessing serum lipidomics, comprising the following steps: Transfer the sample to the first reaction tube; Add methanol to the first reaction tube and shake until homogeneous; Add the internal standard and shake until homogeneous; After adding methyl tert-butyl ether, the first reaction tube is capped; the first reaction tube is shaken and then allowed to stand at a preset temperature. Open the cap of the first reaction tube and add water, then shake well; Close the cap of the first reaction tube and let it stand at the preset temperature; Open the cap of the first reaction tube and transfer the upper lipid layer in the first reaction tube to the second reaction tube; The sample in the second reaction tube was purged with nitrogen. Dichloromethane and methanol were added to the second reaction tube and shaken thoroughly. A specific amount of sample in the second reaction tube is transferred to a sample vial containing an inner tube; Collect the sample bottles and cap them for testing.

[0016] The present invention achieves the following technical effects compared to the prior art: Through highly integrated modular design and collaborative control of the software control module, the complex steps in the pretreatment process for serum sample lipidomics analysis, such as cap opening and closing, pipetting, oscillation mixing, liquid-liquid extraction, and nitrogen blowing concentration, are fully automated. This significantly reduces manual intervention, effectively lowers the risk of errors introduced by human operation and sample cross-contamination, and also reduces the contact between laboratory personnel and organic reagents, ensuring operational safety. By placing the first and second reaction tubes on the first and second heating-cooling-oscillation modules, respectively, precise control of reaction temperature and oscillation frequency is achieved, ensuring the sufficiency and uniformity of protein precipitation and liquid-liquid extraction, thereby guaranteeing the stability of lipid extraction and the accuracy and reliability of detection data. Furthermore, the pipetting module, cap opening and closing module, and nitrogen blowing module can all move flexibly along the three-dimensional direction of the frame. Combined with the programmable custom flow of the software control module, this device possesses a high degree of automation, scalable throughput, and methodological flexibility. It effectively saves the manpower and time costs required for pretreatment, providing strong technical support for high-throughput, standardized, and automated analysis of serum lipidomics, and is particularly suitable for the simultaneous detection and analysis of multiple lipid molecules in serum in large-scale scientific research. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Appendix Figure 1 This is a schematic diagram of the overall structure of the serum lipidomics pretreatment device disclosed in this invention; Appendix Figure 2 This is a schematic diagram of the internal structure of the serum lipidomics pretreatment device disclosed in this invention. The components include: 1. Pipetting module; 2. Cover switch module; 3. Nitrogen blowing module; 4. Second heating, cooling and oscillation module; 5. Reagent storage module; 6. First heating, cooling and oscillation module; 7. Pipe tip box; 8. Waste collection device; and 9. Frame. Detailed Implementation

[0019] 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.

[0020] The purpose of this invention is to provide a serum lipidomics pretreatment device and method, which is a fully automated pretreatment device that integrates liquid transfer, sample and reaction tube capping, shaking and mixing, liquid-liquid extraction and nitrogen blowing concentration. This reduces testing errors caused by human operation, reduces tedious manual operations and the harm of organic solvents to operators, and improves the efficiency of serum lipidomics sample pretreatment and the quality of detection data.

[0021] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0022] refer to Figures 1-2The serum lipidomics pretreatment device disclosed in this embodiment of the invention includes at least a frame 9. The frame 9 is equipped with a pipetting module 1, a cap-switching module 2, a pipette tip box 7, sample cryopreservation tubes, a first heating-cooling-oscillation module 6, a first reaction tube, a reagent storage module 5, a second heating-cooling-oscillation module 4, a second reaction tube, a nitrogen blowing module 3, and a software control module. The pipetting module 1, cap-switching module 2, and nitrogen blowing module 3 can all move along the transverse, longitudinal, and height directions of the frame 9. The pipetting module 1 is used to aspirate and transfer liquids, and the cap-switching module 2 is used to open and close the sample tubes and / or reaction tubes. The system includes a cap; a first reaction tube is mounted on the first heating-cooling-oscillation module 6, and a second reaction tube is mounted on the second heating-cooling-oscillation module 4; a nitrogen blowing module 3 is positioned above the second reaction tube and is used to dry and concentrate the sample with nitrogen. The software control module is communicatively connected to the pipetting module 1, the cap-opening / closing module 2, the first heating-cooling-oscillation module 6, the second heating-cooling-oscillation module 4, and the nitrogen blowing module 3, respectively. It is used to edit and control each module to perform cap-opening / closing, pipetting, oscillation mixing, liquid-liquid extraction, and nitrogen blowing concentration operations in the serum lipidomics pretreatment process. This is achieved through a highly integrated modular design and software... The coordinated control of the control modules enables fully automated operation of complex steps in the pretreatment process of serum sample lipidomics analysis, such as cap opening and closing, pipetting, oscillation mixing, liquid-liquid extraction, and nitrogen blowing concentration. This significantly reduces manual intervention, effectively lowers the risk of errors introduced by human operation and sample cross-contamination, and also reduces the contact between experimental personnel and organic reagents, ensuring operational safety. By setting the first reaction tube and the second reaction tube on the first heating-cooling-oscillation module 6 and the second heating-cooling-oscillation module 4, respectively, precise control of reaction temperature and oscillation frequency is achieved, ensuring the sufficiency and uniformity of protein precipitation and liquid-liquid extraction, thereby guaranteeing the stability of lipid extraction and the accuracy and reliability of detection data. At the same time, the pipetting module 1, cap opening and closing module 2, and nitrogen blowing module 3 can all move flexibly along the three-dimensional direction of the frame 9. Combined with the programmable custom flow of the software control module, the device has a high degree of automation, throughput scalability, and methodological flexibility, which can effectively save the manpower and time costs required for pretreatment. It provides strong technical support for high-throughput, standardized, and automated analysis of serum lipidomics, and is particularly suitable for the simultaneous detection and analysis of multiple lipid molecules in serum in large-scale scientific research.

[0023] It should be noted that the pipette tip box 7 has at least three pipette tip slots, which are used to hold pipette tips of different sizes, including 1000μL, 200μL and 50μL; the reagent storage module 5 includes reagent slots and centrifuge tube slots. The reagent slots are used to hold reagent slots with a capacity of no more than 60mL, and the centrifuge tube slots are used to hold centrifuge tubes with a capacity of no more than 2mL. The centrifuge tubes and reagent slots are used to hold solutions such as internal standards, methyl tert-butyl ether and methanol.

[0024] refer to Figures 1-2 In one implementation, the frame 9 is provided with transverse moving guides corresponding to the pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3. A longitudinal moving guide is slidably mounted on the transverse moving guide, and a height moving guide is slidably mounted on the longitudinal moving guide. The pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3 are respectively mounted on their corresponding height moving guides. That is, three transverse moving guides, three longitudinal moving guides, and three height moving guides are provided, corresponding to the pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3, respectively. By setting transverse moving guides on the frame 9 corresponding to the pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3, and sequentially sliding the longitudinal and height moving guides on the transverse moving guides, the... Each functional module is mounted on its corresponding height-moving guide rail, enabling independent, flexible, and precise positioning of the three core execution modules—pipetting, cap opening / closing, and nitrogen blowing—in three-dimensional space. This three-dimensional motion mechanism not only allows each module to reach its preset working position without interference, achieving highly flexible operation scheduling and avoidance, but also reduces waiting time between functional modules, significantly improving the operational efficiency of the pretreatment process. Furthermore, integrating the three-dimensional motion guide rail system onto the same frame platform results in a compact and rigid overall structure, ensuring the stability and repeatability of each module during high-speed operation and precise positioning. This provides a reliable motion support platform for high-throughput, automated, and standardized serum lipidomics pretreatment operations.

[0025] It should be noted that a slider is provided on the transverse moving guide rail, and the slider is connected to the longitudinal moving guide rail. A slider is provided on the longitudinal moving guide rail, and a height moving guide rail is provided on the slider. A slider is provided on the height moving guide rail. A pipetting module 1, a cover switching module 2, and a nitrogen blowing module 3 are respectively provided on the slider. The slider can slide by means of a telescopic cylinder or an electric telescopic rod, or by means of a screw driven by a motor. The slider and the screw are threadedly connected to achieve sliding. The transverse moving guide rail is located on the top of the frame 9. The pipetting module 1, the cover switching module 2, and the nitrogen blowing module 3 are all suspended upside down.

[0026] As one implementation method, in order to avoid interference between the various moving guides during the movement process, the pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3 can be set on a single moving guide. That is, only one transverse moving guide is set, a longitudinal moving guide is set on the transverse moving guide, a height moving guide is set on the longitudinal moving guide, and the pipetting module 1, the cap switching module 2, and the nitrogen blowing module 3 are simultaneously set on the height moving guide.

[0027] refer to Figures 1-2In one embodiment, the pipetting module 1 includes a pipetting support frame with at least one pipetting channel. A piston is disposed inside the pipetting channel, and a drive device for driving the piston to move within the pipetting channel is also disposed inside the pipetting channel. By disposing of the piston inside the pipetting channel and simultaneously providing a drive device for driving the piston to move within the pipetting channel, precise control of liquid aspiration and dispensing is achieved. The reciprocating motion of the piston inside the pipetting channel forms a closed air chamber, and by utilizing the principle of air displacement, the aspiration and transfer of micro-liter-level liquids can be completed with extremely high precision.

[0028] It should be noted that the pipetting module 1 can also be equipped with a clamping device for holding the pipette tip. The upper end of the pipetting channel is provided with an air passage interface, which is used to connect with an external air passage to provide power for liquid aspiration and dispensing. The clamping device includes a claw or a flexible bayonet for releasably clamping the upper outer wall of the pipetting tip. The pipetting channel drives the pipetting tip to rise and fall to the target position through the clamping device, and completes the aspiration and dispensing of liquid through the air pressure provided by the external air passage.

[0029] With four pipetting channels arranged side-by-side at intervals along the horizontal direction, parallel pipetting operations for multiple samples or reagents are achieved, significantly improving pipetting throughput and pretreatment efficiency compared to single-channel pipetting.

[0030] refer to Figures 1-2 As a preferred method, the pipetting channel is slidably connected to the pipetting support frame, and a spacing adjustment device is provided between adjacent pipetting channels. The spacing adjustment device is used to adjust the spacing between adjacent pipetting channels, so that the relative position between each pipetting channel can be flexibly adjusted according to actual needs, thereby achieving precise adaptation to sample plates of different specifications (such as 96-well plates, 384-well plates) or sample containers with different arrangements.

[0031] refer to Figures 1-2In one implementation, the spacing adjustment device is a guide groove spacing mechanism. The guide groove spacing mechanism includes a spacing plate mounted on a pipetting support frame. The spacing plate has fan-shaped grooves corresponding to the pipetting channels. Sliding blocks are installed within the grooves. The pipetting channels are fixedly connected to their corresponding sliding blocks. A drive mechanism is provided on each sliding block to drive it to move along the groove. By fixing each pipetting channel to the sliding block and embedding it into its corresponding groove, and driving each sliding block along its respective groove via the drive mechanism, the linear or rotary motion of the drive mechanism can be precisely converted into the synchronous convergence or deflection of each pipetting channel in the horizontal direction. This system enables equidistant adjustment of the spacing between adjacent pipetting channels. The guide channel spacing mechanism is compact and has a short transmission chain. It achieves synchronous spacing changes across multiple channels through the cooperation of the spacing plate, chute, and sliding block, eliminating the need for a separate drive source for each pipetting channel. This effectively simplifies the equipment structure, reduces manufacturing costs and control complexity, while improving the synchronicity, consistency, and repeatability of spacing adjustment. It ensures precise alignment of each pipetting channel with the target well position during multi-well plate pipetting operations, providing a simple and responsive spacing adjustment solution for adapting to different specifications of standardized consumables (such as 96-well plates and 384-well plates).

[0032] It should be noted that the spacing adjustment device can also be a lateral drive mechanism that drives the lateral movement of the pipetting channel at each position. The lateral drive mechanism can be a screw motor that drives the screw to rotate, and the screw is threadedly engaged with the moving channel.

[0033] refer to Figures 1-2 In one implementation, the cap-opening module 2 includes a rotary motor, a drive shaft, and a clamp. The output shaft of the rotary motor is connected to the clamp via the drive shaft. The rotary motor drives the clamp to rotate around the axis of the drive shaft. The clamp is equipped with grippers for holding the caps, thus realizing the automated opening and closing of the caps of cryopreservation tubes and first reaction tubes. The cap-opening module 2 has a compact structure and reliable operation. With the help of a three-dimensional motion mechanism, it can realize multi-channel parallel cap opening and closing, which significantly improves the automation level and operating efficiency of cap opening and closing of cryopreservation tubes in the serum lipidomics pretreatment process.

[0034] refer to Figures 1-2 As one implementation method, the nitrogen blowing module 3 includes a nitrogen blowing needle, a proportional valve connected to the nitrogen blowing needle via a pipeline, and a nitrogen storage device connected to the proportional valve. The nitrogen storage device provides a nitrogen source for the nitrogen blowing needle. The proportional valve precisely adjusts the nitrogen flow rate and pressure output to the nitrogen blowing needle according to the control command, thereby realizing precise control of the sample purging and concentration process. The proportional valve can receive analog electrical signals and linearly adjust the nitrogen flow rate through continuous changes in the opening of the internal valve core, so that the nitrogen is stably output at a preset flow rate, which not only ensures the concentration efficiency but also effectively prevents sample splashing or loss due to excessive airflow.

[0035] In one implementation, both the first heating and cooling oscillation module 6 and the second heating and cooling oscillation module 4 include an oscillation module and a heating and cooling module mounted on the oscillation module. The oscillation module includes a rotor motor, an eccentric mechanism, an oscillation plate, a linear guide rail, and a guide rail base. The output shaft of the rotor motor is connected to the eccentric mechanism, which is connected to the oscillation plate to convert the rotational motion of the rotor motor into the reciprocating motion of the oscillation plate in the horizontal direction. The oscillation plate is slidably mounted on the guide rail base via the linear guide rail, which constrains the oscillation plate to slide only in the horizontal direction. The heating and cooling module includes a semiconductor cooling chip, a temperature sensor mounted on the semiconductor cooling chip, and an outer frame for fixing the semiconductor cooling chip. The first reaction tube and the second reaction tube are respectively placed on their respective semiconductor cooling chips, achieving coordinated processing of temperature control and oscillation mixing for the first and second reaction tubes. The oscillation module drives the eccentric mechanism via the rotor motor to... The rotational motion is converted into the horizontal reciprocating motion of the oscillating plate under the constraint of the linear guide rail, ensuring that the sample in the reaction tube is fully and uniformly mixed in the horizontal direction, thus improving the mixing efficiency of the sample and reagent during protein precipitation and liquid-liquid extraction. At the same time, the heating and cooling module precisely controls the temperature of the reaction tube through a semiconductor cooling chip (which can achieve heating or cooling). In conjunction with the temperature sensor set on the semiconductor cooling chip, the temperature is monitored in real time and fed back to the control system to achieve closed-loop PID precise temperature control. This ensures that the sample in the reaction tube is oscillated and mixed under the preset constant temperature conditions, effectively eliminating the influence of external environmental temperature fluctuations on the pretreatment results. The synergistic effect of oscillation and heating and cooling allows key steps such as protein precipitation, liquid-liquid extraction, and phase separation to be completed continuously at the same station without the need to transfer samples between different modules. This reduces operational steps and the risk of contamination, and significantly improves the pretreatment efficiency, temperature control accuracy, and the stability and repeatability of experimental results.

[0036] It should be noted that the eccentric mechanism is a cam mechanism.

[0037] refer to Figures 1-2 As an implementation method, the rack 9 is also equipped with a waste collection device 8 for collecting discarded gun heads.

[0038] This invention also discloses a method for preprocessing serum lipidomics, comprising the following steps: S1: Open the first reaction tube. The cover of the first reaction tube placed on the first heating and cooling oscillation module 6 is opened by the cover switch module 2; S2: Sample is transferred into the first reaction tube. Install the first specification pipette tip (50μL) through the pipetting module 1, transfer 25μL of serum sample into the first reaction tube, and then unload the first specification pipette tip; S3: Add methanol and shake. Install the second-specification pipette tip (1000μL) through the pipetting module 1, transfer 225μL of methanol from the test tube storage module 5 into the first reaction tube, and then unload the second-specification pipette tip; vortex the first reaction tube at a speed of 800r / min for 30s through the first heating, cooling and oscillation module 6. S4: Add internal standard and oscillate Install the first specification pipette tip through the pipetting module 1, transfer 3 μL of internal standard from the test tube storage module 5 into the first reaction tube, and then unload the first specification pipette tip; vortex the first reaction tube at a speed of 800 r / min for 10 s through the first heating, cooling and oscillation module 6. S5: After adding methyl tert-butyl ether, close the cap of the first reaction tube. Install the second-specification pipette tip through the pipetting module 1, transfer 750 μL of methyl tert-butyl ether from the test tube storage module 5 into the first reaction tube, and then unload the second-specification pipette tip; close the cap of the first reaction tube through the cap switching module 2; S6: After shaking the first reaction tube, let it stand. The first reaction tube is vortexed at 800 r / min for 30 s using the first heating and cooling oscillation module 6; the cap of the first reaction tube is closed using the cap switching module 2, and the tube is left to stand at room temperature for 30 min for liquid-liquid extraction. S7: Open the first reaction tube, add water and shake. Open the cap of the first reaction tube using the cap switch module 2; install the third-specification pipette tip (200μL) using the pipetting module 1, transfer 188μL of water into the first reaction tube, and then unload the third-specification pipette tip; vortex the first reaction tube at a speed of 800r / min for 30s using the first heating, cooling and oscillation module 6. S8: Close the first reaction tube and let it stand. The cap of the first reaction tube is closed by the cap switch module 2 and left to stand at room temperature for 15 minutes to allow the system to separate into an upper lipid phase, a middle aqueous phase, and a bottom solid residue. S9: Lipid phase transfer to the second reaction tube The cap of the first reaction tube is opened by the opening and closing module 2; the second specification pipette tip is installed by the pipetting module 1, 600 μL of the upper lipid phase is drawn from the first reaction tube and transferred to the second reaction tube placed on the second heating and cooling oscillation module 4, and then the second specification pipette tip is unloaded. S10: Nitrogen blowing concentration The sample in the second reaction tube was purged with nitrogen using nitrogen purging module 3 at a flow rate of 10 L / min for 15 min, so that the solvent would evaporate and the lipids would concentrate. S11: Redissolve and shake Using the third-specification pipette tip installed in the pipetting module 1, 100 μL of a mixed solution of dichloromethane and methanol (volume ratio 1:1) was transferred into the second reaction tube, and then the third-specification pipette tip was unloaded; the second reaction tube was vortexed at 1200 r / min for 30 s using the second heating-cooling-oscillation module 4 to fully resolvate the concentrated lipids. S12: Transfer to sample vial Install the third-specification pipette tip through the pipetting module 1, draw 90 μL of sample from the second reaction tube, and transfer it to the sample vial placed at the sample loading station. Then unload the third-specification pipette tip. S13: Sample received and ready for testing Collect the sample bottles and cap them, ready for testing.

[0039] It should be noted that, for those skilled in the art, it is obvious that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A serum lipidomics pretreatment device, characterized in that, It includes a frame (9) and a pipetting module (1), a switch cap module (2), a pipette tip box (7), a sample cryopreservation tube, a first heating-cooling-oscillation module (6), a first reaction tube, a reagent storage module (5), a second heating-cooling-oscillation module (4), a second reaction tube, a nitrogen blowing module (3), and a software control module, all mounted on the frame (9). The pipetting module (1), the switch cap module (2), and the nitrogen blowing module (3) can all move along the transverse, longitudinal, and height directions of the frame (9). The pipetting module (1) is used to aspirate and transfer liquids, and the switch cap module (2) is used to open and close the caps of the sample tubes and / or reaction tubes. The first reaction tube is disposed on the first heating and cooling oscillation module (6), and the second reaction tube is disposed on the second heating and cooling oscillation module (4); The nitrogen blowing module (3) is used to dry and concentrate the sample with nitrogen gas; The software control module is connected to the pipetting module (1), the cap switching module (2), the first heating-cooling-oscillation module (6), the second heating-cooling-oscillation module (4), and the nitrogen blowing module (3) respectively, and is used to edit and control each module to perform cap switching, pipetting, oscillation mixing, liquid-liquid extraction and nitrogen blowing concentration operations in the serum lipidomics pretreatment process.

2. The serum lipidomics pretreatment device according to claim 1, characterized in that, The frame (9) is provided with transverse moving guide rails corresponding to the pipetting module (1), the switch cover module (2) and the nitrogen blowing module (3), respectively. A longitudinal moving guide rail is slidably provided on the transverse moving guide rail, and a height moving guide rail is slidably provided on the longitudinal moving guide rail. The pipetting module (1), the switch cover module (2) and the nitrogen blowing module (3) are respectively provided on the corresponding height moving guide rails.

3. The serum lipidomics pretreatment device according to claim 1, characterized in that, The pipetting module (1) includes a pipetting support frame, at least one pipetting channel disposed on the pipetting support frame, a piston disposed inside the pipetting channel, and a driving device for driving the piston to move inside the pipetting channel.

4. The serum lipidomics pretreatment device according to claim 3, characterized in that, The pipetting channel is slidably connected to the pipetting support frame, and a spacing adjustment device is provided between adjacent pipetting channels to adjust the spacing between adjacent pipetting channels.

5. The serum lipidomics pretreatment device according to claim 4, characterized in that, The spacing adjustment device is a guide groove spacing mechanism, which includes a spacing plate disposed on the pipetting support frame, a fan-shaped groove on the spacing plate corresponding to the pipetting channel, and a sliding block disposed in the groove. The pipetting channel is fixedly connected to the corresponding sliding block, and the sliding block is provided with a driving mechanism for driving the sliding block to move along the groove.

6. The serum lipidomics pretreatment device according to claim 1, characterized in that, The switch cover module (2) includes a rotary motor, a drive shaft and a chuck. The output shaft of the rotary motor is connected to the chuck through the drive shaft. The rotary motor is used to drive the chuck to rotate around the axis of the drive shaft. The chuck is provided with grippers.

7. The serum lipidomics pretreatment device according to claim 1, characterized in that, The nitrogen blowing module (3) includes a nitrogen blowing needle, a proportional valve connected to the nitrogen blowing needle via a pipeline, and a nitrogen storage device connected to the proportional valve.

8. The serum lipidomics pretreatment device according to claim 1, characterized in that, Both the first heating and cooling oscillation module (6) and the second heating and cooling oscillation module (4) include an oscillation module and a heating and cooling module disposed on the oscillation module. The oscillation module includes a rotor motor, an eccentric mechanism, an oscillation plate, a linear guide rail, and a guide rail seat. The output shaft of the rotor motor is connected to the eccentric mechanism, and the eccentric mechanism is connected to the oscillation plate to convert the rotational motion of the rotor motor into the reciprocating motion of the oscillation plate in the horizontal direction. The oscillation plate is slidably mounted on the guide rail seat through the linear guide rail, and the linear guide rail is used to constrain the oscillation plate to slide reciprocally in the horizontal direction. The heating and cooling module includes a semiconductor refrigeration chip, a temperature sensor disposed on the semiconductor refrigeration chip, and an outer frame for fixing the semiconductor refrigeration chip. The first reaction tube and the second reaction tube are respectively placed on the corresponding semiconductor refrigeration chip.

9. The serum lipidomics pretreatment device according to claim 1, characterized in that, The frame (9) is also equipped with a waste collection device (8), which is used to collect discarded gun heads.

10. A method for preprocessing serum lipidomics, characterized in that, Includes the following steps: Transfer the sample to the first reaction tube; Add methanol to the first reaction tube and shake until homogeneous; Add the internal standard and shake until homogeneous; After adding methyl tert-butyl ether, the first reaction tube is capped; the first reaction tube is shaken and then allowed to stand at a preset temperature. Open the cap of the first reaction tube and add water, then shake well; Close the cap of the first reaction tube and let it stand at the preset temperature; Open the cap of the first reaction tube and transfer the upper lipid layer in the first reaction tube to the second reaction tube; The sample in the second reaction tube was purged with nitrogen. Dichloromethane and methanol were added to the second reaction tube and shaken thoroughly. A specific amount of sample in the second reaction tube is transferred to a sample vial containing an inner tube; Collect the sample bottles and cap them for testing.

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