Clinical mass spectrometry pretreatment automatic instrument and application

Abstract

The present application relates to a kind of mass spectrum pretreatment automatic instrument, including manipulator module, reaction cup loading module, reagent kit module, ultrasonic mixing module, incubation module, positive pressure module, automatic sampling module and online diluent module, above-mentioned module is respectively installed on platform;Reaction cup in the reaction cup loading module is equipped with three layers of filler, from top to bottom in turn for upper polytetrafluoroethylene filler, middle layer glass fiber filler, lower polytetrafluoroethylene filler, the instrument does not need to disassemble component to realize magnetic solid-phase extraction method and protein precipitation method pretreatment.The present application combines protein precipitation and magnetic bead method on a fully automated pretreatment instrument, reduces the cost of procurement, greatly improves experimental efficiency;Reduce the experimental error due to manual operation in different instruments, save manpower, improve the accuracy of analysis and detection;The present application is suitable for clinical detection.

Description

Technical Field

[0001] This invention relates to an automated instrument, specifically, to an automated instrument for clinical mass spectrometry pretreatment and its application. Background Technology

[0002] Sample preparation of human matrix for clinical mass spectrometry is one of the most time-consuming and labor-intensive steps in the analysis. Over the past few decades, various automated systems have been developed and improved to assist analysts in reporting quantitative results promptly. In automated-assisted mass spectrometry analysis, automated liquid workstations can be used for sample handling steps, including the preparation of calibration standards and quality control samples, sample aliquoting, sample transfer, solvent transfer, and other tasks. These workstations have now largely replaced manual liquid transfer operations (such as 96-well plates) in parallel sample preparation. Automated operation provides higher sample preparation throughput and significantly reduces the monotonous and repetitive work of bioanalysts.

[0003] Automation effectively aids sample preparation in two main ways: precipitation (acid precipitation, organic reagent precipitation, salt precipitation) and extraction (solid-liquid extraction and liquid-liquid extraction). These techniques may require different liquid processing platforms when handling different matrices, solutions, and solvents. For example, automated processing of highly polar substances (e.g., water-soluble vitamins, antibiotics) requires typical automated PPT methods; for moderately polar substances (psychotropic drugs, hormones, fat-soluble vitamins) and ionic compounds (catecholamines), typical magnetic bead methods are needed; and for compounds with low polarity or complex matrices like whole blood samples, auxiliary techniques such as ultrasonic mixing are required. These corresponding techniques ensure optimized pretreatment performance.

[0004] Existing automated instruments are bulky, using multi-well plates in series for multi-channel sample preparation. This makes them suitable only for large medical testing laboratories or regional testing centers. Provincial and municipal hospitals cannot handle the daily throughput required for these instruments. Multi-well plates only allow for centralized sample collection and processing, failing to meet the needs of samples added to clinical practice temporarily. Currently, automation takes many forms and cannot be integrated into a single instrument. Switching between different instrument platforms is necessary for different processing procedures, causing significant inconvenience for operators who need to learn to use various sample preparation instruments. Furthermore, the range of sample preparation services currently available on the market is limited, failing to meet the growing demand. Additionally, current automated instruments primarily focus on serum matrices; no instruments are currently available for samples with complex matrices. Summary of the Invention

[0005] To overcome the technical problems existing in the prior art, this invention provides an automated mass spectrometry pretreatment instrument and its application. By combining protein precipitation and magnetic bead methods into a fully automated pretreatment instrument, procurement costs are reduced, experimental efficiency is greatly improved, experimental errors caused by manual operation of different instruments are reduced, manpower is saved, and the accuracy of analytical detection is improved. This invention is suitable for clinical testing.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] An automated clinical mass spectrometry pretreatment instrument includes a robotic arm module, a reaction vessel loading module, a reagent kit module, an ultrasonic mixing module, an incubation module, a positive pressure module, an automatic sample injection module, and an online diluent module. These modules are mounted on a platform. The reaction vessel in the loading module contains three layers of packing material: an upper layer of polytetrafluoroethylene (PTFE), a middle layer of glass fiber packing, and a lower layer of PTFE packing. The instrument enables pretreatment using magnetic solid-phase extraction (MSE) and protein precipitation methods without requiring disassembly of any components.

[0008] The robotic arm module includes a mounting frame and a robotic arm. It controls the three-dimensional movement of the robotic arm in X, Y, and Z directions to realize the addition, incubation, cleaning, and disposal of reaction cups.

[0009] The reaction cup loading module includes a reaction cup holder and a reaction cup. The reaction cup holder carries the reaction cup forward to the robot arm.

[0010] The reagent kit module includes a reagent kit loading mechanism that holds the reagent kits for providing different reagents.

[0011] The ultrasonic mixing module includes an ultrasonic generator. The module uses ultrasound to agitate the liquid in the reaction vessel, thereby achieving thorough mixing of the reaction solution.

[0012] The incubation module includes an incubation disk and a heating plate, which provide a warm environment for the reaction.

[0013] The positive pressure module uses pressure to direct the liquid in the reaction vessel to a designated location.

[0014] The automatic sample injection module includes a motor drive wheel, a guide rail, and an injection needle, which controls the injection needle to move in and out and reach the designated extraction position.

[0015] The online diluent module dilutes the analytes in the reaction vessel by adding online diluent.

[0016] Preferably, the robotic arm module includes a mounting frame and a robotic arm, and the mounting frame is provided with a track for the movement of the robotic arm.

[0017] Preferably, the ultrasonic mixing module includes a syringe-type reaction cup clamping hole, at which an ultrasonic vibrator is installed to clamp or release the reaction cup. The ultrasonic vibrator is equipped with a power device to drive its movement toward or away from the reaction cup. The ultrasonic vibrator simultaneously achieves clamping and fixing of the reaction cup and non-contact ultrasonic mixing, with an ultrasonic time of 3-5 seconds. Ultrasonic mixing avoids the risk of contamination from contact with the sample; it ensures a more uniform distribution of magnetic beads in the pretreatment system; and ultrasonic mixing can be completed within 3-5 seconds, improving efficiency by more than 50% compared to traditional methods (such as oscillation), significantly shortening the mixing time.

[0018] Existing protein precipitation plates can only handle projects requiring protein precipitation pretreatment, not projects requiring magnetic bead pretreatment. This invention, through the design of a syringe-type reaction cup with a pressing hole, and in conjunction with a syringe-type ultrasonic reaction cup, can simultaneously handle both pretreatment methods, combining them into one instrument.

[0019] Preferably, the reaction cup loading module includes a reaction cup with an ultrasonic vibrator on its outer side; upper and lower layers of polytetrafluoroethylene (PTFE) filler are respectively fused to the cup wall. Fusing PTFE to the cup wall prevents filler dispersion during ultrasonication, thus solving the leakage problem of existing reaction cups.

[0020] The three-layer packing effectively prevents the leakage of reagents such as methanol and acetonitrile, allowing liquid to pass through only under positive pressure. The three-layer packing in the reaction vessel serves two purposes: it secures the filter membrane and prevents liquid leakage even under no pressure.

[0021] Preferably, the upper layer of polytetrafluoroethylene filler with a particle size of 10-30 μm is fired onto the inner wall of the reaction vessel, the middle layer of glass fiber filler with a particle size of 0.1-0.3 μm is filled into the upper and lower layers of polytetrafluoroethylene filler, and the lower layer of polytetrafluoroethylene filler with a particle size of 10-30 μm is fired onto the inner wall of the reaction vessel.

[0022] In existing reaction cups, during non-contact ultrasonic mixing in the mixing module, the packing material inside the cup tends to disperse, causing some protein to leak into the collection liquid. The reaction cup of this invention incorporates polytetrafluoroethylene (PTFE) packing material at both the top and bottom of the packing material. This PTFE packing material is sintered onto the inner wall of the reaction cup, exhibiting strong adhesion and minimizing the risk of leakage during ultrasonication. The middle layer of this reaction cup consists of a 0.1-0.3 μm packing material. Due to the small porosity of the packing material, the liquid in the reaction cup will not flow down under gravity. Furthermore, the PTFE packing layer does not interact with the magnetic beads; only under positive pressure will the liquid flow towards the receiving layer. Automated instruments can achieve both protein precipitation and magnetic attraction functions.

[0023] This invention also provides the application of the above-mentioned automated clinical mass spectrometry pretreatment instrument, which is used for protein precipitation pretreatment or magnetic solid phase extraction pretreatment.

[0024] As a preferred embodiment, the pretreatment process for protein precipitation is as follows: the robotic arm in the robotic arm module picks up the reaction cup in the reaction cup loading module and transfers it to the ultrasonic mixing module; the sample is drawn from the automatic injection module; the self-isotope internal standard and protein precipitant are drawn from the reagent kit module and transferred to the ultrasonic mixing module for ultrasonic mixing; the robotic arm picks up the reaction cup and transfers it to the positive pressure module; the receiving cup receives the filtrate; and the filtrate is transferred to the injector for injection.

[0025] As a preferred method, the pretreatment process for magnetic solid phase extraction is as follows: the robotic arm in the robotic arm module grasps the reaction cup in the reaction cup loading module and transfers it to the ultrasonic mixing module. The sample is then aspirated from the automatic sample injection module. The self-isotope internal standard and dissociation solution are aspirated from the reagent kit module and transferred to the reaction cup. The mixture is then ultrasonically mixed. Magnetic bead solution is aspirated into the reaction cup and ultrasonically mixed again. The cup is then placed in the incubation module (under constant temperature conditions, the adsorption of the analyte and magnetic beads will reach a steady state) and allowed to wait for washing (washing is required for magnetic bead pretreatment to remove proteins not adsorbed by the magnetic beads). The reaction cup is then grasped and transferred to the positive pressure module. After positive pressure and magnetic attraction, the remaining waste liquid is transferred out. Eluent is added and magnetic attraction is used to transfer the remaining waste liquid out. Eluent is added, and the compressor provides gas. The receiving cup receives the filtrate. The online diluent is aspirated from the online diluent module and transferred to the injector for injection.

[0026] Preferably, the automated mass spectrometry pretreatment instrument is used for pretreatment of whole blood samples.

[0027] As a preferred method, the pretreatment process for whole blood tests is as follows: the robotic arm in the robotic arm module grasps the reaction cup in the reaction cup loading module and transfers it to the ultrasonic mixing module; the sample is drawn from the automatic sample injection module; the self-isotope internal standard and dissociation solution are drawn from the reagent kit module and transferred to the reaction cup; after ultrasonic mixing, the magnetic bead solution is drawn into the reaction cup; after ultrasonic mixing, the cup is placed in the incubation module and awaits cleaning; the reaction cup is grasped and magnetically attracted by the positive pressure module, and the remaining waste liquid is transferred out; the eluent is added and magnetically attracted, and the remaining waste liquid is transferred out; after adding the elution solution, the compressor provides gas; the receiving cup receives the filtrate and transfers it to the sample injector for injection.

[0028] In automated instruments, substances in samples are extracted and enriched using protein precipitation and magnetic solid-phase extraction methods. An integrated ultrasonic mixing module (non-contact ultrasound) increases existing mixing efficiency and reduces mixing time. The ultrasonic mixing module incorporates an ultrasonic oscillator that drives an ultrasonic reaction cup to lyse the sample using a non-contact ultrasonic method. This avoids direct contact with the sample, preventing contamination, and the high-frequency ultrasound mode enables ultra-fast sample mixing.

[0029] In automated instruments, substances in samples are extracted and enriched based on protein precipitation and magnetic solid-phase extraction. By improving the ultrasonic reaction cup, a method can be achieved where both protein precipitation and magnetic solid-phase extraction can be performed in the reaction cup.

[0030] The beneficial effects of this invention are as follows:

[0031] 1. This invention enables the pretreatment of protein precipitation and magnetic solid phase extraction to be performed on a single automated instrument, combining protein precipitation and magnetic bead extraction into a fully automated pretreatment instrument. This reduces procurement costs and significantly improves experimental efficiency; it also reduces experimental errors caused by manual operation of different instruments, saves manpower, and improves the accuracy of analysis and detection; this invention is suitable for clinical testing.

[0032] 2. By using ultrasonic mixing, mixing time is reduced and detection throughput is increased; it can be used for pretreatment of low polarity and whole blood samples.

[0033] 3. Before loading the sample, perform pretreatment to reduce damage to the instrument and chromatographic column caused by magnetic beads or trace amounts of protein;

[0034] 4. After processing 2500 samples, the column pressure of this invention is only 2648 psi (compared to 5800 psi for existing protein precipitation plates), which significantly reduces the risk of column damage; the whole blood sample recovery rate is 85%-115%, the inter-batch precision is <8.33%, and the method is robust. Attached Figure Description

[0035] Figure 1 This is a schematic diagram of the structure of the automated clinical mass spectrometry pretreatment instrument of the present invention;

[0036] Figure 2 This is a schematic diagram of the ultrasonic mixing module of the present invention;

[0037] Figure 3 This is a schematic diagram of the structure of the reaction cup of the present invention;

[0038] Figure 4 A comparison graph of column pressure changes between protein precipitation plate treatment and the automated instrument treatment of this invention;

[0039] Illustration: 1-Robotic arm module, 2-Platform, 3-Liquid pump assembly, 4-Peristaltic pump, 5-Positive pressure module, 6-Plunger pump assembly, 7-Online dilution module, 8-Automatic sample injection module, 9-Reagent kit module, 10-Upper layer PTFE packing, 11-Ultrasonic mixing module, 12-Reaction cup loading module, 13-Middle layer glass fiber packing, 14-Incubation module, 15-Lower layer PTFE packing, 16-Reaction cup, 17-Power unit, 18-Ultrasonic vibrator. Detailed Implementation

[0040] The present invention will be further described below with reference to the accompanying drawings and embodiments. It should be understood that the implementation of the present invention is not limited to the following embodiments, and the technical solutions of the present invention will be further described in detail. It should be understood that the implementation of the present invention is not limited to the following embodiments, and any modifications and / or alterations made to the present invention will fall within the protection scope of the present invention.

[0041] In this invention, unless otherwise specified, all parts and percentages are by weight, and the equipment and raw materials used are commercially available or commonly used in the art. Unless otherwise specified, the methods in the following embodiments are conventional methods in the art. Unless otherwise specified, the components or equipment in the following embodiments are general standard parts or components known to those skilled in the art, and their structures and principles can be learned by those skilled in the art through technical manuals or conventional experimental methods.

[0042] Example 1

[0043] Reference Figures 1-3 An automated clinical mass spectrometry pretreatment instrument is disclosed. The instrument includes a robotic arm module 1, a reaction cup loading module 12, a reagent kit module 9, an ultrasonic mixing module 11, an incubation module 14, a positive pressure module 5, an automatic sample injection module 8, and an online diluent module 7. The reaction cup loading module 12, reagent kit module 9, ultrasonic mixing module 11, incubation module 14, positive pressure module 5, automatic sample injection module 8, and online diluent module 7 are respectively installed on a platform 2. The robotic arm module 1 is connected to a liquid pump assembly 3, the positive pressure module 5 is connected to a peristaltic pump 4, and the online diluent module 7 is connected to a plunger pump assembly 6 to achieve precise liquid transfer.

[0044] The robotic arm module includes a mounting frame and a robotic arm. It controls the three-dimensional movement of the robotic arm in X, Y, and Z directions to realize the addition, incubation, cleaning, and disposal of reaction cups.

[0045] The reaction cup loading module includes a reaction cup holder and a reaction cup. The reaction cup holder carries the reaction cup forward to the robot arm.

[0046] The reagent kit module includes a reagent kit loading mechanism that holds the reagent kits for providing different reagents.

[0047] The ultrasonic mixing module includes an ultrasonic generator. The module uses ultrasound to agitate the liquid in the reaction vessel, thereby achieving thorough mixing of the reaction solution.

[0048] The incubation module includes an incubation disk and a heating plate, which provide a warm environment for the reaction.

[0049] The positive pressure module uses pressure to direct the liquid in the reaction vessel to a designated location.

[0050] The automatic sample injection module includes a motor drive wheel, a guide rail, and an injection needle, which controls the injection needle to move in and out and reach the designated extraction position.

[0051] The online diluent module dilutes the analytes in the reaction vessel by adding online diluent.

[0052] The robotic arm module includes a mounting frame and a robotic arm, and the mounting frame is provided with a track for the movement of the robotic arm.

[0053] The ultrasonic mixing module includes a syringe-type reaction cup clamping hole, at which an ultrasonic vibrator 18 is installed to clamp or release the reaction cup. The ultrasonic vibrator is equipped with a power device (motor module) 17 to drive its movement toward or away from the reaction cup. Ultrasonic mixing avoids the risk of contamination from contact with the sample; it ensures a more uniform distribution of magnetic beads in the pretreatment system; and ultrasonic mixing can be completed within 3-5 seconds, significantly shortening the mixing time.

[0054] Existing protein precipitation plates can only handle projects requiring protein precipitation pretreatment, not projects requiring magnetic bead pretreatment. This invention, through the design of a syringe-type reaction cup with a pressing hole, and in conjunction with a syringe-type ultrasonic reaction cup, can simultaneously handle both pretreatment methods, combining them into one instrument.

[0055] The reaction cup loading module includes a reaction cup 16, with an ultrasonic vibrator 18 positioned outside the reaction cup 16. The reaction cup 16 contains packing material, which, from top to bottom, consists of an upper layer of polytetrafluoroethylene (PTFE) packing 10, a middle layer of glass fiber packing 13, and a lower layer of PTFE packing 15. This three-layer packing structure effectively secures the filter membrane and prevents liquid leakage even under pressureless conditions. The ultrasonic reaction cup packing material, with its three-layer structure—an upper layer of PTFE with a particle size of 20 μm, a middle barrier layer of glass fiber with a particle size of 0.22 μm, and a lower layer of PTFE with a particle size of 20 μm—effectively blocks leakage from methanol, acetonitrile, formic acid, sulfosalicylic acid, n-hexane, ethyl acetate, and magnetic bead systems.

[0056] The instrument of this invention has a compact overall size, is suitable for the daily testing throughput of hospitals, and can handle temporary samples that are cut in line.

[0057] Example 2

[0058] Protein precipitation-related items (water-soluble vitamins)

[0059] Using the instrument described in Example 1, the robotic arm in the robotic arm module 1 grasps the reaction cup loading module 12 and transfers it to the ultrasonic mixing module 11. 100 μL of sample is drawn from the automatic sample injection module 8, and 20 μL of its own isotope internal standard and 300 μL of protein precipitant are drawn from the reagent kit module 9 and transferred to the ultrasonic mixing module 11. After non-contact ultrasonic mixing for 10 seconds in the ultrasonic mixing module 11, the positive pressure module 5 compressor provides gas for 10 seconds, the receiving cup receives the filtrate, and the filtrate is transferred to the sample injector for injection.

[0060] Example 3

[0061] Magnetic solid-phase extraction projects (vitamins A, D, E)

[0062] Using the instrument described in Example 1, the robotic arm in the robotic arm module 1 grasps the reaction cup loading module 12 and transfers it to the ultrasonic mixing module 11. 100 μL of sample is drawn from the automatic sample injection module 8, and 50 μL of its own isotope internal standard and 370 μL of dissociation solution are drawn from the reagent kit module 9 and transferred to the reaction cup. After non-contact ultrasonic mixing for 10 seconds, 180 μL of magnetic bead solution is drawn into the reaction cup and mixed non-contactly with ultrasonication for 10 seconds. The cup is then placed in the incubation module 14 for cleaning. The reaction cup is then grasped and magnetically attracted to the positive pressure module 5, and the remaining waste liquid is transferred out. 700 μL of eluent is added and magnetically attracted, and the remaining waste liquid is transferred out. 150 μL of elution buffer is added, and the compressor provides gas for 10 seconds. The receiving cup receives the filtrate and transfers it to the sample injector for injection.

[0063] Example 4

[0064] Complete blood count (vitamins A, D, E)

[0065] Using the instrument described in Example 1, the robotic arm in the robotic arm module 1 grasps the reaction cup loading module 12 and transfers it to the ultrasonic mixing module 11. 100 μL of sample is drawn from the automatic sample injection module 8, and 50 μL of its own isotope internal standard and 200 μL of dissociation solution are drawn from the reagent kit module 9 and transferred to the reaction cup. After non-contact ultrasonic mixing for 10 seconds, 150 μL of magnetic bead solution is drawn into the reaction cup and mixed non-contactly with ultrasonication for 10 seconds. The cup is then placed in the incubation module 14 for cleaning. The reaction cup is then grasped and magnetically attracted to the positive pressure module 5, and the remaining waste liquid is transferred out. 700 μL of eluent is added and magnetically attracted, and the remaining waste liquid is transferred out. 150 μL of elution buffer is added, and the compressor provides gas for 10 seconds. The receiving cup receives the filtrate and transfers it to the sample injector for injection.

[0066] The samples treated in Examples 2 and 4 were subjected to relevant tests, and the test results are as follows:

[0067] Assessment of column damage (comparison between protein precipitation plate and the automated instrument of this invention):

[0068] Automated instruments and nucleic acid extractors are used to process 200 clinical samples daily for on-machine testing. The column pressure change results are as follows: Figure 4 As shown, from Figure 4 It can be seen that the protein precipitation plate has reached the column pressure limit of 5800 psi after 2500 injections, while the present invention has a column pressure of 2648 psi when the number of injections is 2500, which is only 45.65% of the column pressure limit, and can still be used.

[0069] Evaluation of residual magnetic beads (comparison between nucleic acid extractors and automated instruments):

[0070] After sample processing using the magnetic bead method in a nucleic acid extractor, the accumulation of magnetic beads on the stator and rotor of the liquid chromatography six-way valve causes wear. Damage to the instrument can be avoided by using an automated instrument filtration system to trap the magnetic beads.

[0071] Methodological performance validation and evaluation (comparison between nucleic acid extractors and automated instruments):

[0072] Accuracy assessment of whole blood vitamin A, D, and E: The results of accuracy (%) of nucleic acid extraction instrument processing and accuracy (%) of automated instrument processing are shown in Tables 1 and 2.

[0073] Table 1. Nucleic acid extraction instrument processing recovery rate (%)

[0074]

[0075] Table 2. Recovery rate of automated instruments (%)

[0076]

[0077] The recovery rate of vitamin E in whole blood processed by the nucleic acid extractor was less than 85% at high concentrations. The recovery rates of vitamin A and 25-(OH)D3 showed an increasing trend at low, medium, and high concentrations, while the recovery rate of vitamin E showed a decreasing trend at low, medium, and high concentrations. The recovery rates of whole blood processed by the automated instrument were all between 85% and 115%; the recovery rates at low, medium, and high concentrations were relatively evenly distributed.

[0078] Precision assessment of whole blood vitamin A, D, and E: The results of nucleic acid extraction instrument processing and automated instrument processing are shown in Tables 3 and 4.

[0079] Table 3 Nucleic Acid Extraction Instrument Processing

[0080]

[0081] Table 4 Automated Instrument Processing

[0082]

[0083] The inter-batch precision of low-quality control vitamin A, 25-(OH)D2, and vitamin E processed by the nucleic acid extractor exceeded one-third of the allowable total error (8.33%). The quality control of automated instruments was all within 8.33%; the pretreatment methods were robust.

[0084] Methodological performance validation and evaluation (comparison between protein precipitation plates and automated instruments):

[0085] Accuracy assessment of water-soluble vitamins: The results of accuracy (%) of protein precipitation plate treatment and accuracy (%) of automated instrument treatment are shown in Tables 5 and 6.

[0086] Table 5. Protein precipitation plate treatment recovery rate (%)

[0087]

[0088] Table 6. Recovery rate of automated instruments (%)

[0089]

[0090] The recovery rates of water-soluble vitamins processed by protein precipitation plates and whole blood processed by automated instruments were both between 85% and 115%. The recovery rates of samples processed by automated instruments showed little difference among low, medium, and high concentrations, indicating strong method robustness.

[0091] Precision assessment of the water-soluble vitamins project: The results of protein precipitation plate treatment and automated instrument treatment are shown in Tables 7 and 8.

[0092] Table 7 Protein Precipitation Plate Treatment

[0093]

[0094] Table 8 Automated Instrument Processing

[0095]

[0096] The low-quality control VB3 and VB6 precision of protein precipitation plate processing exceeded 1 / 3 of the allowable total error (8.33%). The quality control of automated instrument processing was all within 8.33%; the pretreatment method is robust.

[0097] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An automated clinical mass spectrometry pretreatment instrument, characterized in that, The automated clinical mass spectrometry pretreatment instrument includes a robotic arm module, a reaction vessel loading module, a reagent kit module, an ultrasonic mixing module, an incubation module, a positive pressure module, an automatic sample injection module, and an online diluent module. These modules are mounted on a platform. The reaction vessel in the reaction vessel loading module contains three layers of packing material: an upper layer of polytetrafluoroethylene (PTFE), a middle layer of glass fiber, and a lower layer of PTFE. The automated clinical mass spectrometry pretreatment instrument can perform magnetic solid-phase extraction (MSE) and protein precipitation pretreatment without disassembling any components. The ultrasonic mixing module includes a syringe-type reaction cup clamping hole, and an ultrasonic vibrator is provided at the syringe-type reaction cup clamping hole to clamp or loosen the reaction cup. The ultrasonic vibrator is provided with a power device to drive it to move toward or away from the reaction cup. The ultrasonic vibrator simultaneously achieves clamping and fixing of the reaction cup and non-contact ultrasonic mixing, with an ultrasonic time of 3-5 seconds. An ultrasonic vibrator is provided on the outside of the reaction cup of the reaction cup loading module; the upper layer of polytetrafluoroethylene (PTFE) filler and the lower layer of PTFE filler are respectively fired onto the wall of the reaction cup; the particle size of the upper layer of PTFE filler is 10-30 μm, the particle size of the middle layer of glass fiber filler is 0.1-0.3 μm, and the particle size of the lower layer of PTFE filler is 10-30 μm. The robotic arm module is connected to the liquid pump assembly, the positive pressure module is connected to the peristaltic pump, and the online dilution module is connected to the plunger pump assembly to achieve precise liquid transfer; The robotic arm module includes a mounting frame and a robotic arm, which controls the three-dimensional movement of the robotic arm in X, Y, and Z directions to realize the addition, incubation, cleaning, and disposal of reaction cups; The reaction cup loading module includes a reaction cup holder and a reaction cup. The reaction cup holder carries the reaction cup forward to the robot arm.

2. The automated clinical mass spectrometry pretreatment instrument according to claim 1, characterized in that, The mounting frame is equipped with tracks for the movement of the robotic arm.

3. An application of the automated clinical mass spectrometry pretreatment instrument as described in claim 1, characterized in that, The automated clinical mass spectrometry pretreatment instrument is used for protein precipitation pretreatment and magnetic solid phase extraction pretreatment.

4. The application of the automated clinical mass spectrometry pretreatment instrument according to claim 3, characterized in that, The pretreatment process for protein precipitation is as follows: the robotic arm in the robotic arm module picks up the reaction cup in the loading module and transfers it to the ultrasonic mixing module. The sample is then drawn from the automatic injection module. The internal standard of its own isotope and the protein precipitant are drawn from the reagent kit module and transferred to the ultrasonic mixing module for ultrasonic mixing. The mixture is then ultrasonically mixed. The robotic arm picks up the reaction cup and transfers it to the positive pressure module. The receiving cup receives the filtrate, and the filtrate is transferred to the injector for injection.

5. The application of the automated clinical mass spectrometry pretreatment instrument according to claim 3, characterized in that, The pretreatment process for magnetic solid phase extraction is as follows: The robotic arm in the robotic arm module picks up the reaction cup from the loading module and transfers it to the ultrasonic mixing module. Samples are then drawn from the automatic sample injection module. The self-isotope internal standard and dissociation solution are drawn from the reagent kit module and transferred to the reaction cup. The mixture is ultrasonically mixed. Magnetic bead solution is drawn into the reaction cup and ultrasonically mixed again. The cup is then placed in the incubation module for cleaning. The reaction cup is then picked up by magnetic attraction from the positive pressure module, and the remaining waste liquid is transferred out. Eluent is added and magnetically attracted, and the remaining waste liquid is transferred out. Eluent is added, and gas is supplied by the compressor. The receiving cup receives the filtrate. The online diluent is drawn from the online diluent module and transferred to the sample injector for injection.

6. The application of the automated clinical mass spectrometry pretreatment instrument according to claim 3, characterized in that, The automated clinical mass spectrometry pretreatment instrument is used for pretreatment of whole blood tests.

7. The application of the automated clinical mass spectrometry pretreatment instrument according to claim 6, characterized in that, The pretreatment process for whole blood tests is as follows: The robotic arm in the robotic arm module picks up the reaction cup from the loading module and transfers it to the ultrasonic mixing module. Samples are then drawn from the automated injection module. The reagent kit module draws the self-isotope internal standard and dissociation solution into the reaction cup, which is then ultrasonically mixed. Magnetic bead solution is drawn into the reaction cup, ultrasonically mixed again, and placed in the incubation module for cleaning. The reaction cup is then picked up by magnetic attraction from the positive pressure module, and the remaining waste liquid is transferred out. Eluent is added, and the remaining waste liquid is transferred out after magnetic attraction. Eluent is added, and the compressor provides gas. The receiving cup receives the filtrate and transfers it to the injector for injection.

Images