Complex system separation and multifunctional characteristic synchronous detection system and preparation method and detection method thereof

Abstract

The invention discloses a complex system separation and multifunctional characteristic synchronous detection system which comprises a liquid chromatogram separation unit, a shunting control unit, a mass spectrum / optical detection unit, an electrochemical detection unit and a data analysis unit, an outlet of the liquid chromatogram separation unit is communicated with an inlet of the shunting control unit, an outlet of the shunting control unit is communicated with a first pipeline and a second pipeline, the first pipeline is communicated with the mass spectrum / optical detection unit, and the second pipeline is communicated with the electrochemical detection unit; the mass spectrum / optical detection unit comprises a mass spectrum detection module and an optical detection module, and the mass spectrum detection module, the optical detection module and the electrochemical detection unit are connected in parallel and are all connected with the data analysis unit. Therefore, multi-dimensional synchronous analysis of separation behavior, optical characteristics, structural characteristics and electrochemical properties of target components under single sample introduction can be realized. The invention also discloses a preparation method and a detection method of the system.

Description

Technical Field

[0001] This invention belongs to the field of analytical chemistry and electrochemistry, specifically relating to a system for the simultaneous detection of separation and multifunctional properties of complex systems, as well as its preparation and detection methods. Background Art

[0002] With the deepening of research on natural products, quality control in the food industry, and drug development, the need to simultaneously obtain the chemical structure, content, and physicochemical properties (such as redox activity and pKa value) of each component in complex systems (such as extracts of traditional Chinese medicine, tea, coffee, fruits, and vegetables) is becoming increasingly urgent. These systems have diverse components and significant differences in chemical properties, making it difficult to achieve comprehensive characterization using traditional single analytical methods.

[0003] Currently, the mainstream component analysis technique is the coupling of liquid chromatography with mass spectrometry / optical detectors (e.g., HPLC-UV / MS), which has the advantages of separation capability and structure identification, but has the following significant drawbacks: (1) Functional limitations: Mass spectrometry / optical detectors can only acquire mass spectrometry or spectral information and cannot directly measure the electrochemical properties of components (such as electron transfer ability, antioxidant activity, etc.), which are crucial for understanding the biological activity of compounds (such as drug efficacy mechanism, food preservation, etc.); (2) Redundancy in the process: If it is necessary to supplement electrochemical data, it is usually necessary to collect the chromatographically separated components offline, and then perform electrochemical detection through a traditional electrochemical workstation after preprocessing. This method will lead to problems such as large sample loss, long detection cycle, and poor reproducibility. In addition, the output of electrochemical detection results will also have a certain lag, and it is impossible to achieve synchronous analysis of mass spectrometry detection results, optical detection results and electrochemical detection results. Summary of the Invention

[0004] To address the shortcomings of existing technologies, this invention provides a system for the simultaneous detection of complex system separation and multifunctional properties. By incorporating a flow splitting control unit, the effluent from the liquid chromatography separation unit is precisely controlled and distributed, allowing it to be split as needed to the mass spectrometry / optical detection unit and the electrochemical detection unit. This enables the mass spectrometry, optical, and electrochemical detection modules to simultaneously detect the same separated component, achieving multidimensional simultaneous analysis of the optical, structural, and electrochemical characteristics of the same component. This overcomes the limitation of existing liquid chromatography coupled with mass spectrometry / optical detection techniques in obtaining the electrochemical properties of components. This invention also provides methods for preparing and detecting the system.

[0005] The technical effects to be achieved by this invention are realized through the following technical aspects: In a first aspect, the present invention provides a system for simultaneous detection of separation and multifunctional properties of complex systems, including a liquid chromatography separation unit, a split control unit, a mass spectrometry / optical detection unit, an electrochemical detection unit, and a data analysis unit; The outlet of the liquid chromatography separation unit is connected to the inlet of the split control unit. The outlet of the split control unit is connected to a first pipeline and a second pipeline. The first pipeline is connected to the mass spectrometry / optical detection unit, and the second pipeline is connected to the electrochemical detection unit. The mass spectrometry / optical detection unit includes a mass spectrometry detection module and an optical detection module. The mass spectrometry detection module, the optical detection module, and the electrochemical detection unit are connected in parallel and are all connected to the data analysis unit.

[0006] As a further description of the technical solution of the present invention, the complex system separation and multifunctional characteristic synchronous detection system also includes a collection unit, and the outlet of the diversion control unit is also connected to a third pipeline, the third pipeline being connected to the collection unit, and the diversion control unit including a three-way valve.

[0007] As a further description of the technical solution of the present invention, the electrochemical detection unit includes a microfluidic chip and an electrochemical workstation. The microfluidic chip includes a chip carrier and a plurality of microelectrodes integrated on the chip carrier. The microelectrodes are connected in series or in parallel and each microelectrode is connected to the electrochemical workstation. The microelectrodes include a working electrode, a reference electrode and an auxiliary electrode. The reference electrode and the auxiliary electrode are disposed on the outer periphery of the working electrode.

[0008] As a further description of the technical solution of the present invention, the material of the working electrode is one of nano-graphene, carbon nanotubes or glassy carbon, the material of the reference electrode is Ag or AgCl, and the material of the auxiliary electrode is a carbon atom material.

[0009] As a further description of the technical solution of the present invention, the surface of the working electrode is coated with a modification coating, which is one of metal nanoparticles, organic polymers, enzymes or antibody biosensitive membranes.

[0010] As a further description of the technical solution of the present invention, the chip carrier includes a base plate, an intermediate plate, and a cover plate stacked sequentially from bottom to top. The base plate is provided with a plurality of microelectrode placement slots, and the microelectrodes are placed in the microelectrode placement slots. The bottom of the intermediate plate is provided with a reaction tank corresponding to the position of each microelectrode placement slot. A liquid flow channel is provided above the reaction tank and the liquid flow channel is connected to the reaction tank. The cover plate is provided with a sample inlet channel and a sample outlet channel, and the sample inlet channel and the sample outlet channel are respectively connected to the inlet and outlet of the liquid flow channel.

[0011] As a further description of the technical solution of the present invention, the base plate and the cover plate are made of polymethyl methacrylate, and the intermediate plate is made of polydimethylsiloxane.

[0012] As a further description of the technical solution of the present invention, the microelectrode is provided with a reaction zone and a workstation connection zone at both ends, the working electrode, the reference electrode and the auxiliary electrode all extend to the reaction zone and the workstation connection zone, the reaction zone corresponds to the position of the reaction tank, the workstation connection zone corresponds to the connection between the microelectrode and the electrochemical workstation, and an electrode protective layer is coated between the reaction zone and the workstation connection zone.

[0013] Secondly, the present invention provides a method for preparing a complex system separation and multifunctional property simultaneous detection system, comprising the following steps: The base plate is processed to form a microelectrode placement groove, the middle plate is formed to form a reaction tank and a liquid flow channel through a molding process, and the reaction tank and the liquid flow channel are connected by a punch. The cover plate is processed to form a sample inlet channel and a sample outlet channel. The pre-fabricated microelectrode is placed in the microelectrode placement groove, the intermediate plate is placed on the bottom plate, and the cover plate is placed on the intermediate plate. The bottom plate, intermediate plate and cover plate are bonded and fixed together to obtain the microfluidic chip. The inlet of the split control unit is connected to the outlet of the liquid chromatography separation unit. The split control unit is connected to the mass spectrometry / optical detection unit through the first pipeline. The split control unit is connected to the sample injection channel of the microfluidic chip through the second pipeline. The microelectrode on the microfluidic chip is connected to the electrochemical workstation. The mass spectrometry detection module and optical detection module in the mass spectrometry / optical detection unit, as well as the electrochemical workstation, are connected to the data analysis unit through wired or wireless connections.

[0014] Thirdly, this invention provides a method for the simultaneous detection of complex system separation and multifunctional properties, comprising the following steps: The sample to be tested is separated into analytes by the liquid chromatography separation unit. The analytes flow out with the effluent from the liquid chromatography separation unit and simultaneously enter the mass spectrometry / optical detection unit and the electrochemical detection unit through the regulation of the split control unit. The electrochemical detection unit sets the test items of each microelectrode in the microfluidic chip through the electrochemical workstation. The mass spectrometry detection module and the optical detection module in the mass spectrometry / optical detection unit, as well as the electrochemical detection unit, simultaneously detect the analytes and transmit the detection results to the data analysis unit. The data analysis unit performs synchronous analysis on the various detection data of the analytes.

[0015] In summary, the present invention has at least the following advantages: The complex system separation and multifunctional characteristic simultaneous detection system provided by this invention connects a mass spectrometry detection module, an optical detection module, and an electrochemical detection unit in parallel with a data analysis unit. This enables simultaneous acquisition and analysis of multidimensional data on the chromatographic separation behavior, structural characteristics, optical characteristics, and electrochemical properties (such as pH and antioxidant activity) of the same component in a single injection. By setting up a split control unit to strictly distribute and control the effluent from the liquid chromatography separation unit, the effluent can be split to the mass spectrometry / optical detection unit and the electrochemical detection unit as needed. This ensures that sufficient sample volume is provided to the mass spectrometry / optical detection unit and the electrochemical detection unit while avoiding excessive sample consumption and waste.

[0016] The present invention provides a method for preparing a complex system separation and multifunctional characteristic synchronous detection system. The prepared complex system can realize real-time joint measurement and synchronous analysis of multidimensional data of the structural characteristics, optical characteristics and electrochemical properties of the same separated component under a single injection, which is beneficial to improving the accuracy and efficiency of component detection and analysis in complex systems.

[0017] The present invention provides a method for simultaneous detection of separation and multifunctional properties in complex systems. This method can simultaneously acquire and analyze multidimensional data such as mass spectrometry characteristics, spectral characteristics, redox potential, and electron transfer number of the same component. This enables rapid and accurate qualitative analysis of each component in complex multi-component systems, facilitating the rapid analysis of their functional properties. The detection method of this invention is applicable to scenarios such as active ingredient screening, antioxidant capacity assessment, and metabolite analysis in complex multi-component systems of natural products, food, and pharmaceuticals. It overcomes the technical bottleneck of real-time multi-parameter simultaneous detection in complex multi-component systems, providing a one-stop solution for the structural and functional analysis of unknown substances. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the complex system separation and multifunctional characteristic synchronous detection system according to Embodiment 1 of the present invention; Figure 2 This is a schematic diagram of the microfluidic chip according to Embodiment 1 of the present invention; Figure 3 This is a schematic diagram of the microelectrode structure of Embodiment 1 of the present invention; Figure 4 This is an exploded view of the chip carrier structure according to Embodiment 1 of the present invention; Figure 5 This is a schematic diagram showing the results of electrochemical detection, mass spectrometry detection, and optical detection of the actual sample in Example 3 of the present invention.

[0019] Marked in the image: 1. Liquid chromatography separation unit; 2. Split control unit; 3. Mass spectrometry / optical detection unit; 31. Mass spectrometry detection module; 32. Optical detection module; 4. Electrochemical detection unit; 41. Microfluidic chip; 411. Chip carrier; 4111. Base plate; 4112. Intermediate plate; 4113. Cover plate; 4114. Microelectrode placement slot; 4115. Reaction tank; 4116. Liquid flow channel; 4117. Sample inlet channel; 4118. Sample outlet channel; 412. Microelectrode; 4121. Working electrode; 4122. Reference electrode; 4123. Auxiliary electrode; 4124. Reaction zone; 4125. Workstation connection area; 4126. Electrode protective layer; 5. Data analysis unit; 6. Data collection unit; A. First pipeline; B. Second pipeline; C. Third pipeline. Detailed Implementation

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. The described embodiments are some, but not all, of the embodiments of the present invention.

[0021] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0022] Example 1 refer to Figures 1 to 4 The complex system separation and multifunctional characteristic simultaneous detection system provided in this embodiment includes a liquid chromatography separation unit 1, a split control unit 2, a mass spectrometry / optical detection unit 3, an electrochemical detection unit 4, and a data analysis unit 5. The outlet of the liquid chromatography separation unit 1 is connected to the inlet of the split control unit 2. The outlet of the split control unit 2 is connected to a first pipeline A and a second pipeline B. The first pipeline A is connected to the mass spectrometry / optical detection unit 3, and the second pipeline B is connected to the electrochemical detection unit 4.

[0023] The mass spectrometry / optical detection unit 3 includes a mass spectrometry detection module 31 and an optical detection module 32. The mass spectrometry detection module 31, the optical detection module 32, and the electrochemical detection unit 4 are connected in parallel and are all connected to the data analysis unit 5 via wired or wireless connection. The data analysis unit 5 is used to correlate liquid chromatography retention time, mass spectrometry signal, optical detection signal, and electrochemical response signal, thereby enabling the simultaneous acquisition and analysis of multi-dimensional detection data such as structural characteristics, optical characteristics, and electrochemical properties of the same separated component.

[0024] It is understandable that the mass spectrometry detection module 31 and the optical detection module 32 are connected in parallel in the mass spectrometry / optical detection unit 3. When the sample flows into the mass spectrometry / optical detection unit 3 through the first tube A, it will be divided into two streams and flow into the mass spectrometry detection module 31 and the optical detection module 32 respectively. The mass spectrometry detection module 31 and the optical detection module 32 can detect the same component at the same time. Since the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4 are also connected in parallel, the parallel connection of the mass spectrometry detection module 31, the optical detection module 32 and the electrochemical detection unit 4 is realized, thereby realizing the simultaneous mass spectrometry detection, optical detection and electrochemical detection of the same separated component.

[0025] Because the mass spectrometry / optical detection unit 3 requires a large sample volume for detection, while the electrochemical detection unit 4 requires a smaller sample volume for electrochemical detection, strict control and distribution of the effluent from the liquid chromatography separation unit 1 are necessary. This ensures that the effluent flows into the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4 as needed, achieving a balance in sample supply. By setting up a flow splitting control unit 2, the effluent from the liquid chromatography separation unit 1 can be strictly distributed and controlled, allowing the effluent to be split to the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4 as needed. This ensures that sufficient sample volume is provided to the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4, guaranteeing the authenticity and accuracy of the detection results, while also avoiding excessive sample consumption and waste.

[0026] In some embodiments, the complex system separation and multifunctional characteristic simultaneous detection system further includes a collection unit 6. The outlet of the split control unit 2 is also connected to a third pipeline C, which is connected to the collection unit 6. The collection unit 6 is mainly used to collect excess liquid chromatography eluent. In this embodiment, the split control unit 2 includes a three-way valve, which can be used to control the split of the first pipeline A (corresponding to the mass spectrometry detection module 31 and the optical detection module 32), the second pipeline B (corresponding to the electrochemical detection unit 4), and the third pipeline C (corresponding to the collection unit 6). In practical applications, the sample flow rate can also be controlled by changing the diameter of the first pipeline A, the second pipeline B, and the third pipeline C to precisely control the injection volume of the mass spectrometry detection module 31, the optical detection module 32, and the electrochemical detection unit 4. After the liquid chromatography eluent flows into the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4 as needed, the excess liquid chromatography eluent will flow into the collection unit 6 through the third pipeline C. The eluent flowing into the collection unit 6 can be used for subsequent analysis and detection, such as NMR, if necessary.

[0027] In one implementation, the mass spectrometry detection module 31 is a mass spectrometry detector, and the optical detection module 32 is an ultraviolet detector, specifically a diode array detector (PDA). The electrochemical detection unit 4 specifically includes a microfluidic chip 41 and an electrochemical workstation (not shown in the figure). The microfluidic chip 41 includes a chip carrier 411 and multiple microelectrodes 412 integrated on the chip carrier 411. The microelectrodes 412 are connected in series or in parallel, and each microelectrode 412 is connected to the electrochemical workstation. The microelectrode 412 includes a working electrode 4121, a reference electrode 4122, and an auxiliary electrode 4123. The reference electrode 4122 and the auxiliary electrode 4123 are disposed on the outer periphery of the working electrode 4121.

[0028] It should be noted that different microelectrodes 412 on the microfluidic chip 41 are used to detect different items. The analyte can be subjected to real-time electrochemical detection on each microelectrode 412, such as potential-time scanning or current-time scanning. The electrochemical workstation can simultaneously acquire the potential changes or current changes on each microelectrode 412, thereby obtaining parameter information such as the pH, antioxidant properties, and content of the target analyte. The microelectrodes 412 can be configured in parallel or in series. Parallel configuration can avoid mutual interference between the electrochemical detection behaviors of the microelectrodes 412, while series configuration can reduce the sample flow requirements, avoid affecting the sample supply stability of the mass spectrometry detection module 31 and the optical detection module 32, and ensure the normal and stable operation of mass spectrometry detection, optical detection, and electrochemical detection. In this embodiment, the microelectrodes 412 are connected in series.

[0029] In this embodiment, the working electrode 4121 is made of one of the following materials: graphene nanoparticles, carbon nanotubes, or glassy carbon; the reference electrode 4122 is made of Ag or AgCl; and the auxiliary electrode 4123 is made of carbon atoms. The graphene nanoparticle electrode can be rapidly prepared using laser-induced graphene technology.

[0030] As a further optimization, the surface of the working electrode 4121 can be coated with a modification coating, which can be one of metal nanoparticles, organic polymers, enzymes, or antibody biosensitive membranes. Specifically, the metal nanoparticles can be Au, Pt, or Pd, etc.; the organic polymer can be polyaniline; and the enzyme can be glucose oxidase or amino acid oxidase. Modifying the surface of the working electrode 4121 with metal nanoparticles can improve the electrode's catalytic activity; modifying with polyaniline enables real-time detection of the pH of samples flowing across the electrode surface, thereby obtaining the acid-base properties of the target detection component; modifying with glucose oxidase enables the detection of glucose; modifying with amino acid oxidase enables the detection of amino acids; and modifying with antibody biosensitive membranes enables the detection of specific antigens.

[0031] The chip carrier 411 includes a base plate 4111, an intermediate plate 4112, and a cover plate 4113 stacked from bottom to top. The base plate 4111 is provided with a plurality of microelectrode placement slots 4114, and each microelectrode 412 is placed in a corresponding microelectrode placement slot 4114. The bottom of the intermediate plate 4112 is provided with a reaction tank 4115 corresponding to the position of each microelectrode placement slot 4114. The reaction tank 4115 can provide a reaction space for the electrochemical reaction between the microelectrode 412 and the sample. A liquid flow channel 4116 is provided above the reaction tank 4115 and the liquid flow channel 4116 is connected to the reaction tank 4115. The cover plate 4113 is provided with a sample inlet channel 4117 and a sample outlet channel 4118. The sample inlet channel 4117 and the sample outlet channel 4118 are respectively connected to the inlet and outlet of the liquid flow channel 4116. The sample inlet channel 4117 is connected to the second pipeline B. The sample inlet channel 4117 is used to allow the sample to enter the microfluidic chip 41, and the sample outlet channel 4118 is used to discharge the electrochemical reaction waste liquid.

[0032] It should be noted that when the microelectrodes 412 are connected in series, the liquid flow channel 4116 is set in segments, that is, the sample will flow through each reaction tank 4115 sequentially through the liquid flow channel 4116; when the microelectrodes 412 are connected in parallel, the liquid flow channel 4116 is set in a continuous segment, that is, the sample will be simultaneously diverted to each reaction tank 4115 through the liquid flow channel 4116.

[0033] In this embodiment, the base plate 4111 and cover plate 4113 are made of polymethyl methacrylate (PMMA), and the intermediate plate 4112 is made of polydimethylsiloxane (PDMS). The base plate 4111, intermediate plate 4112, and cover plate 4113 are bonded together. Using PMMA as the material for the base plate 4111 and cover plate 4113 makes it easier to process, reducing processing difficulty and improving processing quality. Using PDMS as the material for the intermediate plate 4112 effectively prevents sample leakage, ensuring the accuracy and reliability of the electrochemical detection results. Alternatively, the base plate 4111 and cover plate 4113 can also be made of other rigid plastics or glass, and the intermediate plate 4112 can be made of other elastomer materials.

[0034] As a further optimization, the microelectrode 412 is provided with a reaction zone 4124 and a workstation connection zone 4125 at both ends. The working electrode 4121, the reference electrode 4122, and the auxiliary electrode 4123 all extend to the reaction zone 4124 and the workstation connection zone 4125. The reaction zone 4124 corresponds to the position of the reaction tank 4115, and the workstation connection zone 4125 corresponds to the connection between the microelectrode 412 and the electrochemical workstation. An electrode protective layer 4126 is coated between the reaction zone 4124 and the workstation connection zone 4125. This can reduce the wear of the microelectrode 412 during the loading and unloading of the microfluidic chip 41, and also prevent sample leakage when the sample flow rate is too high.

[0035] Example 2 refer to Figures 1 to 4 The preparation method of the complex system separation and multifunctional characteristic synchronous detection system provided in Example 1 includes the following steps: The polymethyl methacrylate base plate 4111 is processed by a mold to form two microelectrode placement slots 4114. The polydimethylsiloxane intermediate plate 4112 is formed by a molding process to form two reaction tanks 4115 and corresponding liquid flow channels 4116. The reaction tanks 4115 and the liquid flow channels 4116 are connected by a punch. The polymethyl methacrylate cover plate 4113 is formed by a molding process and a punch to form a sample inlet channel 4117 and a sample outlet channel 4118. Microelectrode 1 412 and microelectrode 2 412 are pre-prepared. The working electrode of microelectrode 1 412 is laser-induced graphene modified with polyaniline, and the working electrode of microelectrode 2 412 is laser-induced graphene. The reference electrodes of microelectrode 1 412 and microelectrode 2 412 are both prepared by drying and curing silver paste, and the auxiliary electrodes are both laser-induced graphene. The pre-prepared microelectrode 1 412 and microelectrode 2 412 are placed in two microelectrode placement slots 4114 respectively. The polydimethylsiloxane intermediate plate 4112 is covered on the polymethyl methacrylate base plate 4111, and the polymethyl methacrylate cover plate 4113 is covered on the polydimethylsiloxane intermediate plate 4112. The polymethyl methacrylate base plate 4111, polydimethylsiloxane intermediate plate 4112 and polymethyl methacrylate cover plate 4113 are bonded and fixed together to obtain the microfluidic chip 41. The inlet of the split control unit 2 is connected to the outlet of the liquid chromatography separation unit 1. The split control unit 2 is connected to the mass spectrometry / optical detection unit 3 through the first pipeline A. The split control unit 2 is connected to the sample injection channel 4117 of the microfluidic chip 41 through the second pipeline B. The microelectrode 412 on the microfluidic chip 41 is connected to the electrochemical workstation. The mass spectrometry detection module 31 and the optical detection module 32 in the mass spectrometry / optical detection unit 3, as well as the electrochemical workstation, are connected to the data analysis unit 5 through wired or wireless connection to complete the preparation of a system for simultaneous detection of complex system separation and multifunctional characteristics.

[0036] Rapid fabrication of microfluidic chips via templates can reduce fabrication costs, and the resulting microfluidic chips possess advantages such as high throughput and high integration. The complex system separation and multifunctional property simultaneous detection system prepared in this embodiment can achieve real-time joint measurement and simultaneous analysis of multidimensional data on the structural characteristics, optical characteristics, and electrochemical properties of the same separated component in a single sample injection, which is beneficial for improving the accuracy and efficiency of component detection and analysis in complex systems.

[0037] Example 3 refer to Figures 1 to 5 The method for simultaneous detection of complex system separation and multifunctional properties provided in this embodiment uses a microfluidic chip 41 with microelectrode 1 412 and microelectrode 2 412 for electrochemical detection. The working electrode of microelectrode 1 412 is laser-induced graphene modified with polyaniline, and the working electrode of microelectrode 2 412 is also laser-induced graphene. The reference electrodes of both microelectrodes 1 412 and 2 412 are prepared by drying and curing silver paste, and the auxiliary electrodes are both laser-induced graphene. The detection method specifically includes the following steps: Sample preparation: Prepare a multi-component mixed solution as the actual sample. The components of the multi-component mixed solution include ascorbic acid, caffeine, sucrose and rutin. Setting up the electrochemical detection items: Set the test mode of microelectrode 412 to obtain the open circuit potential-time curve and the test mode of microelectrode 412 to obtain the constant potential current-time curve through the electrochemical workstation, and set the potential of microelectrode 412 to 1.2V; The actual sample is separated into the analyte by the liquid chromatography separation unit 1. The analyte flows out with the effluent of the liquid chromatography separation unit 1 and simultaneously enters the mass spectrometry / optical detection unit 3 and the electrochemical detection unit 4 through the regulation of the split control unit 2. The mass spectrometry detection module 31 (mass spectrometer detector) and the optical detection module 32 (PDA optical detector) in the mass spectrometry / optical detection unit 3, as well as the electrochemical detection unit 4, simultaneously detect the analyte and transmit the detection results to the data analysis unit 5. The data analysis unit 5 obtains the mass spectrometry detection results, optical detection results and electrochemical detection results of the analyte and performs simultaneous analysis.

[0038] like Figure 5 As shown, the analysis results are as follows: According to the obtained data, the mass spectrum peak with a retention time of about 240 s is sucrose (m / z=341, negative mode), and no detection signal appeared on microelectrode 1, microelectrode 2, or the PDA optical detector. The analysis results of data analysis unit 5 show that the spectral peak and mass spectrum peak with a retention time of about 270 s (m / z=175, negative mode) are ascorbic acid. It shows a positive potential increase peak on microelectrode 1, indicating that it is acidic, and at the same time, it shows a current peak on microelectrode 2, indicating that it also has reducing properties (easily oxidized). The spectral peak with a retention time of about 450 s is caffeine. It has no obvious characteristic peak on the mass spectrum in negative mode, and there is no obvious potential fluctuation on microelectrode 1, but it shows a current peak on microelectrode 2, indicating that it has reducing properties (easily oxidized). The retention time of 880 s is... The spectral peaks around s and the mass spectrum peak (m / z=608, negative mode) are rutin. It shows no obvious potential fluctuation on microelectrode 1, but shows a current peak on microelectrode 2, indicating that it has reducing properties (is easily oxidized).

[0039] As can be seen, the analysis results of each component by the data analysis unit are consistent with the actual characteristics and correspond one-to-one, proving the feasibility, authenticity and reliability of the detection method in this embodiment.

[0040] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0041] Although the description of the invention has been given in conjunction with the specific embodiments described above, it will be apparent to those skilled in the art that many substitutions, modifications, and variations can be made based on the foregoing. Therefore, all such substitutions, modifications, and variations are included within the spirit and scope of the appended claims.

Claims

1. A system for simultaneous detection of complex system separation and multifunctional characteristics, characterized in that, It includes a liquid chromatography separation unit (1), a split control unit (2), a mass spectrometry / optical detection unit (3), an electrochemical detection unit (4), and a data analysis unit (5); The outlet of the liquid chromatography separation unit (1) is connected to the inlet of the split control unit (2). The outlet of the split control unit (2) is connected to a first pipeline (A) and a second pipeline (B). The first pipeline (A) is connected to the mass spectrometry / optical detection unit (3), and the second pipeline (B) is connected to the electrochemical detection unit (4). The mass spectrometry / optical detection unit (3) includes a mass spectrometry detection module (31) and an optical detection module (32). The mass spectrometry detection module (31), the optical detection module (32) and the electrochemical detection unit (4) are connected in parallel and are all connected to the data analysis unit (5).

2. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 1, characterized in that, It also includes a collection unit (6), and the outlet of the diversion control unit (2) is connected to a third pipeline (C), which is connected to the collection unit (6). The diversion control unit (2) includes a three-way valve.

3. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 2, characterized in that, The electrochemical detection unit (4) includes a microfluidic chip (41) and an electrochemical workstation. The microfluidic chip (41) includes a chip carrier (411) and multiple microelectrodes (412) integrated on the chip carrier (411). Each microelectrode (412) is connected in series or in parallel and each microelectrode (412) is connected to the electrochemical workstation. Each microelectrode (412) includes a working electrode (4121), a reference electrode (4122), and an auxiliary electrode (4123). The reference electrode (4122) and the auxiliary electrode (4123) are located on the outer periphery of the working electrode (4121).

4. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 3, characterized in that, The working electrode (4121) is made of one of the following materials: nano-graphene, carbon nanotubes, or glassy carbon; the reference electrode (4122) is made of Ag or AgCl; and the auxiliary electrode (4123) is made of carbon atom material.

5. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 4, characterized in that, The surface of the working electrode (4121) is coated with a modification coating, which is one of metal nanoparticles, organic polymers, enzymes or antibody biosensing membranes.

6. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 3, characterized in that, The chip carrier (411) includes a bottom plate (4111), an intermediate plate (4112), and a cover plate (4113) stacked from bottom to top. The bottom plate (4111) is provided with a plurality of microelectrode placement slots (4114), and the microelectrodes (412) are placed in the microelectrode placement slots (4114). The bottom of the intermediate plate (4112) is provided with a reaction tank (4115) corresponding to the position of each microelectrode placement slot (4114). A liquid flow channel (4116) is provided above the reaction tank (4115) and the liquid flow channel (4116) is connected to the reaction tank (4115). The cover plate (4113) is provided with a sample inlet channel (4117) and a sample outlet channel (4118), and the sample inlet channel (4117) and the sample outlet channel (4118) are respectively connected to the inlet and outlet of the liquid flow channel (4116).

7. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 6, characterized in that, The base plate (4111) and the cover plate (4113) are made of polymethyl methacrylate, and the intermediate plate (4112) is made of polydimethylsiloxane.

8. The system for simultaneous detection of complex system separation and multifunctional characteristics according to claim 6, characterized in that, The microelectrode (412) has a reaction zone (4124) and a workstation connection zone (4125) at both ends. The working electrode (4121), the reference electrode (4122), and the auxiliary electrode (4123) all extend to the reaction zone (4124) and the workstation connection zone (4125). The reaction zone (4124) corresponds to the position of the reaction tank (4115), and the workstation connection zone (4125) corresponds to the connection between the microelectrode (412) and the electrochemical workstation. An electrode protective layer (4126) is coated between the reaction zone (4124) and the workstation connection zone (4125).

9. A method for preparing a complex system separation and multifunctional characteristic synchronous detection system according to any one of claims 6-8, characterized in that, Includes the following steps: The base plate (4111) is processed to form a microelectrode placement groove (4114), the middle plate (4112) is formed to form a reaction tank (4115) and a liquid flow channel (4116) through a molding process, and the reaction tank (4115) and the liquid flow channel (4116) are connected by a punch. The cover plate (4113) is processed to form a sample inlet channel (4117) and a sample outlet channel (4118). The prefabricated microelectrode (412) is placed in the microelectrode placement groove (4114), the intermediate plate (4112) is placed on the bottom plate (4111), and the cover plate (4113) is placed on the intermediate plate (4112). The bottom plate (4111), the intermediate plate (4112) and the cover plate (4113) are bonded and fixed together to obtain the microfluidic chip (41). The inlet of the split control unit (2) is connected to the outlet of the liquid chromatography separation unit (1). The split control unit (2) is connected to the mass spectrometry / optical detection unit (3) through the first pipeline (A). The split control unit (2) is connected to the sample injection channel (4117) of the microfluidic chip (41) through the second pipeline (B). The microelectrode (412) on the microfluidic chip (41) is connected to the electrochemical workstation. The mass spectrometry detection module (31) and the optical detection module (32) in the mass spectrometry / optical detection unit (3), as well as the electrochemical workstation, are connected to the data analysis unit (5) through wired or wireless connection.

10. A method for simultaneous detection of separation and multifunctional properties of complex systems, characterized in that, Includes the following steps: The sample to be tested is separated into components by the liquid chromatography separation unit (1). The components to be tested flow out with the effluent of the liquid chromatography separation unit (1) and simultaneously enter the mass spectrometry / optical detection unit (3) and the electrochemical detection unit (4) through the regulation of the split control unit (2). The electrochemical detection unit (4) sets the test items of each microelectrode (412) in the microfluidic chip (41) through the electrochemical workstation. The mass spectrometry detection module (31) and optical detection module (32) in the mass spectrometry / optical detection unit (3) and the electrochemical detection unit (4) simultaneously detect the components to be tested and transmit the detection results to the data analysis unit (5). The data analysis unit (5) performs synchronous analysis on the various detection data of the components to be tested.

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